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    Effects of Mating Flight on sRNAs Expression in Sexual Matured Virgin Queens (Apis cerana cerana)
    WU Xiao-Bo, WANG Zi-Long, SHI Yuan-Yuan, ZHANG Fei, ZENG Zhi-Jiang
    Scientia Agricultura Sinica    2013, 46 (17): 3721-3728.   DOI: 10.3864/j.issn.0578-1752.2013.17.022
    Abstract1160)      PDF (530KB)(32250)       Save
    【Objective】The objective of this study is to analyze the influence of flight behaviour on sRNAs expression in matured virgin queens (Apis cerana cerana). 【Method】The sexual matured virgin queens of A. cerana cerana were used as flying group that allow to fly in a certain area and the other group was not. The sRNAs expression differences between the two groups were analyzed by using a high-throughput sequencing method. 【Result】Both of the flying matured virgin queens and non-flying counterparts had a complicated sRNAs population, and that the length of sRNAs varied, 22 nt and 27-29 nt being the predominant length, while the frequence percent of the same RNA was different in the two samples. Combining deep sequencing and bioinformatics analysis, the percent of the common sequences in total sRNAs was 92.79%, but the number of unique sRNAs in the flying queens was more than that in non-flying queens. Matching to known miRNA alignment, a total of 25 known miRNAs were significantly different between them, with 1 up-regulated and 24 down-regulated in the flying matured virgin queens while there were 11 target genes of 19 known miRNA which were expressed differentially between them.【Conclusion】There are a large number of sRNAs changes during the flight process of matured virgin queens which may play an important role in adjusting physiology for mating successfully of virgin queens.
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    Cited: Baidu(3)
    Discussion on the Theory and Methods for Determining the Target Yield in Rice Production
    ZOU Ying-bin, XIA Bing, JIANG Peng, XIE Xiao-bing, HUANG Min
    Scientia Agricultura Sinica    2015, 48 (20): 4021-4032.   DOI: 10.3864/j.issn.0578-1752.2015.20.004
    Abstract1054)   HTML77)    PDF (625KB)(26501)       Save
    【Objective】The aim of this study was to discuss the theory and methods for determining the target yield in rice production. 【Methods】The discussion and analysis were based on the results of field experiments conducted in five locations (Chengmai of Hainan Province, Huaiji of Guangdong Province, Binyang of Guangxi Province, Changsha of Hunan Province, and Xingyi of Guizhou Province) in South China with different N application rates (zero N application; moderate N rate: 161—176 kg·hm-2; high N rate: 225 kg·hm-2) and varieties (hybrid varieties Liangyoupeijiu and Y-liangyou 2 and inbred varieties Huanghuazhan and Yixiangyouzhan ) in 2012 and 2013, and the reports of relevant literature in China and abroad.【Results】The field experiments showed that the yield performance of even the same rice variety exhibited significant or extremely significant differences among the five locations. Under N application conditions (moderate and high N rates), Xingyi had the highest average yield (Liangyoupeijiu: 13.20-13.54 t·hm-2, Y-liangyou 1: 13.50-13.78 t·hm-2, Huanghuazhan: 11.26-11.42 t·hm-2, Yuxiangyouzhan: 11.32-11.45 t·hm-2), followed by Changsha, Chengmai, Binyang, and Huaiji had the lowest average yield (Liangyoupeijiu: 6.66-6.71 t·hm-2, Y-liangyou 1: 6.96-7.20 t·hm-2, Huanghuazhan: 6.96-7.11 t·hm-2, Yuxiangyouzhan: 7.35-6.86 t·hm-2). Similarly, the highest average soil-based yield (yield of no N application treatment) was recorded in Xingyi (10.52 t·hm-2), followed by that in Changsha, Chengmai and Binyang, and the lowest average soil-based yield was recorded in Huaiji (4.53 t·hm-2). The rice yield under fertilized conditions (namely fertilized yield) (YF) depended extremely significantly on the soil-based yield (YS). The regression equations under moderate and high N rates were YF=0.814YS+3.337 (R2=0.824) and YF﹦0.864YS+3.094 (R2=0.839), respectively. The contributions of the soil-based yield (the percentage of the soil-based yield in the fertilized yield) ranged from 64.8% to 85.5% on the average of five locations and from 72.7% to 79.3% on the average of four varieties. The analysis of the data (n=315) collected from previous studies also indicated that there was a significant positive linear relationship between the soil-based yield and the fertilized yield (YF=1.031YS+2.421, R2=0.523), and the average contribution of the soil-based yield was 67.7%. In addition, the results showed that yield increased by fertilization was tightly negatively related with soil-based yield contribution; grain yield was significantly quadratically related to plant N uptake and N application rate.【Conclusions】Target yield should be varied from site to site. Soil-based yield comprehensively reflects the paddy soil fertility and the climate productivity, and therefore can be used as the basis to determine the target yield in rice production. The target yield for high yielding cultivation of rice can be determined by the regression equation based on the soil-based yield. Improving soil fertility is an important approach for achieving the target yield.
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    Cited: Baidu(5)
    Scientia Agricultura Sinica    1978, 11 (02): 16-20.   DOI: 10.3864/j.issn.0578-1752.1978-11-02-16-20
    Abstract1173)      PDF (300KB)(22233)       Save
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    Bioinformatics and Expression Analysis of the WRKY Gene Family in Apple
    GU Yan-bing, JI Zhi-rui, CHI Fu-mei, QIAO Zhuang, XU Cheng-nan, ZHANG Jun-xiang, DONG Qing-long, ZHOU Zong-shan
    Scientia Agricultura Sinica    2015, 48 (16): 3221-3238.   DOI: 10.3864/j.issn.0578-1752.2015.16.012
    Abstract1264)   HTML62)    PDF (15602KB)(21099)       Save
    【Objective】In this study, 132 putative WRKY proteins in the apple (Malus domestica Borkh.) genome were identified, so as to provide a basis for studying the theoretical roles of WRKY genes in the regulation of plant stress responses, growth and development, and to provide valuable information for WRKY genes cloning in apple.【Method】WRKY DNA-binding domain (PF03106) downloaded from Pfam protein families database was employed to identify WRKY genes from apple genome using HMMER 3.0. The obtained amino acid sequences were analyzed with the bioinformatics softwares, including DNAMAN 5.0, Weblogo 3, MEGA 5.1, MapInspect and MEME. RT-PCR was used to detect WRKY genes expression in different tissues of apple. 【Result】 Totally 132 apple WRKY genes were found in apple genome. The result of group identification and phylogenetic analysis revealed that apple WRKY genes were classified into Group I, Group II and Group III. Twenty-four MdWRKY proteins with two WRKY domains (group I-N and group I-C) containing CX4CX22-23HXH zinc-finger motif belonged to Group I. Seventy-nine Group II MdWRKY proteins had a single WRKY domain including CX4–5CX23HXH zinc-finger motif and could be further divided into five subgroups (Group II-a: 8 members, Group II-b: 12 members, Group II-c: 31 members, Group II-d: 14 members, and Group II-e: 14 members, respectively), whereas 29 Group III MdWRKY proteins contained a single WRKY domain with CX7CX23–24HXC zinc-finger motif. The results of domain analysis indicated that the WRKY regions contained a highly conserved heptapeptide stretch WRKYGQK at its N-terminus followed by a zinc-finger motif. Chromosome mapping analysis showed that apple WRKY genes were distributed with different densities on 17 chromosomes. The largest number of apple WRKY genes were found on chromosomes 1 and 9 (thirteen genes), followed by chromosome 12 (twelve genes). Only 4 genes located on chromosomes 2, 5 and 14. The results of gene structure analysis revealed that most of the WRKY gene contained 2-5 exons and WRKY gene structure were highly conserved in apple. Conserved motif analysis showed that the conserved motifs 1-6, which specify the WRKY domain, were observed in all apple WRKY proteins, motif 8 and motifs 7 and 9 as the unknown domain were observed in Group II-a and II-b and Group III, respectively. Two WRKY domains were assigned to Group I. RT-PCR results indicated that 12 MdWRKY genes were expressed in roots, stems, leaves, flowers and fruits at various expression levels.【Conclusion】These results suggested that MdWRKY gene family was highly and structurally conserved, and may be involved into the regulation of growth and development processes in apple.
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    Cited: Baidu(22)
    Expression and Function Analysis of the Transcription Factor GmMYB111 in Soybean
    XU Ling, WEI Pei-pei, ZHANG Da-yong, XU Zhao-long, HE Xiao-lan, HUANG Yi-hong, MA Hong-xiang, SHAO Hong-bo
    Scientia Agricultura Sinica    2015, 48 (15): 3079-3089.   DOI: 10.3864/j.issn.0578-1752.2015.15.019
    Abstract804)   HTML34)    PDF (3179KB)(19706)       Save
    【Objective】 A gene encoding MYB transcription factor, designated GmMYB111, was cloned, its basic biological functions and expression pattern were identified in soybean and yeast cells. 【Method】 A MYB transcription factor GmMYB111 was obtained from salt stress-related digital expression profiling (DGEP) data analysis. cDNA sequence of GmMYB111 was isolated and cloned using cDNA from soybean salt-treated roots by RT-PCR method. A homology search was performed using GmMYB111 protein sequence as a query, and protein sequences of high similarity with GmMYB111 from other species were obtained. Using MEGA5.05, multiple sequence alignments between GmMYB111 protein and its homologous ones from other species were done and a phylogenetic tree of homologous species was constructed. The induced expression and tissue-specific expression profiles of target genes in soybean with abiotic stress were detected by real-time fluorescent quantitative PCR. The subcellular localization of GmMYB111 was analyzed using Arabidopsisprotoplast transformation system, and its transcriptional activity and in vivo binding activity were determined by yeast hybrid system. 【Result】GmMYB111 gene, a significantly upregulated gene (27 folds) in response to salt stress, was obtained based on the preliminary digital expression profiling (DGEP) data related to salt stress in authors laboratory. Using RT-PCR method, fragment of this gene was cloned from cultivated soybean root. Sequence alignment revealed that its sequence was consistent with that from the published Williams82 genome database. Bioinformatics analysis indicated that the deduced amino acids had common characteristics of MYB transcription factors with two MYB domains of R2 and R3 at the N-terminal and an acidic amino acid-rich transcriptional activation domain at the C-terminal. Phylogenetic tree analysis suggested that the encoded protein had the closest genetic relationship with GmMYB76, GmMYB12a, and MtMYB61. The expression of GmMYB111 in soybean was induced by high salt, drought, chilling, and ABA treatments, respectively. Real-time fluorescent quantitative PCR detection results showed that the induced GmMYB111 was upregulated by high salt and cold stress, and was first upregulated and followed by a down-regulation by drought stress. There was a wave-like up- and down-regulated expression of GmMYB111 inducted by ABA treatment. Analysis of temporal and spatial expression showed that GmMYB111 was nearly expressed in all detected tissues, and its expression level was relatively high at seedling and low at maturing stage. From the perspective of different tissues, GmMYB111 showed the highest expression in stem, leaf, and flower, relatively low in root, and no expression in pod. Subcellular localization results showed that GmMYB111 was located in the nucleus which belongs to a typical transcription factor, yeast hybrid assay indicated that GmMYB111 had transcriptional activation functions and could bind to the cis-acting element TAACTG motif. 【Conclusion】GmMYB111 is a typical R2R3-MYB transcription factor, with transcriptional activation function and DNA binding activity. Its expression in soybean may be related to the abiotic stress and ABA signal transduction pathway. GmMYB111 is speculated to regulate the soybean response to abiotic stresses by regulating the expression of downstream genes.
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    Effects of Water Stress at Different Growth Stages on the Development and Yields of Winter Wheat in Arid Region
    YAO Ning, SONG Li-bing, LIU Jian, FENG Hao, WU Shu-fang, HE Jian-qiang
    Scientia Agricultura Sinica    2015, 48 (12): 2379-2389.   DOI: 10.3864/j.issn.0578-1752.2015.12.011
    Abstract902)   HTML87)    PDF (544KB)(19399)       Save
    【Objective】 To investigate the influences of water stress at different growth stages on the growth and yields of winter wheat, field experiments were conducted under a rainout shelter during two seasons of 2012-2013 and 2013-2014. The dynamic changes of several eco-physical characteristics of wheat growth were measured and compared, including height, leaf area index, phenology, biomass, and yield. 【Method】 The whole growth season of wheat was divided into five growing stages (wintering, greening, jointing, heading, and grain filling). Water stress occurred at two continuous stages, while irrigations were applied at other stages, which resulted in four different levels of stress period (D1-D4). Two irrigation levels of 40 mm (I1) and 80 mm (I2) were applied. A total of eight treatments, with three replicates for each, followed a split-plot experiment design. An extra control treatment with irrigation at all five stages was arranged beside.【Result】The results showed that normal growth and development of wheat could be obviously influenced by continuous water stress given at vegetative stages. The height, LAI and biomass were the worst for all treatments, when water stress occurred at the stages of wintering and greening. However, the negative influences on wheat growth were not notable when water stress occurred after jointing stage. The average growth rate of height and LAI after jointing was about ten times as that before jointing. There were no notable differences of biomass between all of the treatments until the jointing stage. The biomass values of treatments with water stresses at wintering and greening stages were remarkably lower than other treatments. The irrigation later could not recover these serious biomass losses. Water stress could shorten the whole growth season of wheat, with a maximal 5-day advancing of maturation. At the same irrigation level, the heading and flowering stages could be delayed for 1-3 days for different levels of stress period. For the same irrigation level, relatively higher numbers of productive ears and seeds per ear could be obtained when water stress occurred at the heading and grain filling stages, but with a lower thousand-kernel weight. On the contrary, a relatively higher thousand-kernel weight could be achieved when irrigation was applied at the heading and grain filling stages, but with lower numbers of productive ears and seeds per ear. For irrigation levels of I1 and I2, yields were the lowest when water stress occurred at wintering and greening stages, which was only 42% of the control treatment. However, the treatments with the highest yield were different for different irrigation levels. For I1, it was the treatment with water stress at jointing and heading stages that had the highest yield, or about 63% of the control treatment. For I2, it was the treatment with water stress at greening and jointing stages, which had a yield of about 75% of the control treatment. 【Conclusion】There was a clear interaction between the intensity and occurring stage of water stress. In general, the greening and filling stages were the critical periods of water demand for winter wheat. Reasonable irrigation managements are needed at these two growth stages to guarantee a higher yield of winter wheat in arid region.
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    Cited: Baidu(17)
    Response of Wheat Root Characteristics and Canopy Photosynthesis to Different Irrigation Methods in Lime Concretion Black Soil
    ZHANG Xiang-qian, CAO Cheng-fu, QIAO Yu-qiang, LI Wei, CHEN Huan
    Scientia Agricultura Sinica    2015, 48 (8): 1506-1517.   DOI: 10.3864/j.issn.0578-1752.2015.08.05
    Abstract615)   HTML25)    PDF (396KB)(19228)       Save
    【Objective】The aim of this experiment was to clarify the potential mechanism of the effects of different irrigation methods which was irrigated at early growth stage of wheat on root characteristics, canopy photosynthesis and yield at middle and late growth stages, and to reveal the relationship between root characteristics and canopy photosynthesis. 【Method】 The experiment was conducted in an artificial glass rainproof shed, and the designed ten irrigation treatments in this experiment were CK(adequate water was supplied at early growth stage), W120d (20 d after seedling emerged for 50 mm), W240d (40 d after seedling emerged for 50 mm), W360d (60 d after seedling emerged for 50 mm), W480d (80 d after seedling emerged for 50 mm), W5100d (100 d after seedling emerged for 50 mm), W6120d (120 d after seedling emerged for 50 mm), W720d+60d (20 d after seedling emerged for 25 mm+60 d after seedling emergency 25 mm), W840d+80d (40 d after seedling emerged for 25 mm+80 d after seedling emerged for 25 mm) and W960d+100d (60 d after seedling emerged for 25 mm+100 d after seedling emerged for 25 mm). The effects of different irrigation methods which were irrigated at wheat early growth stage on root characteristics, canopy single leaf area, chlorophyll density, photosynthetic capacity, photosynthetically active radiation, chlorophyll fluorescence parameters and yield of wheat at middle and late growth stages were studied. 【Result】 To appropriate postpone the irrigation date at wheat early growth stage could help to increase total root length, total surface area, total volume, average diameter, total root tips and total root forks, and the difference between W5100d (irrigation was carried out 100 d after seedling emergence) and CK (adequate water was supplied at early growth stage) was insignificant, but they were significantly higher than that of W120d (irrigation was carried out 20 d after seedling emergence) and W6120d (irrigation was carried out 120 d after seedling emergence). The single leaf area of 1, 2 and 3 leaves counted from the top of CK was the highest at booting and flowering stages, but the difference between CK and W5100d was not significant, and the canopy of single leaf area was not significantly affected by irrigation times under the same irrigation amount. When one time of irrigation was made at early growth stage of wheat, the canopy chlorophyll density had a change trend of initial increase and then decrease with the postponement of irrigation date, and the values of W5100d were the highest, while of W120d were the lowest. The canopy photosynthetic rate of W5100d at booting stage, flowering stage and middle of filling stage was significantly higher than that of CK, and was increased by 7.5%, 8.9% and 8.9%, respectively, but the canopy photosynthetic rate was not significantly affected by irrigation times. Under the condition of one time of irrigation, W5100d had the highest values of canopy photosynthetically active radiation at booting stage, flowering stage and middle of filling stage, and when compared to W120d and W6120d , the values were significantly increased by 18.7%, 9.7%, 11.0% and 5.7%, 4.9%, 4.3%, respectively. The chlorophyll fluorescence parameters of Fo, Fm, Fv/Fm, ΦpsII and ETR of W5100d at booting and flowering stages were the highest, and the chlorophyll fluorescence parameters were not significantly affected by irrigation times. The differences in yield and harvest index between W5100d and CK were not significant, and when compared to W120d and W6120d they were increased by 15.4%, 22.1% and 3.2%, 9.2%, respectively.【Conclusion】Too early or too late irrigation at early growth stage had adverse impact on root growth, canopy light utilization and yield formation, while the irrigation date was appropriately postponed, the root characteristics, canopy photosynthesis and yield were better than or equivalent to the treatment of adequate water supply at wheat early growth stage, and the effects of irrigation times were not obvious under the same amount of irrigation water.
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    Cited: Baidu(1)
    Difference Analysis of Post-Anthesis Matter Production and Senescence Characteristics Among Different Nitrogen Efficiency Populations in Wheat Following Rice
    DING Jin-feng, CHENG Ya-mei, HUANG Zheng-jin, LI Chun-yan, GUO Wen-shan, ZHU Xin-kai
    Scientia Agricultura Sinica    2015, 48 (6): 1063-1073.   DOI: 10.3864/j.issn.0578-1752.2015.06.03
    Abstract565)   HTML19)    PDF (492KB)(18428)       Save
    【Objective】This study investigated the post-anthesis characteristics of wheat population in rice-wheat rotation system under high nitrogen utilization efficiency condition. 【Method】 From 2010 to 2012, using wheat variety Yangmai 20 as material, the field experiments were conducted as split-plot design which included 16 combinations of two applications of nitrogen amount (210.0 kg·hm-2 and 262.5 kg·hm-2), two nitrogen applied ratios of basis﹕tillering﹕elongation﹕booting (3﹕1﹕3﹕3 and 5﹕1﹕2﹕2), and four nitrogen topdressing stages (flag leaf stage, booting stage, heading stage and anthesis). In the rice-wheat rotation system, wheat populations at different nitrogen utilization rate (NUR) levels were established by agronomic management. The differences of grain yield, matter production, nitrogen absorption, and flag leaves senescence characteristics after anthesis among different NUR level populations were analyzed, respectively.【Result】The range of NUR in different populations was 31.18%-72.23%. Grain yield of NUR≥60% population (NUR-H) was more than 8 500 kg·hm-2, which was 6.84% higher than NUR 40%-60% population (NUR-M) and was 21.6% higher than NUR≤40% population (NUR-L). There were significant differences among the three populations. NUR was significantly positively correlated with grain yield. And different NUR populations were not distinct in dry matter accumulation amount and nitrogen accumulation amount before the stage of anthesis. With NUR improvement, dry matter accumulation amount after anthesis and at maturity, plant nitrogen content at anthesis, and population nitrogen accumulation amount at maturity increased, which were more than 6 000 kg·hm-2, 17 500 kg·hm-2, 1.50% and 215 kg·hm-2 in NUR≥60% population, respectively. Moreover, along with NUR increasement, decay of population photosynthetic area after anthesis gradually postponed, and net assimilation further increased, as well as flag leaves photosynthetic capacity and anti-oxidant ability after anthesis enhanced, which more obviously performed during the middle and late phase of grain filling. Therefore, photosynthetic material production capacity after anthesis was improved. Decay of leaf area, photosynthetic potential and net assimilation after anthesis in NUR≥60% population was about 0.14 LAI·d-1, 105×104 m2·d·hm-2 and 9.50 g·m-2·d-1, respectively. In summary, grain yield and NUR were relatively high in the treatment of early topdressing panicle fertilizer (flag leaf stage or booting stage) under the condition of appropriate postponing nitrogen application (nitrogen applied ratios= 3﹕1﹕3﹕3). Rather, yield improvement of high nitrogen application (262.5 kg·hm-2) was not obvious, and NUR even was quite low. The grain yield was more than 9 000 kg·hm-2 in experiments carried in two years, and NUR was the highest under the treatment of applied nitrogen amount 210.0 kg·hm-2, nitrogen applied ratios of basis﹕tillering﹕elongation﹕booting= 3﹕1﹕3﹕3 and topdressed nitrogen at flag leaf stage.【Conclusion】Under high nitrogen utilization efficiency condition, plant nitrogen nutrition level was high during the middle and late phases of plant growth and development, nitrogen absorption and accumulation amount increased in wheat following rice, which was better to improve transportation of nitrogen to grain. Additionally, the process of photosynthetic area decay and leaves senescence was postponed during the late phase, and photosynthetic material production capacity was improved, which could be the reason of simultaneous improvement in NUR and grain yield.
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    Cited: Baidu(2)
    Differences in Nitrogen Efficiency and Nitrogen Metabolism of Wheat Varieties Under Different Nitrogen Levels
    WANG Xiao-chun, WANG Xiao-hang, XIONG Shu-ping, MA Xin-ming, DING Shi-jie, WU Ke-yuan, GUO Jian-biao
    Scientia Agricultura Sinica    2015, 48 (13): 2569-2579.   DOI: 10.3864/j.issn.0578-1752.2015.13.009
    Abstract823)   HTML24)    PDF (532KB)(18076)       Save
    【Objective】This study aims to clarify the difference in nitrogen metabolism of wheat varieties with different nitrogen use efficiencies and to achieve optimization of nitrogen application and high-yield.【Method】In this study, Luomai 18 and Yumai 49-198(LH), Xinong 509 and Yunong 202 (LL) were selected from 16 wheat varieties in yield conditions, and GS activity, soluble protein, free amino acids, NO3- and total nitrogen content of leaves were detected under nitrogen levels of N0 (0, CK), N120 (120 kg·hm-2) and N225 (225 kg·hm-2). 【Result】The results showed that GS activity, soluble protein, free amino acids, NO3- and total nitrogen content in leaf, N accumulation of aboveground and grain, yield and NPE of Luomai 18 and Yumai 49-198 (LH) were significantly higher than that of Xinong 509 and Yunong 202 (LL). GS activity, soluble protein, free amino acids, NO3- and total nitrogen content in leaf, N accumulation of above ground and grain, yield increased with the increasing of nitrogen level, and the NPE reduced. But the response of the two types of varieties to nitrogen levels was different, compared with N0, the growing rate of GS activity, soluble protein, free amino acids, NO3- and total nitrogen content in leaf, N accumulation of above-ground of Xinong 509 and Yunong 202 (LL) were significantly higher than that of Luomai 18 and Yumai 49-198(LH), however, the growing rate of yield of Xinong 509 and Yunong 202 (LL) were significantly lower than that of Luomai 18 and Yumai 49-198(LH); the decreasing range of NPE of Luomai 18 and Yumai 49-198 (LH) was significantly higher than that of Xinong 509 and Yunong 202 (LL). 【Conclusion】Compared with Xinong 509 and Yunong 202 (LL), the reason that Luomai 18 and Yumai49-198 (LH) had higher yields and nitrogen use efficiency was that they had higher GS activities. Therefore the ability of nitrogen uptake and assimilation were improved, the nitrogen physiological efficiency in the whole nitrogen metabolism process was increased and better yields were obtained. Increasing the supply of nitrogen, yields of Luomai 18 and Yumai49-198 (LH) were significantly promoted. Luomai 18 and Yumai49-198 (LH) have a stronger tolerance to low nitrogen stress and the potential of yield is greater; Xinong 509 and Yunong 202 (LL) are more sensitive to nitrogen, but their ability in distribution of N is lower.
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    Cited: Baidu(6)
    Agronomic Traits Variation Analysis of Huanghuai Dryland Winter Wheat under Temperature Change Background in China ——Taking Linfen, Shanxi as an Example
    LIU Xin-yue, PEI Lei, WEI Yun-zong, ZHANG Zheng-bin, GAO Hui-ming, XU Ping
    Scientia Agricultura Sinica    2015, 48 (10): 1942-1954.   DOI: 10.3864/j.issn.0578-1752.2015.10.007
    Abstract623)   HTML22)    PDF (507KB)(16654)       Save
    【Objective】Crop variety breeding was result from nature and artificial selection under climate change background. Huanghuai wheat region is the largest wheat region in China, that plays an important role for ensure wheat production and food security in China. The changes of agronomic traits of winter wheat and temperature in Huanghuai dryland in the past 30 years were studied in order to provide a theoretical basis and technological support for dryland wheat breeding to adapt to climate changes in the future. 【Method】 The agronomic traits data of control varieties and temperature data of Linfen in Shanxi province in the past 30 years were analyzed. Linfen is the national representative Huanghuai dryland winter wheat regional test site. The regularity of the main temperature traits of Linfen city, such as the average temperature and over 0 accumulated temperature in winter wheat growth, and agronomic traits of control varieties change trends were analyzed. The relationship between agronomic traits and meteorological elements were also analyzed by correlation, multiple regression and path analysis. 【Result】 The average temperature,over 0 accumulated temperature,the highest and lowest temperature was increased slowly year by year in wheat growth period. In wheat growth period, the average temperature was increased average 0.05 per year, over 0 accumulated temperature was increased average 21.9per year,there was positive relationship between the average temperature and over 0 accumulated temperature. Over 0 accumulated temperature change can reflect temperature change in whole wheat growth period. The highest and lowest temperature was increased 0.02and 0.16per year respectively. The average temperature and over 0 accumulated temperature was increased markedly from sowing to before the winter, the average temperature and over 0 accumulated temperature was increased significantly at vegetative growth stage, but were increased lightly and had a downward trend at reproductive stage. With climate warming, strong winter variety with more tillers was replaced gradually by winter and weak winter variety with moderate tiller. Agronomic traits of control variety were changed not much from 1986 to 1996, and changed greatly from 1997 to 2007, and changed markedly from 2007 to 2014, this change trend consist with temperature change trend in wheat growth period. In evolution of agronomic traits of Huanghuai dryland control varieties, the thousand-kernel-weight and kernels per spike was increased average 1.57% and 3.39% per year respectively, but the effective ear numbers and plant height as well grain yield were decreased average slowly 0.16% and 1.29% per year respectively. There was a significant positive correlation between grain yield and plant height (0.684**), effective ear number (0.531**) and thousand-seed-weight (0.541**). There were 46.73%, 26.17% and 3.26% variation of grain yield was decided by three main factors such as plant height and over 0 accumulated temperature as well as the average temperature from standing to jointing stage respectively. In Huanghuai dryland wheat high-yield breeding evolution, the over 0 accumulated temperature from standing to jointing stage and plant height had a higher positive effect on grain yield, but the average temperature from standing to jointing stage had a negative effect on grain yield. 【Conclusion】 The climate warming had a strong impact on agronomic traits evolution in Huanghuai dryland winter wheat region, selecting new wheat varieties with agronomic traits of middle plant height, middle effective ears numbers, more kernels per spike, and high grain weight is the improvement direction of middle-high yield and drought-resistant varieties in Huanghuai dryland wheat region for adapting to climate change in the future.
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    Study on Producing Area Classification of Newhall Navel Orange Based on the Near Infrared Spectroscopy
    LIAO Qiu-hong, HE Shao-lan, XIE Rang-jin, QIAN Chun, HU De-yu, Lü Qiang1,YI Shi-lai, ZHENG Yong-qiang, DENG Lie
    Scientia Agricultura Sinica    2015, 48 (20): 4111-4119.   DOI: 10.3864/j.issn.0578-1752.2015.20.012
    Abstract616)   HTML31)    PDF (930KB)(16020)       Save
    【Objective】Newhall navel orange (Citrus sinensis L.) fruits from different producing areas in China, exhibit contrasting quality and market values, due to wide-spreading location of various ecologies. Developing a recognition method based on Near-Infrared (NIR) spectroscopy is very important to identify and distinguish fruits from different producing areas. 【Method】One representative orchard was selected from 17 main producing areas distributed throughout southern China, from which one 100 Newhall navel orange samples were collected. NIR spectra were collected with a SupNIR-1500 spectrograph from the surface of fruit equator and shoulder and the filtered juice for each sample, which were further preprocessed through principal component analysis (PCA) for reduced dimensions and noise. By studying artificial neural network (ANN), a classic three-layer ANN model was established with an input layer, a hidden layer of non-linear activation function and an output layer. By studying support vector machine (SVM) with the radial basis function (RBF) being the kernel function and the principal components of NIR spectra being the input, a one-to-one extended SVM model was established with 126 classifiers. Gene algorithm (GA) with excellent natural selection was used to identify the best Genetic character subset from the principal components as inputs of a SVM classifier, thus a GA-optimized SVM model was composed. These three models were used to classify the NIR spectra of filtered juice, thus the production areas of the oranges, the classification accuracies of which decided the best classifier. Furthermore, the accuracies of the best classifier were tested with the NIR spectra from fruit equator and shoulder surface being the inputs. As a comparison, the best NIR spectra could be identified. 【Result】 Producing area classification was implemented with the three-layer ANN classifier with NIR spectra of Newhall orange juice as the input, where the classifying accuracy reached up to 81.45% when there were 11 input neurons and 13 hidden neurons. The studied one-to-one extended SVM classifier with radial basis function being the core function, exhibited higher accuracy of 86.98% when the number of PC was 20, better than ANN classifier. For GA-SVM classifier took into account the interaction of individual inputs, where the PCA-processed results were optimized by GA. During the experiments, classification accuracy hit 89.72% when the population, generation, mating probability, and mutation probability were 200, 100, 0.7 and 0.01 respectively, surpassed ANN and SVM classifier. Subsequent research found the highest accuracy of GA-SVM classifier was 80% when taking the spectra from the fruit equator, and 69% from the shoulder, not good enough as that of orange juice.【Conclusion】Considering the accuracy, GA-SVM classifier was regarded with the most excellence among three investigated classifiers. Spectra of orange juice were selected as the best data to analyze origins traceability. Accuracy of spectra of fruit equator was inferior to juice but superior to the shoulder, thus had the potential for non-destructive origins classification.
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    Identification and Expression Analysis of 1-Aminocyclopropane- 1-Carboxylate Oxidase Gene from Quinclorac-Resistant Barnyardgrass (Echinochloa crus-galli)
    DONG Ming-chao, YANG Xia, ZHANG Zi-chang, LI Yong-feng, GUAN Rong-zhan
    Scientia Agricultura Sinica    2015, 48 (20): 4077-4085.   DOI: 10.3864/j.issn.0578-1752.2015.20.009
    Abstract489)   HTML16)    PDF (4441KB)(15373)       Save
    【Objective】The objective of this study is to clone barnyardgrass (Echinochloa crus-galli) 1-aminocyclopropane-1- carboxylate oxidase gene (EcACO), analyze its expression and test its enzyme activity, and to unravel the quinclorac-resistant mechanism of E. crus-galli to quinclorac.【Method】The partial sequence of EcACO obtained from E. crus-galli transcriptome pyrosequencing was used to design primers for cloning EcACO from quinclorac-resistant and susceptible E. crus-galli. EcACO was then cloned and sequenced. The nucleotide and putative amino acid sequence analysis were compared using DNAman and GenDoc softwares. The transcript levels of EcACO between resistant and susceptible biotype E. crus-galli were determined by real-time quantitative PCR (qRT-PCR) with β-actin gene as the reference. Finally, the open reading frame (ORF) sequences of EcACO from resistant and susceptible biotypes E. crus-galli were inserted into the expression vector pMAL-c5x, respectively. After the recombinant plasmids were transformed into Escherichia coli strain BL21, the fusion proteins were expressed by the induction with 0.4 mmol·L-1 IPTG for 16 h at 18℃. The soluble proteins were purified with MBP column for the measurement of ethylene released from MBP::EcACO fusion protein. 【Result】EcACO was isolated from E. crus-galli with quinclorac-resistant and susceptible biotypes of E. crus-galli. The ORF of EcACO was 936 bp, encoding 311 amino acids, with pI 5.4 and Mw 35 kD. The deduced amino acid sequences shared high identity with other ACO sequences from Setaria italica (93%), Zea mays (92%) and Sorghum bicolor (91%). Compared with EcACO from the susceptible biotype, five site mutations of EcACO were found in the resistant biotype, of which three site mutations were located in the putative conserved domain. Furthermore, qRT-PCR results showed that there was no significant difference in expression level of EcACO between resistant and susceptible biotype. Using the prokaryotic expression system and the measurement of MBP::EcACO activity, the released amount of ethylene in the MBP::EcACO from susceptible biotype was 2.15 folds higher than that from resistant biotype.【Conclusion】EcACO was identified from quinclorac-resistant and susceptible E. crus-galli. Compared with the susceptible biotype, the EcACO from the resistant one had five amino acid mutations, of which three site mutations were in the conserved domain. This might probably contribute to the reduction of released amount of ethylene and result in quinclorac resistance of E. crus-galli.
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    Molecular Cloning and Function of the Yellow Gene from Nilaparvata lugens
    WANG Bo, YAO Yun, XU Ze-wei, LIN Xin-da
    Scientia Agricultura Sinica    2015, 48 (15): 2976-2984.   DOI: 10.3864/j.issn.0578-1752.2015.15.007
    Abstract646)   HTML28)    PDF (5409KB)(14861)       Save
    【Objective】The objectives of this study are to clone NlYellow from the brown planthopper (Nilaparvata lugens), explore the expression profiles during the development in adults and in different tissues, then study its function through RNA interference.【Method】Primers were designed by online version of primer 3 software and NlYellow was cloned, then the nucleotide sequence was translated into amino acid sequence. The alignment of amino acids residues was conducted using N. lugens Yellow and others searched from GenBank. Also a Neighbor-Joining method was used to construct a phylogenetic tree. Quantitative real-time PCR (qRT-PCR) was applied to measure the relative expression levels of N. lugens at different developmental stages (embryo, 1st-5th instar nymphs and adult) and in different tissues (head, thorax, abdomen, leg, wing, midgut, ovary, and testis). NlYellow dsRNA at 0.4 µg was injected into 5th instar nymphs and the adult phenotypes were observed. 【Result】The NlYellow was cloned, and through sequence alignment it was found that the NlYellow was more similar to the ApYellow (98%), but it was less conserved when compared with that of Drosophila melanogaster and Bombyx mori.Phylogenetic analysis showed that it clustered with Acyrthosiphon pisum,too. NlYellow’s expression was variable in the embryo, the expression level in 1st, 3rd, 4th and 5th day were lower than other days. Furthermore, its expression levels were higher in 3rd and 5th instar nymph than others (P<0.05). Tissue-specific expression analysis elucidated that NlYellow expressed in all tissues including head, thorax, abdomen, leg, wing, midgut and testis in male, but the first four tissues had higher expression level (P<0.05). On the contrary, NlYellow was only expressed in head, thorax, leg, and wing in female. Furthermore, its expression level in short wing adult was higher than that of long wing adult (P<0.05). Also in short wing adults, male’s expression level was higher than female (P<0.05). Using RNA interference to knock down the NlYellow expression, it was found that the color of the whole body turned into yellow, especially in thorax, abdomen and legs.【Conclusion】NlYellow might involved in insect pigmentation and change of the body colour.
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    Research on Digitizing Morphological Structure and Growth Process of Grape Tree
    WEN Wei-liang, WANG Yong-jian, LI Chao, WANG Chuan-yu, GUO Xin-yu
    Scientia Agricultura Sinica    2015, 48 (11): 2143-2151.   DOI: 10.3864/j.issn.0578-1752.2015.11.006
    Abstract740)   HTML37)    PDF (934KB)(14233)       Save
    【Objective】 Describing the morphology structure and growth process of plants in a digital way is an effective means of correctly understanding and expressing the development regulation of plants. In order to accurately describe and quantitatively analyze the grape trees with complex morphological structure, the digital expression and processing technologies of grape trees were studied in depth. 【Method】 The basic structure unit was divided and defined according to the morphological structure and growth process of grape tree, including the truck, perennial canes, new shoot and organs on the shoots. The new shoots include bearing and vegetative shoots. Organs on the shoots include buds, inflorescence, ears, secondary shoots, leaves and tendrils. Referencing the experience, habits, and trend of structure naming strategy at home and abroad, combining the actual field observation and following the temporal and space order of growth sequences of each structure unit, a naming scheme for the above ground structure units of grape trees at an annual rate was proposed. The first letters indicate the type and the figures showing the birth order of structure unit. The naming scheme also contains information of the growth and branching source relationship between the relative structure units. It integrated the shoot change process from the initial new shoot to the cane and then to the perennial cane under the perennial mechanism. The new shoots were constituted using an ordered set way in the scheme. On the basis of the naming scheme, the representation method of compound organs and complex operations was given, including representation method of complex organs such as bearing basal shoots and bearing shoot group; digital description of grape tree structure changes by artificial pruning; growth representation of grape trees at an annual rate; growth process backtracking of perennial grape trees; quantitative statistical analysis method of each structure unit. 【Result】 Taking the short shoot pruned V font shape Cabernet Sauvignon grape tree in Zhengzhou National Grape Germplasm Resources Garden and T font shape Xiangfei grape tree in the experimental greenhouse in Institute of Forestry and Pomology of Beijing Academy of Agricultural and Forestry Sciences as an example, the digital expression of the selected two trees was given respectively. The digital expression realized a clear representation of the two trees with complex structures, including the unique naming of each organ and the connection relationship of each other. On the basis of the structure expression, structure units on a new shoot were counted to make a quantitative statistic. The morphological structure change expression such as secondary shoot and tendril pruning and tree annual growth were illustrated as well. The application of digitized grape trees showed the universality and effectiveness of the method for the description of different forms and different growth periods of grape trees. 【Conclusion】 This study contributes to a more intuitive understanding of grape tree morphological structure and growth mode. It has important significance for constructing the functional-structural model, for realizing precisely describing morphological structure of grape trees, visualization expression and quantitative analysis, etc. The study also has important significance for quantitative calculating and simulating the condition, development and evolution of the internal factors of grape trees, and for further prediction and evaluation of these factors. At last, the structure unit definition and naming scheme have reference values to research the woody plants of similar morphological structures.
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    Cited: Baidu(1)
    Overexpression of a Transcription Factor Gene SiNF-YA6 from Millet (Setaria italica) Enhanced the Resistance of Transgenic Plants to Nitrogen Starvation
    FANG Guang-ning, HU Li-qin, WANG Er-hui, XUE Fei-yang, MA You-zhi, XU Zhao-shi, LI Lian-cheng, ZHOU Yong-bin, DIAO Xian-min, JIA Guan-qing, CHEN Ming, MIN Dong-hong
    Scientia Agricultura Sinica    2015, 48 (20): 3989-3997.   DOI: 10.3864/j.issn.0578-1752.2015.20.001
    Abstract682)   HTML18)    PDF (2772KB)(13079)       Save
    【Objective】Nuclear transcription factor Y (NF-Y) belongs to a transcription factor gene family widely existing in eukaryotic. NF-Y-like transcription factors are involved in the process of gene expression regulation of plant growth and development and stress responses. So far, it is never been reported that NF-Y-like transcription factors are involved in plant resistance to low nitrogen stress. In this study, a NF-Y-like transcription factor gene SiNF-YA6 was isolated from millet, and its characteristics and biological function were identified.【Method】A NF-Y-like transcription factor gene SiNF-YA6 highly up-regulated by low nitrogen stress was selected from the results of transcriptome sequencing of millet treated by low nitrogen. DNAMAN and MEGA6.0 software were used to make the phylogenetic tree of NF-YA6. The Real-time PCR was used to analyze expression patterns of SiNF-YA6 under different treatments. SiNF-YA6 was transformed into Arabidopsis protoplast to observe its subcellular localization. SiNF-YA6 was overexpressed in Arabidopsis to identify gene function.【Result】SiNF-YA6 is 714 bp in length and encoding 237 amino acids. SiNF-YA6 contains a CBF conservative domain with 62 amino acids. Phylogenetic tree analysis showed that NF-YA was fell into 6 classes, and SiNF-YA6 belongs to the fifth class. SiNF-YA6 was the highest homology with a rice NF-YA protein (Os03g07880) (77.4%). Expression pattern analysis showed that NF-YA6 was induced by low nitrogen stress, and its expression reach peak after treatment for 5 d that raised about 5 times compared with the expression level at the beginning of treatment. While transcription of SiNF-YA6 under high salt, drought, ABA treatment also showed a trend of increasing. SiNF-YA6 under SA and GA treatments was no obvious different. Tissue specific expression analysis showed that SiNF-YA6 mainly expressed in millet roots. Subcellular localization indicated that SiNF-YA6 was mainly located on the plasmalemma and nucleus in plant cell. Results of low nitrogen stress resistance test showed that the main root length and root surface areaof SiNF-YA6 transgenic Arabidopsis were significantly higher than in wild type Arabidopsis, and the fresh weight of transgenic plants was also significantly greater than the wild type plants (P<0.01) under low nitrogen stress (total nitrogen content of 1 mmol·L-1), and the growthof SiNF-YA6 transgenic Arabidopsis showed no obvious difference with wild-type Arabidopsis under SA and GA treatments. Those results indicated that overexpression of SiNF-YA6 gene significantly enhanced transgenic plants resistance to low nitrogen stress. The downstream genes, including some nitrogen transporter genes (NRT2.1 and NRT2.4), were up-regulated in SiNF-YA6 transgenic Arabidopsis. However, SiNF-YA6 transgenic Arabidopsis showed no obvious difference with wild-type Arabidopsis under high salt and drought stresses.【Conclusion】Overexpression of SiNF-YA6 enhanced transgenic plants tolerance to low nitrogen stress by up-regulated expression of several nitrogen transporter genes, which will give new candidate for improvement of the nitrogen use efficiency in crops.
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    Biochemical and Molecular Characteristics of Glutamic Decarboxylase from Bactrocera dorsalis
    WEI Dong, WANG Tao, DOU Wei, WANG Jin-Jun
    Scientia Agricultura Sinica    2014, 47 (16): 3184-3194.   DOI: 10.3864/j.issn.0578-1752.2014.16.007
    Abstract537)   HTML15)    PDF (722KB)(12956)       Save
    【Objective】 The study aimed to determine the γ-aminobutyric acid (GABA) content and glutamic acid decarboxylase (GAD) specific activity of Bactrocera dorsalis, and clone the complete sequence of a GAD gene (BdGAD1). The changes of GABA content, GAD specific activity, and expression of BdGAD1 in different developmental stages and body tagmata of adults after avermectin stimuli provided basic data of the resistance mechanism of avermectin mediated by GABA. 【Method】 The content of GABA in B. dorsalis was determined through the method of high performance liquid chromatography, and the dose and time effects of avermectin stimuli on GABA content were determined. The change of specific activity of GAD in B. dorsalis was determined via the microplate method with the substrate of glutamate. According to the screened GAD gene sequence fragment from the transcriptome data of B. dorsalis, the complete sequence of cDNA was amplified using rapid amplification of cDNA ends (RACE). The open reading frame (ORF), deduced amino acid sequence, and molecular weight were predicted, and a phylogenetic tree with GAD genes from other insects was constructed using maximum likelihood method to clarify its phylogenetic relationship. Besides, the RNA was extracted from different developmental stages (egg, larva, pupa, and adult) and different tagmata (head, thorax, and abdomen) of adult. Based on the reference evaluation, α-Tubulin was used as housekeeping gene for qPCR to analyze the expression profiles of different developmental stages, tagmata, and stimulated by avermectin. 【Result】 The GABA contents of B. dorsalis increased under the stimuli of avermectin, and there was a positive correlation between GABA content and the avermectin dose and treatment duration, suggesting that B. dorsalis may mediate the content of GABA to avoid the damage of avermectin. Moreover, the specific activity of GAD in B. dorsalis also increased with the increase of treatment doses. A complete sequence of BdGAD1 was cloned by RACE amplification with a full length of 1 755 bp, and ORF of 1 197 bp encoding 398 amino acids. The GenBank accession number was KC763804. This gene exhibited a close relationship with the gene from Anopheles gambiae based on maximum likelihood phylogenetic tree. The amino acid identity was up to 97%. The expression level of BdGAD1 was the highest in larva among different developmental stages, and was the highest in abdomen among different tagmata. Under the stimuli of avermectin, the expression of BdGAD1 was also unregulated.【Conclusion】 Avermectin could increase GABA content by increasing the expression level of BdGAD1 and specific activity of GAD resulting in more GABA synthesis. This might be one reason for the resistance of B. dorsalis against avermectin.
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    Expression, Function and Regulation of Chitin Synthase 2 Gene in Locusta migratoria
    LIU Xiao-Jian, CUI Miao, LI Da-Qi, ZHANG Huan-Huan, YANG Mei-Ling, ZHANG Jian-Zhen
    Scientia Agricultura Sinica    2014, 47 (7): 1330-1340.   DOI: 10.3864/j.issn.0578-1752.2014.07.010
    Abstract616)   HTML20)    PDF (903KB)(11101)       Save
    【Objective】Chitin synthase is one of the key enzymes responsible for chitin synthesis in insects. As this enzyme is absent in higher animals, it could be served as a potential target for developing safe and effective insecticides. In our earlier research, the cDNA of chitin synthase 2 gene (LmCHS2, GenBank accession number: GU067731) in Locusta migratoria was cloned. The objectives of this paper are to further study the expression, function and regulation of LmCHS2, and to provide a scientific basis for effective pest control using RNAi methods.【Method】Based on the nucleotide sequence of LmCHS2, a pair of specific expression primers was designed, the expression patterns of LmCHS2 were studied in eggs, nymphs and adults by RT-qPCR. The dsRNA of LmCHS2 was synthesized in vitro, and then injected into the female or male adults on day 1, respectively. The midguts dissected from the injected insects on day 5 were pooled for each RNA extraction. cDNA synthesis and RT-qPCR were performed to determine the down-regulation of LmCHS2. After dissected the whole gut, the midgut changes and integrity of peritrophic matrix (PM) were observed to explore the biological functions of this gene in L. migratoria adults. Locusts were maintained with no food in different times, and feeding again, to observe the changes of guts. Then the transcript levels of LmCHS2 were detected by RT-qPCR. 【Result】 LmCHS2 was almost undetectable during the early and middle embryogenesis, but dramatically up-regulated in late eggs. It was consistently expressed throughout the nymphal and adult stages. After dsCHS2 was injected into the female or male adults on day 1, significantly reduced transcript of LmCHS2 was observed as compared with that of the controls, and resulted in a decreased feeding and a high mortality of insects (78% for female and 85% for male adults). After dissection, it was found that there was virtually no food contained in dsCHS2-injected insects and the average length of midguts and gastric caeca was shorter than that of the control. Furthermore, histological observation of midguts showed that the control locusts contained a fully developed PM, however, locusts injected with dsCHS2 exhibited a disrupted PM or even absence of the PM. Locusts were treated under starvation for 48 h, the midguts hardly contained food and the average length of midguts was significantly shorter than that of the control midguts. From the H & E stained results, it was found that the PM was almost absent in non-fed midguts while the PM of control midguts was well-structured, which was very similar with the RNAi. But after fed again, the insects contained a fully developed PM. When locusts were maintained with no food for 24 h and 48 h, the transcript levels of LmCHS2 were suppressed significantly. When locusts were fed for another 0.5 h period, the transcript levels increased to the control level rapidly, which suggested that feeding affected the expression of LmCHS2. 【Conclusion】LmCHS2 is responsible for chitin biosynthesis of peritrophic matrix of the midgut and plays a key role for the development of L. migratoria. The decreased expression of this gene affected the integrity of the PM, thus hindered the food absorption and led to the mortality of the locusts. In addition, feeding regulated the expression of LmCHS2.
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    Screening of Putative Proteins in Vector Psammotettix alienus L. that are Interacted with Coat Protein of Wheat dwarf virus by a Split-ubiquitin Yeast Membrane System
    ZHAO Yi-ze, LIU Yan, WANG Xi-feng
    Scientia Agricultura Sinica    2015, 48 (12): 2354-2363.   DOI: 10.3864/j.issn.0578-1752.2015.12.008
    Abstract568)   HTML18)    PDF (2842KB)(10459)       Save
    【Objective】To investigate the interaction between the leafhopper (Psammotettix alienus L.) and Wheat dwarf virus (WDV), a cDNA library of leafhopper was constructed using a split-ubiquitin yeast membrane system. The protein interaction analysis was done by using WDV CP as bait protein to screen a cDNA library of P. alienus. 【Method】Total RNA of leafhopper was isolated from 2 g of insects. Poly A+ RNA was enriched from 100 ng of total RNA and double-stranded cDNA was synthesized using SMART technology. After digestion with the Sfi I enzyme, the fragmented cDNA was ligated to prey vector pPR3-N, and then also digested with Sfi I enzyme to construct the split-ubiquitin yeast membrane system cDNA library. The full-length gene, WDV CP, amplified from wheat leaves infected by WDV was ligated into bait fusion vector, pDHB1. After functional assay, pDHB1-WDV CP vector was co-transformed into NMY51 with empty library vector in order to get an optional concentration of 3-AT. Then using the split-ubiquitin yeast membrane system, proteins interact with the bait pDHB1-WDV CP were screened from the cDNA library of P.alienus. Gene ontology (GO) and pathway information of proteins were analyzed from Uniprot and KEGG websites.【Result】Detection of the cDNA library showed that the unamplified library contained 2.0×106 independent clones, the titer of the amplified library was 1.3×106 cfu. The recombination rate was above 97%. The sizes of most inserts were above 1 kb in the cDNA library. The correct ligated fusion bait vector pDHB1-WDV CP was verified by restriction enzyme digestion analysis and sequencing. Functional assay showed that the fusion protein was functionally correctly expressed in the yeast and suited to this system. In library screen test, 280 clones were got from the cDNA library of P. alienus. Twelve proteins were selected for further research based on the functional analysis in terms of GO. Finally, 9 proteins confirmed by β-galactosidase assay were interacted with WDV CP. GO annotation analysis showed 9 putative proteins were involved in 10 biological processes including protein dephosphorylation, carbohydrate metabolic process, transport, etc. Molecular functions included metal ion binding, phosphate ion carrier activity, folic acid transporter activity, protein complex binding, etc. These proteins also were involved in ubiquitin mediated proteolysis, endocytosis, arachidonic acid metabolism, cAMP signaling pathway, PPAR signaling pathway.【Conclusion】A high-quality cDNA library was constructed and 9 proteins were interacted with WDV CP, which could be used for insect vector and WDV interaction analysis.
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    Effect of Root Layer Regulations on Soil Nitrate of the Rotation System of Sweet Corn-Cucumber in Greenhouse Vegetable Production System
    HAO Xiao-ran, PENG Ya-jing, ZHANG Li-juan, WANG Lin, JU Xiao-tang, JI Yan-zhi, REN Cui-lian
    Scientia Agricultura Sinica    2015, 48 (12): 2390-2400.   DOI: 10.3864/j.issn.0578-1752.2015.12.012
    Abstract615)   HTML20)    PDF (447KB)(9838)       Save
    【Objective】 Taking sweet corn as a catch crop, the accumulation of nitrate in soil profile under different root layer regulation and control measures and their influence on growth of succession cucumber were studied. 【Method】 Sweet corn was planted in the fallow season of the traditional greenhouse in the North China Plain. Using traditional cultivation as the control, a field plot experiment was conducted with 2 root layer regulation and control measures: adding soil conditioner and straw returning to field. The experiment was carried out from May 2008 to May 2011, a total of three sweet corn-cucumber crop rotation, six crops were planted. Sweet corn was planted from early June to September every year, and cucumber seedlings were reared from the early October to the end of January next year. Cucumber plants were planted in early February the same year. In the sweet corn season, three treatments were conducted and arranged at random with three replications. The plot size was about 4 m×2 m, plot interval was 0.3 m, and 1 m of guard rows were laid out between plots. 【Result】In the sweet corn planting seasons, the highest grain yields were obtained in soil conditioner treatment and the yields were 6.2, 7.4 and 7.9 t·hm-2 in 2008, 2009 and 2010. The total N uptake rate in soil conditioners and straw returning to field treatments was higher than the traditional planting. After planting sweet corn for three seasons, straw and soil conditioner treatments promoted the growth and development of root system in 20-60 cm soil layer and enhanced root to absorb nutrients from deeper soil. Both the two treatments reduced the NO3--N of soil profile, especially the subtraction of NO3--N in 100-200 cm of the crop root zone, the NO3--N subtractive trend roughly was soil conditioner>straw returning of field>the traditional planting. After cucumber harvest in three seasons, the differences in cucumber yield, the biomass and nitrogen uptake were not significant, the average soil NO3--N residue of three seasons in 0-200 cm was in the order of straw returning of field<soil conditioner<the traditional planting. After three cropping seasons, the nitrogen surplus in 0-200 cm soil layer in the traditional planting, soil conditioner and straw returning to field treatments was, respectively, 1 911.6, 1 966.83, and 1 930.2 kg·hm-2, and conditioning treatment was significantly higher than that of traditional planting. 【Conclusion】In the greenhouse soil with high nitrate accumulation, along with the increasing length of planting, adding soil conditioner to soil and appropriate straw returning to field, the subtraction of NO3--N in 100-200 cm of the crop root zone was more than traditional planting. Planting catch crops significantly decreased the effects of soil NO3--N in soil of the succession cucumber. Soil conditioners and straw returning to field can significantly improve the ability of sweet corn to decrease soil NO3--N, slow soil NO3--N leaching, and improve economic returns.
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    Cited: Baidu(1)
    Molecular Characterization and Function of Chitinase 10 Gene (OcCht10) from Oxya chinensis
    LI Da-Qi-1, WANG Yan-1, ZHANG Jian-Qin-1, LI Tao-1, SUN Yi-2, ZHANG Jian-Zhen-1
    Scientia Agricultura Sinica    2014, 47 (7): 1313-1320.   DOI: 10.3864/j.issn.0578-1752.2014.07.008
    Abstract556)   HTML7)    PDF (642KB)(9657)       Save
    【Objective】 The objectives of this study are to obtain cDNA sequence of chitinase 10 gene (OcCht10) from Oxya chinensis, analyze its functional domain and phylogenetic relationship with chitinases from other known insect species, investigate its expression patterns and biological function during molting process, and to provide a new candidate gene for pest control.【Method】 cDNA fragments of OcCht10 were searched from O. chinensis’ transcriptome database. After blast analysis, the cDNA sequence of OcCht10 was assembled and translated, the functional domains of OcCht10 were predicted by bioinformatics methods. Phylogenetic analysis was performed with other insect chitinase 10 amino acid sequences. The first-stranded cDNAs were synthesized by using RNA isolated from integument of each day of 5th instar nymphs and various tissues of the 6th day in 5th instar nymphs. Reverse transcription quantitative PCR (qPCR) was carried out to analyze the gene expression patterns. Biological function of OcCht10 was studied by RNA interference method. The dsRNA primers were designed for dsOcCht10 synthesis in vitro. The dsRNAs were injected into the 2nd day of 5th instar nymphs for RNA interference, integument was dissected for silencing efficiency detection at 24 h after injection by using qPCR method. The phenotype was carefully observed and mortality was calculated till control insects molted to adults.【Result】 The obtained cDNA (9 318 bp) of OcCht10 contained an open reading frame of 8 613 bp, encoding 2 870 amino acid residues and a non-coding region of 705 bp at 3′ end. There were about 500 bp lost in 5′ end. The deduced amino acid sequence included five chitinase catalytic domains and six chitin binding domains. Phylogenetic analysis showed that OcCht10 belonged to chitinase group Ⅱ, the genes from this group were crucial for insect molting based on references. Tissue specific expression analysis of OcCht10 showed that it was predominately expressed in the integument, foregut and hindgut, which developed from ectoderm. The results suggested that OcCht10 may be involved in chitin metabolism of insect integument. Developmental expression patterns showed that OcCht10 was highly expressed before and after molting stages, lower in middle stages of 5th instar nymphs, which implied that OcCht10 could digest chitin of integument during molting process. RNA interference results indicated that the corresponding transcript level was silenced by 70% after OcCht10 dsRNA injection. Compared with the dsGFP injected control group, the nymphs injected with OcCht10 dsRNA displayed slow development and failed to detach old cuticle during molting, the mortality reached 100%.【Conclusion】 The partial cDNA sequence of OcCht10 was obtained from O. chinensis, the mRNA expression of OcCht10 was higher in the integument before molting; OcCht10 is involved in O. chinensis molting process, and dsOcCht10 injection can effectively silence mRNA expression of this gene and result in the block of ecdysis and even death of O. chinensis.
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    Cited: Baidu(2)
    Effect of Irrigation and Fertilizer Coupling on Greenhouse Tomato Yield, Quality, Water and Nitrogen Utilization Under Fertigation
    XING Ying-ying, ZHANG Fu-cang, ZHANG Yan, LI Jing, QIANG Sheng-cai, WU Li-feng
    Scientia Agricultura Sinica    2015, 48 (4): 713-726.   DOI: 10.3864/j.issn.0578-1752.2015.04.09
    Abstract1047)   HTML26)    PDF (550KB)(9571)       Save
    【Objective】Water and fertilizer are two major factors for limiting crop yield increase. Unreasonable application of irrigation and nitrogen can hinder yield increase, but increase the soil nitrate accumulation, and reduce the crop quality, water and nitrogen use efficiency. The study was aiming at dealing with the problems of irrigation and fertilization on greenhouse vegetables in semi-arid northwest areas, which through the effects of irrigation and fertilizer coupling on greenhouse tomato yields, quality, and water and nitrogen use efficiency under fertigation. The purpose of this study was to research a high-yield, good-quality and high-efficiency fertigation system for drip-irrigated greenhouse tomato. 【Method】Greenhouse tomato plot experiments were conducted to analyze the responsive rules of irrigation amount and fertilizer rate on tomato growth, yield, quality, distribution of soil NO3--N, water and nitrogen absorption and utilization. The experiments consisted of a furrow fertigation treatment (control treatment, 100% ET0, N240-P2O5120-K2O150 kg·hm-2), three drip irrigation levels (high irrigation, W1, 100% ET0; medium irrigation, W2, 75% ET0; low irrigation, W3, 50% ET0) and three fertilizer levels (high fertilizer, F1, N240-P2O5120-K2O150 kg·hm-2; medium fertilizer, F2, N180-P2O590-K2O112.5 kg·hm-2; low fertilizer, F3, N120-P2O560-K2O75 kg·hm-2), and total of 10 treatments.【Result】The results showed that tomato yield, dry matter accumulation and total nitrogen absorption in drip fertigation treatments were 31.04 t·hm-2,  3 208 kg·hm-2 and 73.13 kg·hm-2 higher than that in the furrow fertigation treatment, increased by 46.9%, 54.0% and 82.4%, respectively. In addition, the fruit Vc content, water use efficiency (WUE) and nitrogen use efficiency (NUE) increased by 61.8%, 46.4% and 76.5%, respectively, but the NO3--N content in the soil decreased. In drip fertigation treatments, the highest total dry matter accumulation of W1F2 treatment was 9 248 kg·hm-2, both yield and plant nitrogen uptake were positively related to the amount of irrigation and fertilizer. The fertilization was found to exert more impact on the yield compared with irrigation. The treatment W1F2 had thelargest range of yield increase and nitrogen uptake. The WUE decreased and the NUE raised gradually with the increase of irrigation amount and decrease of fertilizer rate. The treatment W3F1 produced the largest WUE (47.7 kg·m-3) and the W1F3 treatmentof the largest NUE (65.6 %), respectively. The WUE of W3F2 and the NUE of W1F2 had much more greater increasing scale than the other treatments. The nitrate nitrogen content in soil was significantly affected by irrigation, fertigation and irrigation-fertigation interactive effects. Nitrate nitrogen content in the soil increased first but then decreased with the increase of irrigation water. Nitrate nitrogen content consistently increased as the increase of fertilizer. Significant nitrate nitrogen accumulation was identified at the lateral boundary of the wetted volume rather than under the dripper. The treatment W1F2 with high irrigation water and medium fertilizer produced lower nitrate nitrogen content and more uniform distribution of nitrate nitrogen in the soil than other treatments. Increases in irrigation water significantly reduced fruit Vc, lycopene and soluble sugar contents and their nutrition accumulation, but the quality indicators’ content and nutrition accumulation were increased first then decreased with the increase of fertilizer. The treatment W3F2 had the largest Vc and lycopene content as well as nutrition accumulation, and the largest soluble sugar contents as well as larger nutrition accumulation. 【Conclusion】The results indicated that the drip fertigation technology could achieve the goal of high yield, good quality and high efficiency for greenhouse tomatoes. The treatment W1F2 with high water and medium fertilizer (100%ET0, N180-P2O590-K2O112.5 kg·hm-2) was preferable for higher yield and NUE interest, as well as lower nitrate nitrogen content in soil; but the treatment W3F2 with low water and medium fertilizer (50%ET0, N180-P2O590-K2O112.5 kg·hm-2) was more appropriate in terms of higher WUE and the largest Vc, lycopene and soluble sugar contents.
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    Effects of Water and Nitrogen Coupling on Root Growth and Single Fruit Weight of Greenhouse Muskmelon
    YUE Wen-jun, ZHANG Fu-cang, LI Zhi-jun, WU Li-feng
    Scientia Agricultura Sinica    2015, 48 (10): 1996-2006.   DOI: 10.3864/j.issn.0578-1752.2015.10.012
    Abstract541)   HTML8)    PDF (489KB)(9152)       Save

    【Objective】The objective of this experiment is to study the effects of different water and nitrogen levels on root growth and a fruit weight of greenhouse muskmelon, to explore the relationships between root growth, single fruit weight and water and nitrogen supply, and to provide a scientific basis for water and reasonable nitrogen application of local greenhouse muskmelon.【Method】According to the environment parameters (light, temperature, humidity, etc.) inside the solar greenhouse, ‘Yi pin tian xia 208’ of muskmelon was chosen as the test cultivar, irrigation application amount was determined by using modified Penman- Monteith equation. A total of 9 treatments were designed. Muskmelon plants were subjected to three irrigation treatments, that wass 70%, 100% and 130% of crop ETc (Evapotranspiration) and three N levels, including low nitrogen level N1 (70 kg N·hm-2), traditional nitrogen level N2 (130 kg N·hm-2) and high nitrogen level N3 (180 kg N·hm-2). Treatments were completely arranged randomly. A greenhouse muskmelon crop was grown under mulched drip irrigation conditions to investigate the effects of different water and nitrogen levels on root growth, distribution and single fruit weight.【Result】 Most of the total root lengths were concentrated on the 0-30 cm soil layer. Total root length increased slowly as soil layer increased. Total root length, single fruit weight and water use efficiency of muskmelon descended with the rising of irrigation water amount at the same nitrogen level. The highest values of total root length and single fruit weight were observed in W2N2, which were 6 625.48 cm and 818.94 g, respectively. Total root length and single fruit weight of muskmelon showed a trend of decreasing after the first increasing with the rising of irrigation water amount at the same nitrogen level. Water use efficiency descended with the rise of irrigation application rate, PFPN descended with the rise of nitrogen application rate. There was a significant relationship between root length with less than 2 mm, root dry weight and yield,and the correlation reached a significant level, the more root growth, the higher yield would increase, reasonable amount of irrigation water and nitrogen fertilizer could promote root system to absorb water and nutrients, thus increasing production.【Conclusion】The change rule of total root length in the depth of root vertical direction could be simulated by the equation y=A(1-Bx), the equation determination coefficient R2 reached more than 0.9. The principle component analysis method was used to evaluate the root growth of muskmelon, and the results showed that comprehensive principle component could represent 97.27% total root information, the highest comprehensive evaluation was obtained in W2N2. Unreasonable irrigation and nitrogen applications could result in reducing single fruit weight, characteristic parameters of muskmelon root, water use efficiency and partial factor productivity from applied N. It was concluded that the irrigation level W2 and nitrogen level N2 could be recommended as the best combination of water and nitrogen, which promoted the root growth, improved single fruit weight, water and nitrogen use efficiency of muskmelon in the muskmelon production under drip irrigation with plastic film mulching in the experimental area.

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    Cited: Baidu(3)
    Effects of Exogenous Nitric Oxide on Main Growth and Physiological Characteristics of Seedlings of Cotton Under NaCl Stress
    LI Cui-Fang, LIU Lian-Tao, SUN Hong-Chun, ZHANG Yong-Jiang, ZHU Xiu-Jin, LI Cun-Dong
    Scientia Agricultura Sinica    2012, 45 (9): 1864-1872.   DOI: 10.3864/j.issn.0578-1752.2012.09.023
    Abstract1073)      PDF (591KB)(7539)       Save
    【Objective】 The purpose of this study is to clarify the effects of nitric oxide (NO) on cotton seedlings growth and leaf physiological characteristics under NaCl stress. 【Method】 In a hydroponic experiment, two NaCl concentrations (50 mmol•L-1 and 100 mmol•L-1), four sodium nitroprusside (SNP) concentrations (0, 50, 100 and 200 μmol•L-1) for each NaCl level were designed, and at the same time a conventional culture was set. The growth and leaves physiological characteristics of cotton seedlings were measured after 10 days of treatment.【Result】 The results showed that the treatment of NaCl stress inhibited the growth of cotton seedlings. The internodes of the cotton seedlings under NaCl stress were inhibited further by SNP which is an exogenous nitric oxide donor. The relative water content of cotton seedlings, especially the relative water content of the stem and root were decreased in SNP treatments under NaCl stress. The treatments of SNP added in 100 mmol•L-1 NaCl stress increased the contents of soluble protein and chlorophyll, and the ratio of chlorophyll a/b, the effects of 50 μmol•L-1 SNP was the best. Under both of the NaCl stresses, SNP improved the activities of SOD and POD significantly, while the activities of CAT decreased.【Conclusion】 Under this experimental condition, exogenous nitric oxide inhibited internodes elongated, alleviated chlorophyll degradation, enhanced antioxidant capacity and improved leaves quality of cotton seedlings under NaCl stress. The SNP concentration from 50 μmol•L-1 to 100 μmol•L-1 could alleviate NaCl stress for cotton seedlings, the cotton seedlings under NaCl stress were seriously injured by high concentrations (200 μmol•L-1)of SNP.
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    Cited: Baidu(4)
    Advances in Research of Transcriptional Regulatory Network in Response to Cold Stress in Plants
    LIU Hui, LI De-jun, DENG Zhi
    Scientia Agricultura Sinica    2014, 47 (18): 3523-3533.   DOI: 10.3864/j.issn.0578-1752.2014.18.001
    Abstract709)   HTML9)    PDF (864KB)(6591)       Save
    Cold stress seriously influences plant growth, development, and crop yield. In order to survive, plants have evolved complex and high-efficiency regulatory networks to respond and adapt to cold stress. Among these regulatory networks, transcriptional regulation plays crucial roles. Transcription factors can regulate a set of genes by binding to cis-acting regulatory elements in the promoter regions, and play crucial roles in abiotic stress-responsive transcriptional regulatory network in plants. In this review, the authors comprehensively summarized the transcription factors involved in regulating plant response to cold stress, including AP2/ERF (APETALA2/ethylene responsive factor), MYB (myeloblastosis), bHLH (basic helix-loop-helix), NAC (NAM, ATAF1, ATAF2 and CUC2), ZFP (zinc finger protein), WRKY, VOZ (vascular plant one zinc-finger protein), CAMTA (calmodulin-binding transcription activator), and EIN3 (ethylene-insensitive 3). Their structure characteristics were simply summarized, while their functions and regulatory mechanisms were emphatically introduced. Based on the relationships among transcription factors, a transcriptional regulatory network diagram of plant response to cold stress was draw. In this network, CBF (C-repeat binding factor) transcription factors are considered as master molecular switches. CBFs specifically bind to the DRE/CRT (dehydration-responsive element/C-repeat element, A/GCCGAC) cis-acting regulatory element of the promoter region of the cold-responsive genes, such as COR (cold regulated), LTI (low-temperature induced), DHN (dehydrin), RD (responsive to dehydration), etc, and activate their expression. The expression of CBFs are positively regulated by ICE1/2 (inducer of CBF expression 1/2), CAA1 (circadian clock-associated 1), LHY (late elongated hypocotyl), MYB56, ZFP1/182, and CAMTA1/2/3, whereas negatively regulated by MYB15, MYBS3, ZAT12, PIF4/7, WRKY34, and EIN3. The transcriptional activity of ICE1 protein is strictly regulated by post-translational modification, including SIZ1 (SAP and Miz 1)-mediated SUMO (small ubiquitin-related modifier) modification and HOS1 (high expression of osmotically responsive gene 1)-mediated ubiquitination. The HOS1–SIZ1 system strictly regulates and perfectly fine-tunes the expressions of ICE1–CBFs and their targets to cope with temperature change. Moreover, the expression of ICE1 is negatively regulated by JAZ1/4 and positively regulated by ERF2. Except the CBF-dependent cold-response pathway, some transcription factors regulate responses of plant to cold stress through CBF-independent pathways, such as JERF3, MYB2/4/96/3R-2, WRKY19/21/76, NAC1/2, SAP1/8, etc. Uncovering transcriptional regulatory network in response to cold stress in plants provide a theoretical basis for improving the cold tolerance of crops through transgenic technology. Transgenic studies indicate that some transcription factors possess potential usefulness in genetic improvement of cold stress tolerance in plants, such as AtCBF1AtCBF3, AtICE1, AtCCA1α, TaCBF14/15, TaNAC2, TaWRKY19, VrCBF1/4, MdCIbHLH1, PtrbHLH, OsMYB2, GmNAC20, JERF3, and ZFP182. Furthermore, the main problems in current researches and the key points in future studies were also proposed, in hope of providing reference for further revealing the molecular mechanisms underlying plant adaptation to cold stress.
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    Cited: Baidu(6)
    Progress in Research of FLP/FRT Site-Specific Recombination System in Higher Eukaryotes
    ZHAO Ai-Chun, LONG Ding-Pei, TAN Bing, XU Long-Xia, XIANG Zhong-Huai
    Scientia Agricultura Sinica    2011, 44 (15): 3252-3263.   DOI: 10.3864/j.issn.0578-1752.2011.15.021
    Abstract1765)      PDF (505KB)(6147)       Save
    FLP/FRT site-specific recombination system derived from 2 μm plasmid of yeast has been widely used in Arabidopsis thaliana, Oryza sativa Linnaeus, Mus musculus, Drosophila melanogaster, Caenorhabditis elegans and other higher eukaryotic organisms, and gradually become one of the powerful tools of genetic manipulation in transgenic animals and plants research areas. This review introduced the recombination principles of FLP/FRT system and its application in higher eukaryotes, and systematically summarized the main achievements of the system in transgenic plants, mammals, insects and other higher eukaryotic model organisms. In addition, the main problem, application prospect and developmental trend of the FLP/FRT system were discussed in this review.
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    Cited: Baidu(7)
    Research Advances of Applying Virus-Induced Gene Silencing in Vegetables
    LI Jie,LUO JiangHong,YANG Ping
    Scientia Agricultura Sinica    2021, 54 (10): 2154-2166.   DOI: 10.3864/j.issn.0578-1752.2021.10.011
    Abstract607)   HTML35)    PDF (542KB)(6128)       Save

    Recently, the virus-induced gene silencing (VIGS) as a reverse genetics tool is used for gene function analysis. Due to its advantages of simple construction, low cost and short cycle, VIGS technology has been extensively and deeply studied in the field of functional genomics. VIGS technology, as a fast, effective, high-throughout new technology, has played an important role in research of vegetable functional genes in plant development processes, disease resistance, stress resistance, biosynthesis and metabolic regulation. Herein, it is of great significance to excavate new genes and identify the function of disease resistance, stress resistance genes, crop improvement and molecular breeding by using VIGS technology. Many VIGS systems with virus as vector have been successfully established in vegetable crops, but they still have some shortcomings. With the in-depth exploration of the mechanism of VIGS and the continuous development of virus vectors, VIGS has been applied to a wider range of vegetable crops. This paper reviewed the current status and research progresses of gene function of eggplant, melons and leafy vegetables based on VIGS technology in recent years, and the mechanism of VIGS technology, the application of virus vector and the progress of VIGS technology was briefly analyzed. Meanwhile, the advantages and disadvantages of VIGS technology, RNA interference (RNAi) and current CRISP/CAS9 technology were compared and analyzed. It focused on the application of VIGS technology in vegetable fruit development and disease resistance, and the latest progresses of VIGS technology in vegetable crop metabolic regulation, hormone regulation, biotic and abiotic stress responses were summarized. The cases of studying target genes function and silencing phenotypes of solanaceous, melon, leafy and legume vegetables by VIGS were listed. Finally, the problems and deficiencies of VIGS technology in studying gene function of vegetable crops were summarized, such as lack of suitable VIGS vector, lack of effective virus vector infection method, difficulty in systematic silencing in some tissues, low silencing efficiency, inherent limitations of VIGS, etc. At the same time, the future research directions of VIGS technology in the development of virus vectors with higher specificity and stability, selection of efficient gene fragments, and establishment of virus vectors suitable for more host range were proposed. The application foreground of gene function analysis, improvement, molecular breeding of vegetable crops and production not carrying exogenous gene of vegetable varieties by VIGS technique was prospected. This review would provide a guidance and give ideas for future studies on the growth and development of vegetable crops, secondary metabolism and adversity stress related gene function research and breakthrough in the key factors restricting VIGS technique.

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    Cloning and Function Analysis of Salicylic Acid Binding Protein Gene GmSABP2 from Soybean
    JIA Ya-jun, WANG Xiao-ting, XU Na, GUO Na, XING Han
    Scientia Agricultura Sinica    2015, 48 (18): 3580-3588.   DOI: 10.3864/j.issn.0578-1752.2015.18.002
    Abstract531)   HTML7)    PDF (3325KB)(5967)       Save
    【Objective】The aim of this study is to clone and analyze soybean protein gene GmSABP2, which is binded with salicylic acid, and transform Arabidopsis for analyzing salt tolerance and drought tolerance, and further understand the molecular mechanism of salt tolerance and drought-resistance of the gene. 【Method】 Using SABP2 as a probe, the soybean genome database was searched, and the highest sequence homology was picked out and named as GmSABP2. The gene GmSABP2 was cloned by using electronic cloning technology. The DNAMAN program was used to analyze the amino acid sequence alignment and the conserved domain amino acid by the CD-search conducted NCBI. The MEGA program was applied to make the phylogenetic analysis. The phenotypic variation of soybean seedlings under salt and drought stress was analyzed. The expression of the characteristics of GmSABP2 under salt and drought conditions was analyzed by Real time-PCR of soybean seedlings. Gateway technology was used to build plant expression vector pEarleyGate103-GmSABP2, shifted into Agrobacterium tumefaciens EHA105, infected Arabidopsis by utilizing flower dip method, then the homozygous transgenic plants were obtained by resistance screening and finally the salt and drought tolerance was analyzed. The wild-type plants and transgenic plants were treated under salt and drought stresses, and both the seed germination, root length and mature plants were counted under stress conditions. 【Result】The cDNA sequence of GmSABP2 was obtained and the open reading frame is 786 bp and total length of the sequence is 1 235 bp, encoding 261 amino acids. And molecular weight is 29.15 kD, an isoelectric point is 5.58. The amino acid sequence alignment and phylogenetic analysis showed that GmSABP2 and tobacco SABP2, Rauvolfia serpentina PNAE had the highest similarity. Using the CD-search of NCBI, it was found that the Abhydrolase_6 (pfam: 12697) as conserved domain hydrolases. Soybean SABP2 protein belongs to SABP2 α/β hydrolase superfamily. Using MEGA program to build a system of species multiple phylogenetic tree, it was found that SABP2 of soybeans, Theobroma cacao SABP2 and Solanum lycopersicum SABP2 have a close genetic relationship, but has a distant genetic relationship with Arabidopsis SABP2. The phenotype of soybean seedlings under salt and drought conditions was analyzed and it was found that there were significant stress effects. Real time-PCR analysis showed that GmSABP2 under salt and drought conditions were upregulated expression. Arabidopsis thaliana resistance analysis showed that under normal culture conditions, the wild-type plants and transgenic plants could germinate and grow. Under 150 mmol·L-1 NaCl treatment conditions, seed germination rate of wild-type plants was 38%, seedling root length after 12 days was 0.4 cm and the survival rate of mature plants was 49%; The seed germination rate of transgenic lines was 67%, seedling root length after 12 days was 1.1 cm and mature plants survival was 78%. Under 20% PEG6000 treatment conditions, the seed germination rate of wild-type plants was 31%, seedling root length after 12 days was 0.5 cm and mature plants survival rate was 36%; The seed germination rate of transgenic lines was 57%, seedling root length after 12 days was 1.0 cm and mature plants survival rate was 66%.【Conclusion】 The GmSABP2 gene increases the resistance of Arabidopsis plants under salt and drought conditions.
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    Structural Stability and Degradation Mechanisms of Anthocyanins
    Scientia Agricultura Sinica    2009, 42 (3): 996-1008.   DOI: 10.3864/j.issn.0578-1752.2009.03.031
    Abstract1319)      PDF (467KB)(5885)       Save

    Anthocyanins in plants are water-soluble pigments exhibiting bioactive functions, which can be used in food, pharmacy and cosmetics, but they are not stable. Main factors having impacting anthocyanins, degradation mechanisms and stabilizing methods of anthocyanins are reviewed in this article, it is of significance to enhance investigations on the anthocyanins degradation and stability.

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    Scientia Agricultura Sinica    1995, 28 (02): 58-65.   DOI: 10.3864/j.issn.0578-1752.1995-28-02-58-65
    Abstract1317)      PDF (295KB)(5800)       Save
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    Research Progress of Soil Microbial Mechanisms in Mediating Plant Salt Resistance
    KONG YaLi,ZHU ChunQuan,CAO XiaoChuang,ZHU LianFeng,JIN QianYu,HONG XiaoZhi,ZHANG JunHua
    Scientia Agricultura Sinica    2021, 54 (10): 2073-2083.   DOI: 10.3864/j.issn.0578-1752.2021.10.004
    Abstract862)   HTML74)    PDF (464KB)(5470)       Save

    Soil salinization has seriously hindered the sustainable agricultural production. Remediation of salt affected areas with efficient, low cost and adaptable method is a challenging goal for scientists. Soil microorganisms play important roles in regulating rhizosphere environment of plants, enhancing plant development and productivity. Adaptation of plants to stress driven by soil microbes has been attracted extensive attention. The identification and exploitation of soil microorganisms that interact with plants in alleviating salt stress provides a new strategy for the improvement of saline area, as well as new approaches to discover mechanisms involved in stress tolerance. Knowledge of the underlying physiological mechanisms by which diverse microbes mediate stress tolerance, is critical to the effective use of these microbes to assure sustained agricultural production. This paper reviewed the recent studies about the mechanisms of soil microorganisms mediated in plant salt stress tolerance from the aspects of plant nutrient absorption, osmosis balance, hormone levels and antioxidant function. The beneficial effects and lack of current researches related to soil microorganism in the regulation of plant salt tolerance were evaluated, and the directions of the future research were also proposed. At present, improving nutrient and water uptake efficiency to maintain plant ion homeostasis under salt stress, increasing auxin synthesis and reducing ethylene release to regulate plant hormone levels seem to be promising target processes for soil biota-improved plant salt tolerance. However, limited success has been obtained in application of microorganism to large-scale agricultural production, due to the competition of introduced single microbial strains with native soil microbial communities which resulted in many bacterial strains has little colonization efficiency. The researches related to microbial mediated plant salinity tolerance should break through the single microbial inoculation, further clarify the mechanism of plant-microbial interaction at the community level, and solve the key problems of microbial utilization in agricultural production.

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    Effects of Seawater Irrigation on Fruit Quality of Cabernet Sauvignon
    HUANG LiPeng, ZHANG XiuYuan, WANG yang, FU YanDong, ZHAI Heng, SHAO XiaoJie
    Scientia Agricultura Sinica    2017, 50 (18): 3581-3590.   DOI: 10.3864/j.issn.0578-1752.2017.18.014
    Abstract343)   HTML8)    PDF (1274KB)(5416)       Save
    【Objective】This study was carried out to investigate the effects of seawater irrigation on grape fruit quality and to explore the reasonable application of sea water in improving grape quality.【Method】The Cabernet Sauvignon grapes from two bases of Penglai Guobin and COFCO Changcheng were taken as test materials for 4 consecutive years in 2013-2016 in Penglai. Additionally, the different seawater treatments were applied in 2016, including 10% magnetized sea water, and 10% sea water plus fertilizer magnetization, 1 and 2 years 10% water. Some quality parameters were determined, including fruit soluble solids, titratable acid, total phenolics, tannins, anthocyanins, reductive Vitamin C and aroma. 【Result】 The results showed that an experiment of 10% seawater irrigation was conducted at Guobin base, which significantly increased the soluble solids content of Cabernet Sauvignon fruit and decreased titratable acid content, but had no significant effect on the color difference of pericarp in 3 and 4 years. Four seawater irrigation treatments including 10% magnetized water, 10% water fertilizer, 10% water for 1 year and 10% water for 2 years significantly reduced the titratable acid, sugar acid ratio increased at COFCO Changcheng base. Bacterial manure and magnetization seawater significantly increased the content of sugar compared with the control. In contrast, significant difference in fruit sugar components was found among different seawater treatments. Fruit fructose contents from 1- and 2-year seawater irrigation were significantly enhanced by 35.3% and 42.7%, respectively. Glucose increased by 66.7% and 70.7%. The contents of total phenols, anthocyanin content, reductive Vitamin C were increased by seawater treatments. Among them, 4 kinds of seawater treatment significantly increased the Vitamin C content of the fruit by 46.8%, 60.8%, 57.2% and 79.7%, respectively, and adding bacterial fertilizer and magnetized water significantly reduced the content of tannin in fruits. There were 38 kinds of aroma components in Cabernet Sauvignon grape fruits were detected, including alcohols, aldehydes, esters, ketones and other substances. There were 24 aromas in fruits of seawater irrigation treatments and control. Seawater irrigation increased the total content of aroma components of 14 kinds of aroma. Fruit aroma was significantly increased, and total aroma amount was increased by 92.2%, 75%, 25.9%, and 38.8% in the treatments of 10% sea water and bacterial fertilizer, 10% magnetized seawater, one-year 10% seawater and two-year 10% seawater, respectively. Among the main aroma components of (E)-2-Hexen-1-ol, 2- hexenal, (E)-2-hexenal, nonanal and other substances, the highest content of 2- hexenal in the fruits of magnetized seawater and magnetized seawater plus bacterial fertilizer treatments in, increased by 46.9%, 95.6%, 0.5%, 19.3% as compared with the control. 【Conclusion】 It was concluded that 10% seawater irrigation could promote the quality of wine grape, and seawater irrigation increased water soluble solids content and (E)-2-Hexen-1-ol, 2- hexenal aroma components in fruits.
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    Identification and Comparative Analysis of Metabolites in Grape Seed Based on Widely Targeted Metabolomics
    SHENG HongJie, LU SuWen, ZHENG XuanAng, JIA HaiFeng, FANG JingGui
    Scientia Agricultura Sinica    2023, 56 (7): 1359-1376.   DOI: 10.3864/j.issn.0578-1752.2023.07.013
    Abstract377)   HTML37)    PDF (4037KB)(4964)       Save

    【Objective】 Grape seeds have high bioactivity because they are rich in many metabolites. The objective of this study was to comprehensively identify the metabolite components in grape seeds, to compare and to analyze the differences of metabolites among different varieties, and to explore the relationship between metabolites in grape seeds and skin color and variety origin, so as to provide a reference basis for further development and utilization of grape seeds. 【Method】 The mature seeds of purple-skinned Kyoho (V. labruscana: V. labrusca × V. vinifera, JFS), pink-skinned Wink (V. vinifera, WKS), and yellow-skinned Italia (V. vinifera, YDS) were used for widely targeted metabolomics analysis by UPLC-MS/MS. The metabolites were identified and compared by multivariate statistical methods. 【Result】 The quality of metabolomics data was good, and the data of samples within groups was repetitive and the differences in the data of samples among groups were existed. A total of 514 metabolites were identified in the seeds of three grape varieties, including 6 primary metabolites, such as amino acids and lipids, and 20 secondary metabolites, such as proanthocyanidins and resveratrol. Among different varieties, the metabolite components were similar but the metabolite contents were significantly different. The relative contents of most metabolites were high in the dark variety Kyoho, followed by the light variety Wink, but low in the colorless variety Italy, indicating that the metabolite contents in grape seeds might be positively correlated with the skin color. The relative contents of metabolites in the seeds of Wink and Italia were similar, while they were greatly different from those of Kyoho, indicating that the metabolite contents in grape seeds might be related to the variety origin. The differential metabolites among different varieties were mainly involved in phenylpropane biosynthesis, anthocyanin synthesis, lipid metabolism, etc. pathway. There were many phenolic compounds in the differential metabolites and the metabolites with the large difference were mainly flavonoids. Grape seeds were rich in phenols and lipids. In addition to monomeric flavane-3-ols and their polymers, the relative contents of other phenolic compounds such as flavones and flavonols were also high. There was no significant difference in the relative contents of resveratrol among the three varieties. The relative contents of glyceryl phosphatide such as lysophosphatidylcholine were high, while those of linolenic acid were low. There was little difference in the relative contents of lipids among different varieties. 【Conclusion】 The metabolite components in grape seeds of different varieties were similar, while the metabolite contents were related to the skin color and the variety origin. Phenols and lipids were important components of metabolites in grape seeds and could be used as good sources for food and other processing industries.

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    Physiological Mechanisms of Abiotic Stress Priming Induced the Crops Stress Tolerance: A Review
    WANG Xiao, CAI Jian, ZHOU Qin, DAI TingBo, JIANG Dong
    Scientia Agricultura Sinica    2021, 54 (11): 2287-2301.   DOI: 10.3864/j.issn.0578-1752.2021.11.004
    Abstract1492)   HTML72)    PDF (1536KB)(4712)       Save

    Abiotic stress factors, including heat stress, cold stress, drought stress, waterlogging stress etc., are the limiting factors for plant growth and crop production. Moreover, the frequency, extent and duration of abiotic stresses have predicted to be increased with global climate change. Therefore, improving crops resistance to abiotic stress or finding strategy to reduce the adverse effects of abiotic stress on crop yield and quality is of great significance for ensuring stable crop production and food security. “Priming” by exposing plants to moderate stress in the early growth stage can induce plant resistance to a later severe stress episode which happened during critical stage of the plant growth. The priming is an adaptive strategy that primed plants could effectively mount a faster and/or stronger defense response and actively improve the defensive capacity of plants under stress, compared with non-primed plants. According to the reoccurred stress types, the priming can be generally separated to four types, including in-generational stress tolerance (the priming stimulus and the stress occurred at later stage are the same stress type), intra-generational cross tolerance (the priming stimulus and the stress occurred at later stage are different stress types), transgenerational stress tolerance (the priming stimulus and the stress occurred at next generations are the same stress type), and transgenerational cross tolerance (the priming stimulus and the stress occurred at next generations are different stress types). In this review, the main physiological mechanisms were discussed, including plant photosynthetic response mechanism, antioxidant mechanism, and osmotic regulation, signal transduction mechanisms (plant hormones, Ca2+, hydrogen peroxide, nitric oxide, etc.), and epigenetic modification mechanisms (DNA methylation, histone modification, etc.). Then, the perspectives for the further research on understanding the underlying mechanisms of stress priming and the application of priming effects in the crop production were suggested. After revealing the mechanisms of priming, the candidate genes and proteins which play key regulatory roles in the acquisition of crop stress tolerance are found. And then, we can stimulate the related genes and protein expression which can actively induce the formation of stress tolerance in the critical crops growth stage, thereby effectively alleviating the adverse effects of abiotic stress on crop yield during the critical stage, which is meaningful for the crop production.

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    Envirotyping and Its Applications in Crop Science
    XU Yunbi
    Scientia Agricultura Sinica    2015, 48 (17): 3354-3371.   DOI: 10.3864/j.issn.0578-1752.2015.17.004
    Abstract2145)   HTML24)    PDF (2735KB)(4693)       Save
    Global climate changes have increasing impacts on worldwide environments. Crop productivity is largely determined by interaction between the genotype a crop has and the environment surrounding the crop plants. With modern biotechnologies, genotypic contribution to a phenotype can be dissected at molecular level into individual genetic components. However, the environmental factors that have significant impacts on crops have not been dissected individually, and thus their contribution to phenotype can be only inferred by their integrative effect under different types of environments, or described for the whole experimental plot by comparing pairwise major environmental factors. The author proposed a concept of environmental assay for the first time by coining a word “etyping”, which represents “envirotyping”, a more suitable word used in this article. The term “envirotype” is used to describe all internal and external environmental factors and their combinations that affect plants across growth and developmental stages. The external environmental factors include moisture, fertilizers, air, temperature, light, soil properties, cropping system and companion organisms. Envirotyping refers to dissecting and measuring all these environmental factors. Environmental information can be collected through various approaches, including multi-environmental trials with environmental data accumulated related to trial locations; geographic and soil information systems containing environmental data for climate, weather, and soil; and small weather stations that collect factors related to weather, precipitation, temperature and air. Using remote sensing and other instruments, many external environmental factors can be measured for plant canopy, plant surroundings, and even for single plots or individual plants. Environmental information will be increasingly used for environment characterization, genotype-by-environment interaction analysis, phenotype prediction, disease epidemic prediction, near iso-environment construction, understanding of the response of plants to specific environmental factors, agronomic genomics, and precision farming. In the future, envirotyping needs to be improved to zoom into specific plots and individual plants across growth and developmental stages, along with the development of integrative information system and decision support tools to bring genotypic, phenotypic and envirotypic information together. Envirotypic information will finally contribute as a third dimension to the crop research and development system involving genotype-phenotype-envirotype complex. Such efforts will help establish a high-efficient crop breeding and production system based on the concept of the three-dimensional profile.
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    Review on Research in Plant Nutrition and Fertilizers
    BAI You-lu
    Scientia Agricultura Sinica    2015, 48 (17): 3477-3492.   DOI: 10.3864/j.issn.0578-1752.2015.17.014
    Abstract777)   HTML9)    PDF (481KB)(4587)       Save
    This article reviewed the major achievements and influential events in plant nutrition and fertilizer since 1840. In plant nutrition physiology and biology fields, the essential plant nutrition elements, the process of nutrient absorption and translocation, the nutrient transfer and re-utilization, the role of mycorrhizal in plant nutrition, and plant nutrition and disease resistance were reviewed. In nutrition diagnosis field, available nutrient elements extraction and determination in chemical diagnosis, and spectrum diagnosis were reviewed. In the fertilization technology field, the fertilization models including nutrient classification model, fertilizer effecting function model, nutrients balance model and DRIS method were reviewed. The article also described the development of precision fertilization and irrigation technology. The pollutions caused by fertilization, including greenhouse gas emissions, groundwater pollution and surface water pollution were summarized. This article also introduced current main fertilizer products and fertilizer resources worldwide. In this article, it was pointed out that first, the key to solve fertilizer related problems is to improve fertilizer use efficiency. Secondly, to solve the problem of excessive fertilization on vegetables, new and improved theory and technology on vegetable fertilization are needed. Thirdly, the selection of nutrient efficiency gene should have to be taken place at common nutrient conditions in field. Fourthly, new fertilization technology is an important step to improve the fertilizer efficiency. Lastly, fertilizer industry development needs to be based on currently available fertilizer resources. The development of plant nutrition and fertilizer science is closely related to the population, resources and environmental problems that the people are facing. Fertilizer secures food security, but at the same time causes tremendous pressure to the environment. “Get more food with scientific use of fertilizer” is the direction of plant nutrition and fertilizer research in the future.
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    Research Progress of PPR Protein in Plant Abiotic Stress Response
    LI Cheng, LU Kai, WANG CaiLin, ZHANG YaDong
    Scientia Agricultura Sinica    2023, 56 (24): 4801-4813.   DOI: 10.3864/j.issn.0578-1752.2023.24.001
    Abstract560)   HTML74)    PDF (499KB)(4446)       Save

    Abiotic stress is one of the main factors causing global grain yield reduction. It is of great significance to study the function and response mechanisms of plant stress-related proteins to improve crop stress resistance. Pentatricopeptide repeat (PPR) proteins, belong to the largest family of nuclear coding proteins in higher plants and are named because they contain highly specific PPR motifs. Depending on motif type and arrangement, PPR proteins can be classified as P and PLS, and PLS proteins can be further classified as PLS, E, E+, DYW, and other subclasses based on their carboxyl-terminal domains. PPR proteins are widely distributed in terrestrial plants, mainly in chloroplasts and mitochondria, and a few in the nucleus. As sequence-specific RNA binding proteins, PPR proteins are involved in multiple aspects of plant RNA processing, including RNA editing, splicing, stabilization, and translation. PPR protein plays a variety of important roles in the whole life process of plants, but the mechanism of its action in plant stress resistance is not well understood. Based on the localization and function of PPR proteins related to abiotic stress reported, the mechanism of PPR proteins involved in regulation of abiotic stress, including post-transcriptional regulation and retrograde signaling, was reviewed and discussed in this paper. Post-transcriptional regulation is related to the role of PPR proteins in the modification of RNA after transcription. It is generally believed that PPR affects stress resistance in plants by regulating the expression of stress-related genes via binding RNA and by regulating the metabolism of organelle RNA. In terms of retrograde signaling, damage to PPR proteins can lead to impaired mitochondrial or chloroplast function, and then produce various retrograde signals (such as ROS), thereby regulating the expression of related genes and resisting adversity. However, since plastid signaling is affected by many environmental factors, some of which are still unclear, the mechanism of the PPR protein in retrograde signaling remains to be clarified. In addition, PPR proteins are pleiotropic and some have important effects on plant growth and reproduction while acting on stress resistance. Finally, this paper further analyzed the current research status of PPR protein as an RNA editing tool, discussed the remaining problems and research prospects of PPR protein in the direction of abiotic stress, and pointed out the key points and difficulties that need to be paid attention to in future research, to provide references for further research on PPR protein and crop abiotic stress resistance breeding.

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    Research Progress on Machine Learning for Genomic Selection in Animals
    LI MianYan, WANG LiXian, ZHAO FuPing
    Scientia Agricultura Sinica    2023, 56 (18): 3682-3692.   DOI: 10.3864/j.issn.0578-1752.2023.18.015
    Abstract745)   HTML59)    PDF (570KB)(4169)       Save

    Genomic selection is defined as using the molecular marker information that covered the whole genome to estimate individual’s breeding values. Using genome information can avoid many problems caused by pedigree errors so as to improve selection accuracy and shorten breeding generation intervals. According to different statistical models, methods of estimated genomic breeding value (GEBV) can be divided into based on BLUP (best linear unbiased prediction) theory, based on Bayesian theory and others. At present, GBLUP and its improved method ssGBLUP have been widely employed. Accuracy is the most used evaluation metric for genomic selection models, which is to evaluate the similarity between the true value and the estimated value. The factors that affect the accuracy can be reflected from the model, which can be divided into controllable factors and uncontrollable factors. Traditional genomic selection methods have promoted the rapid development of animal breeding, but these methods are currently facing many challenges such as multi-population, multi-omics, and computing. What’s more, they cannot capture the nonlinear relationship between high-dimensional genomic data. As a branch of artificial intelligence, machine learning is very close to biological mastery of natural language processing. Machine learning extracts features from data and automatically summarizes the rules and use to make predictions for new data. For genomic information, machine learning does not require distribution assumptions, and all marker information can be considered in the model. Compared with traditional genomic selection methods, machine learning can more easily capture complex relationships between genotypes, phenotypes, and the environment. Therefore, machine learning has certain advantages in animal genomic selection. According to the amount and type of supervision received during training, machine learning can be classified into supervised learning, unsupervised learning, semi-supervised learning, and reinforcement learning. The main difference is whether the input data is labeled. The machine learning methods currently applied in animal genomic selection are all supervised learning. Supervised learning can handle both classification and regression problems, requiring the algorithm to be provided with labeled data and the desired output. In recent years, the application of machine learning in animal genomic selection has been increasing, especially in dairy and beef cattle. In this review, machine learning algorithms are divided into three categories: single algorithm, ensemble algorithm and deep learning, and their research progress in animal genomic selection were summarized. The most used single algorithms are KRR and SVR, both of which use kernel tricks to learn nonlinear functions and map data to higher-dimensional kernel spaces in the original space. Currently commonly used kernel functions are linear kernel, cosine kernel, Gaussian kernel, and polynomial kernel. Deep learning, also known as a deep neural network, consists of multiple layers of connected neurons. An ensemble learning algorithm refers to fusing different learners together to obtain a stronger supervised model. In the past decade, the related literature on machine learning and deep learning has shown exponential growth. And its application in genomic selection is also gradually increasing. Although machine learning has obvious advantages in some aspects, it still faces many challenges in estimating the genetic breeding value of complex traits in animals. The interpretability of some models is low, which is not conducive to the adjustment of data, parameters, and features. Data heterogeneity, sparsity, and outliers can also cause data noise for machine learning. There are also problems such as overfitting, large marks and small samples, and parameter adjustment. Therefore, each step needs to be handled carefully while training the model. This paper introduced the traditional methods of genomic selection and the problems they face, the concept and classification of machine learning. We discussed the research progress and current challenges of machine learning in animal genomic selection. A Case and some application suggestions were given to provide a certain reference for the application of machine learning in animal genomic selection.

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    Changes of the Photosynthetic Pigment and Differential Expression of the Correlated Genes in a Chlorophyll-Deficient Cucumber Mutant (Cucumis sativus L.)
    MIAO Han,GU Xing-fang,ZHANG Sheng-ping,WANG Xiao-wu,FANG Zhi-yuan,ZHANG Zhen-xian
    Scientia Agricultura Sinica    2010, 43 (19): 4027-4035.   DOI: 10.3864/j.issn.0578-1752.2010.19.015
    Abstract789)      PDF (438KB)(4107)       Save

    【Objective】 Cucumber chlorophyll-deficient mutant is an ideal material for studies on the photosynthetic system and genetic breeding. To make clear on the mechanism of the leaf colour transformation in yellow leaf mutant will provide a theoretical basis for quantitative trait loci gene mapping and gene cloning. 【Method】 A cucumber chlorophyll-deficient mutant named 9110Gt and its wild type named 9110G were used as materials in this study. The photosynthetic pigment components, contents and Pchl contents in 9110Gt and 9110G were analysed. cDNA-AFLP and cDNA-Ad-SRAP were employed to detect the differentially expressed genes. 【Result】 Spectral analysis showed that there was no significant difference in the pigment components between 9110Gt and 9110G, but the contents of photosynthetic pigment and protochlorophyll in 9110Gt were lower than in wild type in yellow period. Nine candidate transcript derived fragments (TDFs) were obtained. The candidate transcript derived fragments had highly identities with cucumber mitochondrial and chloroplast genome sequence. 【Conclusion】 It was concluded that there was a blockage in the protochlorophyllide biosynthesis, then leading to lower chlorophyll content and make changes in the proportion of photosynthetic pigment of mutant. The leaf colour mutant was may be due to gene mutations in chloroplast development and chlorophyll biosynthesis.

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    Progress of Genetic Research for Protein Content in Rice
    Scientia Agricultura Sinica    2008, 41 (1): 1-8.   DOI: 10.3864/j.issn.0578-1752.2008.01.001
    Abstract1854)      PDF (318KB)(3935)       Save
    The protein content was one of main index that measures the rice nutrition value, so that improving the protein content was important target for nutrition quality breeding in rice.This paper described the protein type, amino acid’s constitute and the nutrition value in rice. Meanwhile the paper summarized the difference among genotype of rice and different part or organization of brown rice, the enviroment’s influence on the protein content, the genetic and molecular mechnasim research of the protein content, which will provide the basis for the protein research and prebreeding for high protein rice .
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    Progress in Research of CRISPR/Cas9 System in Genome Targeted Editing in Plants
    XIE Li-nan, SONG Feng-yan, ZHANG Yang
    Scientia Agricultura Sinica    2015, 48 (9): 1669-1677.   DOI: 10.3864/j.issn.0578-1752.2015.09.01
    Abstract936)   HTML15)    PDF (1085KB)(3931)       Save
    When exogenous DNA was imported into plant cell by transgenic technology, DNA fragment will integrate into the genome by homologous recombination or nonhomologous recombination. In addition, the plants seedling will achieve corresponding target traits. Homologous recombination occurred when the exogenous DNA and the same or similar sequences in receptor cells recombined and integrated to the receptor cell’s chromosomes, so the sequence will be possible to precisely modified and transformed. However, in some cases, to avoid the fracture caused by DNA or chromosomal DNA degradation or the impact on the vitality, the two double-stranded DNA break ends will be joined without considering the sequence similarity by error-prone nonhomologous end joining. As a result, precise mutations control is more difficult to achieve because insertion and/or deletion and other variety of mutations are high-frequently occurred in non-homologous recombination than in homologous recombination. Unfortunately, the frequency of homologous recombination is very low in plants which results the undesirable genome editing. Site-specific nucleases make genome editing more efficient and more precise by the great improvement of the efficiency in the homologous recombination. Such nucleases, zinc finger nuclease (ZFN) and transcription activator-like effector nucleases (TALENs), have been demonstrated to efficiently produce a DNA double-strand break at target site and to induce genome modification in a variety of organisms including plants. However, some defects found in the specific application, such as off-target effects, specific combined site with genome affected by chromosomal location and adjacent sequence, furthermore, technical complexity and time-consuming for assemble limit its application. The clustered, regularly interspaced, short palindromic repeats(CRISPR) system is a prokaryotic adaptive immune system which widely exists in bacteria and archaea. As the result of the long-term evolution, the system can defense against the degradation of RNA to guide the invasion of the virus or phage DNA. Recent advances in the study demonstrated CRISPR/Cas typeⅡ system was a promising system of genome editing strategy with high efficiency, affordability and easy to engineer compared with ZFNs and TALENs. Many precise genome editing cases by CRISPR/Cas system were found successfully in human cells, zebra fish, mice and bacterial, including gene insertion, deletion, mutation in multiple sites or in specific locus. Although the application in plants is still in a relatively limited range, there is an outstanding prospect of CRISPR/Cas system in plant genetic engineering. A brief summary of the composition and principles was presented firstly. Then, the authors emphasis on citing numerous cases involved its application of exogenous and endogenous genes editing in cotyledon and dicotyledonous plants which demonstrate that the CRISPR/Cas9 system has become a powerful tool in plant genome engineering. Finally, the future of the genome technology application in agriculture and plant genetic engineering was discussed which will provide a reference for researchers in genetic modification.
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    Characteristics and Succession of Rhizosphere Soil Microbial Communities in Continuous Cropping Watermelon
    GUO HanYue, WANG DongSheng, RUAN Yang, QIAO YiZhu, ZHANG YunTao, LI Ling, HUANG QiWei, GUO ShiWei, LING Ning, SHEN QiRong
    Scientia Agricultura Sinica    2023, 56 (21): 4245-4258.   DOI: 10.3864/j.issn.0578-1752.2023.21.009
    Abstract275)   HTML36)    PDF (3190KB)(3899)       Save

    【Objective】The aim of this study was to investigate the effects of continuous cropping on the construction and potential functions of bacterial and fungal communities in the rhizosphere soil of watermelon, and to clarify the adaptability of rhizosphere microorganisms to environmental changes, so as to provide a theoretical basis for ecological control of watermelon continuous cropping obstacles and healthy maintenance of farmland.【Method】In this study, the rhizosphere soil of watermelon without continuous cropping (CK), continuous cropping for 2 times and continuous cropping for 6 times was used as the research object. 16S rRNA and ITS high-throughput sequencing were used to analyze the effects of continuous cropping on the bacterial and fungal communities of rhizosphere soil of watermelon.【Result】With the increasing continuous cropping times, the bacterial diversity index in the rhizosphere soil of watermelon showed a trend of first decreasing and then increasing, while the fungal diversity index decreased significantly. At the bacterial genus level, multiple times of continuous cropping decreased the relative abundance of Sphingomonas and Lysobacter in watermelon rhizosphere soil; at the fungal genus level, the relative abundance of Fusarium increased with the continuous cropping times. Compared with CK, the network complexity of continuous cropping was higher, but the stability of network structure was lower. In addition, compared with CK, the relative abundance of biodegradation pathways of harmful substances and metabolic pathways of amino acids could be significantly reduced after continuous cropping for 6 times; the relative abundance of pathogenic fungi significantly increased in the fungal community after continuous cropping for 6 times. During community succession, the stochastic processes dominated rhizosphere bacterial community construction in watermelon under continuous cropping, while the deterministic processes dominated rhizosphere fungal community construction in watermelon.【Conclusion】Continuous cropping caused changes in community characteristics, functional composition and succession process of rhizosphere bacteria and fungi. The decrease of key functions of bacterial community, the increase of pathologic fungi and the decrease of stability of microbial community network might be the important factors leading to occurrence of watermelon continuous cropping obstacles.

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    Deoxynivalenol Contamination in Wheat and Its Management
    SHI Jian-rong, LIU Xin, QIU Jian-bo, JI Fang, XU Jian-hong, DONG Fei, YIN Xian-chao, RAN Jun-jian
    Scientia Agricultura Sinica    2014, 47 (18): 3641-3654.   DOI: 10.3864/j.issn.0578-1752.2014.18.012
    Abstract627)   HTML6)    PDF (733KB)(3894)       Save
    Deoxynivalenol (DON), one of the type B trichothecene mycotoxins, also known as vomitoxin, can cause sickness such as vomiting, feed refusal, bloody diarrhea, dermatitis, hemorrhaging, and even death because of inhibition of DNA, RNA and protein synthesis at the ribosomal level in nonruminant animals, especially swine. DON is the metabolite of Fusarium graminearum species complex(FGSC) which causes wheat head blight around the world. In the last 10 years, FHB has become a severely epidemic for many times in China (especially in Jiangsu, Anhui, Hubei, Henan and Shandong), north America and European countries, causing that DON becomes severe food safety issue for wheat grains. This paper presents the DON contamination of wheat, distribution and chemotype of DON-produced FGSC, the factors which influence the DON production, and the management of DON contaminated grains to facilitate the risk evaluation and surveillance of DON-contaminated wheat in this country.
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    Structure Characteristics of Stomata in Leaves and Vascular Bundles in Culms of Transgenic Rice Expressing C4 Photosynthesis Enzymes
    ,,,,
    Scientia Agricultura Sinica    2006, 39 (05): 902-909.   DOI: 10.3864/j.issn.0578-1752.at-2005-5653
    Abstract2387)      PDF (1872KB)(3883)       Save
    【Objective】The aim of this study is to discover the biological structure basis of transgenic rice expressing C4 photosynthesis enzymes and photosynthesizing highly and efficiently. 【Method】Using four transgenic rice lines over-expressing maize photosynthesis enzymes of phosphoenolpyruvate carboxylase (PEPC), pyruvate, othophosphate dikinase (PPDK), PEPC + PPDK, and rice nicotinamide adenine dinucleotide phosphate-malic enzyme (NADP-ME) as materials, and an untransformed wild type Kitaake as the control, the structures of stomata in leaves and vascular bundles in culms of the seedlings were observed with a scanning electron microscope and mesophyll cells were observed with a transmission electron microscope. 【Result】 Compared with the control, the density and the area of the stomata in leaves of the transgenic rice lines were greater, with the PPDK line greatest among the four lines. The density of stomata in the leaves of transgenic rice with two polymerizing genes (PEPC + PPDK) was increased, but the area of stomata was reduced. The stacking degree of chloroplast thylakoids in mesophyll cells was denser, and some thylakoids were arranged more orderly along the long axis of chloroplast for the transgenic rice lines than those for the WT. The transgenic rice lines exhibited thicker and stronger culms as compared to WT. Except for PPDK, all the transgenic rice lines had larger areas of the tissue structure performing substance-transporting function, such as the outer and inner vascular bundles and their catheter and sieve tubes in culms, as compared with WT. 【Conclusion】 More number and larger area of stomata in leaves and more developed vascular bundles in culms of C4 transgenic rice lines, all these structure characteristics were the basis of its, photosynthesizing highly and efficiently, and closely associated with the greater dry matter accumulation of seedlings.
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    Apomixis——The Process of Asexual Seed Formation

    Scientia Agricultura Sinica    2009, 42 (2): 377-387.   DOI: 10.3864/j.issn.0578-1752.2009.02.001
    Abstract1506)      PDF (375KB)(3709)       Save

    Apomixis is a natural trait of reproduction in which embryo is formed without the union of male and female gametes. It is defined as the asexual formation of a seed from the maternal tissues of the ovule, avoiding the processes of meiosis and fertilization, leading to embryo development. According to this international prevalent idea, only unreduced apomixis, nor reduced apomixis, is referred to it. Apomictic processes have been observed in more than 400 plant species spanning 40 different families in angiosperms and are most common in the Poaceae, Asteraces, Rosaceae and Rutaceae, but appears to be absent in important crops. This paper is concerned with some aspects of apomixis, such as cytoembryoic mode, distribution in angiosperms, new species or straits with trait of apomixes. The progress of genetics, molecular mechanism of apomixis, and its application prospect in agriculture and the ecological significance were discussed.

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    Research on the Mechanisms of Pre-Harvest Sprouting and Resistant Breeding in Wheat
    DONG HuiXue, CHEN Qian, GUO XiaoJiang, WANG JiRui
    Scientia Agricultura Sinica    2024, 57 (7): 1237-1254.   DOI: 10.3864/j.issn.0578-1752.2024.07.003
    Abstract982)   HTML62)    PDF (692KB)(3683)       Save

    Pre-harvest sprouting (PHS) refers to the germination of cereal crops on the spike in high humidity conditions before grain harvest. Wheat PHS is a significant problem that affects both the yield and quality of wheat. Seed dormancy level is a major factor influencing the resistance of wheat PHS, and domesticated crops often exhibit reduced seed dormancy levels, making cultivated wheat more prone to PHS compared to its wild ancestors. Wheat PHS is mainly regulated by external environmental factors such as temperature and humidity, as well as internal plant hormones (GAs, ABA, IAA, MeJA, ET, BR). Researchers have identified a range of materials resistant to PHS, cloned key genes regulating PHS resistance, such as PM19, MFT, MKK3, Myb10-3D, Vp1. New wheat materials resistant to PHS have been successfully developed through molecular marker-assisted selection, artificial synthesis of wheat, and CRISPR/Cas9 gene editing technology. This article reviews the genetic mechanism of PHS resistance in wheat and the latest progress in PHS resistance breeding research. In the future, it is necessary to continue exploring key genes related to PHS resistance, and employ biotechnological breeding methods to cultivate new PHS-resistant wheat varieties.

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    The Development of CRISPR/Cas9 System and Its Application in Crop Genome Editing
    JING Run-chun, LU Hong
    Scientia Agricultura Sinica    2016, 49 (7): 1219-1229.   DOI: 10.3864/j.issn.0578-1752.2016.07.001
    Abstract1371)   HTML43)    PDF (1232KB)(3679)       Save
    In this review, the steps of development of the CRISPR/Cas9 genome editing system and its applications in various plant genomes were highlighted. The CRISPR/Cas9 genome editing technology originates from the prokaryotic adaptive immune systems that confer resistance to foreign genetic elements such as plasmids and phages. The natural CRISPR/Cas systems show extensive structural and functional diversity. Based on the Cas protein contents and amino acid sequences, the natural CRISPR/Cas systems have been classified into three major classes, Type I, TypeⅡ and Type III. The TypeⅡCRISPR/Cas system is the engineered one for targeted genome editing purpose in most of cases so far, as it needs optimization of the Cas expression and design of the sgRNA only. In 2013, the first applications of CRISPR/Cas9 genome editing technology in plants were published. Since then, the CRISPR/Cas9 system has been used in various plant species for targeted genome editing. Like ZFNs and TALENs, CRISPR/Cas9 system uses engineered nuclease to generate double-strand breaks (DSBs) on the targeted DNA site, and subsequently to stimulate cellular DNA repair mechanisms by exploiting either NHEJ pathway or HDR pathway to generate small insertions/deletions/genome modifications. CRISPR/Cas9 technology allows researchers to perform targeted mutagenesis on target genes of different crops, precisely and easily changing the sequences and functions of particular genes at exact chromosomal locations in different plant genomes. Compared with ZFNs and TALENs technologies, CRISPR/Cas9 genome editing system is based on RNA-guided engineered nucleases, and is easier to manipulate. Furthermore, CRISPR/Cas9 is capable of introducing DSBs at multiple sites. The potential of multiplexing provides practical advantages over ZFNs and TALENs technologies, to edit multiple target genes in the same pathway simultaneously. Due to the practical advantages of CRISPR/Cas9 over the other genome editing technologies, it establishes a prosperous outlook in gene discovery and trait development in crop genetic improvement and breeding studies. In this review, the possible applications of CRISPR/Cas9 genome editing technique in various aspects of plant genetics and breeding were also discussed, except the targeted genome editing. CRISPR/Cas9 genome editing technology is another stepping stone in utilizing genetic manipulation in genetic studies and breeding, after genetic modification. Unlike genetic modification, CRISPR/Cas9 genome editing technique generates phenotypic variation that is indistinguishable from that obtained through natural means or conventional mutagenesis. This ambiguity challenges the current GMO regulatory definitions, and provides a potential barrier for further use of CRISPR/Cas9 genome editing technique in crop genetics and breeding.
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    Effects of Caragana korshinskii Kom. on Serum Indexes, Rumen and Colon Microbiota of Tan Sheep
    HAN Jing, WANG XiaoQi, DUAN ZiYuan
    Scientia Agricultura Sinica    2023, 56 (14): 2812-2827.   DOI: 10.3864/j.issn.0578-1752.2023.14.015
    Abstract192)   HTML9)    PDF (1783KB)(3622)       Save

    【Objective】 This study aimed to examine the effects of Caragana korshinskii Kom. (CK) at the rate of 10% of the diet on blood lipids, immune indicators, and rumen and colon microbiota of Tan sheep, so as to provide a theoretical basis for CK being partial Tan sheep forage. 【Method】 Twelve Tan sheep (4-month-old) with an initial average weight of 27.5 ± 3.32 kg were randomly divided into 2 groups (6 in each group): NC group was fed with 30% concentrate and 70% alfalfa; CK group, with 30% concentrate, 60% alfalfa and 10% CK, and drinking water ad libitum. The nutrient composition of the two groups were similar, but the fiber content of CK was slightly higher than that of alfalfa. The preparatory experiment was in 10 days and the formal experiment was lasted for 60 days. At the end of the 60 days, jugular vein blood was collected to measure blood biochemical indexes (blood lipids and immune factors) and rumen fluid, and colon content were collected to detect the changes of flora in rumen and colon, respectively, by 16S rRNA sequencing method. Through construction of the recombinant plasmid, such as Selenomona, Prevotella, Blautia, Desulfovibrio, Ruminococcus and Bacteroides, the absolute quantitative of important genera were detected by real-time quantitative PCR (qPCR) comparing to its standard curve, respectively. 【Result】 At the end of the trial period, the end weight and daily average feed intake of Tan sheep were similar between two groups. Compared with NC group, the indexes related to the lipid absorption and digestion in CK group decreased, such as the content of serum triglyceride (TRIG, P<0.05) and very low density lipoprotein (VLDL, P<0.01) decreased markedly, while the concentration of low density lipoprotein (LDL) tended to decrease significantly (0.05<P<0.1). The inflammatory and lipolytic factor, such as interleukin-6 (IL-6), increased significantly (P<0.05) whereas the inflammatory factor, γ-Interferon (IFN-γ) marked decreased tendentiously (P=0.058). By analyzing microflora of rumen fluid and colon contents, adding CK to diet increased community Shannon diversity index of colonic bacteria communities significantly (P<0.01), while tended to decrease the Chao1 index in rumen (0.05<P<0.1). Compared with NC group, the relative abundance of Oscillospira under CK decreased (P=0.07), while that of Akkermansia notable under CK increased (P<0.05) in rumen. The relative abundance of harmful bacteria, like Treponema, and carbohydrate degradation related Bacteroides significantly decreased (P=0.059, P=0.061), but that of Oscillospira, which a butyric acid producing bacteria related to obesity, constipation and immunity, gained markedly (P<0.05) in colon under CK. It was evidently deduced that the bacterial communities in rumen and colon had significant difference on the capacities for lipid metabolism and biosynthesis of other secondary metabolite between two groups (P<0.05). Based on Pearson correlation analysis, it was showed that the level of serum TRIG was positively correlated with rumen and colonic Pseudobutyrivibrio (P<0.05), which was both fiber degrading bacteria and butyric acid producing bacteria. 【Conclusion】 There was no significant effect of CK partial substitution for alfalfa on the growth performance of Tan sheep, but could reduce the concentrations of serum TRIG, LDL and VLDL through affecting the gastrointestinal tract microbiota, and then participate in the lipid metabolism of the host.

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    Recent Advances and Prospects on the Technology to Increase Fertilizer Use Efficiency
    Xiang YAN Ji-yun Yun Ming-zao LIANG
    Scientia Agricultura Sinica    2008, 41 (2): 450-459.   DOI: 10.3864/j.issn.0578-1752.2008.02.019
    Abstract2200)      PDF (351KB)(3568)       Save
    How to increase fertilizer use efficiency (FUE)and decrease environment pollution caused by nutrient losses has been concerned by the whole world for a long time. It is very important to increase FUE in China for its low FUE and serious losses of nutrient. The recent advances of the technologies to increase FUE are reviewed in this article, which include applying site-specific nitrogen management and real-time nitrogen management to fertilizer management, development of new style slowly release and controlled release fertilizer to control nutrient release,technology of site specific nutrient management and using urease inhibitor or nitrification inhibitor to decrease nitrogen losses. And some technologies to increase FUE are previewed in the future.
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    Effect of Light Quality on Photosynthesis and Chlorophyll Fluorescence in Strawberry Leaves
    ,,
    Scientia Agricultura Sinica    2005, 38 (02): 369-375.   DOI: 10.3864/j.issn.0578-1752.at-2004-2003
    Abstract1282)      PDF (297KB)(3552)       Save
    Chlorophyll contents, maximal photochemical efficiency of PSII (Fv/Fm) , Fm/Fo, amount of inactive PSⅡreaction centers (Fi-Fo) and rate of QA reduction were correlated with red/blue ratios positively, but chlorophyll (a/b) ratios were correlated with red/blue ratios negatively. Carotenoid contents were in the order: blue film >green film> red film, white film and yellow film, which were correlated with red/far-red ratios negatively. The apparent quantum efficiency (AQY), photorespiratory rate(Pr)and carboxylation efficiency (CE) also were strongly affected by light quality. The net photosynthetic rate(Pn)of strawberry leaves under green film was significantly lower than that of under all other film treatments.
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    Degradation of Antibiotics and Passivation of Heavy Metals During Thermophilic Composting Process
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    Scientia Agricultura Sinica    2006, 39 (02): 337-343.   DOI: 10.3864/j.issn.0578-1752.at-2004-2884
    Abstract3351)      PDF (423KB)(3516)       Save
    【Objective】The problem that there are various antibiotic medicines and heavy metal elements in excrements of livestock and poultry have been concerned on the farm utilization extensively. The techniques for the antibiotics degradation and heavy metals passivation were studied in the experiments.【Method】As composting is a new alternative bioremediation technology for treatment of organic pollutants and heavy metals, two experiments were conducted to evaluate the effect of using thermophilic composting to break down the content of tetracyclines(TTC, OTC, CTC) and H2O extractible of heavy metals(Cu, Zn, Cr, As). 【Results】The result of TCs degradation test indicated that the degradation of TTC, OTC and CTC using P+S and C+S in all treatments was effective, but in comparision, the addition of BM bacterium for TCs degradation in treatment was more effective, and its biodegradation rate was higher than that of P+S+TCs and C+S+TCs treatment. The biodegradation rate of OTC was the lowest in all treatments, and the rate of C+S+OTC treatment was 40.23%. The order of the three TCs biodegradation rate was TTC> CTC> OTC. The air-slaked coal has a passivation effect on the heavy metal elements in excrements of livestock and poultry. The result of heavy metal passivation test showed that the effect of addition of air-slaked coal to H2O extractible content of heavy metals, such as Cu, Zn, Cr and As, were obvious, the passivation effect of ari-slaked coal treatment was better than that of control. The H2O extractible content of heavy metals of Cu, Zn, Cr and As, after composting of pig manure, reduced by 6.17%, 6.40%, 4.17% and 1.83%, respectively, meanwhile, the H2O extractible contents after composting of chicken manure reduced by 7.07%, 5.69%, 5.50% and 2.07%, respectively. 【Conclusion】With the different thermophilic composting conditions, tetracyclines antibioticses can be degraded, and the addition of foreign beneficial bacterium can reduce residues in antibiotic medicine. Thermophilic composting also can lower the effects of heavy metals on plants. Air-slaked coal has passivation function on extractible heavy metals.
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    Cited: Baidu(106)