Scientia Agricultura Sinica

Application of Yr52 Gene in Wheat Improvement for Stripe Rust Resistance

FANG TaoHong,ZHANG Min,MA ChunHua,ZHENG XiaoChen,TAN WenJing,TIAN Ran,YAN Qiong,ZHOU XinLi,LI Xin,YANG SuiZhuang,HUANG KeBing,WANG JianFeng,HAN DeJun,WANG XiaoJie,KANG ZhenSheng

Scientia Agricultura Sinica. 2022, 55(11): 2077-2091. doi:10.3864/j.issn.0578-1752.2022.11.001

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【Objective】 The objective of this study is to evaluate the application value of the high temperature adult plant (HTAP) resistance gene Yr52 in wheat production for improving stripe rust resistance. And wheat lines with good agronomic characters and high disease resistance were developed and selected. It laid a foundation for making full use of the existing HTAP resistance resources and improving the yield related traits.【Method】 The stripe rust resistance gene Yr52 was introgressed to Lunxuan 987 (LX987), Bainong Aikang 58 (AK58) and Han 6172 (H6172) by backcrossing and self-crossing combined with marker-assisted selection breeding. Adult-plant resistance of donor parent, receptor cultivars and their progeny lines were evaluated in the disease nursery fields at Mianyang, Sichuan and Yangling, Shaanxi by mixed endemic physiological races CYR32, CYR33 and CYR34. In comparison to the Chinese Spring reference genome, the flanking SSR markers Xcfa2040 (6.8 cm-Yr52) and Xbarc182 (1.2 cm-Yr52) of Yr52 were combined to search for markers of 35K SNP chip in the physical interval of target genes, and developed into derived cleaved amplified polymorphic sequences (dCAPS) and kompetitive allele specific PCR (KASP) markers. The resistance gene Yr52 was detected in BC₂F_5:6 progeny lines.【Result】 The evaluation of adult plant resistance and agronomic traits indicated that nineteen BC₂F_5:6 lines with LX987 background was obtained: Among of them, 11 were high resistance (IT=0-3, DS=1%-20%), 8 were moderate resistance (IT=4-6, DS=15%-30%), the average thousand kernel weight (TKW), kernels per spike (KPS), productive tiller number per line (PTN), plant height (PH) and spike length (SL) was 45.33 g, 46, 7, 113.26 cm and 10.05 cm, respectively. Four BC₂F_5:6 families with AK58 background: all showed high resistance (IT=0-3, DS=5%-25%); the average TKW, KPS, PTN, PH and SL were 44.67 g, 48, 7, 96.54 cm and 10.17 cm, respectively. Five BC₂F_5:6 lineages with H6172 background showed high resistance to stripe rust (IT=0-3, DS=5%-20%). The average TKW, KPS, PTN, PH and SL were 43.74 g, 49, 8, 109.72 cm and 10.06 cm, respectively. The detection rate of three simple sequence repeat (SSR) molecular markers Xbarc182, Xcfa2040 and Xwmc557 linked to Yr52, in offspring population were 78.57%, 66.67% and 66.67%, respectively. One dCAPS marker Xdcaps-Yr52-1 and one KASP marker Xkasp-Yr52-1 were successfully developed, and the detection rates were 73.68% and 41.67%, respectively. The agronomic traits of lines with high (IT=0-3) and medium (IT=4-6) resistance levels were compared. The results showed that the average TKW (P>0.05), PTN (P>0.05) and KPS (P<0.05) of lines with IT=0-3 were higher than those of families with moderate resistance level lines (IT=4-6). Five lines with disease resistance and stable agronomic traits were selected by evaluated of PH=80-105 cm, PTN≥6,KPS≥45, TKW≥42 g, SL≥8 cm.【Conclusion】 Yr52 was found to be resistant to all of the present predominant races in the adult plant stage. After introgression Yr52 into the main susceptible Chinese wheat varieties, the progeny lines with good disease resistance and agronomic characters could be used for breeding resistance varieties with multi-gene polymerization, it is enriching for the diversity of disease resistance genes and achieving durable utilization. The development of molecular markers will facilitate detect the utilization of Yr52 gene in resistance identification of germplasm in the future.

Accurate Identification and Comprehensive Evaluation of Panicle Phenotypic Traits of Landraces and Cultivars of Sorghum bicolor (L.) Moench in China

XU Xiao,REN GenZeng,ZHAO XinRui,CHANG JinHua,CUI JiangHui

Scientia Agricultura Sinica. 2022, 55(11): 2092-2108. doi:10.3864/j.issn.0578-1752.2022.11.002

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【Objective】 Through the research on the phenotypic diversity and genetic variation of the sorghum germplasm resources in the panicle, we will screen for superior sorghum germplasm, enrich the genetic information of sorghum panicle-related traits, and provide a reference for the conservation and efficient use of existing germplasm resources and the selection and breeding of new varieties.【Method】 Using 320 sorghum accessions from different parts of China as test materials, we accurately identified 12 panicle traits (grain length, grain width, thousand-grain weight, grain hardness, grain density, corneous endosperm rate, kernel weight per panicle, main panicle length, panicle neck length, panicle neck diameter, primary branches length, primary branches number) in two different ecological environments. A comprehensive evaluation of sorghum germplasm resources using correlation analysis, principal component analysis, cluster analysis, and other methods. We screened elite sorghum germplasms with different outstanding characteristics according to the comprehensive evaluation value F and target traits. 【Result】 The frequency distribution of each quantitative trait showed a trend of high in the middle and low on both sides. The two-year frequency distribution and curve trend of grain hardness, kernel weight per panicle and grain density, and corneous endosperm rate were similar at the Baoding and Jinzhong test sites. Most of the traits only showed normal distribution in one year or a single location. Except for the main panicle length and number of primary branches, the other traits differed between the two test sites in the same year. The mean diversity index (H') distribution of the 12 panicle traits ranged from 1.72 to 2.11, among which the average diversity index of grain hardness was the highest, and the average diversity index of primary branches length was the lowest. The coefficients of variation of grain hardness, corneous endosperm rate, kernel weight per panicle, primary branches length, and the number of primary branches were all higher than 30.00%. The cumulative contribution rate of the extracted four principal components was 65.39%. Cluster analysis classified the 320 accessions into three groups, class I can be used as the germplasm for screening process (broom) sorghum; class II is suitable for selecting excellent germplasm for grain (brewing) sorghum; class III was the germplasm with poor panicle traits. We screen 29 superior germplasm with outstanding characteristics according to the comprehensive score value F and target traits. 【Conclusion】 The variability of sorghum germplasm resources in panicle traits was rich and diverse; the coefficient of variation of corneous endosperm rate and primary branches length was high; grain length, grain width, grain hardness, grain density, and kernel weight per panicle were significantly affected by environmental conditions. We screened 29 superior germplasm.

Establishment of High Efficient Extraction and Detection Technology of Sesamin and Screening of High Sesamin Germplasm

ZHONG YanPing,SHI LiSong,ZHOU Rong,GAO Yuan,HE YanQing,FANG Sheng,ZHANG XiuRong,WANG LinHai,WU ZiMing,ZHANG YanXin

Scientia Agricultura Sinica. 2022, 55(11): 2109-2120. doi:10.3864/j.issn.0578-1752.2022.11.003

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【Objective】 Establish an efficient extraction and detection technology for sesamin content evaluation in sesame seeds, then apply it to examine sesamin content variation and identify high-sesamin content materials in sesame germplasm, which will lay the foundation for advancing basic research and breeding of sesame with high sesamin content. 【Method】 Using sesame seeds as raw materials, the single factor tests, including diameters of steel balls, crushing time, crushing amplitude, and seeds weight, were investigated. The seeds were extracted with 80% ethanol via ultrasonic mechanical crushing and cavitation, and the sesamin content was determined by High Performance Liquid Chromatography (HPLC). Based on the Box-Behnken center combined test design principle, we carried out the response surface test with four factors and three levels and established the quadratic polynomial regression equation model with sesamin content as response value. We drew the response surface diagram and contour diagram and determined the main factors affecting sesamin content and the interaction between factors. Finally, the optimal extraction conditions of sesamin were determined, and 1 151 local resources and innovative germplasm were analyzed. The accessions with high sesamin content≥9 g·kg^-1 were selected and identified at the Oil and Product Quality Supervision and Testing Center of the Ministry of Agriculture and Rural Affairs. 【Result】 The regression model established in this experiment was extremely significant (P<0.05), and the lack of fit was insignificant (P=0.1768). The model had a good fitting degree and could be used to predict sesamin content. The influence of the four factors on sesamin content was as follows: crusher amplitude>diameters of steel balls>crushing time>seeds weight. The interaction between the crusher amplitude and diameters of steel balls was significant, while that between crusher amplitude and crushing time was close to the significance level. The optimal conditions for ultrasonic-assisted extraction of sesamin were as follows: steel ball diameter of 6.5 mm, crushing time of 225 s, crushing amplitude of 1 335 r/min, and seeds weight of 0.20 g. Under these conditions, the sesamin content detected was 4.601 g·kg^-1, which was consistent with the predicted value of 4.633 g·kg^-1. Fifteen varieties with specific high sesamin content were identified from 1 151 sesame germplasm for breeding purposes. The highest content was 14.36 g·kg^-1, and the average content was 12.35 g·kg^-1.【Conclusion】 An efficient extraction technology of sesamin was established,compared with the traditional method, the method only needed 0.2 g seeds, which improved the extraction efficiency and reduced sample consumption, the operation was simple, and the sesamin content in sesame seeds could be accurately detected.

Effects of Water-Saving Irrigation on Grain Yield and Quality: A Meta-Analysis

MENG Yi,WENG WenAn,CHEN Le,HU Qun,XING ZhiPeng,WEI HaiYan,GAO Hui,HUANG Shan,LIAO Ping,ZHANG HongCheng

Scientia Agricultura Sinica. 2022, 55(11): 2121-2134. doi:10.3864/j.issn.0578-1752.2022.11.004

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【Objective】 Compared with continuous flooding, the water-saving irrigation can increase water use efficiency. However, the effects of water-saving irrigation on yield and quality in rice paddies have not been clearly defined. The objective of this study was to identify the systematic effects of water-saving irrigation on rice yield and quality through Meta-analysis techniques. 【Method】 In the present study, a total of 34 studies that adapted a water-saving treatment and continuous flooding as the control involving 263 paired observations were included across this dataset. The meta-analysis was conducted to identify the responses of yield and quality to water-saving irrigation as affected by experimental type, water-saving irrigation type, cropping system, rice type, the period of water-saving irrigation, soil total nitrogen (N), soil texture, N rate, and the number of N application. 【Result】 Overall, the water-saving irrigation did not significantly affect grain yield and quality relative to continuous flooding. In terms of water-saving irrigation type, the moderate water-saving irrigation increased brown rice rate (+0.9%), milled rice rate (+1.5%), and head milled rice rate (+2.3%), but did not affect grain yield, chalkiness percentage, chalkiness degree, length/width ratio, amylose content, gel consistency, and protein content relative to continuous flooding. However, the severe water-saving irrigation significantly decreased grain yield (-22.1%), brown rice rate (-2.7%), milled rice rate (-2.7%), and head milled rice rate (-3.6%), and increased chalkiness percentage (+28.0%) and chalkiness degree (+46.7%), while no marked differences were observed on length/width ratio, amylose content, gel consistency, and protein content. Furthermore, compared with continuous flooding, the water-saving irrigation reduced protein content (-9.8%) of late rice, but did not affect that of early rice, middle rice, and single rice. 【Conclusion】 Compared with continuous flooding, the moderate water-saving irrigation could improve rice milling quality, and did not affect grain yield, appearance quality, cooking and eating quality, and nutrition quality. The severe water-saving irrigation significantly reduced rice yield, milling quality, and appearance quality, while no significant effects were found on cooking and eating quality and nutrition quality. The results provided an insight to evaluate the responses of grain yield and quality to water-saving irrigation.

Photo-Temperature Potential Yield of Spring Wheat at Different Accumulated Temperature Ranges and Its Response to Climate Change in Qinghai-Tibet Plateau

ZHANG ZeMin,LÜ ChangHe

Scientia Agricultura Sinica. 2022, 55(11): 2135-2149. doi:10.3864/j.issn.0578-1752.2022.11.005

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【Objective】 The aim of this study was to assess accurately the potential yield of spring wheat and its responses to climate change, which was of great significance to exploit the agricultural resources and to ensure the food security in the Qinghai-Tibet Plateau (QTP). 【Method】 This paper firstly calibrated the WOFOST model based on published research data, and then simulated the photo-temperature potential yield (Yp) of spring wheat at 113 stations in the QTP based on the model and the daily meteorological data during 1958 to 2017. Further, the response of spring wheat potential yield to climate change in three classified accumulated temperature ranges was analyzed by using Theil-Sen Median slope (Sen’s slope), Pearson’s correlation and stepwise multiple linear regression (SMLR) methods. 【Result】 During 1958 to 2017, the annual average potential yield at stations throughout the QTP was between 3.20 and 8.68 t·hm^-2. The potential yield was relatively high in regions with the accumulated temperature range of 1 600-3 400℃·d, including the Yijiang Lianghe Region, the Hehuang River Valley and the northern parts of Ganzi, showing a slightly increase trend. In the regions with accumulated temperature below 1 600℃·d and above 3 400℃·d, the potential yield was relatively low and showed a significantly increase and decrease trend, respectively (P<0.01). For the whole QTP, the daily average temperature (Tave), maximum temperature (Tmax) and minimum temperature (Tmin) in the growing season of spring wheat showed significantly increase trends (P<0.01), and the increase rate of Tmin was higher than that of Tave and Tmax, while temperature diurnal range (Td) and solar radiation (Ra) decreased at rates of 0.08℃ and 8.96 MJ·m^-2 per decade, respectively. The change trends of climatic factors differed obviously among the three accumulated temperature ranges: with increase of accumulated temperature, the increase rates of Tmax, Tmin and Tave and the decrease rate of Td were reduced, while the decrease rate of Ra was increased. There was a significantly positive correlation between changes of Ra and Yp in the QTP and in the three accumulated temperature ranges (P<0.01), but the influence of Tave and Tmax became weak with the increase of accumulated temperature. Furthermore, the influence of Tmin changed from positive to negative, and both of the positive effects of Td and Ra increased firstly and then decreased. At the stations with accumulated temperature below 1 600℃·d, Tave was the critical factor determining potential yield of spring wheat, and its increase of 1℃ could result in 885.71 kg·hm^-2 increase in the Yp (P<0.01). At stations with accumulated temperature range of 1 600-3 400℃·d, Ra played a decisive role, i.e., the potential yield of spring wheat increased by 3.42 kg·hm^-2 for increase of 1 MJ·m^-2 (P<0.01). At stations with accumulated temperature above 3 400℃·d, the potential yield of spring wheat decreased by 398.65 kg·hm^-2 and increased by 3.07 kg·hm^-2 when Tmin and Ra increased by 1℃ and 1 MJ·m^-2 (P<0.01), respectively. 【Conclusion】 The potential yield of spring wheat and its responses to radiation and temperature changes showed a great difference in different ranges of accumulated temperature. The results revealed the potential yield level of spring wheat and identified its response to climate changes, and thus provided supports to exploit the yield increase potential of spring wheat in different regions of the QTP.

Cuticle Protein Genes TcCP14.6 and TcLCPA3A are Involved in Phosphine Resistance of Tribolium castaneum

CHEN ErHu,MENG HongJie,CHEN Yan,TANG PeiAn

Scientia Agricultura Sinica. 2022, 55(11): 2150-2160. doi:10.3864/j.issn.0578-1752.2022.11.006

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【Objective】 Cuticle proteins (CPs) are the main components of insect cuticle, and numerous studies have confirmed that the CP genes were involved in insecticide resistance. The objective of this study is to clarify the roles of TcCP14.6 (cuticle protein CP14.6) and TcLCPA3A (larval cuticle protein A3A) in phosphine resistance of Tribolium castaneum.【Method】 The FAO (Food and Agriculture Organization of the United Nations)-recommended bioassay method was used to determine the phosphine resistance levels of five different T. castaneum populations. The TcCP14.6 and TcLCPA3A sequences were downloaded from the T. castaneum genome data, and their encoded amino acid sequences, signal peptides and conserved domains were predicted via the online services. The total RNAs were extracted from different tissues (the cuticles of head, thorax and abdomen, wing, leg, gut, Malpighian tubules and fat body), different phosphine resistance levels, as well as phosphine induction of T. castaneum, respectively. With TcRPS and TcRPL as internal reference genes, the RT-qPCR was used to analyze the expression patterns of TcCP14.6 and TcLCPA3A in different tissues, different phosphine resistance levels, and in response to phosphine induction. Lastly, the RNA interference (RNAi) technology and bioassay method were used to explore the relationship between the two CP genes and phosphine resistance in T. castaneum.【Result】 The bioassay analysis showed that Jiangsu (JS, RR=1.7) and Yunnan (YN, RR=3.0) belonged to susceptible populations, Hunan (HN, RR=20.2) belonged to moderately resistant population, Sichuan (SC, RR=395.4) and Guangdong (GD, RR=862.7) belonged to highly resistant populations. The sequence analysis demonstrated that both TcCP14.6 and TcLCPA3A proteins consisted of signal peptides and chitin binding domains. The RT-qPCR analysis suggested that TcCP14.6 and TcLCPA3A all had a higher expression in the peripheral tissues (the cuticles of head, thorax and abdomen, wings and legs) of T. castaneum, and with a lower expression in the internal tissues, such as fat body, gut and Malpighian tube. Besides, with the increase of phosphine resistance levels in T. castaneum, the expression level of TcCP14.6 was up-regulated, while the expression level of TcLCPA3A was down-regulated. After phosphine induction for 6 h in T. castaneum, the expression levels of TcCP14.6 and TcLCPA3A were up-regulated and down-regulated, respectively. The injection of dsRNA could significantly inhibit the expression of TcCP14.6 and TcLCPA3A in phosphine resistance (GD) and susceptible (YN) populations of T. castaneum. When treated with phosphine (LC₃₀), the mortality significantly increased after the TcCP14.6 was silenced. Instead, the mortality significantly decreased after the TcLCPA3A was silenced.【Conclusion】 The two CP genes TcCP14.6 and TcLCPA3A are involved in phosphine resistance of T. castaneum.

Analysis on the Status of Insecticides Registered on Small Insects of Fruits and Vegetables in China Based on Recommended Dosage

MAO LianGang,GUO MingCheng,YUAN ShanKui,ZHANG Lan,JIANG HongYun,LIU XinGang

Scientia Agricultura Sinica. 2022, 55(11): 2161-2173. doi:10.3864/j.issn.0578-1752.2022.11.007

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【Objective】 The objective of this study is to provide scientific data for the registration, administration and reduction of insecticides on small insects based on the status analysis of insecticides registered on small insects of fruits and vegetables in China.【Method】 The information of insecticides registered on China Pesticide Registration Information Website for controlling ten kinds of major small insects, tobacco whitefly, greenhouse whitefly, thrip, spider mite, aphid, coccid, leaf miner, striped flea beetle, psyllid, and rust mite, in the fields of fruits and vegetables in China were inquired on June 30, 2020. In this paper, the numbers of single insecticides and mixtures registered on small insects of fruits and vegetables were analyzed, and the active ingredient dosage of single insecticides and their registered numbers of different formulations were also analyzed. Moreover, the active ingredients with high dosages were selected and analyzed in terms of formulation and control target. Finally, six active ingredients, five crops and five control targets with the largest dosage were selected as possible combinations for further analysis.【Result】 Entries of insecticides registered for controlling aphid and spider mite were significantly more than the other small insects. The top five small insects with highest dosage, mean dosage and dosage range of single insecticides were as followed: spider mite, coccid, aphid, greenhouse whitefly, and rust mite. Fifteen active ingredients with high dosages were selected and the potential problems such as large recommended dosage differences and abnormal high value points, were found in lime sulfur, dinotefuran, azocyclotin, imidacloprid, dichlorvos, and sodium pimaric acid. A higher active ingredient dosage of single insecticides was observed in the formulation of crystallization (powder), aqueous solution, water soluble powder, emulsion, smoke generator, and granule and for the crops of apple trees, citrus trees, cucumber (protected field), cucumber and cruciferous vegetables. There were obvious problems, large recommended dosage differences and abnormal high value points in the combinations of azocyclotin-citrus trees-spider mite and lime sulfur-apple trees-spider mite, which were attributed to the differences of formulations and companies.【Conclusion】 In order to reduce the dosage of insecticides and slow down the development of small insect resistance to insecticides, it is suggested that insecticide varieties with abnormal high dosages of active ingredients should be reduced for registration and the high efficiency and low risk insecticide varieties with lower dosages of active ingredients will be recommended based on the study of the minimum effective dose of insecticides.

Chemical Composition and Bioactivity of Dissolvable Organic Matter in Biochars

BIAN RongJun,LIU XiaoYu,ZHENG JuFeng,CHENG Kun,ZHANG XuHui,LI LianQing,PAN GenXing

Scientia Agricultura Sinica. 2022, 55(11): 2174-2186. doi:10.3864/j.issn.0578-1752.2022.11.008

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Volatile organic components escape from the solid feedstock when biomass pyrolyze, which is re-absorbed onto the surface of biochar in the form of dissolvable organic matter (DOM) following cooling. The condense Biochar DOM has a complex chemical composition, mainly containing small molecular weight organics, polycyclic hydrocarbons rich in functional groups. With high activity of chemical reaction and biological stimulation, the biochar DOM may alter the form and availability of nutrients and pollutants, mediating microbial abundance and community structure, and regulating plant growth and metabolism. These activities and chemical compositions vary with types of biomass feedstock and pyrolysis conditions. The role of biochar DOM in biological regulation is generally recognized as bio-promotion though a few DOM molecules is known of potential toxicity to plants, animals and microbes. In order to reach a differential and valorized utilization, the biochar DOM could be extracted for producing commercial liquid fertilizer. The identification of potential bio-toxic molecules in biochar DOM in relation to feedstock and pyrolysis condition deserve further studies, which is fundamental for optimizing production of biochar and biochar-derived products, and minimizing environmental risks following land allocation.

Effects of Combined Application of Organic Fertilizer and Chemical Fertilizer on Root Characteristics and Yield of Cotton Under Different Water Conditions

WANG Ning,FENG KeYun,NAN HongYu,ZHANG TongHui

Scientia Agricultura Sinica. 2022, 55(11): 2187-2201. doi:10.3864/j.issn.0578-1752.2022.11.009

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【Objective】 The aim of this study was to explore the effects of combined application of organic and chemical fertilizer on the growth and development of cotton root system, dry matter accumulation and yield under different water conditions, so as to provide a theoretical basis for rational utilization of water and fertilizer resources in arid areas.【Method】 The fixed position experiments were conducted in 2018 and 2019. The experiment was designed by split block, the main plot treatment consisted of severe water deficit (W1), mild water deficit (W2) and normal water deficit (W3), and the soil water content was 32%-46%, 51%-62% and 67%-81% of the field capacity, respectively. The split-plot treatment was composed of four fertilizer treatments: no fertilizer (F0), single application of chemical fertilizer (F1), 30% organic fertilizer with 70% chemical fertilizer (F2) and 70% organic fertilizer with 30% chemical fertilizer (F3). The effects of fertilizer on root morphological characteristics, vertical distribution of root length, root activity, dry matter accumulation in shoot and yield traits of cotton at flowering and boll stage under different water conditions were analyzed.【Result】 The results showed that the growth of cotton root at the flowering and boll stage was inhibited by water deficit, and the root length, root surface area, root volume and root vitality were significantly reduced compared with normal water. The dry weight of the shoot decreased and the root-shoot ratio increased, then the number of bolls and boll weight per plant decreased, finally leading to a decline in seed cotton yield, while W1 had the most serious negative effect among the water treatments. The effects of various fertilizer treatments on root morphological characteristics under different water conditions were significantly different: the root length, root surface area and root volume were increased by fertilizer treatments significantly, and the combined application of organic and chemical fertilizer were significantly higher than that of single fertilizer F1 under W2 and W3. Under the condition of W2, the root length, root surface area and root volume were improved with the increase of the proportion of organic fertilizer, and F3 had an average increase of 18.1%, 12.2% and 35.0% in 2 years compared with F1, respectively. Under the condition of W3, F2 had the most significant promotion effect on root morphology, which was 7.6%, 17.0% and 20.1% higher than that of F1 on root length, root surface area and root volume, respectively. Under W1, the treatments of fertilizer had an inhibitory effect on root length and root surface area, among which F1 had the most significant inhibitory effect. The combined application of organic and chemical fertilizer could promote the growth of roots in the 20-40 cm soil layer under W2 and W3, and significantly increase the distribution ratio of root. Furthermore, compared with single application of chemical fertilizer, it could significantly increase the root activity of each soil layer, promote the increase of shoot and root dry weight, reduce the root-shoot ratio, and increase the number of bolls per plant and the seed yield of cotton. Under the water condition of W2, the boll number per plant and seed cotton yield under F3 was the highest one, which were 13.2% and 17.2% higher than that under F1, respectively. However, under the condition of W3, the boll number per plant and seed cotton yield under F3 was the highest one, which was 16.1% and 9.2% higher than that under F1, respectively. 【Conclusion】 Combined application of organic and chemical fertilizer could optimize root morphology and spatial distribution, improve root activity, promote dry matter accumulation in shoot and increase lint yield, and alleviate the effects of mild water deficit on cotton root growth and development. With the combined application of organic and chemical fertilizer under different water condition, the optimal fertilizer treatments were 70% organic fertilizer with 30% chemical fertilizer (F3) under mild water deficit, and 30% organic fertilizer with 70% chemical fertilizer (F2) under normal water condition.

Peptidome Analysis of Mesocarp in Melting Flesh and Stony Hard Peach During Fruit Ripening

LI Ang,MIAO YuLe,MENG JunRen,NIU Liang,PAN Lei,LU ZhenHua,CUI GuoChao,WANG ZhiQiang,ZENG WenFang

Scientia Agricultura Sinica. 2022, 55(11): 2202-2213. doi:10.3864/j.issn.0578-1752.2022.11.010

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【Objective】 The aim of this study was to explore the differences between melting flesh and stony hard peaches at the peptide level and precursor protein level during fruit ripening, so as to provide a theoretical basis for mining the key peptides of determining or regulating the ripening process.【Method】 The characteristics of peptides and precursor protein functions in CN13 (melting flesh, MF) and CN16 (stony hard, SH) peaches were analyzed by peptidome, the relative contents of precursor proteins and peptides during MF and SH peach fruit ripening were compared, and the precursor proteins of different peptide segments were analyzed by function enrichment.【Result】 In this study, the peptides of CN13 and CN16 (S3 and S4III) were extracted for mass spectrometry. A total of 473 precursor proteins were identified, including 2 580 specific peptide sequences. The molecular weight, isoelectric point and cleavage sites of the peptide were summarized. In addition, the high-abundance precursor proteins corresponding to endogenous peptides were explored by COG function annotations and pathway enrichment analysis, and the results showed that the precursor proteins were mainly involved in the processes of general function prediction, post-translational modification, protein turnover, energy production and conversion, carbohydrate transport and metabolism. The enrichment analysis showed that the differential peptide precursor proteins of CN13 were related to biological processes, such as oxidation reduction, oxygen and oxygen and electron transport chain, which were mainly involved in glycolysis/gluconeogenesis, pentose phosphate pathway and RNA transport; the differential peptide precursor proteins in CN16 were related to biological processes, such as response to metal ion, response to inorganic substance, and response to cadmium ion, which were mainly involved in microbial metabolism in diverse environments, spliceosome and RNA transport; the differential peptide precursor proteins at the same stage in CN13 and CN16 at S4III were related to gene expression, translation and cellular macromolecular biological processes, which were mainly involved in RNA degradation, RNA transport and splicing.【Conclusion】 There were significant differences in peptides between CN13 and CN16 during fruit ripening. The precursor proteins of differential peptide were involved in starch/sucrose metabolism, glycolysis and ribosome synthesis, and it was suggested that these metabolic pathways were closely related to peach fruit ripening, which provided a theoretical reference for further exploring the key peptides of regulating peach fruit ripening and senescence.

Co-Expression Network and Transcriptional Regulation Analysis of Sulfur Dioxide-Induced Postharvest Abscission of Kyoho Grape

YANG ShengDi,MENG XiangXuan,GUO DaLong,PEI MaoSong,LIU HaiNan,WEI TongLu,YU YiHe

Scientia Agricultura Sinica. 2022, 55(11): 2214-2226. doi:10.3864/j.issn.0578-1752.2022.11.011

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【Objective】 Sulfur dioxide (SO₂) treatment can effectively prevent Botrytis cinerea and postharvest decay, but it can lead to berry abscission, the purpose of this study was to explore the molecular mechanism of grape berry abscission induced by SO₂. 【Method】 Kyoho grapes were treated with SO₂, samples were collected at 2, 4 and 6 d, respectively, and the grape berry abscission rates of the control group (CK) and SO₂ treatment group. The samples of ‘Kyoho’ grapes under CK control and SO₂-treated were sequenced at 2, 4 and 6 d after harvest by high-throughput sequencing technique, the grape genome was used as the reference genome for sequence alignment, and the gene expression was calculated by TPM algorithm, the transcriptome data were systematically analyzed by gene set enrichment analysis (GSEA), gene co-expression network (GCN) and transcription regulation network prediction, and the expression was verified by using quantitative real-time fluorescence PCR (qRT-PCR). 【Result】 SO₂ treatment could significantly induce grape berry abscission, the grape berry abscission treated with SO₂ was 9.88% at 2 d and 19.24% at 4 d, which were significantly higher than those in the control group, the abscission rate reached 38.25% at 6 d, while the abscission rate of the control group was only 11.85%. GSEA analysis showed that the GO biological process enriched in CK group was mainly related to oxidative stress response, cell wall metabolism and phenylalanine metabolic pathway, and CK group was enriched in plant cell wall tissue, pectin metabolism, cell wall modification, polygalacturonic acid and other pathways. The GO biological process enriched in SO₂ group is mainly related to energy metabolism pathway, and it is enriched to photosynthesis, tetrapyrrole metabolism, precursor metabolite and energy production, glucose metabolism and other pathways in SO₂ group at 2 d. The KEGG metabolic pathways enriched in CK group mainly include the mutual conversion of pentose and glucuronic acid, galactose metabolism, plant hormone signal transduction, citric acid cycle (TCA cycle), etc. SO₂ group includes photosynthesis, citric acid cycle (TCA cycle), glycolysis, etc. GCN divided the leading genes in GSEA analysis into 12 levels: level 4-9 were enriched in energy metabolism and glucose metabolism-related pathways, while level 4 was enriched in hormone response and oxidative stress response. The transcriptional regulation prediction analysis of GCN key level gene promoter sequences showed that there were 987 pairs of regulatory relationships among 95 transcription factors (TFs). WRKY14, WOX8, KUA1 were continuously downregulated under SO₂ treatment, wihle MYB60, MYB73, ANL2, ERF2, DOF3.6, GATA25, WRKY57, KAN2, ATHB6 were continuously upregulated under SO₂ treatment. In addition, MYB15, WRKY11, WRKY33, WRKY40, WRK75 were first adjusted upwards and then downwards. The transcriptional regulatory networks of ERF2, MYB60 and WRKY40 revealed that the regulated target genes were involved in cell wall metabolism, sugar metabolism and other related pathways. The qRT-PCR results showed that the up-regulated expression trend of PME36 and ERF2 was similar, and GAUT7, MYB60 and UGE3 had similar up-regulated expression trends. In addition, WRKY40 was induced to be up-regulated at 2 and 4 d of SO₂ treatment, PPME1 and COMT1 expression was consistently down-regulated, and LAC15 was significantly up-regulated at 4 d of SO₂ treatment. 【Conclusion】 SO₂induced the expression of genes related to nutrient metabolism, energy metabolism and cell wall metabolism pathway, which was regulated by a variety of transcription factors, and eventually leads to the grape berry abscission.

Effects of Cellulose Replacing Starch on the Gel Properties of Myofibrillar Protein

WANG LiJian,LUO Cheng,PAN XueFeng,CHEN Xia,CHEN YinJi

Scientia Agricultura Sinica. 2022, 55(11): 2227-2238. doi:10.3864/j.issn.0578-1752.2022.11.012

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【Objective】 The effects of replacing starch with cellulose on the gel properties of myofibrillar protein were investigated to provide a theoretical basis for the application of replacing starch with cellulose in sausage.【Method】 Different proportions starch/cellulose-myofibrillar protein were set as the simulation system to study the water holding capacity, color, texture properties, and stress and strain during fracture deformation, to analyze the dynamic rheological properties and creep recovery properties, and to observe the spatial distribution and three-dimensional network structure of starch/cellulose-myofibrillar protein.【Result】 By increasing the proportion of starch and cellulose, the water holding capacity and storage modulus of the composite gel were improved. When the addition proportion of starch and cellulose increased from 0 to 2.0%, the hardness and chewiness of composite gel gradually increased to the maximum; compared with the control group, the hardness was increased by 29.47% and 43.69% respectively, and the chewiness was increased by 34.82% and 41.58%, respectively; however, L* and whiteness had a tendency to decrease. The maximum stress (9 681.86 Pa) and the minimum strain (1.14) were obtained when 2.0% cellulose was added. The value of the creep modulus of the composite gel decreased gradually with the increase of the proportion of starch and cellulose, while the value of the composite gel in the cellulose group decreased more obviously under the same proportion. Paraffin sections showed that starch and cellulose were simply embedded in the gel network structure, but did not cross-link with the protein, and the cellulose formed larger irregular regions in the myofibrillar protein gel system. Based on microstructure analysis, the surface of the control group was rough and there were more cavities. When starch and cellulose were added, the composite gel became uniform and compact, and the cavities were reduced. The protein network structure of cellulose group had better uniformity and compactness under the same addition ratio.【Conclusion】 The addition of starch and cellulose to myofibrillar protein could improve the water holding capacity, color, texture properties, stress and strain during fracture deformation, rheological properties and microstructure of the composite gel, but cellulose had a more significant improvement effect on the composite gel. Therefore, the application of cellulose as starch substitutes in gelatinous sausage was feasible.

Association Analysis of the ADIPOQ Variation with Sheep Growth Traits

LIANG Peng,ZHANG TianWen,MENG Ke,SHAO ShunCheng,ZOU ShiFan,RONG Xuan,QIANG Hao,FENG DengZhen

Scientia Agricultura Sinica. 2022, 55(11): 2239-2256. doi:10.3864/j.issn.0578-1752.2022.11.013

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【Objective】 The aim of the study was to explore the effects of genetic variation of ADIPOQ on growth traits of sheep, and to find the molecular genetic markers related to growth traits in Ningxia high-quality mutton sheep breeding, so as to achieve the purpose of molecular assisted breeding. 【Method】 The mutation sites of ADIPOQ in Dupo sheep, Tan sheep and Small-Tailed Han sheep were obtained by Allegro Targeted Genotyping. At the same time, the ear tissues of 383 different hybrid progenies of three breeds were collected. The SNPs were genotyped by Sequenom Mass ARRAY®SNP. Haploview was used to analyze linkage disequilibrium and construct haplotypes of the polymorphic loci, as well as association analysis between the SNPs in ADIPOQ with growth traits of newborn and 3-month-old sheep.【Result】 A total of 7 SNPs were screened, and 7 SNPs showed polymorphism in the hybrid population. The dominant genotypes of SNP1-SNP7 were CC, GG, GG, CT, AG, GG and AA, and the dominant alleles were C, G, G, C, G, G and A. X² test showed that all loci were in Hardy Weinberg equilibrium (except SNP7 site deviated from the equilibrium in all individuals). SNP1 (except F₂), SNP4, SNP5 and SNP6 were moderately polymorphic (0.25≤PIC<0.50) in all hybrids and all individuals, while SNP2 and SNP3 were moderately polymorphic (0.25≤PIC<0.50) in F₂ and SNP7 were moderately polymorphic (0.25≤PIC<0.50) in H₁, and were low polymorphic (PIC<0.25) in other populations. The results of linkage imbalance analysis showed that SNP2-SNP3 and SNP5-SNP6 formed two strong linkage, each of which constructed three haplotypes, and formed 4 and 6 genotypes after combination, respectively, among which the dominant genotype was H1H1 and H4H6. 13 genotypes were produced after the haplotypes formed by SNP2-SNP3 and SNP5-SNP6 were recombined, and the dominant genotype was H1H1H4H6. Single SNP correlation analysis revealed that: SNP1 site in F₁ population, the primary chest of GG genotype was significantly higher than that of CG genotype (P<0.05); in H₂ population, the primary body length of CC genotype was significantly higher than that of GG genotype (P<0.05). SNP2 site in H₁ population, the primary body height of CC genotype was significantly lower than that of CG and GG genotypes (P<0.05). SNP3 site in F₁ population, the three months body weight of AG genotype was significantly higher than that of GG genotype; in H₁ population, the primary body height of AA genotype was significantly lower than AG and GG genotypes (P<0.05). SNP4 site in F₁ population, the three months weight of CC genotype was significantly higher than that of CT and TT genotypes (P<0.05); in F₂ population, the primary weight and primary chest of CT genotype were significantly higher than that of TT genotype (P<0.05), which of TT genotype was significantly lower than that of CC and CT genotype (P<0.05) in H₁ population; in H₂ population, the primary body height and primary body length of TT genotype were significantly higher than that of CT genotype (P<0.05). SNP5 site in F₂ population, AA genotype had significantly higher primary body height and primary body length than GG genotype (P<0.05). There were significant differences in primary weight, body height, body length, three months body height and chest among different genotypes of SNP6 (P<0.05). SNP7 site in H₂ population, AA genotype had significantly higher primary body length than GA genotype (P<0.05). When combined with all groups, it was found that: the primary weight and body length of CG genotype at SNP2 were significantly higher than that of GG genotype (P<0.05); the primary weight of TT genotype at SNP4 was significantly lower than CC and CT genotypes (P<0.05); the primary weight of AA genotype at SNP6 was significantly higher than GG and GA genotype (P<0.05); and the primary chest was significantly higher than GG genotypes (P<0.05). There was no significant difference in growth traits among the other five genotypes (P>0.05). The results of haplotype association analysis showed that the primary weight and primary body length of H2H3 genotype were significantly higher than those of other genotypes (P<0.05), while the primary weight of H5H5 genotype was significantly higher than that of H5H6 and H6H6 (P<0.05), the primary body height was significantly higher than that of H5H6 (P<0.05), and the primary chest was significantly higher than that of H6H6 (P<0.05); the three months weight of H4H4 genotype was significantly higher than that of H5H5 genotype (P<0.05), the body weight and body length at three months were significantly higher than those of H4H6 genotype (P<0.05), and the chest of three months was significantly higher than that of H4H5, H5H6 and H5H5 genotype (P<0.05). After SNP2-SNP3 and SNP5-SNP6 haplotypes were recombined, the primary weight, body height, body length and chest of H2H3H4H4 genotype were the highest, which were different from other genotypes, and the body length at three months of H1H1H4H6 genotype were significantly lower than that of H1H2H4H4 and H2H2H4H4 genotypes (P<0.05).【Conclusion】 The results showed that the different SNPs and combined genotypes in ADIPOQ had effect on sheep different growth traits, the seven SNPs in this study could be used as potential molecular markers for growth traits in Ningxia high quality mutton sheep breeding.

The Response of Leymus chinensis Cloned Offspring to Mowing

GUO FengHui,DING Yong,JI Lei,LI XianSong,LI XiLiang,HOU XiangYang

Scientia Agricultura Sinica. 2022, 55(11): 2257-2264. doi:10.3864/j.issn.0578-1752.2022.11.014

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【Background】 Grazing could alter the morphological and photosynthetic physiological characters of the Leymus chinensis cloned offspring, but whether grazing influence the adaptability of that to grazing is unclear. 【Objective】 The aim of this study was to investigate whether grazing history enhanced the adaptability of Leymus chinensis cloned offspring to livestock grazing.【Method】 The Leymus chinensis cloned offspring with different grazing histories (enclosed in 1983 VS long-term free grazing) were used to conduct pot experiment in the greenhouse, and their adaptability to simulated livestock grazing (clipping) was compared in terms of individual traits, ramet number, biomass, and biomass allocation. 【Result】 (1) There was a significant interact effect between maternal grazing history and clipping treatment. The individual height and biomass of grazing (GZ) were more resistant to clipping treatment than that of nograzing (NG), while the response of ramet number to clip was not influenced by maternal grazing history. (2) Grazing history altered the responses of aboveground, root, rhizome and total biomass to the clipping treatment. There was significant interaction between grazing history and mowing treatment in terms of the rhizome biomass, while the interaction on other three indicators were not significant. However, the other three indicators of NG had the larger plasticity index and absolute reduction to clipping treatment. Therefore, the maternal grazing experience enhanced the adaptability of L. chinensis cloned offspring to clipping treatment. (3) The cutting biomass of NG was significantly lower than that of GZ, but the cutting degree of NG was significantly higher than that of GZ.(4) The responses of GZ biomass allocation to clipping treatment were not significant, while rhizome biomass allocation of NG significantly decreased under clipping treatment. 【Conclusion】 The grazing disturbance could enhance the adaption of Leymus chinensis cloned offspring to grazing. The maternal grazing history did not enhance the adaptability of L. chinensis cloned offspring to livestock grazing by altering biomass allocation. The response of leaf photosynthetic physiology and grazing avoidance might be the reasons for the enhancement of grazing fitness. In this study, the environmental disturbance factors, such as soil factors, were excluded through control experiments, and the response of L. chinensis cloned offspring traits to grazing was studied from the perspective of the plant itself. Thus, this study provided a new perspective for fully understanding the process of grazing degradation in grassland ecosystem.

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