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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
    Abstract1052)      PDF (530KB)(32067)       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
    Abstract948)   HTML62)    PDF (625KB)(26383)       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
    Abstract1122)      PDF (300KB)(22200)       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
    Abstract1101)   HTML47)    PDF (15602KB)(20939)       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
    Abstract711)   HTML30)    PDF (3179KB)(19594)       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
    Abstract779)   HTML75)    PDF (544KB)(19219)       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
    Abstract556)   HTML20)    PDF (396KB)(19214)       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
    Abstract532)   HTML15)    PDF (492KB)(18402)       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
    Abstract696)   HTML18)    PDF (532KB)(18047)       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
    Abstract558)   HTML16)    PDF (507KB)(16540)       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
    Abstract526)   HTML22)    PDF (930KB)(15976)       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
    Abstract417)   HTML11)    PDF (4441KB)(15307)       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
    Abstract571)   HTML22)    PDF (5409KB)(14739)       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
    Abstract611)   HTML30)    PDF (934KB)(14117)       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
    Abstract628)   HTML11)    PDF (2772KB)(12954)       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
    Abstract479)   HTML10)    PDF (722KB)(12941)       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
    Abstract544)   HTML15)    PDF (903KB)(11072)       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
    Abstract523)   HTML15)    PDF (2842KB)(10449)       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
    Abstract539)   HTML14)    PDF (447KB)(9764)       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)
    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
    Abstract960)   HTML21)    PDF (550KB)(9538)       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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    Cited: Baidu(27)
    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
    Abstract479)   HTML2)    PDF (642KB)(9514)       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)
    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
    Abstract471)   HTML3)    PDF (489KB)(9007)       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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    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
    Abstract1030)      PDF (591KB)(7445)       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)
    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
    Abstract474)   HTML2)    PDF (3325KB)(5813)       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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    Scientia Agricultura Sinica    1995, 28 (02): 58-65.   DOI: 10.3864/j.issn.0578-1752.1995-28-02-58-65
    Abstract1287)      PDF (295KB)(5789)       Save
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    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
    Abstract1660)      PDF (505KB)(5485)       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)
    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
    Abstract2321)      PDF (1872KB)(3744)       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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    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
    Abstract840)   HTML9)    PDF (1085KB)(3653)       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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    Cited: Baidu(5)
    Degradation of Antibiotics and Passivation of Heavy Metals During Thermophilic Composting Process
    ,,,,
    Scientia Agricultura Sinica    2006, 39 (02): 337-343.   DOI: 10.3864/j.issn.0578-1752.at-2004-2884
    Abstract3278)      PDF (423KB)(3375)       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)
    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
    Abstract1276)   HTML42)    PDF (1232KB)(3367)       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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    Cited: Baidu(4)
    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
    Abstract1242)      PDF (467KB)(3348)       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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    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
    Abstract2053)      PDF (351KB)(3163)       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
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    Scientia Agricultura Sinica    2005, 38 (02): 369-375.   DOI: 10.3864/j.issn.0578-1752.at-2004-2003
    Abstract1191)      PDF (297KB)(3154)       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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    The Possible Effects of Global Warming on Cropping Systems in China Ⅰ. The Possible Effects of Climate Warming on Northern Limits of Cropping Systems and Crop Yields in China
    YANG Xiao-guang,LIU Zhi-juan,CHEN Fu
    Scientia Agricultura Sinica    2010, 43 (2): 329-336.   DOI: 10.3864/j.issn.0578-1752.2010.02.013
    Abstract1514)      PDF (476KB)(3058)       Save
    【Objective】 Increasing temperature in China significantly since the 1980s has become a consensus in the context of global climate change. How the change affected the agriculture or even the cropping system has attracted more and more attention of the government and scientists. In this study, the possible effects of climate warming on the countrywide northern limits of cropping systems, the northern limits of winter wheat and double rice, and the stable-yield northern limits of rainfed winter wheat - summer maize rotation in China from 1981 to 2007 were analyzed. And the possible changes of crop yield due to the changes of planting limits during the period from 1950s to 1981 and the period from 1981 to 2007 were compared and discussed. 【Method】 A recognized calculation method of agro-meteorological indicators were used in the study. The countrywide northern limits of cropping systems, the northern limits of winter wheat and double rice and the stable-yield northern limits of rainfed winter wheat–summer maize rotation were draw by ArcGis for two periods from 1950s to 1980 and 1981 to 2007, according to the indices of the climate zoning index of cropping system, winter wheat and double rice northern limits and stable-yield northern limits of rainfed winter wheat-summer maize rotaion. 【Result】 Compared with the results during the period from 1950s to 1980, the northern limits of the two-cropping system from 1981 to 2007 occurred significant spatial displacement in Shaanxi, Shanxi, Hebei, Beijing and Liaoning provinces and city. The northern limits of the three-cropping system occurred largest spatial displacement in Hunan, Hubei, Anhui, Jiangsu, and Zhejiang provinces. Without the consideration for variety and social economic changing, grain yield per hectare of main cropping pattern could increase about 54%-106% if the one-cropping system changed to two-cropping system, while increase about 27%-58% if two-cropping system is changed to three-cropping. The northern limits of winter wheat moved northwards and westwards to different degrees in Liaoning, Hebei, Shanxi, Shaanxi, Inner Mongolia, Ningxia, Gansu and Qinghai provinces,comparing with the results during the 1950s-1980 period. For example in Hebei province, northern limits of winter wheat moved north,leading to about 25% grain yield increase in the changing area if the cropping system is changed from spring wheat to winter wheat in one year. The northern limits of double rice cropping area in Zhejiang, Anhui, Hubei, and Hunan provinces moved northwards. Considering the heat resource, the change of limits can increase the grain yield. The stable-yield northern limits of rainfed winter wheat-summer maize moved southeastwards in most regions, mainly caused by the decreased rainfall during recent years in the study areas.【Conclusion】 During the past 50 years, the climate warming caused the northwards movement of the northern limits of cropping system, and the northern limits of winter wheat and double rice. The changes may increase the unit grain yield in the changing area. However, the stable-yield northern limits of rainfed winter wheat–summer maize rotation moved southeastwards due to the decreasing rainfall.
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    Characteristics of Nutrient Input/Output and Nutrient Balance in Different Regions of China
    LI Shu-Tian, JIN Ji-Yun
    Scientia Agricultura Sinica    2011, 44 (20): 4207-4229.   DOI: 10.3864/j.issn.0578-1752.2011.20.009
    Abstract1573)      PDF (667KB)(3032)       Save
    Clearance of nutrient input/output and balance in farmland in different regions of China is critical for nutrient management, scientific distribution and application of fertilizer resources, and improving nutrient use efficiency, etc. Based on existed literatures this paper systematically analyzed and discussed the current nutrient resources and application status, estimated the amount of manures, crop straws and their potential amount of nutrients NPK. The characteristics of nutrient input, output and balance in different regions of China were analyzed and evaluated.  Strategies and suggestions on nutrient management were discussed based on the existed issues of nutrient input and output.
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    Cited: Baidu(82)
    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
    Abstract1305)      PDF (375KB)(2993)       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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    Advances and Prospects in Research of Biochar Utilization in Agriculture
    CHEN Wen-Fu, ZHANG Wei-Ming, MENG Jun
    Scientia Agricultura Sinica    2013, 46 (16): 3324-3333.   DOI: 10.3864/j.issn.0578-1752.2013.16.003
    Abstract1047)      PDF (610KB)(2861)       Save
    Biochar has a bright prospect due to its good structure, physicochemical properties and broad raw materials of its production. It has already been a hotspot in the fields of agriculture, energy and environment. The influences of biochar were reviewed comprehensively on soil, crops, agricultural eco-system, and its important roles in food security of China. The application value and industrialization of biochar in agriculture from the low-carbon, recycle and sustainable point of view were discussed. Utilization of biochar would play much more important roles in improving soil obstacles and increasing crop production capacity of soil, which will benefit the sustainable development of agriculture and the national food security of China. At the end, the prospective of biochar industrialization and development in China were proposed, which will provide relevant references for the well development of biohcar industry.
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    Cited: Baidu(80)
    Transcriptomic Analysis of Sesame Development
    WEI Li-Bin, MIAO Hong-Mei, ZHANG Hai-Yang
    Scientia Agricultura Sinica    2012, 45 (7): 1246-1256.   DOI: 10.3864/j.issn.0578-1752.2012.07.002
    Abstract946)      PDF (727KB)(2807)       Save
    【Objective】 To enrich sesame transcriptome data, the transcriptome sequencing and bioinforamtics analysis on sesame growth and development and seed formation processes were performed in this study. 【Method】 Six transcriptome sequencing libraries for developing sesame plants and seeds were constructed and sequenced using Illumina RNA sequencing technique, and the global transcriptome information was analyzed subsequently.【Result】8.80 Gb available trancriptome data were acquired as the adaptor sequences, duplication sequences and low-quality reads were removed from the original 12.69 Gb solexa sequencing data. And 26 837 uni-transcripts, longer than 100 bp, were obtained by de novo assembly method (http://www. ncbi.nlm.nih.gov/genbank/TSA.html, GenBank ID: JP631635-JP668414). The total scaffold sequence length reached 18.35 Mb with the 1 006 bp of N50, the average uni-transcript length was 683 bp. Annotation analysis of uni-transcripts indicated that 25 331 transcripts had homolog in public protein database; however, 1 506 sequences were no hit and might be sesame-specific. With COG and GO functional classifications, all uni-transcripts were grouped into 24 and 42 function categories, respectively, in which many functional categories, such as material and energy metabolism, signaling, transcription regulation and defense reactions, etc. were included. Furthermore, compared with plant growth and development transcripts, 1 277 sequences expressed more than 10-fold low in seed formation process, and the transcript level of 990 uni-trancripts could not be measured. In addition, 660 sequences were found with more than 10-fold high expression level during seed development, in which 296 sequences seemed as seed-specific. 【Conclusion】With the transcriptome sequencing of Sesamum radiatum and several cultivars of S. indicum L., this study gave a global insight into the characteristics of sesame development transcriptome and thousands of transcript sequences with important function were acquired for future genes expression and regulation research about sesame growth and development.
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    Cited: Baidu(20)

    Advances in Biosynthesis Pathways and Regulation of Flavonoids and Catechins

    XIA Tao,GAO Li-ping
    Scientia Agricultura Sinica    2009, 42 (8): 2899-2908.   DOI: 10.3864/j.issn.0578-1752.2009.08.031
    Abstract1121)      PDF (324KB)(2797)       Save

    Flavonoids are the major secondary metabolites with diverse biological activities in high plant. As one of the main subgroups of flavonoids, catechins usually account for 12%-25% dry weight in fresh leaves of tea plant [Camellia sinensis (L.) O. Kuntze]. Catechins are not only the most important components in tea flavor, but also possess a lot of physiological functions, such as antioxidant activity, antimutagenic and anticarcinogenic potential ,anti-cardiovascular diseases, anti-ultraviolet radiation and so on. This paper reviewed the progresses in the flavonoids or tea catechins branch pathway and regulation, subcellular localization and organization of flavonoid enzymes, and may provide additional insights into regulating gene expression and engineering the catechins biosynthesis in tea plant.

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    The Physiochemical Characteristics of Amylopectin and Their Relationships to Pasting Properties of Rice Flour in Different Varieties
    ,,,,,
    Scientia Agricultura Sinica    2006, 39 (06): 1122-1129.   DOI: 10.3864/j.issn.0578-1752.at-2005-5617
    Abstract1205)      PDF (501KB)(2764)       Save
    【Objective】The objective of this paper is to explore the relationship between the physicochemical properties of amylopectin and pasting character of rice-flour. 【Method】Eight amylopectins from different rice varieties were used as materials, and the physicochemical properties such as blue value (BV) indicating iodine binding capacity and maximum absorption (λmax) and the chromatogram of debranched amylopection were analyzed by Sephadex G75.【Result】The results showed that the differences of BV and λmax.Ap of amylopectin among all cultivars were significant. BV and λmax.Ap of amylopectins from indica varieties were higher than that from Japonica varieties except Baxiluodao. A positive correlation (r=0.995**) between BV and λmax.Ap (except waxy cultivars) was observed. The average chain length of FrⅠ, FrⅡand FrⅢ separated by Sephadex G75 was greater than 100 glucose unite, 44-47 glucose unite and 10-17 glucose unite, respectively. The weight percentage of chain length distribution varied with typically different cultivars. Indica waxy cultivar contained more long chains than Japonica waxy cultivar, Indica cultivar consisted of more long branched chains than Japonica cultivars except Baxiluodao, and amylopectin from hybrid combination had more long chains than that from conventional cultivar. There was a positive correlation between the proportion of short chain FrⅢ and characters of RVA profile such as peak viscosity and breakdown (r=0.969**, r=0.949**). Correspondingly, the negative correlation of the proportion of long chain Fr (Ⅰ+Ⅱ) with peak viscosity and breakdown was discovered (r=-0.969**, r=-0.949**). 【Conclusion】The results suggest that the chain length distribution of debranched amylopection has a relation to rice quality. The higher ratio of Fr(Ⅰ+Ⅱ) in amylopectins from the non-waxy varieties, the more long branched chain, the texture of which shows that hardness and cohesiveness are high, and that looseness and adhesiveness are low, the taste sensory is delicious.
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    Cited: Baidu(53)

    Identification and Effect Analysis of QTLs Conferring Salt Tolerance at Seedling Stage Using Solanum pennellii LA716 Introgression Lines in Tomato

    YU Qing-hui,LIU Lei,WANG Bai-ke,YANG Sheng-bao,YANG Tao,LI Jun-ming,HOU Xi-lin
    Scientia Agricultura Sinica    2010, 43 (4): 761-768.   DOI: 10.3864/j.issn.0578-1752.2010.04.013
    Abstract1002)      PDF (285KB)(2731)       Save

    【Objective】 The introgression line population, which was developed from a salt-sensitive parent S. lycopersicum M82 and a salt-tolerance wild species S. pennellii LA716, was employed in this paper to identify QTLs conferring salt tolerance at the seedling stage. Meanwhile the primary genetic effect and interaction of some QTLs were also analyzed. 【Method】 Seedlings with four fully developed true leaves were transferred into hydroponic tanks containing 50 L of half-strength modified Hoagland solution. The same amount of salts was added every day to achieve a final concentration of 700 mmol?L-1NaCl + 70 mmol?L-1 CaCl2. After the final salt concentration was reached, the plants were evaluated using a scale of 0 to 10. Dunnett test was used for ANOVA analysis. 【Result】 Seven unambiguous QTLs (Stq2a, Stq2b, Stq6a, Stq7a, Stq7b, Stq8 and Stq10) located respectively on chromosome 2, 6, 7, 8 and 10 were identified, which were responsible for controlling salt tolerance at seedling stage and derived from wild relative S. pennellii LA716. The survival percentage increased by 18.9%-83.8% as compared to the control M82 under this salt stress. QTL possibly involved in IL6-4 and IL6-3 needed to be confirmed in the further experiments. Genetic analysis of some QTLs showed a significant dominant effect with the exception of Stq6b. The interaction of QTLs presented a typically less-than-additive but two QTLs located on chromosome 7 gave a reduced effect. 【Conclusion】 This is the first report to identify QTLs conferring salt tolerance from wild species S. pennellii LA716 at the seedling stage. These identified QTLs would provide a sound base for tomato breeding with salt tolerance.

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    Analysis of the Source Areas of the Early Immigration of White-Backed Planthopper in Northeast Guangxi of China
    QI Hui-Hui, ZHANG Yun-Hui, JIANG Chun-Xian, SUN Ming-Yang, YANG Xiu-Li, CHENG Deng-Fa
    Scientia Agricultura Sinica    2011, 44 (16): 3333-3342.   DOI: 10.3864/j.issn.0578-1752.2011.16.005
    Abstract1247)      PDF (1102KB)(2713)       Save
    【Objective】The objective of this study is to research on the immigratory peak periods and the source areas of the white-backed planthopper (WBP), Sogatella furcifera (Homoptera: Delphacidae), in the northeast Guangxi of China and to provide a basis for early warning and effective control. 【Method】 The Jiaduo light trap was used to monitor the immigratory peak periods of the WBP in Xing’an country, Guangxi from 2007 to 2010. And the Hysplit_4, a software system was used to analyze their migratory trajectory for ascertaining source areas of the WBP. 【Result】 The immigratory peak periods of the WBP in Xing’an country lasted mainly from the mid-May to the end of June. Immigratory abundance of the WBP population in 2007 and 2010 was higher than that in 2008 and 2009. The highest immigratory peak period went back with time from 2007 to 2010. The trajectory analysis showed that the source areas of the WBP were mainly in the north of the Vietnam, and the part of Hainai Island, the south of Guangxi, the southwest of Guangdong. 【Conclusion】 The low level airflow carried large-scale immigration of the WBP from the source areas to the northeast of Guangxi, while heavy rainfall caused mass descent of the migratory population into the areas.
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    Cited: Baidu(18)
    Gene Cloning and Expression Analysis of 5-HT Receptors in Silkworm (Bombyx mori)
    LI Hai-yin, LI Yan, CHEN Xi, CHEN Peng, CHEN Ping
    Scientia Agricultura Sinica    2015, 48 (5): 987-1001.   DOI: 10.3864/j.issn.0578-1752.2015.05.17
    Abstract460)   HTML3)    PDF (20736KB)(2707)       Save
    【Objective】The objective of this study is to clone four kinds of 5-HT receptor genes and investigate their expression in different tissues of Bombyx mori, and to provide basic knowledge for further functional studies of these 5-HT receptor genes.【Method】Four 5-HT receptors were cloned based on genome database and using RT-PCR techniques. The bioinformatic method was used to analyze 5-HT receptor genes of B. mori and homology between species. The expression profiles of these genes in different tissues of larvae and adults were investigated by using semi-quantitative real-time PCR.【Result】Based on the predicted gene sequences, special primers were designed to clone the 5-HT receptor genes. Four 5-HT receptor genes 5-HT1ABm, 5-HT1BBm, 5-HT2Bm and 5-HT7Bm (GenBank accession number: KM236100-KM236103) were cloned. The open reading frame (ORF) of 5-HT1ABm, 5-HT1BBm, 5-HT2Bm and 5-HT7Bm was 1 395, 1 341, 1 881 and 1 497 bp, which encoded a polypeptide of 464, 446, 626 and 498 amino acids, respectively. They were typical of G protein-coupled receptors with seven transmembrane protein domains. The sequences were aligned and the phylogenetic tree of four 5-HT receptors was analyzed with other insects and vertebrates. Similarity of the amino acid sequence of 5-HT1ABm, 5-HT1BBm, 5-HT2Bm and 5-HT7Bm was only 30.4%. While, the similarity of 5-HT1A, 5-HT1B, 5-HT2 and 5-HT7 were 45.4%, 61.4%, 48.4%, and 54.1%, respectively in insects. In addition, 5-HT receptors had high homology and more conservative transmembrane region than non-transmembrane region in insects and vertebrates. The phylogenetic tree indicated that the same type receptors from different species got together, then the same receptor formed branch as species genetic relationships. The evolutionary relationships of 5-HT receptors in silkworm were close to Manduca sexta. The results of semi-quantitative real-time PCR showed that 5-HT1Abm, 5-HT1BBm expressed in all tissues in larvae, 5-HT2Bm only had expression in head, ventral chain and testis in larvae, 5-HT7Bm had expression in head, ventral chain, midgut, fat body, testis and ovary in larvae. 5-HT1ABm, 5-HT1BBm,and 5-HT7Bm expressedin other tissues except head in male of adults, whereas 5-HT7Bm in the reproductive system expression of male adults was significantly higher than other tissues. 5-HT2Bm did not express in adult.【Conclusion】Four 5-HT receptor genes were cloned in silkworm. They have more conservative in insects and vertebrates, and homologous relationships are close to M. sexta. The results of semi-quantitative real-time PCR showed that the tissue expression patterns were diverse.
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    Progresses in Research of Genome-Wide Association Studies in Livestock and Poultry
    WANG Ji-Ying, WANG Hai-Xia, CHI Rui-Bin, GUO Jian-Feng, WU Ying
    Scientia Agricultura Sinica    2013, 46 (4): 819-829.   DOI: 10.3864/j.issn.0578-1752.2013.04.016
    Abstract627)      PDF (609KB)(2706)       Save
    Genome-wide association study (GWAS) is a new research method for complex traits developed in recent years. In the last several years, many GWAS were performed focusing on economically important traits, genetic defect diseases, resistant or susceptible ability to complex diseases, breed characteristics of the main livestock and poultries. These studies not only enlarged molecular markers used in the marker-assisted selection, but provided important information for elaborate the genetic mechanism of these traits. In this study, the populations, main statistical methods and results of the GWAS in main livestock and poultry were reviewed. Furthermore, some prospect of GWAS in the future was made. This review will provide reference for further research on genetic background of the important traits of livestock and poultry by GWAS.
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    Cited: Baidu(13)
    Function, Research, and Application of Antimicrobial Peptides
    DAN An-Shan, MA De-Ying, FENG Xing-Jun, MA Qing-Quan, DONG Na, WANG Liang, 吕Yin-Feng , ZHU Xin
    Scientia Agricultura Sinica    2012, 45 (11): 2249-2259.   DOI: 10.3864/j.issn.0578-1752.2012.11.014
    Abstract1170)      PDF (631KB)(2658)       Save
    Antimicrobial peptides (AMPs) distribute widely in all organisms. As an important part of the non-specific immune functions of organisms, AMPs process the properties of various biological functions and low tendency of developing drug resistance and have a potential application prospect in many fields. This review deals with classification, biofunction and antibacterial mechanism of AMPs, particularly summurizes the research progess in the exploitation and application of AMPs in animals in recent years.
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    Cited: Baidu(14)
    Advances in Study on Mechanism of Foliar Nutrition and Development of Foliar Fertilizer Application

    Scientia Agricultura Sinica    2009, 42 (1): 162-172.   DOI: 10.3864/j.issn.0578-1752.2009.01.020
    Abstract1559)      PDF (344KB)(2652)       Save

    Advances in study of mechanisms of foliar nutrition and foliar fertilizer application were reviewed. Plant leaves can absorb nutrients as roots do. There are two basic pathways for nutrients entering into the plant tissue: one is through the stomata and the other is via cuticle hydrophilic pores. Some plant leaves also can absorb nutrients actively through plasmodesmata. So the foliar uptake of nutrients depends upon plant types and the state of plants growth, composition of foliar fertilizer, characters of nutrients and environmental factors such as temperature, illumination and so on. Foliar application to dicotyledon is better than to monocotyledon because of different leaf types and structures of surface wax, and of different varietals plants of homogeneity. It is necessary to spray right fertilizers at right stage of plant growth. Some assistants, such as surfactants are required to enhance nutrients absorption in foliar fertilizers, however, most of the surfactants have a poor solubility among compositions of foliar fertilizers as they are pesticides surfactants. The development of foliar application was promoted by the research on mechanism of foliar nutrition, and now foliar application is becoming an important fertilization method. But soil application can not be replaced by foliar application of fertilizers. Foliar spray can be only as an efficiency assistant method to soil application because the quantity of nutrients supplied is limited. In recent years, foliar fertilizers developed promptly both in quantity and variety, but the quality is poor in China. So the techniques of foliar application need to be improved. Scientists should intensify their research on the mechanism of foliar nutrition and the utilization of assistants in foliar fertilizers. In future, it is necessary to enhance the quality of foliar fertilizers and foliar fertilization, study on optimization of specialized foliar fertilizers and the techniques of foliar application.

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    Analysis of Genetic Diversity and Genetic Basis of Chinese Rapeseed Cultivars (Brassica napus L.) by Sequence-Related Amplified Polymorphism Markers
    YanCheng Wen,,,,
    Scientia Agricultura Sinica    2006, 39 (02): 246-256.   DOI: 10.3864/j.issn.0578-1752.at-2005-5414
    Abstract1837)      PDF (578KB)(2575)       Save
    【Objective】In order to investigate the Genetic diversity and genetic basis of rapeseed (B. napus L.) in China, 【Method】Total 130 accessions developed in different years since 1949 were analyzed using SRAP (sequence-related amplified polymorphism) markers. 【Result】A total of 509 amplified fragments and 123 polymorphic fragments were detected by applying 25 SRAP primer combinations. The polymorphic fragment percentage was 24%. The number of amplified fragments and polymorphic fragments per primer combination were 20.4 and 4.9, respectively. 130 B. napus accessions were divided into four groups of A, B, C and D at genetic distance of 0.12. About 78.5% of total accessions were classified into group C. Group C could also be divided into I, II, III, IV and V sub-groups at genetic distance of 0.10. About 58.5% of total accessions were classified into sub-group I, indicating the genetic diversity of 58.5% accessions of total was poor. The results demonstrated that the genetic basis of B. napus L. accessions released before 1980 was the narrowest while those released in 1980s reached the widest. In 1990s, the genetic distances of B. napus L. accessions declined again. The genetic basis of B. napus L. accessions narrowed further after 2000. Though the difference of mean genetic distance between accessions bred in 1990s and after 2000 did not reach a significant level, the difference in mean genetic distance in different periods was at 0.01 or 0.05 significant level. The difference in mean genetic distance between accessions bred in China and introduced from abroad reached a significant level at 0.01. 【Conclusion】All these results showed that SRAP markers were economic, effective, and reliable.
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    Progresses in Research of Litchi Floral Differentiation
    CHEN Hou-Bin, SU Zuan-Xian, ZHANG Rong, ZHANG Hong-Na, DING Feng, ZHOU Bi-Yan
    Scientia Agricultura Sinica    2014, 47 (9): 1774-1783.   DOI: 10.3864/j.issn.0578-1752.2014.09.012
    Abstract599)   HTML6)    PDF (545KB)(2552)       Save
    The commercial production of litchi (Litchi chinensis Sonn.), a subtropical evergreen fruit tree, has been restricted in the two narrow areas of 17o-32o latitude in the north and south hemispheres. Within the multiple cultivars-based industrial structures, unreliable flowering has taken place in almost all litchi cultivars. The flowering status of varieties, flower bud differentiation stage performance, influence factors, flower bud differentiation mechanism and flowering regulation are reviewed in this paper. It is considered that, litchi trees must have fully matured terminal shoots in autumn and be florally induced under chill conditions for a quite long period during the winter. Primordial inflorescence initiated upon the ascending temperatures and with release of water stresses during late winter or early spring, the inflorescence and flowers developed at mild conditions with warm days and cool nights. Any one unsatisfactory factor during the phase changes might cause failure of flower formation. The major factors influencing litchi floral development are state of terminal shoots, temperatures and water. Carbohydrate accumulation in shoots and the endogenous hormones affect flowering. In order to solve the instability problem in production of flowering and ensure good flowering under extreme and marginal conditions, the molecular mechanism about litchi floral differentiation and why young shoots and leaves could not be florally induced with chills, how the chill signals perceived and transduced, stimulate the floral transition of the terminal buds, when and how the flower sex is determined should be studied, and to explore nutrient, water management, growth regulators and other mechanical measures according to the phase changes of floral differentiation.
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    Cited: Baidu(13)
    Progress in Plant Small RNA Research via High-Throughput Sequencing
    WEI Bo,ZHANG Rong-zhi,LI Ai-li,MAO Long
    Scientia Agricultura Sinica    2009, 42 (11): 3755-3764.   DOI: 10.3864/j.issn.0578-1752.2009.11.001
    Abstract1141)      PDF (327KB)(2526)       Save

    Small RNAs are 20-30 nucleotides long non-coding RNAs. Small RNA-mediated post-transcriptional regulation represents a new mechanism of gene regulation and plays important roles in plant growth and development as well as responses to different environmental stresses. In plants, there are tremendous amount of small RNAs of various types. The advance of high- throughput sequencing technology has significantly facilitated their discoveries. In this review, in addition to a brief introduction of small RNA features, the authors attempted to summarize recent progresses in small RNA research using next generation sequencing technology and aim to highlight the new insights in their genome organization, function and evolution in plants.

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    Advances in Research of Molecular Ecology of Carbon Fixation Microorganism
    YUAN Hong-Chao, QIN Hong-Ling, LIU Shou-Long, NIE San-An, WEI Wen-Xue, WU Jin-Shui
    Scientia Agricultura Sinica    2011, 44 (14): 2951-2958.   DOI: 10.3864/j.issn.0578-1752.2011.14.012
    Abstract992)      PDF (308KB)(2503)       Save
    Alleviating the global climate change is one of the most important environmental challenges human faced. Autotrophic microbes have a strong adaptability to environmental changes and potential to CO2 fixation, thus it is important for researchers to study on molecular mechanisms of CO2 &#64257;xation by autotrophic microbes. Most of the autotrophic microbes fix CO2 via Calvin cycle, although there are another four alternative pathways. The crucial reaction of this pathway is catalyzed by the enzyme ribulose-1, 5-bisphos-phatecarboxylase/oxygenase (RubisCO). Therefore, the RubisCO and the encode gene can be used as functional gene markers to investigate the community and diversity of the CO2 fixation microorganisms in different ecological systems. The previous molecular analysis of microbial carbon fixation has emphasized on aquatic systems, revealing the characteristics of carbon fixation microbial communities and the response to the different habitats. Recent studies on the terrestrial systems have found out that CO2 fixation microorganism also numerous and widespread in soils whereas the functional implication of those autotrophies is not clear, and need to be future identified. Domestic researches on the molecular ecology of soil CO2 &#64257;xation microorganism as yet have not been reported. In the present paper, the species of carbon fixation microorganism, the dynamics of CO2 &#64257;xation and the current status concerning the carbon fixation by microorganism were reviewed, the main problems and further researches of the molecular ecology of carbon fixation in agricultural ecosystems were also discussed. The aim of this study is to provide useful information for Chinese molecular ecology research of CO2 fixation microorganism.
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    Cited: Baidu(13)