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    16 December 2021, Volume 54 Issue 24
    CROP GENETICS & BREEDING·GERMPLASM RESOURCES·MOLECULAR GENETICS
    Construction of High-Density Genetic Map and QTL Analysis of Grain Shape in Rice RIL Population
    ZHANG YaDong,LIANG WenHua,HE Lei,ZHAO ChunFang,ZHU Zhen,CHEN Tao,ZHAO QingYong,ZHAO Ling,YAO Shu,ZHOU LiHui,LU Kai,WANG CaiLin
    Scientia Agricultura Sinica. 2021, 54(24):  5163-5176.  doi:10.3864/j.issn.0578-1752.2021.24.001
    Abstract ( 926 )   HTML ( 106 )   PDF (1849KB) ( 351 )   Save
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    【Objective】Rice grain shape is an important agronomic trait directly related to yield, which affects the appearance quality and commercial value of rice. Research on new rice grain shape genes is of great value for revealing the genetic mechanism of rice grain shape, and it can provide some new genetic resources for molecular breeding. 【Method】In the present study, a RIL population which constructed by an extra-large grain japonica rice variety TD70 and a small-grain indica rice variety Kasalath was used as the research material. The phenotypic data of grain shape, such as grain length, grain width, grain thickness and thousand grain weight were investigated. Using the Genotyping-By-Sequencing approach to re-sequence the parents and RILs to obtain SNP information. The sliding window method (the number of SNP/InDel is 15) was used for genotype calling and recombination breakpoint determination. Based on these results, a high-density Bin map was constructed. Meanwhile, the compound interval mapping method of QTL IciMapping software was used to map the QTLs related to grain shape. 【Result】A high-density genetic map containing 12 328 Bin markers was constructed. The number of Bin markers on each chromosome is 763 to 1367, and the average physical distance between markers was 30.26 kb. The frequency distribution of each trait for RIL population was continuous, which were consistent with the characteristics of quantitative characters, so it was suitable for the detection of QTL. QTL analysis of RIL population in 2018 showed that 40 grain-shape QTL were detected, including 12 grain length QTL, 9 grain width QTL, 8 grain thickness QTL, and 11 thousand-grain weight QTL. QTL analysis was performed of RIL population in 2019, and 56 grain-related QTL were detected, including 15 grain length QTL, 11 grain width QTL, 13 grain thickness QTL, and 17 thousand-grain weight QTL. Based on the two-year mapping results, we have mapped a total of 96 grain shape QTL. We found that 11 QTL could be detected for two consecutive years; among them, 7 QTL have been cloned and 4 new QTL were distributed on 1, 3, 4 and 5 chromosomes. Among the 4 new QTL, qGL-1-2 and qGL-5-2 was related to grain length, qGT-3-2 related grain thickness and qGW-4-1 related to grain width. 【Conclusion】We constructed a molecular genetic linkage map containing 12 328 Bin markers and used the map to analyze the grain shape loci of extra-large grain rice resources. Four new QTLs related to grain shape were obtained, which can be used for subsequent fine mapping, cloning and functional studies.

    Analyzing Genetic Effects for Plant Height and Panicle Traits by Means of the Mixed Inheritance Model of Major Gene Plus Polygene in Foxtail Millet
    GUO ShuQing,SONG Hui,YANG QingHua,GAO JinFeng,GAO XiaoLi,FENG BaiLi,YANG Pu
    Scientia Agricultura Sinica. 2021, 54(24):  5177-5193.  doi:10.3864/j.issn.0578-1752.2021.24.002
    Abstract ( 638 )   HTML ( 47 )   PDF (2625KB) ( 250 )   Save
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    【Objective】Plant height and panicle traits are key yield-dependent traits in foxtail millet. The objective was to probe into inheritance patterns of plant height and panicle traits and provide a reference basis for genetically improving related traits and mapping their genes. 【Method】Yugu 18, a high performing foxtail millet variety, was arranged as the male parent to cross two foxtail millet varieties, Huangruangu and Hongjiugu, and thus two F7 populations of which each was composed of recombinant inbred lines involving 250 family lines(YYRIL and YRRIL)were established. Phenotypic data of Five agronomic traits of the two populations, plant height, panicle length, internode length under panicle, spikelet number per panicle and grain weight per panicle, were genetically examined in two different environments using the mixed inheritance model of major gene plus polygene. 【Result】In these two environments, all the five agronomic traits showed continuous variations with their kurtosis and skewness values standing at the absolute value of less than 1 and thus presenting a distribution close to a normal distribution, were characterized by typical inheritance of quantitative traits; some of these traits saw super-parent separation phenomena. The correlation analysis among the traits showed that the plant height appeared significantly and positively correlated with the panicle length, and an extremely significantly positive correlation between spikelet number per panicle and grain weight per panicle was also found in the two environments. The analysis by the inheritance model showed that the best inheritance models for the plant height of the YYRIL and YRRIL population were the PG-AI and PG-A polygene models, and the heritability of the polygenes standing at 95.15% and 91.27%, respectively. The best inheritance models for the spikelet number per panicle of the two populations were the PG-AI, with the heritability of the polygenes standing at 70.07%-71.58%. The best inheritance models for the internode length under panicle of the two populations were the 4MG-CEA and 3MG-CEA of which both were models for equally additive major genes. In YYRIL, the heritability of the major genes for the internode length under panicle stood at 9.69%, and the four pairs of major genes had an equal additive effect value of -0.34, taking negative effect; and in the YYRIL, the heritability of the major genes for the internode length under panicle stood at 45.78%, and the 3 major gene pairs in question had an equal additive effect value of 1.17, taking positive effect. In the YYRIL, the best inheritance model for the panicle length was the MX2-ED-A, a model for two pairs of dominant epistatic major genes and additive polygenes, with the heritability of the major genes and polygenes standing at 43.56% and 50.56%, respectively. the two pairs of panicle length-dependent major genes separately had the additive effect values of -1.21 and 1.68 and the polygenes had a lower additive effect value of -0.0017; in the YRRIL, the best inheritance model for the panicle length was the MX2-AE-A, a mixed inheritance model for two pairs of accumulative effect major genes and additive polygenes; the major genes and polygenes for the panicle length had heritability values standing at 46.40% and 46.91%, respectively. The first pair of panicle length-dependent major genes had an additive effect value of 1.53, taking positive effect; The additive and epistatic interactions effect value of the first×the second pairs of major genes were 0.60. The polygenes had an additive effect value of -0.47, taking the lower negative inheritance effect. In the YYRIL, the best inheritance model for the grain weight per panicle was the MX2-ED-A; the grain weight per panicle followed the inheritance model for two pairs of dominant epistatic major genes + additive polygenes with the heritability of the major genes and polygenes standing at 69.09% and 12.08%; the additive effect values of the two pairs of grain-weight per panicle-dependent major genes were separately 0.58 and 5.82, with the additive effect of the second pair of major genes dominating, and the additive effect value of polygenes stood at a value of -3.81. In the YRRIL, the best inheritance model for the grain weight per panicle was the 3MG-PEA, an inheritance model for three pairs of partially equal additive major genes; the heritability of the grain weight per panicle-dependent major genes stood at 81.10% and the additive effect values of the three pairs of major genes separately were -2.68, -2.68and 2.66, all taking negative effect. 【Conclusion】In foxtail millet, the plant height and spikelet number per panicle had similar inheritance models, were all under polygenic control with a higher heritability and environmentally affected to a slight content; the inheritance of the internode length under panicle was genetically controlled by major genes, which had a lower heritability and were environmentally affected to a great extent, and thus environmental factors should be taken into full account in production; the panicle length was genetically controlled jointly by major genes and polygenes; the grain weight per panicle was genetically controlled by major genes with a high heritability in both of the two population and probably carried major QTL.

    TILLAGE & CULTIVATION·PHYSIOLOGY & BIOCHEMISTRY·AGRICULTURE INFORMATION TECHNOLOGY
    Effects of Straw Mulching and Phosphorus Application on Wheat Yield, Phosphorus Absorption and Utilization in Hilly Dryland
    XIANG XiaoLing,CHEN SongHe,YANG HongKun,YANG YongHeng,FAN GaoQiong
    Scientia Agricultura Sinica. 2021, 54(24):  5194-5205.  doi:10.3864/j.issn.0578-1752.2021.24.003
    Abstract ( 385 )   HTML ( 5 )   PDF (655KB) ( 195 )   Save
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    【Objective】 The main limiting factors of winter wheat production are less rainfall in winter and spring, low content of soil organic matter and serious lack of available phosphorus in hilly dryland of Sichuan province. In order to provide a technical scheme for the high and stable yield of wheat and phosphorus efficient utilization of dryland wheat in hilly region of Sichuan province, this experiment studied the effects of straw mulching in idle season and phosphorus application on dryland winter wheat yield, phosphorus absorption and utilization. 【Method】 A two-year experimental design from 2018 to 2020 in Renshou, Sichuan province was as follows: straw mulching (SM) or without straw mulching (NSM) as the main-plot treatment, and three kinds of phosphorus application of 0 (P0), 75 (P75) and 120 (P120) kg·hm-2 as the sub-plot treatment. The differences of dry matter accumulation and transport, yield traits and phosphorus uptake and utilization of wheat were analyzed under straw mulching and phosphorus application. 【Result】 The yield increasing effect of straw mulching was higher than that of phosphorus application. In 2018-2019 and 2019-2020, compared with the non-mulched treatments, the effective spike, grain number per spike and yield of wheat in the mulched treatments were increased by 17.7% and 8.48%, 15.6% and 11.2%, 18.6% and 13.5%, respectively. Compared with no P application, the effective spike, grain number per spike and yield of wheat in 75 kg·hm-2 P application for the two seasons of wheat increased by 18.2% and 8.79%, 21.1% and 6.09%, 30.2% and 16.1%, respectively. Similarly, compared with no P application, the effective spike, grain number per spike and yield of wheat in 120 kg·hm-2 P application increased by 21.2% and 9.53%, 20.2% and 4.03%, 31.8% and 17.9% respectively. In addition, the straw mulching significantly increased the dry matter and phosphorus accumulation of wheat at anthesis and maturity stages, which were increased with the increase of phosphorus application. Additionally, straw mulching and phosphorus application significantly enhanced the amount of pre-anthesis dry matter and phosphate translocation, and improved the dry matter and phosphate accumulation of grain. Straw mulching also significantly increased the contribution rate of pre-anthesis dry matter transporting to grain, but did not increased the contribution rate of pre-anthesis phosphorus transport to grain. While phosphate application significantly increased the contribution rate of pre-anthesis dry matter and phosphorus transport to grain. Straw mulching before sowing promoted the absorption and utilization of phosphorus, the absorption efficiency of phosphorus fertilizer increased by 27.3% and 23.7%, respectively, and the partial productivity of phosphorus fertilizer increased by 17.8% and 14.7%, respectively. 【Conclusion】 Straw mulching was more beneficial to promote dry matter and phosphorus transport before anthesis, but also to promote phosphorus accumulation after anthesis and to improve the absorption and utilization efficiency of phosphorus fertilizer. The yield increase was achieved by increasing effective spike and grain number per spike. Overall, the straw mulching combined with 75 kg·hm-2 phosphorus fertilizer was a high-yield and high-efficiency cultivation measure and phosphorus fertilizer management plan for wheat in hilly dryland in Sichuan.

    Effects of Tillage Practices and Nitrogen Fertilizer Application Rates on Grain Yield, Protein Content in Winter Wheat and Soil Nitrate Residue in Dryland
    HUANG Ming,WU JinZhi,LI YouJun,FU GuoZhan,ZHAO KaiNan,ZHANG ZhenWang,YANG ZhongShuai,HOU YuanQuan
    Scientia Agricultura Sinica. 2021, 54(24):  5206-5219.  doi:10.3864/j.issn.0578-1752.2021.24.004
    Abstract ( 410 )   HTML ( 5 )   PDF (756KB) ( 222 )   Save
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    【Objective】 The objective of the present study was to clarify the suitable combined pattern of tillage and nitrogen (N) rate for dryland wheat to achieve the target of high-yield, high-quality and environment-friendly production. 【Method】 In the dry year 2016-2017 and wet year 2017-2018, a field experiment was carried out in the typical dryland in the western region of Henan province. In the experiment, the two tillage practices, including subsoiling (ST) and ploughing (PT), were set as main treatment, and the four N fertilizer application rates of 0 (N0), 120 (N120), 180 (N180) and 240 kg·hm-2 (N240), respectively were set as secondary treatment. The subsoiling operation in ST was interval one year and conducted about two weeks after the previous wheat harvest, and the ploughing operation in PT was carried out each year around late July to early August after once heavy precipitation. The grain yield, grain protein content and its yield, and plant N absorption and utilization in wheat were tested, as well as the nitrate residue in the 0-200 cm soil layer in dryland. 【Result】 The plant N accumulation after jointing stage, grain yield, protein yield and N use efficiency in wheat and the nitrate residue in 0-200 cm soil layer at harvest could be significantly regulated by annual precipitation type, tillage practice and N rate, and the interaction of tillage practice and N rate. Compared with PT, ST increased the shoot N accumulation after jointing and the pre-anthesis N translocation under all the four N treatments, and the N harvest index under N240, as well as increasing the N uptake efficiency, N agronomy efficiency, N recovery efficiency and N partial factor productivity by 8.6%-15.3%, 23.9%-86.5%, 8.1%-26.1% and 9.1%-20.3% averaged across different N treatments, respectively. Therefore, compared with PT, the grain yield under ST was significantly increased by 11.9% and 12.4%, respectively, and the grain protein content was kept no significant change, while the average protein yield was increased by 12.4% and 13.5%, but the average nitrate residue was respectively reduced by 11.9% and 25.4% in 0-200 cm soil layer averaged across all the four N treatments in the dry year and the wet year. With the increase of N rate, the shoot N accumulation, pre-anthesis N translocation amount, contribution rate of post-anthesis N accumulation to grain, and grain protein content in wheat and the soil nitrate residue at harvest were significantly increased, and there was a significant decrease on the contribution rate of pre-anthesis N translocation to grain, N grain production efficiency, N uptake efficiency and N partial factor productivity, but the changes in N agronomy efficiency, N recovery efficiency, grain yield and protein yield varied with annual precipitation type and tillage practice. The STN240 had the highest shoot N accumulation in the two years. In addition to no significant difference of grain and protein yield between ST240 and ST180 in the dry year and also of the protein content between ST240 and PT240 in the wet year, the grain yield, protein content and protein yield in ST240 were significantly higher than the other treatments in the two experimental years, the N recovery efficiency of and N agronomy efficiency in the wet year under ST240 were not less than or even significantly higher than that in the N application treatments under PT, and therefore decreased the nitrate residue in 0-200 cm soil at harvest by 16.4% compared with PT240. In general, the N rate at 180 kg·hm-2under PT could reach the highest grain yield and the optimal protein yield, N agronomy efficiency and N recovery efficiency. Compared with other treatments, the N rate at 240 kg·hm-2under ST was the best combination, which could increase the N efficiency and reduce soil nitrate residue via subsoiling during summer fallow period, as well as improving the protein content through the increased N fertilizer rate, and finally increased the grain yield and protein yield by 2.6%-45.0% and 7.3%-81.4%, respectively. 【Conclusion】 Subsoiling tillage could help synchronously to improve the grain yield, protein yield and N efficiency and reduce soil nitrate residue. The suitable N application rate for subsoiling tillage should be higher than that for ploughing tillage. The PTN180 was an optimal combination of tillage practice and N rate for high-yield and high-efficiency, and the STN240 was an optimal model for realizing the collaborative target of high-yield, high-quality, high-efficiency, and low-nitrate residue in dryland wheat production system.

    PLANT PROTECTION
    The Resistance Mechanism of Apple Transcription Factor MdWRKY40b to Powdery Mildew
    SHA RenHe,LAN LiMing,WANG SanHong,LUO ChangGuo
    Scientia Agricultura Sinica. 2021, 54(24):  5220-5229.  doi:10.3864/j.issn.0578-1752.2021.24.005
    Abstract ( 349 )   HTML ( 36 )   PDF (4179KB) ( 154 )   Save
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    【Objective】The research aims to explore the mechanism of apple resistance to powdery mildew regulated by transcription factor gene MdWRKY40b, and to provide a theoretical basis for apple powdery mildew resistance breeding. 【Method】The 552 bp specific fragment of MdWRKY40b was cloned using the cDNA obtained from the ‘Gala’ leaf tissue, the forward and reverse sequence of this gene specific fragment was constructed into the RNAi expression vector pB7GWIWG2(Ⅱ) by the Gateway technique, and then transgenic plants were obtained by an Agrobacterium-mediated method. Notably, the young leaves at the stem tips of tissue culture seedlings were chosen as the materials for Agrobacterium-mediated infection experiments. The expression levels of MdWRKY40b, superoxide dismutase gene (SOD), catalase gene (CAT), peroxidase gene (POD), and β-1,3-glucanase in transgenic plants were analyzed by RT-qPCR technology. Furthermore, using one-year-old plants as materials, the resistance of transgenic plants to powdery mildew was analyzed through the inoculation test of the powdery mildew pathogen (Podosphaera leucotricha). The leaves of transgenic and control plants at 0, 5, 10, 15, and 20 d after inoculation with P. leucotricha were used as materials, and the accumulation of superoxide anion and hydrogen peroxide in plant materials during P. leucotricha infection was analyzed by staining with nitrotetrazolium chloride solution (NBT) and diaminobenzidine (DAB). Furthermore, the activities of SOD, POD, CAT, and β-1,3-glucanase in plants were determined during P. leucotricha infection. 【Result】Three MdWRKY40b gene silencing lines were identified by PCR detection. The RT-qPCR results showed that the gene silencing efficiency of RNAi-1, RNAi-2, and RNAi-3 was 95.2%, 92.2%, and 79.8%, respectively. The expression level of SOD, CAT, and β-1,3-glucanase in transgenic plants was significantly up-regulated compared with the wild-type. Under the condition of P. leucotricha infection, compared with wild-type plants, the area of powdery disease spots in the leaves of transgenic plants decreased significantly, the accumulation of superoxide anion and hydrogen peroxide was reduced significantly, and the activities of antioxidant enzymes such as SOD, CAT, and POD regulating superoxide anion metabolism and β-1,3-glucanase related to disease resistance were significantly enhanced. 【Conclusion】The silencing of MdWRKY40b enhances the resistance of apple plants to powdery mildew. It is speculated that the up-regulated expression level of SOD, CAT, and β-1,3-glucanase in transgenic plants improves the basic resistance of apple plants to powdery mildew, and increases the scavenging ability of plants to superoxide anions. It can maintain a low concentration of superoxide anion content in response to P. leucotricha infection, so as to reduce the damage of high concentrations of superoxide anion to plants.

    Investigation and Analysis of Weed Community Succession in Winter Wheat Field of Shandong Province
    GAO XingXiang,ZHANG YueLi,AN ChuanXin,LI Mei,LI Jian,FANG Feng,ZHANG ShuangYing
    Scientia Agricultura Sinica. 2021, 54(24):  5230-5239.  doi:10.3864/j.issn.0578-1752.2021.24.006
    Abstract ( 486 )   HTML ( 22 )   PDF (694KB) ( 131 )   Save
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    【Objective】In the past 10 years, the species, quantity and occurrence area of weeds in winter wheat field of Shandong Province have changed significantly. The objective of this study is to clarify the succession law and reason of weed community, and to provide a theoretical basis for making precise control strategy of weeds in wheat field. 【Method】A total of 300 investigation sites were investigated in the winter wheat field of Shandong Province, and three areas were selected for each investigation site. The weed community in winter wheat field of Shandong Province for each area was investigated three times from 2009 to 2019 using inverted W-pattern sampling method, and the distribution and change rule of weed community in seven regions of winter wheat field of Shandong Province were analyzed. 【Result】The characteristics of weed community changes in winter wheat field of Shandong Province were: (1) Descurainia sophia and Capsella bursa-pastoris had always been the dominant weeds in winter wheat field of Shandong Province, and the relative abundance of D. sophia always ranked first in the three surveys, which was 20.68%, 22.49% and 22.24%, respectively, and C. bursa-pastoris always ranked in the top three, with the relative abundance of 15.49%, 15.77% and 14.51%, respectively. (2) There were more and more species of dominant weeds, and the degree of weed damage was more and more serious. In the 2009-2010 annual survey, there were only two species D. sophia and C. bursa-pastoris with an relative abundance of ≥10.00%. In the last two surveys, there were three species (increased Bromus japonicus) and five species (increased B. japonicus, Aegilops squarrosa and Galium aparine). (3) The occurrence and harm degree of gramineous weeds increased gradually. In 2009-2010, B. japonicus and A. squarrosa were mainly distributed in northwest plain region, north coastal region and central mountain region, in 2013-2014, they spread to the southern mountain region and plain regions of middle Shandong, and in 2018-2019, they had spread all over the province. The relative abundance of B. japonicus was 7.66%, 16.39% and 17.94% in the three surveys, and that of A. squarrosa was 3.08%, 7.61% and 11.38%, respectively. Although the total dominance of Lolium multiflorum, Alopecurus myosuroides and Avena fatua was not high, but it increased gradually. (4) The distribution of broad-leaved weeds such as Veronica persica and G. aparine was increasing. The relative abundance of G. aparine in the three surveys was 8.29%, 8.94% and 10.00%, respectively, which increasing year by year, and the distribution area was gradually expanding. Now it was widely distributed in all regions of Shandong Province, especially in the southwest plain region, the southern mountain region and central mountain region, and also in the northwest plain region. The relative abundance of V. persica in the three surveys was 1.08%, 1.18% and 2.05%, respectively, V. persica mainly distributed in the southwest plain region and southern mountain region, and had also been distributed in other regions. (5) The largest change of weed community was occurred in hill regions of eastern Shandong, and the species diversity showed a rising trend. In 2009-2010, the Shannon Wiener index, which reflected species diversity, was only higher than the north coastal region, ranking sixth. In 2013-2014, it surpassed the northwest plain region and plain regions of middle Shandong, ranking fourth. In 2018-2019, it ranked second, only was lower than that of the southern mountain region. 【Conclusion】The change of farming system, the spread and invasion weeds and the continuous application of single herbicide in large area obviously promote the change of weed community in winter wheat field of Shandong Province. According to the results of weed change in winter wheat field of Shandong Province, the comprehensive weed control strategies of chemical herbicide and agronomic measures, precise control technology and alternative use of herbicides with different mechanisms should be promoted.

    SOIL & FERTILIZER·WATER-SAVING IRRIGATION·AGRICULTURAL ECONOMY & MANAGEMENT
    Application Effect of Fungi Promoting Secondary Fermentation in Composting
    WEI QiHang,FENG Yao,MA QianQian,LI YanLi,LIU YuanWang,LI ZhaoJun,REN YanFang
    Scientia Agricultura Sinica. 2021, 54(24):  5240-5250.  doi:10.3864/j.issn.0578-1752.2021.24.007
    Abstract ( 465 )   HTML ( 6 )   PDF (1980KB) ( 230 )   Save
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    【Objective】Secondary fermentation is indispensable to improve the functional value of chicken manure or other breeding wastes. This study applied the screened fungi in the secondary fermentation stage of chicken manure composting to explore the effect of the fungi on promoting the secondary fermentation, and the fungi were identified, so as to provide the theoretical basis and support for the high value and resource utilization of chicken manure. 【Method】The products of chicken manure after primary fermentation were used as raw materials, and two strains of fungi ( FCM1 and FCM3 ) were previously screened in the laboratory and were added before secondary fermentation. Three treatments were set up, including raw materials mixed with 1% (V:W,L·kg-1) sterile liquid medium (CK), raw materials mixed with 1% liquid medium containing FCM1 (F1), and raw materials mixed with 1% liquid medium containing FCM3 (F3), and some indexes were detected, including temperature, pH, electroconductibility (EC), ammonium nitrogen, nitrate nitrogen, GI and humic acid in different secondary fermentation stages. 【Result】Compared with CK, the addition of fungi could promote the temperature rise and humification process during the secondary fermentation stage. Compared with the CK, F3 treatment reached the mesophilic period 8 days earlier, but F1 treatment delayed the second fermentation of chicken manure composting. As for F3 treatment, the thermophilic stage (>50℃) lasted 13 days, GI reached 80% at 25 days earlier than that under CK, and the humification degree was relatively higher (PHA=74.58%, PHA: the ratio of humin and total humus). The correlation analysis showed that GI had significant positive correlation with pH and PHA, and negative correlation with EC. In addition, FCM3 was identified as Alternaria sp. by morphology and ITS rDNA sequencing. 【Conclusion】A strain of fungus FCM3 could promote the secondary fermentation of chicken manure composting, and improve the humification of composting materials, which was of great significance to promote the high value utilization of chicken manure.

    The Spatial and Temporal Evolution, Regional Correlations and Economic Coordinated Development Effect for Chinese Agricultural Science and Technology Level: Taking Provincial Public Agriculture Research Institutions as an Example
    YUE HuiLi,ZHANG Zhao,ZHANG HuiJie,LIU ShengPing,ZHANG Jie
    Scientia Agricultura Sinica. 2021, 54(24):  5251-5265.  doi:10.3864/j.issn.0578-1752.2021.24.008
    Abstract ( 383 )   HTML ( 4 )   PDF (3947KB) ( 112 )   Save
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    【Objective】In the long run, agricultural science & technology and agricultural economics should show a dynamic and mutually beneficial collaborative development trend, which is more conducive to accelerating the development of modern agriculture. To study further the spatial and temporal evolution trends and spatial distribution characteristics of science and technology development of pubic agriculture research institutions, and the coordinated development condition between agricultural science & technology and regional economics, contributes to correctly understand the coordinated development mechanism of agricultural science & technology innovation and regional economy among provinces, and analyze the potential reasons for the poor coordinated development degree of agricultural science & technology and agricultural economy, and explore to find the way to improve the agricultural science & technology innovation level and resolve the problem of “wide gap between agriculture S&T and agriculture economy”. 【Method】Based on numbers of agricultural researchers, the patent grants and numbers of published papers, the GIS technology and ESDA methods are applied for the study of spatial and temporal evolution trends and spatial distribution characteristics of science and technology output by provincial public agriculture research institutions, and coordinated development condition between agricultural science & technology and agricultural economics is revealed. 【Result】 The results show that: During ten years, the science and technology output of provincial public agriculture research institutions are decreasing gradually from the eastern coastal areas to the surroundings; the patent grants are increasing obviously for most provincial agriculture research institutions, while the quantity growth of publication of papers and the agricultural researchers are indistinctive. From the perspective of spatial distribution characteristics, notwithstanding the science and technology output of provincial public agriculture research institutions present the characteristics of local spatial autocorrelation, large-scaled spatial agglomeration effects are not formed. From the perspective of coordinated development condition between agricultural science & technology and agricultural economics, the Eastern cities by Shandong, Jiangsu, Guangdong, Fujian etc. had good performance, the central regions is average and the coordinated development condition between agricultural technology and agricultural economics for the most provinces in Northwest, Southwest and Northeast China are weak. Hereby some policy proposals are put forwarded in aspect of improving the level of science and technology of provincial public agriculture research institutions, and promoting the coordinated development condition between agricultural science & technology and agricultural economics. 【Conclusion】During 2008-2018, the science & technology level of provincial public agriculture research institutions gradually decreased from the eastern coastal areas to the surroundings, showing the features of spatial heterogeneity, and large-scaled spatial agglomeration effects has not been formed. The coordinated development degree between agricultural science & technology level of provincial public agriculture research institutions and provincial agricultural economics is lower, consequently, it is necessary to accelerate the construction of the spatial agglomeration pattern of the coordinated development of agricultural science & technology and agricultural economics, which provides support for solving the problem of “wide gap between agriculture S&T and agriculture economics”.

    HORTICULTURE
    The Role and Mechanism of Tomato SlNAC29 Transcription Factor in Regulating Plant Senescence
    WANG Ping,ZHENG ChenFei,WANG Jiao,HU ZhangJian,SHAO ShuJun,SHI Kai
    Scientia Agricultura Sinica. 2021, 54(24):  5266-5276.  doi:10.3864/j.issn.0578-1752.2021.24.009
    Abstract ( 502 )   HTML ( 36 )   PDF (2708KB) ( 185 )   Save
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    【Background】 Tomato(Solanum lycopersicum)is an important horticultural crop with continuous flower bud differentiation and fruiting. Premature senescence seriously limits tomato plants growth period, crop yield and fruit quality. NAC (NAM, ATAF1/2 and CUC2) transcription factor family regulates leaf senescence process in Arabidopsis, rice and other plants. Nevertheless, the roles of tomato NAC transcription factor in the regulation of leaf senescence have not been well understood. SlNAP2 (NAC-like, activated by apetala3/pistillata) is known to be involved in the regulation of tomato leaf senescence. 【Objective】 SlNAC29 transcription factor is the homologous gene of SlNAP2 in tomato, while its function remains largely unclear. In this study, the role of SlNAC29 and its underlying mechanism in leaf senescence was investigated, which can provide some scientific basis for tomato senescence regulation and germplasm innovation. 【Method】Condine Red (CR) was used as the wild-type background in this study. qRT-PCR was used to analyze the relative expression of SlNAC29 in tomato leaves at different senescence stages. Slnac29 homozygous mutant lines and the OE: SlNAC29 stable overexpression lines were generated through CRISPR/Cas9 gene editing and over-expression approaches, respectively. Using these lines, plant growth phenotypes, chlorophyll content, leaf photosynthesis, transcription of senescence- and chlorophyll degradation- related genes were analyzed under both natural and dark-induced senescence conditions. The clustering heat map was used to analyze the relative expression of 29 genes, including senescence-, chlorophyll degradation- and ABA biosynthesis/signaling-associated genes. Based on gene expression profiles, four of them were selected to electrophoretic mobility shift analysis (EMSA) to identify the SlNAC29-target gene during senescence process. 【Result】The relative expression of SlNAC29 was significantly up-regulated in early senescent and senescent leaves, as compared with young and mature leaves. Under natural growth condition, the Slnac29 mutant lines showed no differences with the wild-type in terms of plant growth phenotypes and photosynthetic rate. By contrast, the height of OE: SlNAC29 plant was shorter than wild-type plants, OE: SlNAC29 plants also showed lower chlorophyll content and photosynthetic rate, which were only 25% and 50% of the wild-type control, respectively. Under dark-induced senescence condition, the leaves of wild-type plants turned yellow and the chlorophyll content decreased significantly. The senescent phenotypes were alleviated in Slnac29 mutant lines, which not only have significant higher chlorophyll content, but also showed higher transcript level of senescence-associated genes (SAGs) and chlorophyll degradation-related genes. On the contrary, the dark-induced senescence effect was aggravated in OE: SlNAC29. Cluster analysis showed that several genes, especially SAGs and chlorophyll degradation-related genes SlSAG12, SlAGT1, SlSGR1 and SlNYC1, were significantly up-regulated in OE: SlNAC29 plants. The EMSA analysis showed that SlNAC29 could directly bind to the promoter of SlAGT1 (Glyoxylate aminotransferase), a member of SAGs. Moreover, the relative expression of SlAGT1 in OE: SlNAC29 was significantly higher than that of wild-type and Slnac29 plants. 【Conclusion】 SlNAC29 transcription factor is involved in the regulation of leaf senescence in tomato plants, which promotes the senescence process under dark conditions. SlNAC29 may directly bind to the promoter region of the senescence-related gene SlAGT1 to regulate its transcriptional expression.

    Accumulation of Sugar and Flavonoids as Well as Their Association with Changes of Light Intensity During Fruit Development of Rosa roxburghii
    FAN WeiGuo,PAN XueJun,HE ChunLi,CHEN Hong,ZHOU YuJia
    Scientia Agricultura Sinica. 2021, 54(24):  5277-5289.  doi:10.3864/j.issn.0578-1752.2021.24.010
    Abstract ( 478 )   HTML ( 30 )   PDF (811KB) ( 107 )   Save
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    【Objective】The accumulation of sugar and total flavonoids and their association with changes of light intensity during fruits development of Rosa roxburghii Tratt. were studied, so as to provide a scientific basis for the fruit quality control of Rosa roxburghii. 【Method】 Four-year-old ‘Guinong 5’ fruit-bearing trees of Rosa roxburghii was used as the material in the experiment, the natural light intensity was used as control (R0, CK), and three treatments were set up with the light intensity reduced by 20% (R20), 40% (R40) and 60% (R60), respectively. The accumulations of soluble sugar and total flavonoids as well as activities of related anabolic enzymes in fruits were analyzed, and the relationship between them and light intensity was determined. 【Result】Sugar and total flavonoids were accumulated continuously during the growth and development of Rosa roxburghii fruit, but their accumulation of different development periods had significant differences. The sugar in fruits began to accumulate rapidly after the slow growth stage, and the accumulation of total soluble sugar and sucrose reached the highest at maturity stage, in which the accumulation of sucrose accounted for 36.97% of the total soluble sugar. The accumulation of glucose and fructose in the rapid expansion period was the largest, but only accounted for 10.50% and 18.18% of the maximum accumulation of total soluble sugar at fruit maturity stage, respectively. Rosa roxburghii fruit was sucrose accumulation type. The total flavonoids in fruits were rapidly accumulated from young fruit stage to rapid fruit expansion stage, and then the accumulation of total flavonoids did not increase significantly. During the development of Rosa roxburghii fruit, the accumulation of sugar and total flavonoids under different light intensities was significantly different, while the decrease of light intensity was not conducive to the accumulation of sugar and total flavonoids in fruits. Sucrose synthase (SS), sucrose phosphate synthase (SPS), invertase (IVR) were the key enzymes affecting the metabolism of sugar in fruit. Alanine ammonia lyase (PAL), cinnamic acid-4-hydroxylase (C4H), 4-coumaric acid coenzyme A ligase (4CL) and chalcone synthase (CHS) were the key enzymes affecting the metabolism of flavonoids in fruit. The changes of light intensity were closely related to the accumulation of sugar, total flavonoids and the activities of related anabolic enzymes in fruits, while the activities of SS, SPS, IVR, PAL, C4H, 4CL and CHS were significantly inhibited by the decrease of light intensity. SS and SPS were sensitive to the decrease of light intensity during the whole fruit development of Rosa roxburghii. There was a significant positive correlation between sugar and total flavonoids accumulation in fruits, while the accumulation of sugar and total flavonoids and the activities of SS, SPS, IVR, PAL, C4H, 4CL and CHS were significantly or extremely significantly positively correlated with the changes of light intensity. 【Conclusion】Rosa roxburghii fruit were sucrose accumulation type, and the sugar in fruits began to accumulate rapidly after the slow growth stage, while the total flavonoids in fruits were rapidly accumulated from young fruit stage to rapid fruit expansion stage. The decrease of light intensity was not conducive to the accumulation of sugar and flavonoids in fruits. The contents of sugar and flavonoids in fruits could be increased by improving light conditions in production, which was conducive to improving the quality of Rosa roxburghii fruit.

    FOOD SCIENCE AND ENGINEERING
    Effects of Trolox on Proliferation and Differentiation of Pig Muscle Stem Cells
    HU RongRong,DING ShiJie,GUO Yun,ZHU HaoZhe,CHEN YiChun,LIU Zheng,DING Xi,TANG ChangBo,ZHOU GuangHong
    Scientia Agricultura Sinica. 2021, 54(24):  5290-5301.  doi:10.3864/j.issn.0578-1752.2021.24.011
    Abstract ( 392 )   HTML ( 25 )   PDF (3538KB) ( 104 )   Save
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    【Objective】The objective of this study was to investigate the regulatory effects of Trolox, an antioxidant water-soluble vitamin E analogue, on the proliferation and differentiation of pig muscle stem cells, which possibly was affected by reactive oxygen species. The study could provide a theoretical fundament for further optimizing the proliferation and differentiation process of cultured meat seed cells in vitro. 【Method】Initially, 0, 50, 100 and 200 μmol·L-1 of Trolox was added to pig muscle stem cells during their proliferation culture for 3 d, respectively. The blood cell counter and CCK8 technology were both used to detect the influence of Trolox on the cell proliferation. RT-qPCR was employed to measure the expression level of PAX7 gene for the further characterizing cellular stemness induced by Trolox. Meanwhile, Western Blotting was also used to testify the expression level of PAX7 protein. The intracellular reactive oxygen species was stained by CellROX fluorescent dye, and the High-throughput High-content Live Cell Confocal Imaging System was adopted to evidence the regulatory effect of Trolox on reactive oxygen species. Trolox was additionally used to the in vitro myogenic differentiation of pig muscle stem cells. RT-qPCR was used to detect the expression levels of early differentiation marker genes MYOG, CAV-3 and terminal differentiation marker gene myosin heavy chain (MyHC). Western Blotting was applied to detect the expression of MyHC protein, whereas Immunofluorescence technique stained MyHC and counted the proportion of MyHC positive cells. 【Result】The cell proliferation fold statistics indicated that the proliferation fold of pig muscle stem cells in 50 or 100 μmol·L-1 Trolox treatment groups was significantly higher than that under the control group (P<0.05); CCK8 test showed that the absorbance value under 50 or 100 μmol·L-1 Trolox treatment groups on the 3rd day was significantly higher than that under the controls (P<0.05); Trolox concentrations at 100 or 200 μmol·L-1 significantly up-regulated the expression of PAX7 gene (P<0.05), but had no dominant effect for the improvement of PAX7 proteins with no statistical difference (P>0.05). The level of reactive oxygen species in the cells was significantly reduced (P<0.001) when Trolox applied. Moreover, after the addition of Trolox to the cells in their differentiation process, MYOG and CAV-3 in the predifferentiation stage as well as MyHC genes in the terminal differentiation stage were dramatically up-regulated (P<0.05). However, Western Blotting results exhibited that the expression of MyHC protein had no a huge change (P>0.05), while the immunofluorescence results displayed that the proportion of MyHC positive cells had an increasing trend but there was no statistical difference (P>0.05). 【Conclusion】Trolox promoted the proliferation and differentiation of pig muscle stem cells via reducing reactive oxygen species in the cells.

    ANIMAL SCIENCE·VETERINARY SCIENCE·RESOURCE INSECT
    Genetic Diversity and Origin Characteristics of Chicken Species Based on Mitochondrial DNA D-loop Region
    TANG XiuJun,FAN YanFeng,JIA XiaoXu,GE QingLian,LU JunXian,TANG MengJun,HAN Wei,GAO YuShi
    Scientia Agricultura Sinica. 2021, 54(24):  5302-5315.  doi:10.3864/j.issn.0578-1752.2021.24.012
    Abstract ( 345 )   HTML ( 24 )   PDF (4094KB) ( 128 )   Save
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    【Objective】 The purpose of this study was to investigate the genetic diversity and origin characteristics of the whole sequence of mitochondrial DNA D-loop region in chicken breeds (complete set line) with different growth rates, so as to provide a theoretical basis for breeding and traceability of broiler breeds. 【Method】 15 broiler breeds with different growth rates were used as research materials, including eight yellow-feathered broiler lines (5 medium-fast and 3 slow lines), two local chicken breeds (Gushi chicken and Tibetan chicken), two introduced chicken breeds (Recessive White and Anka), one white-feathered broiler (Ross 308), 817 hybrid broiler and one commercial layer line (Dawu Brown Eggshell Hens). Chicken blood was collected, and DNA was PCR amplified. The full sequences of mtDNA D-loop region of 683 individuals from 15 chicken breeds were sequenced, and the genetic diversity and haplotype characteristics of each chicken breed were analyzed using DnaSP 5.10 software. The genetic distance between breeds was calculated using MEGA 4.0 software, and then a phylogenetic tree was constructed between different haplotypes and the red original chicken. 【Result】The full sequence size of the D-loop region of 15 chicken breeds ranged from 1 231 to 1 232 bp, and individuals with sequence length of 1 231 bp had C-base deletion at 859 bp. 45 variant loci were detected in 683 individuals, which were combined into 53 haplotypes and could be divided into four haplotype groups, including A, B, C and E. Among them, the medium-fast broilers, 817 hybrid broilers and high-yielding laying hens were all haplotype E as the dominant haplotype (≥48.89%); the dominant haplotypes were B haplotypes for Hongguang black chickens, A haplotypes for Jinghai yellow chickens, and four haplotypes were relatively balanced for Xueshan chickens (the proportion of E haplotypes ≤38.46% for three chicken breeds); the haplotypes of local chicken breeds were A and C haplotypes for Gushi chickens and A and B haplotypes for Tibetan chickens. The genetic diversity of the 15 chicken breeds ranged from 0.496 to 0.853 for Hd and from 0.00146 to 0.00673 for Pi. The relatively rich genetic diversity was found in the Xinxing Dwarf Yellow Chicken, Xueshan Chicken, Jinghai Yellow Chicken and Ross 308; the relatively low genetic diversity was found in the Tibetan Chicken, the High Laying Chicken, the Anka Chicken, the Xinxing partridge Chicken No. 4 and the Xugang Yellow Chicken No. 1. The range of Kiumura two-parameter distance of 15 chicken breeds was 0.0016-0.0113, among which the intra-breed genetic distance of Ross 308 was the largest while the intra-breed genetic distance of 817 hybrid broiler and high-yielding chicken was the smallest; the inter-breed genetic distance was the largest between high-yielding chicken and Tibetan chicken, and the smallest between high-yielding chicken and 817 hybrid broiler; the genetic distance between medium-fast broiler was relatively small while the genetic distance between medium-fast broiler, slow-fast and local chicken breeds was relatively large; the genetic distance between Jinghai and local chicken breeds was relatively large. Cluster analysis showed that the haplotypes A, B, and gallus gallus spadiceus were clustered in one group. Haplotype E and gallus gallus murghi were clustered in another group. Haplotype C was clustered with four subspecies of jungle fowl, including gallus gallus murghi, gallus gallus spadiceus, gallus gallus gallus and gallus gallus bankiva. 【Conclusion】The genetic diversity of mitochondrial D-loop region varied among different chicken breeds; E haplotypes were strongly correlated with broiler growth rate, and E haplotypes were the dominant haplotypes in all medium and fast populations, while the proportion of E haplotypes in slow populations was less than 40%; the national chicken population had multiple red proto-chicken maternal origins, indicating that it was domesticated under neutral selection. The results of the study provided a theoretical basis for broiler breed selection and tracing as well as resource exploitation.

    Advance in Genome-Wide Scan of Runs of Homozygosity in Domestic Animals
    ZHANG PengFei,SHI LiangYu,LIU JiaXin,LI Yang,WU ChengBin,WANG LiXian,ZHAO FuPing
    Scientia Agricultura Sinica. 2021, 54(24):  5316-5326.  doi:10.3864/j.issn.0578-1752.2021.24.013
    Abstract ( 933 )   HTML ( 62 )   PDF (728KB) ( 267 )   Save
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    Runs of homozygosity (ROH) is a long tract of homozygous genotypes commonly found in individuals and populations, which generates on the offspring’s genome inherited identical haplotypes from each parent. ROH contains a wealth of genetic information about populations, which makes it a useful tool for providing information to study how populations change over time. Moreover, ROH can estimate the genetic relationships between individuals to minimize the inbreeding mating rates. In addition, ROH can expose harmful mutations in the genome. The frequencies, sizes and distributions of ROHs in the genome are influenced by natural and artificial selection, recombination, linkage disequilibrium, population history, mutation rate and inbreeding level. Recently, with the use of high-throughput genotype technology and the reduction of second-generation sequencing costs, livestock and poultry breeding have entered into the genomic era. The selection intensity of the elites in livestock and poultry significantly increase to improve their performances, but it will increase inbreeding and cause inbreeding depression as well. Based on ROH molecular information, it is more accurately to detect past and nearest in close relative mating. The ROH-based inbreeding coefficient (FROH) can obtain an individual's true inbreeding coefficient, i.e. the realized inbreeding coefficient, and the pedigree-based FPED is the expectation value of inbreeding coefficient. In the absence of genealogical information, FROH can be used to infer information about a group's history and the inbreeding levels. Meanwhile, the selection reshapes ROH patterns in different regions of the genome. In addition, the selection can increase the homozygosities around the target point, and harmful mutations are thought to occur more frequently in the ROH region, which can be detected by ROH to reduce the risk of complex diseases. After long-term selection, one ROH appeared in multiple individuals’ genomes in the same population, resulting in ROH islands. It has confirmed the correlation between ROH and the selected genomic region. The candidate genes related to economic traits can be annotated on the ROH islands by means of biological information. In addition, ROH also provides a new perspective for assessing the genetic diversity in domestic animals. Genome-wide ROH detection on the population can used to investigate the genetic structure of this population, and FROH can evaluate the impact of inbreeding in the current breeding program, which can adjust breeding plans to protect the genetic diversity of varieties. Therefore, ROH has gradually become an important index to explore the historical population structure, the level of inbreeding, candidate gene identification. There are mainly two kinds of methods to identify ROH: observation genotype counting method and model-based analysis. Commonly used softwares include PLINK, GERMLINE, BEAGLE, GARLIC, etc. In practical applications, PLINK is the most common ROH detection tool. Since the SNP chip for cattle was firstly used in domestic animals, the cattle population was firstly conduct genome-wide ROH detection. Now, studies on ROH are becoming more popular in pigs, sheep and other domestic animals. This review mainly described the principle of ROH formation and its detection methods, as well as progress of its application in livestock and poultry, so as to provide reference for the genetic breeding of livestock and poultry.