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    16 July 2019, Volume 52 Issue 14
    CROP GENETICS & BREEDING·GERMPLASM RESOURCES·MOLECULAR GENETICS
    Breeding of New Wheat Varieties with Early Maturity and High Yield Under Late Sowing
    GAO DeRong,WANG Hui,LIU Qiao,ZHU DongMei,ZHANG Xiao,LÜ GuoFeng,ZHANG XiaoXiang,JIANG Wei,LI Man
    Scientia Agricultura Sinica. 2019, 52(14):  2379-2390.  doi:10.3864/j.issn.0578-1752.2019.14.001
    Abstract ( 463 )   HTML ( 30 )   PDF (400KB) ( 347 )   Save
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    【Objective】The objectives of this study were to investigate the growth characteristics during the period from sowing to elongation and from anthesis to maturity in wheat cultivars (varieties), and to explore the possibility of breeding new cultivars with the feature of rapid growth at both early and late stages to achieve high yield and early maturity under late sowing condition in the middle and lower reaches of the Yangtze river. 【Method】 In the seasons of 2012-2013 and 2013-2014, experiments were conducted to compare leaf number, stem and tiller number and the parameters of grain filling among 18 new wheat lines and cultivars for screening the cultivar with rapid development at both early and late growth stages. In the season of 2014-2015, the differences of stem and tiller number, leaf area index (LAI), dry matter accumulation, yield and yield components were analyzed among Yangmai 16 and other 4 cultivars under the conditions of the normal (5th Nov.) and late (19th Nov.) sowing time to clarify the growth mechanics of high-yield about Yangmai 16 with rapid development at both early and late growth stages under late-sowing treatment.【Result】 Fu F101 showed a rapid development at early growth stage with a significantly higher number of leaves and tillers (before winter and during winter period) than Zhen 10216 in the 2012-2013, Ning 09-118 and Zhen 10216 had longer grain filling and rapid development at late growth stage with higher maximum grain filling rate (Gmax) and mean grain filling rate (Gmean) thus higher grain weight than other cultivars. The 2013-2014 trial showed similar results with Fu F101 performing rapid development at early stage and Zhen 10216 at late stage. Yangmai 16 produced even more tillers than Fu F101 before winter and similar tillers to Fu F101 during winter period. Meanwhile, both Gmean and Gmax of Yangmai 16 were similar to Zhen 10216 with Gmax being more than 2.0 mg/(grain·d). LAI, stem and tiller number and dry matter accumulation of Yangmai 16 in winter period were significantly higher than Ningmai 13 and other cultivars. In 2014-2015 trial, the yield of Yangmai 16 was lower than Yangmai 22 and similar to Yangmai 20, Yangmai 23 and Ningmai 13 under normal sowing time. However, under late sowing, the yield of Yangmai 16 was significantly higher than Yangmai 20 and Ningmai 13 and had the lowest yield reduction (5.2%) compared to other cultivars.【Conclusion】Significant differences were found among varieties in the rate of development at early and late growth stages. The rapid development at both early and late stages of Yangmai 16 makes it an excellent variety with not only a reasonable yield under normal sowing date but minimum yield reduction under late sowing. In summary, rapid development at both early and late stages can be used as a key criterion for selecting wheat varieties suitable for late sowing.

    Phenotypic Variation and Genome-wide Association Analysis of Root Architecture at Maize Flowering Stage
    ZHANG XiaoQiong,GUO Jian,DAI ShuTao,REN Yuan,LI FengYan,LIU JingBao,LI YongXiang,ZHANG DengFeng,SHI YunSu,SONG YanChun,LI Yu,WANG TianYu,ZOU HuaWen,LI ChunHui
    Scientia Agricultura Sinica. 2019, 52(14):  2391-2405.  doi:10.3864/j.issn.0578-1752.2019.14.002
    Abstract ( 441 )   HTML ( 28 )   PDF (5010KB) ( 441 )   Save
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    【Objective】The root system as an important organ absorbing water and nutrients for plants, is essential for growth and grain yield of maize. More understanding of the genetic mechanism of the root architecture of maize is of great significance for the practice of high-yield breeding of maize.【Method】In this study, 111 maize elite inbred lines were used as an association population. In 2017, six belowground nodal root-related traits, i.e. nodal root layer number (RLN), total nodal root number (TRN), nodal root angle (RA), nodal root area (RS), nodal root volume (RV) and nodal root dry weight (RDW), were measured under four environments including Beijing, Yongshou of Shanxi province, Dingxiang of Shanxi province and Yuanyang of Henan province. The average of the four environments were used as the phenotypic data of the six root-related traits. The statistical analysis and correlation analysis were carried out on six traits, and the differences of six root-related traits for inbred lines developed in different eras and for inbred lines in different heterotic groups were also analyzed. Based on 152352 high-quality SNP markers obtained in this population, the FarmCPU model was used for genome-wide association analysis to obtain significantly associated SNP loci, and candidate genes were predicted based on the LD interval sequence of these significant associated SNPs, and a functional enrichment analysis of candidate genes was carried out.【Result】The phenotypic analysis showed that the six belowground nodal root traits exhibited a normal distribution and high level of heritability. The correlation analysis showed that RLN and TRN are negatively correlated with RA and RS; RA, RS, RV and RDW are significantly positively correlated with each other. With the advance of maize breeding era, RLN and TRN had a decreasing trend, and RA and RS had an increasing trend. There were no significant differences for RDW and RV among inbred lines of different eras. The belowground root structure of those inbred lines from different maize heterotic groups also showed differences, and the six traits of the Lüda Red Cob group were all higher than those of other groups. Genome-wide association study (GWAS) yielded 26 significantly associated SNPs (P<0.00001) referring to RLN, TRN, RV and RDW, of which 11 SNPs located in root-related QTLs previously reported, and 2 SNPs were detected to be significant correlation with RLN and TRN. A total of 177 candidate genes were found based on those significantly associated SNPs, of which 135 genes have functional annotation, the gene Zm00001d037368 was one pleiotropic gene influencing RLN and TRN. The results of enrichment analysis of candidate genes mainly involved in metabolic regulation, the response to stress, transporter activity, catalytic activity, binding protein and cellular components in plants.【Conclusion】The root architecture of maize inbred lines from different eras and different heterotic groups had differences with various degree. Root-related genetic loci and candidate genes identified by the genome-wide association analysis, a total of 26 loci associated with root-related traits were identified.

    TILLAGE & CULTIVATION·PHYSIOLOGY & BIOCHEMISTRY·AGRICULTURE INFORMATION TECHNOLOGY
    Study of Straw-Biochar on Utilization Potential, Industry Model and Developing Strategy in Northeast China
    ZHANG WeiMing,CHEN WenFu,MENG Jun,JIN Liang,GUO Wei,ZHAO HongLiang
    Scientia Agricultura Sinica. 2019, 52(14):  2406-2424.  doi:10.3864/j.issn.0578-1752.2019.14.003
    Abstract ( 324 )   HTML ( 22 )   PDF (898KB) ( 562 )   Save
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    As a strategic emerging industry, straw-biochar technology and its industrialization are beneficial to systematically and comprehensively solve the prominent problems of straw burning, straw utilization and soil degradation in Northeast China, which has large cultivated land and crop straws resource, thus become a hot research topic. Based on the systematic analysis of nearly 20 years in the Northeast China of straw resources dynamic change, availability utilization resource and development trend, this paper discussed the resource base, technic approach and development potential of straw-biochar utilization, and from an industrialization perspective to comprehensive analyze the resources, technology, conditions, advantages, markets and development potentials of the biochar-industrialization development in Northeast China. Our research results showed that the straw-biochar industrialization had rich available utilization biomass resources, mature industrialization conditions, and huge development potential and space in Northeast China, which had great significance and value on promoting the straws resource utilization, the cultivated land quality improvement and agricultural sustainable development in Northeast China. Finally, based on the actual situation of agricultural, economic and social development, we constructed the straw-biochar industry model including the whole industrial chain, and put forward the development strategy suggestions to the biochar industrialization in Northeast China, aiming at providing reference for solving the practical problems of straws processing and agricultural production, and promoting the healthy and stable development of biochar industrialization.

    Spectral Response Analysis of Canopy Water Content of Winter Wheat Under Different Irrigation Conditions
    SUN Qian,GU XiaoHe,SUN Lin,WANG Miao,ZHOU LongFei,YANG GuiJun,LI WeiGuo,SHU MeiYan
    Scientia Agricultura Sinica. 2019, 52(14):  2425-2435.  doi:10.3864/j.issn.0578-1752.2019.14.004
    Abstract ( 371 )   HTML ( 31 )   PDF (1406KB) ( 486 )   Save
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    【Objective】 Rapid and non-destructive diagnosis of canopy water content is of great significance for monitoring winter wheat growth, drought assessment and variable irrigation. The response of canopy spectrum to canopy water content under different irrigation treatments was analyzed in this study.【Method】Based on field variable irrigation experiments, the influence of growth stage and irrigation water on canopy water content of winter wheat were analyzed. The response rule of canopy spectrum to canopy water content under different irrigation treatments was explained. The canopy equivalent water thickness (EWTc) was used as the characterization index. Based on continuous wavelet transform (CWT), a spectral diagnostic model of EWTc of winter wheat was constructed. The accuracy of the model was verified by independent samples. 【Result】 The results showed that the EWTc of winter wheat increased with the increase of irrigation water in the later growth stage, and decreased with the advance of growth process. The canopy spectral reflectance of winter wheat decreased with the progress of the growth process. The canopy spectral reflectance of winter wheat at different irrigation treatments in near infrared and mid-infrared bands were as follows: 1 water > 0.5 water > 0 water. Compared with the original canopy spectral reflectance, the correlation between wavelet coefficients after continuous wavelet transform and EWTc was improved in different degrees at the decomposition scales of 1, 2, 3, 5, 6 and 7. In addition, the increase ranged from 8.40% to 26.20%. The spectral diagnostic model of canopy equivalent water thickness constructed at 2 400 nm in scale 6, 1 596 nm in scale 2, and 2 397 nm in scale 7 was better in stability and accuracy. The verification sample determined the coefficient R 2=0.5411, and RMSE=0.0127 cm.【Conclusion】The canopy water content of winter wheat changed regularly with irrigation time and irrigation amount, and showed obvious spectral response characteristics in the water sensitive band. Continuous wavelet transform technology could effectively improve the correlation between canopy spectral parameters and canopy equivalent water thickness. The spectral diagnosis of canopy water content of winter wheat was realized. It could provide technical support for variable irrigation decision-making in winter wheat field.

    PLANT PROTECTION
    Expression of Stilbene Synthase Gene and Resistance to Powdery Mildew Analysis of Chinese Wild Vitis quinquangularis
    LIU MengQi,WU FengYing,WANG YueJin
    Scientia Agricultura Sinica. 2019, 52(14):  2436-2449.  doi:10.3864/j.issn.0578-1752.2019.14.005
    Abstract ( 435 )   HTML ( 14 )   PDF (5684KB) ( 334 )   Save
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    【Objective】 Grapevine is an important fruit crop in the world. European grape is one of the most important cultivated grape varieties for its high quality and high yield. However, it is susceptible to fungal diseases, especially the powdery mildew. Resveratrol is a metabolite of stilbene synthase (STS) in grape. It is known as a phytoalexin and plays an important role in disease resistance. Besides, resveratrol in fruits has a health-care effect on humans. Chinese wild Vitis quinquangularis ‘Danfeng-2’ displays resistance to powdery mildew and high resveratrol content. The objective of this study is to analyze the function of STS from Chinese wild V. quinquangularis ‘Danfeng-2’, and to apply it to disease-resistance breeding to improve European grapes’ disease-resistance and stilbenoids content. 【Method】 VqSTS26 and VqSTS32 were obtained by homologous cloning from Chinese wild V. quinquangularis ‘Danfeng-2’, and the pCAMBIA35S:: VqSTSs:: GFP overexpression vector was constructed. The meristem callus of V. vinifera Thompson Seedless was induced from the organogenesis, which was used as the receptor material. The transgenic grapes were obtained by agrobacterium-mediated genetic transformation. The expression of STS at transcriptional level and the content of stilbenoids at metabolite level were compared between transgenic plants and wild-type Thompson Seedless under natural growth conditions and Uncinula necator induction. The growth and development of U. necator on transgenic and wild-type grapes’ leaves were observed by microscope. The spore germination, hyphal growth and conidiophore formation were counted, and the powdery mildew resistance of transgenic plants was analyzed. 【Result】 Eight transgenic plants with VqSTS26 and five transgenic plants with VqSTS32 were successfully obtained by PCR detection and Western blot identification. Real-time quantitative PCR analysis (qRT-PCR) showed that the expression of STS was significantly increased in transgenic grapes under natural condition, and the expression of its upstream gene PAL and downstream gene RSGT was up-regulated, while the expression of CHS was down-regulated due to the substrate competition with stilbene synthase. Liquid chromatographic analysis showed that the stilbenoids content in transgenic grapes was higher than that in wild-type Thompson Seedless, mainly existed in the form of trans-piceid. The expression of STS and its product synthesis were induced by U. necator. The expression of STS in transgenic grapes was significantly increased at 1-2 dpi (days post induction) under U. necator induction and was highest at 7 dpi, while trans-resveratrol and viniferin were added from original trans-piceid. The STS expression and its products’ content in transgenic plants displayed excellently higher levels than those in wild-type Thompson Seedless under U. necator induction. U. necator developmental process on grape leaves was observed by microscope. It was found that the growth of U. necator on transgenic grapes was inhibited compared with that of wild-type Thompson Seedless, and the development of hyphae was later than that on wild-type Thompson Seedless. The number of conidiophore on transgenic grape leaves at 7 dpi was lower than that on wild-type Thompson Seedless. 【Conclusion】 Overexpression of VqSTS26 and VqSTS32 from ‘Danfeng-2’ in Thompson Seedless can improve the expression of STS, promote the formation and accumulation of stilbenoids, and inhibit the growth of U. necator on transgenic Thompson Seedless leaves. Therefore, Chinese wild V. quinquangularis ‘Danfeng-2’ and its carrying STS, as well as its products are important germplasm resources and genetic resources for the improvement of resistance to powdery mildew and content of stilbenoids in European grape varieties.

    Proposition, Development and Application of the Integrated Microbiome Agent (IMA)
    Bo LIU,QianQian CHEN,JiePing WANG,ChuanQing RUAN,YanPing CHEN,JiangPing XIA,JianMei CHE,Zheng CHEN,ZhiZhen PAN,Xiao WEN,YuJing ZHU,HaiFeng ZHANG,XueFang ZHENG
    Scientia Agricultura Sinica. 2019, 52(14):  2450-2467.  doi:10.3864/j.issn.0578-1752.2019.14.006
    Abstract ( 402 )   HTML ( 17 )   PDF (2603KB) ( 597 )   Save
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    【Objective】Using microbial fermentation bed as fermentation tank, pig manure nitrogen was continuously added to achieve aerobic fermentation in medium temperature. Microbiome with high bacterial content identified by metagenomic technology was named as integrated microbiome. Integrated microbiome agent (IMA) was produced as plant disease biocontrol agent.【Method】The production process is as follows: material preparation→fermentation→nitrogen feeding→aerobic fermentation control→product processing→product packaging, etc. The fully manufacturing processes of IMAs were as follows: the litters consisting of soybean cake powders (30%) and Pleurotus eryngii mushroom substrates (70%) were added onto the pig microbial fermentation bed that had been used for more than one year with 10 cm thick; the aerobic fermentation was conducted for 20 days by ploughing the litters mixing with pig manures one time per day; then, the upper 20 cm litters were removed to produce IMAs by drying, crushing, screening and packaging.【Result】The physicochemical properties of the IMA products were moisture content 29.74%, pH 7.56, organic matter content 44.46%, total nitrogen content 2.23%, humic acid content 11.20%, crude fiber content 14.06%, and the amount of bacteria was 145×10 8cfu/g. The results of metagenomic determination showed that the average number of the short sequences (reads) was 99 701.75 in the IMA samples, and the IMA samples contained 39 phyla, 96 classes, 189 orders, 383 families, 786 genera, and 1 281 species of bacteria per gram. Among bacteria, 46 Bacillus-like species were detected, and 9 species were the new records in China, namely: (1) Bacillus aerophilus, (2) Bacillus eiseniae, (3) Bacillus filamentosus, (4) Bacillus kochii, (5) Bacillus rhizosphaerae, (6) Lysinibacillus macroides, (7) Oceanobacillus caeni, (8) Ornithinibacillus scapharcae, (9) Virgibacillus oceani. Importantly, no bacterial pathogen of pig was found in the IMA products. The total content of the cultural Bacillus-like species was 2.062×10 8cfu/g, and they accounted for 1.42% of total bacterial abundance in the metagenomic data. Therefore, the total amount of effective bacteria was estimated to be 145×10 8cfu/g in the IMA products based on the cultural Bacillus-like species content and total abundance of metagenomic detecting. The germination rate of mung bean in the IMA group was 96.67%, which was not significantly different from that in the clear water control group (P>0.05), but the radicle length was 58.08% higher than that in the control group. When the seedling-raising substrate was mixed with 5%-10% of IMAs, the emergence rate and plant height of tomato were increased by 3.0% and 25.1%, respectively, and the corrected control efficacy on tomato bacterial wilt was 79.41%. According to the national quality standards for the bio-organic fertilizers (NY884-2012), those of the IMA products were initially set as followings: organic matter content≥40%, moisture content≤30%, pH 5.5-7.5, fecal coliform count≤100 cfu/g, mortality of ascarid egg>95%, expiry date>6 months; The contents of heavy metals: As<15 mg·kg -1, Cd≤15 mg·kg -1, Pb ≤15 mg·kg -1, Cr≤15 mg·kg -1, and Hg≤15 mg·kg -1; The amount of living bacteria was adjusted to the total living bacteria amount≥30×10 8cfu/g, Bacillus-like species amount≥2×10 8cfu/g.【Conclusion】The concept and product technical standard of the integrated microbiome agents (IMAs) were proposed. The IMA can promote the growth of seed root and have a good control efficacy on tomato bacterial wilt.

    SOIL & FERTILIZER·WATER-SAVING IRRIGATION·AGROECOLOGY & ENVIRONMENT
    Response of Different Forms of Nitrogen Migration in Typical Red Soil to Long-Term Different Fertilization Systems
    SHEN FengMin,JIANG GuiYing,ZHANG YuJun,LIU Fang,LIU ShiLiang,LIU KaiLou
    Scientia Agricultura Sinica. 2019, 52(14):  2468-2483.  doi:10.3864/j.issn.0578-1752.2019.14.007
    Abstract ( 319 )   HTML ( 14 )   PDF (559KB) ( 300 )   Save
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    【Objective】 This study was aimed to explore the characteristics of different nitrogen forms migration under different fertilizations in typical red soil, so as to provide theoretical basis for rational amount of applied nitrogen in red soil region. 【Method】 Based on the long-term experiment site located at Jinxian, Jiangxi province, four typical treatments were chosen as: (1) no fertilization (CK); (2) mineral nitrogen, phosphorus, potassium (NPK); (3) NPK combined with straw (NPKS); (4) NPK combined with pig manure and straw (NPKSM). Four levels of soil samples of 0-10 cm, 10-20 cm, 20-40 cm and 40-60cm were collected from each treatment. Soil total nitrogen (TN), alkali-hydrolyzable nitrogen (AN), nitrate nitrogen (NO3 --N), ammonium nitrogen (NH4 +-N), dissolved organic nitrogen (DON) and microbial biomass nitrogen (SMBN) were measured and analyzed. 【Result】 The results showed that the content of nitrogen forms under different treatments decreased with soil depth, while the different nitrogen forms demonstrated different characteristics in different soil layers. The available nitrogen forms (e.g. AN, NO3 --N, NH4 +-N, DON and SMBN) were concentrated at 0-20 cm soil layers, and their content obviously decreased in 20-60 cm compared with 0-20 cm under all treatments. The TN content in 0-40 cm layers was no significantly change. The fertilizer treatments (i.e. NPK, NPKS and NPKSM) could improve all nitrogen forms content compared with CK, which was the highest under NPKSM treatment. The TN content was little change under the same treatment in 0-40 cm, while the TN content was ordered as NPKSM>NPKS>NPK>CK in 0-60 cm. The AN content under the same treatment was obviously decreased with soil depths. Therein, compared with the AN content in 10-20 cm, which in 20-40 cm was decreased by 42% (CK), 50% (NPK), and 44% (NPKS, NPKSM), respectively. The NO3 --N and NH4 +-N content were highest under NPKSM treatment in each soil layers. Therein, the drop of NH4 +-N between 40-60 cm and 0-10 cm was higher than that of NO3 --N, which ordered as 54% (NPKS)>51% (CK)>48% (NPK)>36% (NPKSM). The DON content was significantly different among the treatments in 0-20 cm, and which was higher under the treatment with mineral fertilizer combined with organic fertilizer. The DON content under CK and NPK treatments in 40-60 cm was increased slightly, while significantly decreased under NPKS and NPKSM treatments. The SMBN was significantly different among treatments in 10-20 cm layer compared with other soil layers, and ordered as NPKSM>NPKS>NPK>CK. The proportion of each form of nitrogen to total nitrogen was decreased with soil depth. There was significantly positive correlation between TN, AN, NO3 --N, DON and SMBN at 0-20 cm (P≤0.05), among which TN, DON, AN and SMBN existed very significant positive correlation (P≤0.01). The early, late rice grain yield, straw yield, total biomass, and their nitrogen accumulation were significantly higher under fertilization treatments (i.e. NPK, NPKS, and NPKSM) than that under CK treatment, which was the highest under NPKSM treatment. Moreover, the residual mineral nitrogen and apparent nitrogen loss were also highest under NPKSM treatment. 【Conclusion】 In red soil, the effect of fertilization on nitrogen form focused on the upper layer (e.g. 0-20 cm). All the nitrogen forms content under all treatments decreased with soil depths. All forms of nitrogen content at 0-60 cm was improved under NPKSM. Meanwhile, the crop yield and their nitrogen accumulation were significantly increased under NPKSM treatment, so did with the residual mineral nitrogen and apparent nitrogen loss.

    Effects of Various Paddy-Upland Crop Rotations and Nitrogen Fertilizer Levels on CH4 Emission in the Middle and Lower Reaches of the Yangtze River
    LIU ShaoWen,YIN Min,CHU Guang,XU ChunMei,WANG DanYing,ZHANG XiuFu,CHEN Song
    Scientia Agricultura Sinica. 2019, 52(14):  2484-2499.  doi:10.3864/j.issn.0578-1752.2019.14.008
    Abstract ( 399 )   HTML ( 10 )   PDF (554KB) ( 477 )   Save
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    【Objective】The study was carried out to evaluate the effects of various paddy-upland systems and nitrogen fertilizer levels on CH4 emissions from paddy fields in the middle and lower reaches of the Yangtze River. 【Method】 Field CH4 emissions were collected during the rice growing season based on the long-term paddy-upland crop rotation experiments (2003-by now), including rice-fallow (RF), rice-green manure (Chinese milk vetch; RC-G), rice-wheat (RW) and rice- potato with rice straw mulch (RP), with three nitrogen levels (N0), N1 (142.5 kg N·hm -2) and N2 (202.5 kg N·hm -2)) from 2016 to 2017. 【Result】 (1) The results showed that the effect of crop rotation and nitrogen fertilizer on CH4 emission in paddy fields was significant mainly on the early stage of tillering (from 7 to 30 days after transplanting), which accounted for 51.9%-72.3% of the cumulative CH4 emission of the whole growth period. (2) In addition, both crop rotations and nitrogen levels affected the CH4 emission. Rotations with winter crops (including RP, RW and RC-G) significantly increased CH4 cumulative emissions in rice season at N0 level compared to the RF, being 74.1%-145.1%, 68.5%-109.9% and 56.4%-108.6% higher in RP, RW, and RC-G, respectively. (3) The response of CH4 emissions to rotations was different along with increasing nitrogen fertilizer (N0 to N1 and N2). CH4 emissions increased along with the increase of nitrogen fertilizer application under RF, RP and RW, at N2 level, CH4 cumulative emissions of RP, RW and RF were 51.2-55.8 g·m -2, 45.3-51.5 g·m -2 and 25.0-30.5 g·m -2, respectively, with 23.0%-38.4%, 26.7%-33.7% and 35.3%-43.5% higher than that of N0 level, and 9.9%-19.7%, 20.8%-23.1% and 17.4%-18.8% higher than that of N1 level. While decreased or kept consistent in RC-G, CH4 cumulative emissions under N1 and N2 decreased by 20.7%-42.4% and 10.6%-16.6%, respectively compare with N0. (4) Analyses of functional microbial in related to CH4 emission during early tillering stage showed that rotations with full return of straw and/or green manure could significantly increase the abundance of both methanogens and methane oxidizing bacteria under N0. The response mechanism of related microorganisms to nitrogen fertilizer varied with crop rotation pattern, and the application of nitrogen fertilizer promoted the proliferation of methanogens, but inhibited the proliferation of methane oxidizing bacteria, but the extent of the change varied with crop rotations. With the increase of nitrogen application, the mcrA gene abundance of RP, RW and RF increased by 191.4%, 160.6% and 143.3%, respectively, while RC-G only increased by 62.6%. (5) In addition, the ratio of mcrA/pmoA in RF, RP and RW increased along with the increase of nitrogen application, which increased 71.4%-141.1%, 197.1%-258.2% and 84.6%-165.5%, respectively. The RC-G showed a downward trend, which declined 26.8%-42.3%. The change rule was basically consistent with CH4 emission. 【Conclusion】 Combining with the properties of straw returning in winter, the amount of straw returning in RW and RP was significantly higher than that under RC-G in this study, while the ratio of C/N in RP and RC-G was significantly lower than that under RW. Therefore, the relative amount of C/N returned from straw might be the key to interfere with the effect of nitrogen fertilizer level on CH4 emission from paddy field systems. When carbon was abundant in the system, the relevant microbial activity was restricted by available nitrogen in the soil, and the input of inorganic nitrogen could reduce nitrogen limitation and significantly increase CH4 emission. When the carbon was insufficient and the inorganic nitrogen continues to be invested, the related microbial reproduction was inhibited by the limited carbon source in the soil, and the CH4 emission was relatively reduced.

    HORTICULTURE
    Genome-Wide Identification and Expression Analysis of SAP Family in Grape
    DING Lan,GU Bao,LI PeiYing,SHU Xin,ZHANG JianXia
    Scientia Agricultura Sinica. 2019, 52(14):  2500-2514.  doi:10.3864/j.issn.0578-1752.2019.14.009
    Abstract ( 458 )   HTML ( 20 )   PDF (2981KB) ( 335 )   Save
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    【Objective】The aim of this paper was to identify the Stress Associated Protein (SAP) family genes of grape (Vitis vinifera L.) genome and to know the profile of SAP family regarding the number and structure of genes. The biological rhythm of SAP family and its expression pattern under different hormones and abiotic stress were analyzed by qRT-PCR. The study would provide theoretical basis for further exploring the roles of SAP family genes in grapevine. 【Method】SAP15, a gene selected from the transcriptome database of Vitis amurensis cv. ‘Shuangyou’ and Vitis vinifera cv. ‘Red Globe’, had a significantly up-regulated expression under the cold treatment. According to the conserved structural domains, the SAPs in grape genome-wide were identified by BLAST of NCBI and grape genome database. DNAMAN5.0, MEME, GSDS2.0, ExPASy and MEGA6 software were used for various bioinformatics analysis of VvSAPs, including the study of nucleotide sequences, gene structure, protein structure, physical and chemical properties, chromosome localization and phylogeny. Quantitative real-time PCR (qRT-PCR) was employed to detect the circadian rhythm of VvSAPs and their relative expression levels under hormone and abiotic stress.【Result】In total, 15 SAPs with conservative domains of AN1 were identified from grape genome and most of them contained A20 conservative domain. They could be classified into Class Ⅰ, Class Ⅱ and Class Ⅲ based on their conservative structure domain and the location on chromosome in grape. Analysis of physical and chemical properties of VvSAP family showed that the numbers of amino acids were between 109 and 293, and the theoretical equivalence points were between 7.99 and 9.68. Localization of genes on the chromosome revealed that 15 VvSAPs were distributed on 9 chromosomes of the grape, and there were 3 SAPs on chromosome 8. By the sub-cellular location analysis, we noticed that VvSAPs were mainly expressed in nucleus, chloroplast and cytoskeleton in grape. The secondary structure of VvSAP were mainly irregular crimp and α-helix. The gene structures were highly conserved, and VvSAP1-VvSAP12 had no intron, while VvSAP13-VvSAP15 contained one intron. The results of qRT-PCR analysis showed that VvSAP1 and VvSAP9 had extremely low expression or no expression under hormone and abiotic treatment, so they were preliminarily identified as pseudogenes. While VvSAP10 was proved to be down-regulated under 400 mmol·L -1NaCl treatment, the remaining genes were up-regulated under 50 μmol·L -1 ABA, 100 μmol·L -1SA and 400 mmol·L -1NaCl treatment, respectively. VvSAP10-VvSAP14 were highly responsive to 50 μmol·L -1ABA treatment, and their relative expressions were improved by 37.19, 36.63, 21.69, 58.34 and 267.35 times, respectively, after 24 hours treatment. After 4 hours under NaCl treatment, VvSAP8 and VvSAP11 had the highest relative expression level, which were 13.16 and 12.42 times higher than those without NaCl treatment, respectively. The VvSAP15 had the highest response towards 4℃ treatment, and its relative expression level was 35.90 times after 8 hours treatment comparing to the 0 hour.【Conclusion】15 VvSAPs family were identified from the grape genome, of which VvSAP1 and VvSAP9 were preliminarily identified as pseudogenes. These members were localized across 9 chromosomes of the grape, and the evolution of VvSAP was highly conservative. All members of VvSAP family responded to adversity and had circadian rhythm, However, the expression pattern of VvSAPs was varied under different stress.

    Establishment of Appraisal System for the Stem and Branch Characteristics and Varieties Evaluation of Spray Cut Chrysanthemum
    MA WanRu,FANG WeiMin,WANG HaiBin,ZHANG Fei,CHEN SuMei,CHEN FaDi,GUAN ZhiYong
    Scientia Agricultura Sinica. 2019, 52(14):  2515-2524.  doi:10.3864/j.issn.0578-1752.2019.14.010
    Abstract ( 348 )   HTML ( 4 )   PDF (1446KB) ( 158 )   Save
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    【Objective】Since the characteristics of stems and branches were important commercial traits of cut chrysanthemum, the present study was to establish an appraisal system for selecting stem and branch characteristics of spray cut chrysanthemum, which provided theoretical guidance for screening superior varieties. 【Method】Observing and counting 9 factors related to the characteristics of stems and branches of 300 spray cut chrysanthemum varieties in the consecutive two years, an appraisal system was established by means of expert consultation, analytic hierarchy process and K-Means cluster method to carry out variety evaluation.【Result】The weight of the 9 factors was determined by analytic hierarchy process. Flower lateral convergence, flower vertical convergence, flower branch angle, stem filling degree had the most important impact, and weight was 0.219, 0.219, 0.123 and 0.121, respectively. Then combined the score criterion, 300 varieties were classified into 4 grades by K-Means cluster method. There were 62 varieties graded excellent, 114 graded good, 80 graded general and 44 graded poor, the proportion was 20.7%, 38%, 26.7% and 14.6%, respectively.【Conclusion】The system constructed by expert survey method, analytic hierarchy process and K-Means cluster analysis method could effectively evaluate the stem and branch characteristics of spray cut chrysanthemum varieties, and the selected varieties could be promoted in production.

    ANIMAL SCIENCE·VETERINARY SCIENCE·RESOURCE INSECT
    Transcriptome Analysis of Differentially Gene Expression Associated with longissimus doris Tissue in Fuqing Goat and Nubian Black Goat
    LIU Yuan,LI WenYang,WU XianFeng,HUANG QinLou,GAO ChengFang,CHEN XinZhu,ZHANG XiaoPei
    Scientia Agricultura Sinica. 2019, 52(14):  2525-2537.  doi:10.3864/j.issn.0578-1752.2019.14.011
    Abstract ( 366 )   HTML ( 27 )   PDF (2399KB) ( 637 )   Save
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    【Objective】The aim of this study was to analyze the transcriptome differentially genes expression of longissimus doris tissue in Fuqing goat and Nubian Black goat.【Method】 Two goat breeds, including Fuqing goat and Nubian Black goat, were used as experimental animal, and fatten of castrated ram were used as experimental samples. We determined the samples’ADG in 12 months, and IMF content in longissimus doris tissue at the age of 12 months. Transcriptome sequencing of longissimus doris tissue in Fuqing goat and Nubian Black goat were performed by using the Illumina HiSeq TM 2500 platform with 3 biological replicates per goat breed, and verified by quantitative real-time PCR (qRT-PCR). Differentially expressed genes (DEGs) were selected and enriched based on GO and COG and KEGG database. 【Result】 By fatten of castrated ram, the ADG of Nubiya Black goats in 12 months was higher that of Fuqing goats (Sig.=0.000). But, the IMF content in longissimus doris tissue of Fuqing goats at age of 12 months was higher than that of Nubiya Black goat (Sig.=0.003). A total of 44.76 Gb clean data were obtained in six samples. We found 608 DEGs between Fuqing goat and Nubian Black goat, including 61 DEGs up-regulated genes and 547 DEGs down-regulated genes. Moreover, 518 DEGs and 148 DEGs and 418 DEGs were enriched by GO and COG and KEGG database, respectively. KEGG pathway analysis showed that DEGs annotated to 222 metabolic pathways, and 44 pathways were enriched significantly, such as regulation of actin cytoskeleton, and Jak-STAT signaling pathway, and MAPK signaling pathway and Type Ⅰdiabetes mellitus associated with meat quality and growth traits in goat. The results showed that 8 DEGs (IGF1, ACSL5, PCK2, PPARGC1A, JAK2, STAT4, IRF8 and MAP4K1) might relate to meat quality and growth traits of goat were screened by GO functional enrichment and KEGG Pathway analysis. In addition, 707 new genes or transcripts were found by BLAST, including 15 DEGs between two breeds. By qRT-PCR verification, the pattern of selected genes was consistent with the results of transcriptome sequencing, which showed the sequencing results were reliable. 【Conclusion】Totally, 608 DEGs and 707 new genes of longissimus doris tissue in Fuqing goat and Nubian Black goat were screened by transcriptional analysis, and revealed that four pathways might play an important role in goat meat quality and growth traits.

    Cloning and Identification of Long-Chain Non-Coding RNA Linc24063 and Its Correlation with the Expression Level of miRNAs in Yak
    WANG Hui,CHAI ZhiXin,ZHU JiangJiang,ZHONG JinCheng,ZHANG ChengFu,Xin JinWei
    Scientia Agricultura Sinica. 2019, 52(14):  2538-2547.  doi:10.3864/j.issn.0578-1752.2019.14.012
    Abstract ( 273 )   HTML ( 21 )   PDF (984KB) ( 234 )   Save
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    【Objective】The aim of this study was to clone and to identify the protein coding potential of the long-chain non-coding RNA Linc24063 in yak, and then to analyze its correlation with the expression of miRNAs in mammary gland, which might provide basis for studying the function of Linc24063 in yak. 【Method】Twelve healthy female yaks at 4.5-year old in the first lactation period were selected as experiment animal. After fasting slaughter, the tissue samples of cerebrum, mammary gland, kidney, heart, liver and ovary were collected for total RNA extraction. Firstly, the sequence of Linc24063 was cloned by 5' RACE and 3' RACE, and the sequence conservation, chromosomal location and coding potential were analyzed by bioinformatics, then the prokaryotic expression assays were used to analyze its protein coding ability. Its expression levels in cerebrum, mammary gland, kidney, heart, liver and ovary tissue were determined by using RT-qPCR. Secondly, the conserved miRNAs interacting with Linc24063 were predicted by using the miRNA database of cattle and sheep, combined with miRanda and mireap software. The same miRNAs were obtained from the published differentially expressed miRNAs during different stages of lactation in cattle and the conserved miRNAs, and then GO enrichment and KEGG pathway analysis were performed on their target genes. Finally, Pearson correlation coefficient was used for analyzing the correlation between the expression of Linc24063 and miRNAs in the mammary gland of yak. 【Result】The length of Linc24063 5'RACE and 3'RACE fragments were 476 bp and 356 bp, respectively. Sequencing analysis showed that Linc24063 was 758 bp in length and located in the Dlk1-Dio3 imprinting domain of 21 chromosomes in yak. Bioinformatics predicts results showed that its coding potential was lower, and prokaryotic expression experiments further demonstrated that Linc24063 had no protein coding ability, suggesting that Linc24063 was a real lncRNA. Tissue expression profiling showed that Linc24063 had highest expression in mammary gland and lowest expression in liver and ovary. After bioinformatics analysis, Twenty-one conserved miRNAs interacting with Linc24063 were screened, among which 13 miRNA were differentially expressed in the mammary gland reported previously. The 13 miRNAs targets were enriched in TGF-β, PI3K-Akt, insulin and other signaling pathways, suggesting that Linc24063 might participate in the biosynthesis of milk fat and protein via these signaling pathways. In mammary gland tissue, the expression of Linc24063 was significantly negatively correlated with miR-200a (P=0.001) and miR-141 (P=0.02), and significantly positively correlated with miR-27a (P=0.023), but no correlation with miR-24 (P=0.601). 【Conclusion】The results showed that Linc24063 was located in the Dlk1-Dio3 imprinting domain and expressed higher in mammary gland, and might play an important roles in the biosynthesis of milk fat and protein via miR-200a, miR-141 and miR-27a, which would be beneficial for revealing the regulatory mechanism of Linc24063 in milk fat and protein synthesis.

    RESEARCH NOTES
    The Correlation Between the Stress Relaxation Characteristics and the Quality of ‘Haiwode’ Kiwifruit
    LU Dan,ZHAO WuQi,ZENG XiangYuan,WU Ni,GAO GuiTian,ZHANG QingAn,ZHANG BaoShan,LEI YuShan
    Scientia Agricultura Sinica. 2019, 52(14):  2548-2558.  doi:10.3864/j.issn.0578-1752.2019.14.013
    Abstract ( 279 )   HTML ( 15 )   PDF (414KB) ( 300 )   Save
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    【Objective】This study aimed to extract the quality properties of Haiwode kiwifruit, and to make further effort to investigate the correlation between its stress relaxation and the quality properties. In addition, a rapid analysis method was considered for quickly predicting the quality properties of kiwifruit by using the mechanical characteristics of kiwifruit.【Method】This study measured the nutrition compositions, color, texture and other indicators of the kiwifruit at the different storage periods, then the factor analysis was applied to select the real quality properties of the kiwifruit, and the model of the stress relaxation was established and the correlation analysis was conducted between the model of stress relaxation and the real quality properties of the kiwifruit. Finally, the mathematical model of stress relaxation predicting the quality features of kiwifruit was obtained by the ridge regression with a final verification. 【Result】The Maxwell model with seven components could better describe the stress relaxation of the kiwifruit, and the coefficient of determination of the fit reached 0.999. The Vc, soluble solids and ΔE had significantly positive correlation with elastic parameters and viscosity coefficients (P<0.05). Vc and E0, ΔE and E0 had a correlation coefficients of 0.901 and 0.813, respectively. The texture indexes of hardness, elasticity and cohesiveness were also highly correlated with elastic parameters and viscosity coefficients (P<0.05). The correlation coefficients between the hardness and η1, the springiness and E0 were 0.807 and 0.951, respectively. The decision coefficients of the Vc, soluble solids, ΔE, hardness (TPA), cohesion and springiness using in the prediction model were of 0.906, 0.717, 0.883, 0.709, 0.708 and 0.851, respectively. And all of them had a great statistical significance (P<0.05). The results show that the decision coefficients between the model predicted values and the measured values to the validation collection were of 0.882, 0.880, 0.869, 0.690, 0.733 and 0.814, respectively. The t-test values were all less than 2.145, indicating that there were no significant differences between the predicted values and the measured values. 【Conclusion】 The results showed that the stress features of the Haiwode kiwifruit were significantly correlated with its quality features. Thus, the mathematical model of stress relaxation constructed by the ridge regression method could accurately predict the quality features of kiwifruit, providing a theoretical basis for the evaluation of the kiwifruit quality during storage period.