【Objective】 In order to provide more information about pollen fertility regulation in thermo-photo-sensitive genic male sterile (TGMS) wheat BNS and BNS366, the relationship between male sterility and the changes of endogenous hormones during microsporogenesis was analyzed.【Method】BNS and BNS366 were used as experimental materials, and Aikang58 and Zhengmai366 (Near-isogenic line of BNS366) were used as controls, respectively. Four different types of wheats were sowed at October 10^th (normal sowing) and December 2^nd (late sowing). I₂-KI method was used to measure the frequency of fertile pollen grains. National and international methods were used to determine self-seeds rate. Indirect enzyme-linked immunosorbent assays (ELISA) method was used to detect 6 kinds of endogenous hormone contents in leaves from the pistil and stamen differentiation stage to the trinuclear stage, young panicles from the pistil and stamen differentiation stage to the tetrad stage and anthers from the uninucleate stage with small-vacuole to the trinuclear stage.【Result】Under normal sowing conditions, the frequency of fertile pollen grains, the national and international rate of self-seeds in both BNS and BNS366 wheats were zero that were completely male sterility, while under late sowing conditions, the three indexes were 34.74%, 43.12% and 48.48% in BNS that was partial male sterility, as well as 92.63%, 55.37 % and 67.94% in BNS366 that was normal fertility. However, the three indexes in Aikang58 and Zhengmai366 wheats under normal and late sowing conditions were 82.56%-94.00%, 73.90%-82.31% and 96.54%-139.26%, respectively, that were normal fertility and no significant difference were found between the two sowing conditions. Moreover, the increased frequency of fertile pollen grains-∆ [(frequency under normal conditions-frequency under late sowing conditions)×100%/ frequency under late sowing conditions] in BNS and BNS366 were lower than those in Aikang58 and Zhengmai366. Multiple comparisons of the hormone’s levels were carried out in the two experiment groups, respectively. We found that the increased hormones levels [(hormones levels under normal conditions - hormones levels under late sowing conditions)×100%/hormones levels under late sowing conditions] differed in all four different varieties. The correlation analytic method was carried on the statistical analysis to the endogenous hormone content and the frequency of fertile pollen grains, and to the endogenous hormone content-∆ and the frequency of fertile pollen grains-∆. For both of BNS and BNS366, there were common characters in dynamic changes of endogenous hormone contents during developmental stages of wheat young panicles in sterile lines. The IAA (indole-3-acetic acid) contents were insufficient in the leaves of the tetrad (BNS) and the pistil primordia differentiation stage (BNS366), and insufficient in the anthers of the uninucleate stage with small-vacuole (BNS) and the uninucleate stage with big-vacuole (BNS366), too, but sufficient in the anthers of the binucleate stage. The gibberellic acid (GA) contents were insufficient in young panicles of the tetrad stage and in anthers of the uninucleate stage with small-vacuole. There were no differences of the zeatin riboside (ZR) contents among the materials. The abscisic acid (ABA) contents tilted towards short supply (BNS) and that was inadequate really (BNS366) in young panicles of the tetrad stage. There were also no differences among the materials in the brassinosteroid (BR) contents. The methyl jasmonate (MeJA) contents were generally insufficient in leaves of the pistil and stamen differentiation stage, the tetrad stage, the uninucleate stage with small-vacuole or the uninucleate stage with big-vacuole, and the trinucleate stage, and in young panicles at the pistil and stamen differentiation stage (BNS) or at the tetrad stage (BNS366), respectively, and in anthers at the uninucleate stage with big-vacuole.【Conclusion】Under normal sowing conditions,the lack of IAA, GA, ABA, especially MeJA before the uninucleate stage with big-vacuole in BNS and BNS366 might promote the occurrence of male-sterility.

【Objective】In this study, the purpose was to study the response of cotton seeds to chilling stress, and to identify the chilling tolerance of different test varieties during the germination period in Gossypium hirsutum. We were also committed to analyzing the cold-tolerance characteristics of tolerant and sensitive varieties at the physiological levels, and to provide a reference for further research on the molecular mechanism of cold tolerance during the germination period. 【Method】Fifty-three upland cotton varieties were used to determine the imbibition rate and relative imbibition rate under chilling stress. In addition, we also measured the cold tolerance indexes such as germination index, vigor index, mean germination time, average germination speed, germination potential, germination rate, seed germination coefficient, radicle fresh weigh, radicle dry weight, embryo fresh weight, embryo dry weight, productive efficiency and material growth rate under chilling stress. We used correlation analysis, principal component analysis, membership function analysis and cluster analysis to comprehensively evaluate the 15 morphological indexes in the imbibition and germination stage. We also measured the changes in antioxidant enzyme activities, osmotic adjustment substance concentration and the expression of antioxidant enzyme genes between different cold-tolerant varieties under chilling stress. 【Result】Under chilling stress, the relative water absorption and water absorption rate of cotton seeds showed a downward trend, and the cold tolerance indexes of 53 cotton varieties showed significant differences. There was a strong correlation between imbibition rate and relative imbibition rate during the imbibition stage, and their correlations with other germination indicators were not significant or negatively correlated. In addition, the correlation analysis also found that indicators such as radicle fresh or dry weigh, vigor index, average germination speed and mean germination time could better reflect the cold tolerance of different varieties. Principal component analysis showed that three principal components could be obtained through simplification, with contribution rates of 55.17%, 18.27% and 8.79%, respectively. The results of membership function and clustering analysis showed that the 53 varieties could be divided into four categories according to comprehensive evaluation value: high tolerant (5 varieties), tolerant (13 varieties), null-tolerant (26 varieties) and sensitive (9 varieties), of which Xinluzhong4 and Zhong103026 are the most resistant and sensitive variety to cold stress, respectively. After chilling treatment, the activity of SOD, POD and CAT in the seeds of Xinluzhong4 could be restored to the level close to or exceeding the control in a short time, the soluble protein concentration in Xinluzhong4 was significantly higher than that in Zhong103026. Gene expression analysis showed that the expression of GhPrx53 and GhCSD1 tended to be consistent with the changes of POD and SOD enzyme activity, respectively. 【Conclusion】The identification indexes of cold tolerance in upland cotton at the germination stage are diversified. The radicle fresh weight, radicle dry weight and vigor index can be used as positive indicators for cold tolerance, while the mean germination time and average germination speed can be used as negative indicator for cold tolerance during the germination period. Soluble protein concentration and the enzyme activities of POD, SOD and CAT can be used as physiological indicators for cold tolerance identification during cotton germination.

【Objective】The soluble sugar content in sweetpotato storage root is an important indicator of edible quality and processing performance. The study on the characteristics of soluble sugar components and their relationship with edible quality can help to understand the changes of soluble sugar components and their influence on taste during processing, providing a basis for the selection of varieties for table use and processing, and breeding and utilization of germplasm resources for special use. 【Method】 The content of soluble sugar components in raw and steamed storage roots of 102 sweetpotato germplasm were determined by high performance liquid chromatography with evaporative light scattering detector (HPLC-ELSD). The characteristics were analyzed according to dry matter content types, and correlation and stepwise linear regression were used to analyze the relationship between soluble sugar components and taste and their contribution to edible quality. 【Result】 There were four soluble sugars of fructose, glucose, sucrose and maltose in both raw and steamed storage roots. In the raw storage root, the sucrose content was the highest, with an average content of 25.79 mg·(g·FW)^{- 1}, accounting for 45.31% of the soluble sugar. The fructose and glucose content were similar and closely related, and the fitting equation for fructose content (y) and glucose content (x) was y = 0.807x + 1.275. The maltose content was the lowest, with an average content of 6.79 mg·(g·FW)^-1, which only accounts for 11.92% of the soluble sugar. The content of soluble sugar in raw storage root mainly depended on the fructose and glucose content. Generally, the varieties with low dry matter content had higher content of soluble sugar and fructose in raw storage root, and the biochemical sweetness was better. During the process of steaming, the changes of soluble sugar content were mainly due to the production of a large number of maltose, which increased from 0.96-24.67 mg·(g·FW)^-1 of raw root to 14.80-136.16 mg·(g·FW)^-1. The content of soluble sugar in steamed storage root was determined by the maltose content, and the variety types with medium or high dry matter content had higher content of soluble sugar and maltose. The improvement of taste quality mainly attributed to the increasing soluble sugar content during the process of steaming. Maltose, fructose and sucrose were the important factors affecting the taste quality, and maltose contributed nearly 50% to the increase of taste quality, particularly to the flavor and texture. Fructose content contributed the most to the viscosity while sucrose contributed more to the texture than fructose. 【Conclusion】 The content of soluble sugar and fructose in raw storage root was an important indicator to evaluate the sweetness of raw sweetpotato. The content of soluble sugar and maltose in steamed storage root was an important indicator to evaluate the edible quality of sweetpotato. Maltose, fructose and sucrose were the important soluble sugar components that determined the edible quality and processing performance of sweetpotato. In this study, 11 sweetpotato germplasm with specific soluble sugar in storage root was selected.

【Objective】 This study was expected to explore the relationship between growth and development characteristics and yield formation of summer maize varieties differing in maturities, providing theoretical references for the selection of new maize varieties with early maturity, high yield and adaptability to grain mechanical harvesting. 【Method】 In 2017, the early maturity hybrids Denghai 518 (DH518), Hengzao 8 (HZ8) and the mid-late maturity hybrids Denghai 605 (DH605) and Zhengdan 958 (ZD958) were used as test materials. In 2018, the early maturity hybrids Denghai 518 (DH518), Jingnongke 728 (JNK728), and the mid-late maturity hybrids Denghai 605 (DH605) and Zhengdan 958 (ZD958) were used as test materials. The planting density was 75 000 plants/hm². The leaf establishment law, ear development characteristics, and the difference in solar radiation and accumulated temperature requirements at different growth stages were analyzed. 【Result】 The solar radiation and accumulated temperature requirements of different maize varieties at various growth stages were: R1-R6>VE-R1>sowing-VE. The demand for accumulated temperature of different varieties after flowering was higher than that before flowering. The difference in demand for accumulated temperature of mid-late maturity hybrids and early-maturing varieties was mainly in V6-R1, which was specifically manifested in the 9-16th leaf establishment period; There was no significant difference in the demand for accumulated temperature after flowering. Correlation analysis showed that the yield of summer maize varieties at different maturity stages had extremely significant positive correlations with spike height and spike height coefficient. The development characteristics of tassel in summer maize cultivars at different maturity stages were different. The length of tassel in late-maturing cultivars was significantly lower than in early-maturing cultivars, but the number of tassel branches, total tassel spikelet, spikelet abortion rate and effective spikelet of late-maturing varieties were significantly higher than those of early-maturing varieties; The differentiation of female ears to grains was greatly affected by the genotype of the variety. 【Conclusion】The growth and development characteristics of summer maize varieties differing in maturities were significantly different, and the yield of the mid-late maturity hybrids was significantly higher than that of the early maturity hybrids. The typical characteristics of early-grain mechanical harvesting corn varieties were fewer total leaves, shorter V6-R1 time and faster dehydration rate.

【Objective】The aim of this study was to clarify effects of elevated temperature and CO₂ concentration on the photosynthetic characteristics of maize under intercropping with peanut, so as to provide the theoretical basis for the green high-yield and high-efficient cultivation of maize intercropping peanut (maize||peanut) under the condition of climate change in the future.【Method】In this study, maize intercropping peanut 2﹕4 pattern was taken as the research object. In 2018, ambient temperature ambient CO₂ concentration (TC) and elevated temperature and CO₂ (+T+C) were set up in the experiment, and elevated temperature ambient CO₂(+TC) was added in 2019 to conduct the experiment, under two phosphorus levels of P₀ (0) and P₁₈₀ (180 kg P₂O₅·hm^-2), respectively. The effects of elevated temperature and CO₂ on photosynthetic pigment content, SPAD value, and photosynthetic response curves to light intensity and CO₂ concentration and the related parameters of intercropped maize were studied.【Result】 (1) Compared with TC, +TC increased the content of chlorophyll b and chlorophyll a+b of intercropping maize at 34 days after seedlings, while this treatment reduced the chlorophyll a/b value. Under the condition of phosphorus application at 55 days after seedling, SPAD value, AQY, CE, A_max, V_c,max, J_max, and TPU of maize were increased by 7.80%, 18.18%, 18.86%, 13.34%, 13.33%, and 20.14%, respectively, and maize grain yield increased by 19.2% to 28.1%. Compared with +TC, +T+C improved the AQY, but reduced LCP of intercropping maize at 55 and 65 days after seedling; The CE, A_max, V_c,max, J_max, and TPU of intercropping maize were increased by 13.58%-32.96%, 21.31%-11.61%, 9.35%-14.5%, 9.52%-15.13% and 8.82%-26.16% at 55 days after seedling, respectively; The yield were increased by 5.25%-18.70%. All reached significant difference levels (P<0.05). (2) Compared with TC, +T+C increased the SPAD value of intercropping maize at big bell mouth and filling stages, which were increased by 4.68%-12.91% and 7.88%-18.37%, respectively, while decreased by 8.63%-12.72% at dough stage; chlorophyll a, b, and a+b of intercropping maize increased by 17.58%-19.54%, 52.55%-59.55%, and 26.08%-28.47%, respectively, at 35 days after seedlings; chlorophyll a/b were decreased by 23.04%-25.18%; AQY and LSP_n were increased by 30.30%-75.76% and 16.87%-19.44%, respectively, at 55 days after seedlings; CE, A_max, V_c,max, J_max and TPU increased by 15.72%-36.78%, 24.91%-32.66%, 20.77%-29.83%, 20.93%-30.48%, and 27.16%-30.74%, respectively; The yield increased by 7.24%-52.0%. All reached significant difference levels. (3) Compared with no phosphorus application, phosphorus application increased the content of chlorophyll b of intercropping maize under TC, +TC and +T+C by 24.15%, 18.64% and 22.04%, respectively, at 85 days after seedling; LSP_n was increased by 13.30%, 17.0% and 9.86%, respectively, at 34 days after seedling; and the yield was increased by 24.2%-67.2%, 55.6% and 27.8%-38.0%, respectively. All reached significant difference levels. 【Conclusion】 Elevated temperature or elevated temperature and CO₂ could increase chlorophyll content and net photosynthetic rate of intercropping maize at early growth stage, showing a positive synergy, while elevated CO₂ could alleviate the negative effects of elevated temperature at the middle and late growth stage. Elevated temperature and CO₂ could increase the yield of intercropping maize, the key lies in the improvement of chlorophyll content, carboxylation efficiency, maximum electron transfer rate and triose phosphate utilization rate at the early and middle growth stage. The supply of phosphorus had an obvious positive effect.

【Objective】The objective of this study is to identify the fungal pathogens causing the main diseases of soybean plants in Hebei Province, screen the fungicides which can effectively inhibit the identified pathogens, and to provide a basis for chemical control of soybean disease in Hebei Province.【Method】The main pathogens of soybean collected from Hebei Province were isolated and purified according to the disease symptom. The morphology of colony, mycelium and spore was observed and identified by using 3D ultra depth of field digital microscope and Olympus Microscope. The rDNA-ITS regions of five pathogenic strains were amplified by PCR and sequenced by universal primers ITS1-ITS4. Neighbor-joining method of software MEGA 7.0 was used to construct phylogenetic trees and analyze the relationship among different pathogens. The pathogens with specific primer sequences were identified and verified. The main pathogen kinds of soybean plants in Hebei Province were identified by the combination of macromorphology, microscopic characteristics and molecular biology techniques. The method of mycelium growth rate and the control effects of different fungicides on the major diseases in vitro leaves or young stems of soybean were used to screen the fungicides.【Result】Based on the analysis of macromorphology, microstructure and sequence, the pathogens isolated from the main soybean producing areas in Hebei Province were Fusarium equiseti, Colletotrichum chlorophyti, Phoma herbarum, Alternaria alternata and Exserohilum rostratum. The corresponding diseases were soybean root rot, anthracnose, leaf spot of P. herbarum, black spot and leaf spot of E. rostratum. The results of virulence test showed that the above pathogens were all sensitive to the triazole fungicides, such as difenoconazole, triflumizole, metconazole and Qoi fungicide pyraclostrobin. F. equiseti was sensitive to Qoi fungicide trifloxystrobin, the pathogens except F. equiseti were sensitive to amide succinate dehydrogenase inhibitors fluopyram, C. chlorophyti and E. rostratum were sensitive to alkyl polyamines fungicide xinjunan, C. chlorophyti was sensitive to organic sulfur bionic fungicide ethylicin, the EC₅₀ values of the inhibitors were less than 10 μg·mL ^-1. The control results of different fungicides on the major soybean diseases showed that the fungicides of difenoconazole, triflumizole, metconazole, pyraclostrobin, trifloxystrobin significantly inhibited above diseases, fluopyram significantly inhibited the diseases except soybean root rot, xinjunan and ethylicin significantly inhibited anthracnose and leaf spot of E. rostratum, the control effects were above 90%.【Conclusion】Five kinds of pathogen and disease of soybean in Hebei Province are identified. The triazole sterol biosynthesis inhibitors difenoconazole, triflumizole, metconazole and Qoi respiration inhibitor pyraclostrobin were recommended as the optimal fungicides for soybean fungal diseases in Hebei Province.

【Background】Conyza canadensis, a pernicious weed, is harmful and lack of the effective management strategy. The caprylic acid can efficiently inhibit the photosynthesis of C. canadensis, which is a potential bio-herbicidal compound. Light harvesting chlorophyll a/b-binding protein (LHC) is the key complex protein in PSI, and plays an important role in photosynthesis.【Objective】The objective of this study is to explore the LHC gene in response to caprylic acid, and to provide a theoretical basis for the development of bio-herbicide.【Method】The open reading frame (ORF) of CcLhca-J9 was cloned from C. canadensis leaf by homologous PCR and RACE. The nucleotide sequence was analyzed by DNAMAN. The phylogenetic tree was constructed with the high similarity nucleotide from NCBI by neighbor-joining method and MEGA software. The molecular weight, isoelectric point and protein structure were analyzed using SWISS-MODEL and ExPaSy websites. Using homologous modeling results as a model, the affinity of caprylic acid and CcLhca-J9 protein was analyzed using AutoDock 4.2 software. RT-qPCR was used to analyze the difference of CcLhca-J9 expression level of caprylic acid, water and ferulic acid treatments.【Result】The CcLhca-J9 ORF is 744 bp, encoding 247 amino acid residues with a molecular weight of 26.766 kD and theoretical pI of 6.43, which belongs to Chloroa b-bind family protein. The phylogenetic analysis showed that the CcLhca-J9 had the most closed relationship with the Lhca protein of Tanacetum cinerariifolium (GEW73959.1) and Artemisia annua (PWA35049.1), and was in the same branch of Asteraceae group, with consistency of more than 85%, indicating that the gene family was conserved. The second structure of CcLhca-J9 protein had alpha helix, random coil, extended strand, beta turn. The homologous three-dimensional model of CcLhca-J9 protein was compared by 4y28.1.O (2.80Á), the tertiary structure was a monomolecular object with six chlorophyll a ligands, which was a typical light harvesting chlorophyll a/b-binding protein. Molecular docking result showed that hydrogen and p-π bond interaction were produced between caprylic acid and CcLhca-J9 amino acid residues Gly68, Phe67, Phe69 and Arg197. The result of RT-qPCR showed that the expression level of CcLhca-J9 was significantly decreased after 0-8 h caprylic acid treatment. Compared with ferulic acid control and water treatment, caprylic acid treatment inhibited the expression of CcLhca-J9.【Conclusion】CcLhca-J9 has the typical function of light harvesting chlorophyll a/b-binding protein, which may be involved in the process of inhibiting the growth of C. canadensis leaves and is a herbicidal target with the potential to develop herbicides.

【Objective】 Anhui is a national major grain production province. Clearance of the nutrient resource quantity of main crop straw and utilization potential under straw returning in Anhui province could provide reference data for reasonable utilization of straw nutrient resources and synthetic fertilizers reduction. 【Method】 Based on statistical data and literature review, the capacity of main crop straw, the nutrient resource quantity of straw and potential for straw incorporation to substitute nutrient inputs were estimated for different cities and regions of Anhui province in this research. 【Result】 The amounts of crop straw from rice, wheat, maize and rapeseed were 17.58, 22.51, 7.12 and 2.75 million tons per year in Anhui province during the period of 2016-2018, respectively. The crop straws were mainly produced in North area of Huai River, and Area between Yangtze River and Huai River, accounting for 47.0% and 33.6% of the total provincial crop straw yields, respectively. The main crop straws contained 0.40 million tons of N, 0.13 million tons of P₂O₅ and 0.85 million tons of K₂O, respectively. The straw nutrient resources from rice, wheat, maize and rapeseed accounted for 44.6%, 34.4%, 13.9% and 7.1% of the total provincial main crop straw nutrient resources, respectively. In the main crop-cultivating areas of Anhui province, the amounts of synthetic fertilizers substituted by rice straw incorporation per unit area of arable land were N 29.5-35.0 kg·hm^-2, P₂O₅ 13.6-16.2 kg·hm^-2 and K₂O 132.9-157.5 kg·hm^-2. The amounts of synthetic fertilizers substituted by wheat straw incorporation were N 24.1-33.5 kg·hm^-2, P₂O₅ 8.4-11.7 kg·hm^-2 and K₂O 83.5-115.8 kg·hm^-2. For maize, straw incorporation could substitute N 24.7-32.7 kg·hm^-2, P₂O₅ 12.5-16.5 kg·hm^-2 and K₂O 59.7-79.0 kg·hm^-2. In the main rapeseed-planting areas, rapeseed straw incorporation could substitute N 24.1-34.0 kg·hm^-2, P₂O₅ 11.2-15.8 kg·hm^-2 and K₂O 101.3-142.7 kg·hm^-2. 【Conclusion】 Using crop straws properly could satisfy the basic potassium requirement for crop production and partly substitute the inputs of synthetic nitrogen and phosphorus fertilizers. Making full use of nutrient resources from main crop straws may provide an effective solution for synthetic fertilizers reduction under straw returning in Anhui province.

The biogas slurry (BS) is a kind of residue liquid of biogas produced by anaerobic fermentation with livestock and poultry manure as the main raw materials. The biogas project in China began in the 1970s. With the development of livestock and poultry farming, and anaerobic digestion technology of livestock manure, the production of anaerobic fermentation BS from livestock manure has greatly increased. The BS from livestock and poultry manure is also a kind of sewage, thus its safe disposal or treatment has become an issue that must be faced and solved to prevent from environmental pollution resulted from agricultural waste in China. At present, the BS has been mainly returned to agricultural land directly without any harmless treatment, as a main disposal method of BS. However, several harmful pollutants (including heavy metals and antibiotics) in livestock manure can still remain in the BS in the process of anaerobic fermentation, and to some extent, even exceed the limits of national water quality standards, endangering farmland ecological environment safety. In this paper, a large number of research data published in the last decade were collected, and the concentrations of heavy metals and antibiotics in animal feeds and manure (including pig, cattle, and chicken) were analyzed, and the impacts of the BS returned into the farmlands on soil quality, water environment and agricultural product safety were evaluated. The research results were showed as follows. (1) The BS was at a risk of seriously exceeding the limits for water quality standard. For example, the concentrations of pollutants (including Hg, Cd, As, Pb, Cr, Cu, Zn and Cl) detected in BS mostly exceeded the limits of the Standards for Irrigation Water Quality (GB 5084—2005), and exceeded the limits (Class IV-V) of the Standards for Groundwater Quality (GB/T 14848—2017). In addition, several antibiotic residues were detected in BS, in which tetracycline antibiotics were higher than other antibiotics, exceeded the threshold value for water quality (10 ng·L^-1) set by European Medicines Agency (EMEA). (2) The BS was at a risk of agricultural land pollution. For example, the concentrations of pollutants (including Cd, As, Pb, Ni, Cr, Cu and Zn) detected in the soils applied with BS exceeded the Risk Screening Values for Soil Contamination of Agricultural Land in the Soil Environmental Quality Risk Control Standard for Soil Contamination of Agricultural Land (GB 15618—2018). In addition, it was reported that several antibiotic residues (such as tetracyclines and quinolones, with the concentration of 3.9 and 14.3 mg·kg^-1, respectively) were detected in the soils applied with BS for 6 years, exceeding the allowable limit of soil antibiotic residues (0.1 mg·kg^-1) stipulated by the Steering Committee of the Veterinary International Committee on Harmonization (VICH). (3) The BS was at a safety risk of agricultural products. As some field experiments reported, several pollutants detected in crops applied with BS (such as Cd, Pb, Ni, Cr, Zn and As in vegetables) exceeded the Safety Qualification for Agricultural Product-Safety Requirements for Non-environmental Pollution Vegetable (GB/T 18406.1—2001), and Maximum Levels of Contaminants in Food (GB 2762—2017). The results indicated that because the rates of BS in agricultural land were usually high, a large amount of BS directly used into farmlands by waste disposal method might lead to the risk of excessive pollutants accumulated in the soils, even exceeding GB 15618—2018 limits in a short term. In order to ensure the safety of soil, water and agricultural products, it is imperative to make pre-treatments for removal of pollutants (including heavy metals and antibiotics) in BS, not exceeding the limit standards for BS returning in farmlands. Therefore, the further research and application on BS treatment technologies for removal of heavy metal and antibiotic pollutants are still needed to strengthen to ensure BS safe utilization in agricultural land.

【Objective】 The ways and laws of soil nutrient loss and surface loss in slope farmland in karst area under different rainfall intensities and underground holes (fissures) were studied in order to provide a theoretical basis for the control of soil nutrient loss and agricultural non-point source pollution control in karst slope farmland. 【Method】 Taking the slope farmland with a slope of 15° in the karst area as the research object, the surface morphology and underground pore (fissures) characteristics of karst slope farmland were simulated under different rain intensities (30, 50, 70, and 90 mm·h^-1) and underground pore (fissures) gap degrees (1%, 3%, and 5%), with a total of 36 rainfalls in the cross-test, in order to explore the characteristics of surface and underground nutrient loss. 【Result】 (1)Rain intensity had a significant effect on runoff and sediment production on slope farmland in karst area (P<0.05). Surface and underground runoff and sediment production increased with the increase of rain intensity, and its critical rain intensity might be between 30 and 50 mm·h^-1, when the runoff and sediment production were from the underground to the ground; with the increase of the underground pore (fissures) gap, the amount of underground runoff and sand production increased, and the surface was in the opposite pattern. (2)The loss of runoff nutrients in karst slope farmland was mainly through the surface form, which was through the form of underground pores (fissures) under the light rain intensity; the loss and modulus of TN, TP and TK in runoff increased with the increase of rain intensity, and the effect of rain intensity on the nutrient loss concentration of each runoff was not obvious. In addition, the amount of nutrient loss and the loss modulus of the ground surface decreased with the increase of the underground pore (fissures) gap, and vice versa and the increase of underground pore (fissures) gap made the proportion of underground runoff nutrient loss gradually increase. (3)Nutrients would be lost through the form of attachment to the sediment, of which the surface sediment was mainly lost. The average loss concentration, loss amount and loss modulus of each sediment nutrient on the surface and underground increased with the increase of rainfall intensity, and the average concentration and loss amount of TK loss were significantly higher than TN and TP under different rainfall intensities. Under the same rain intensity, the average loss concentration, loss amount and loss modulus of surface nutrients showed a decreasing trend with the increase of pore (fissures) porosity, and the opposite was underground, but the loss amount was mainly from the surface to the surface and underground. Both were equally important. (4)Correlation analysis showed that rainfall intensity had a significant positive correlation with runoff and sediment loss. The impact of rain intensity on nutrient runoff was higher than sediment, and surface runoff was most affected by rain intensity. The effect of underground pore (fissures) gap on the loss of sediment nutrients was higher than that of runoff. Among the sediment nutrients, the loss of underground sediment nutrients was greatly affected by it. 【Conclusion】The surface was the main form of nutrient loss in karst slope farmland, and the effect of underground pores (fissures) on nutrient loss could not be ignored. In the prevention and control of nutrient loss on slope farmland, the surface could slow down the runoff and sediment yield of slope farmland by increasing vegetation coverage and adding litter, etc. The underground soil was fixed by the roots of the vegetation, thus reducing the leakage of nutrients to the underground through the fissure, thereby reducing the soil nutrient loss of the slope farmland.

【Objective】In our previous study, Ethylene-responsive factors (ERFs) involved in abiotic stress responses in cauliflower were identified. In the present study, one of these ERF transcription factors with unknown function, named BraERF023a, was cloned. The expression profiles and function of BraERF023a under salt and drought stresses were further explored. 【Method】BraERF023a was cloned by RT-PCR in cauliflower. The sequence characteristics of BraERF023a were analyzed by biosoftwares such as MEGA 6. The expression profiles of BraERF023a under salt and drought stresses were explored by qRT-PCR. BraERF023a overexpression vector was then constructed and genetically transformed into Arabidopsis. The phenotypes and survival rate of overexpression BraERF023a transgenic Arabidopsis under salt and drought stresses were observed and analyzed.【Result】Sequence analysis confirmed that BraERF023a coding region was 597 bp in length and encoded a protein containing 198 amino acids with an AP2 conserved domain. BraERF023a was highly conserved in Brassica. The expression level of BraERF023a significantly increased under both salt and drought stresses in cauliflower. Functional analysis indicated that comparative with the wild-type controls, overexpression BraERF023a transgenic Arabidopsis lines exhibited better growth vigor and higher plant survival rate under salt or drought stress.【Conclusion】 BraERF023a played important roles in positive response to salt and drought stresses in cauliflower. Overexpression of BraERF023a in Arabidopsis could significantly improve the tolerance of transgenic lines to salt and drought stresses.

【Objective】The content and composition of free and glycosidically-bound monoterpenes during low temperature storage were investigated, and the changes of aroma components for Muscat flavor from the perspective of more comprehensive metabolites were interpreted, so as to provide a theoretical basis for the better study of the change of grape flavor quality and the establishment of the best storage condition.【Method】Two new grape varieties RuiduHongmei and RuiduZaohong were used as materials. The healthy fruits were precooled to -1 - 0℃ firstly, and then were transferred into PE grape fresh-keeping film. The film was subsequently sealed and put into cold storage ((2±1)℃, 90% RH). The samples were taken every 15 days. The physical, chemical and appearance quality indexes of the fruits were determined by conventional methods. The contents of free and glycosidically-bound monoterpenes in grape berries were determined by headspace solid phase microextraction combined with gas chromatography and mass spectrometry (SPME-GC-MS).【Result】In the process of low temperature storage, the content of soluble solids showed a decreasing trend, while the changes of titratable acid showed slightly different in two grape cultivars; the berry weight loss, percentage of the decayed, berry drop ratio and the browning index were increased with the extension of low temperature storage time, but berry retention force exhibited a decrease; the compounds analysis results showed that the main free monoterpenes in the two grape varieties included linalool, β-Myrcene, β-cis-Ocimene, limonene, cis-furan linalool oxide and geraniol; and the major glycosidically-bound monoterpenes included linalool, β-Myrcene, geraniol, geranial, β-cis-Ocimene, nerol oxide and so on. During low temperature storage, the content of total free monoterpenes decreased significantly; the change patterns of 28 free monoterpenes were grouped into four types; compared to the initial storage stage, the content of most of the free type compounds was decreased sooner or later in the process of storage. Principal component analysis showed that free monoterpenes, such as geranic acid, nerol oxide, linalool and 4-Terpineol (M17) could be used as the major contributions for different storage time samples. Each glycosidically-bound monoterpene also showed different change trends during low temperature storage in two grape cultivars. The content of the total glycosidically-bound monoterpenes showed opposite trends to that of free monoterpenes. The glycosidically-bound rose oxides could be used as marker monoterpenes to distinguish the two grape varieties.【Conclusion】RuiduHongmei had a better low temperature storage characteristics than RuiduZaohong. The contents of free monoterpenes decreased significantly in low temperature storage, the free geranic acid, nerol oxide and linalool could be used as the main components for different storage time, and the bound oxidized rose could be used as the marker for variety differentiation.

【Objective】 Dry aging meat is popular among consumers because of its unique product characteristics. Thus, the objective of this study was to clarify the regularity of water migration in meat during dry aging by exploring the effects of dry aging on the water-holding capacity of lamb gigot and water migration in it, thereby provide a theoretical foundation for high quality meat production.【Method】The hind legs of twenty-six Small-tail Han sheep (male, carcass weight (23.4±1.09) kg, 6-7 months old) were obtained and assigned into three groups randomly which were wet aging group, relative humidity (80±5) % dry aging group (RH80 dry aging group) and relative humidity (60±5) % dry aging group (RH60 dry-aging group) at (2±2)℃. The drying loss, water content, cooking loss, water distribution and protein secondary structure of lamb gigots were measured at aging 0, 7, 14, 21, and 28 d.【Result】The drying loss of gigots in dry-aging groups was significantly higher than that in wet-aging group (P<0.05). The water content of RH60 dry-aging gigots was significantly lower than that of wet-aging gigots after aging 7 days (P<0.05), and no significant difference of water content was observed between RH80 and RH60 dry-aging group (P>0.05). According to the results of the water-holding-capacity (WHC) test, the cooking loss in dry-aging groups was significantly lower than that in wet-aging group (P<0.05), except RH80 dry-aging group at aging 14 d. Additionally, at aging 14 d, the cooking loss of samples had no difference from that at 0 d (P>0.05), which indicated that the WHC of dry-aging groups was better than that of wet-aging group and the WHC of the sample was improved at dry-aging 14 d. The results of protein secondary structure of gigots demonstrated that compared with that at aging 7 d, the disordered structure of protein in wet-aging group, RH80 dry-aging group and RH60 dry-aging group reduced by 9.2%, 14.1%, and 17.26% at aging 14 d, respectively, which indicated a better protein stability and WHC of gigots at aging 14 d. According to the results of low-field nuclear magnetic resonance (LF-NMR), at aging 21 d, transverse relaxation time of free water (T₂₂) in gigots of three aging groups increased significantly compared with that at aging 14 d, which indicated an increase in the freedom degree of water and a decrease in WHC. In all of the three aging groups, immobilized water (P₂₁) had the largest proportion among the total water in gigots. At the early stage of aging, the proportion of P₂₁ decreased at the beginning and then increased while the proportion of bound water (P_2b) increased first and then decreased. This indicated that immobilized water transferred to bound water first and then bound water transferred back to immobilized water at the later stage of aging, the P₂₁ decreased while proportion of free water (P₂₂) increased, which meant that immobilized water converted into free water at this aging stage with a decrease of WHC.【Conclusion】Dry aging gigots was lower in water content and higher in WHC, compared with wet aging gigots. During dry aging of gigots, there was a conversion relationship between bound water and immobilized water, and between immobilized water and free water. Besides, the increase of immobilized water meant an increase in WHC of gigots, while an increase of free water indicated the decrease of gigots WHC. During a long aging time of gigots, the different relative humidity had a significant effect on drying loss and cooking loss of dry-aging gigots, however, which did not significantly affect its water content and water migration.

【Objective】The objective of this study was to investigate the effects of germination and extrusion process on volatile flavor compounds of brown rice by analyzing changes of the volatile flavor compounds of brown rice, so as to provide a reference for evaluating and improving the volatile flavor of brown rice. 【Method】The volatile flavor compounds of raw brown rice (RBR), extruded brown rice (EBR), germinated brown rice (GBR) and extruded germinated brown rice (EGBR) were analyzed by headspace solid phase microextraction coupled with gas chromatography-mass spectrometry (HS-SPME-GC-MS). The qualitative and relative quantitative analyses of the volatile flavor components were carried out by NIST14 database, retention index (RI), and internal standard. Gas chromatography olfactory (GC-O) was used to analyze the composition spectrums of the active odor compounds and their overall flavor profile as well as the volatile flavor characteristics of brown rice samples. 【Result】 A total of 28 volatile flavor compounds were identified, including aldehydes, alcohols, terpenes, esters, heterocycles and arenes. After extrusion, the contents of aldehydes, heterocycles and arenes in brown rice were increased significantly, while the contents of alcohols and terpenes decreased significantly; moreover, new esters were detected after extrusion. After germination treatment, the content of alcohols was increased, while the contents of aldehydes, terpenes, heterocycles and arenes decreased; however, esters were undetected after germination treatment. After extrusion, the content of aldehydes, terpenes, heterocycles and arenes in germinated brown rice were increased, furthermore, while the content of alcohols decreased; moreover, new esters were detected. What’s more, the contents of aldehydes, terpenes, esters and heterocyclic compounds in germinated brown rice were significantly higher than those in brown rice after extrusion. The principal component analysis showed that the increases of the contents of compounds, such as heptanal, nonanal, benzaldehyde, D-limonene, toluene and 2-AP, were highly correlated with extrusion, while the increases in contents of 2-ethyl-1-hexanol and 3-methylbutanol were highly correlated with germination, and the increases in contents of ethyl acetate, butyl acetate, dimethyl sulfide and pyridine were highly correlated with germination-extrusion treatment. GC-O results showed that 19 kinds of active odor compounds were detected, with the odor intensity value (OIV) of some compounds’ over ≥ 3, such as hexanal, heptanal, hexanol, 1-octene-3-ol, 2-acetyl-1-pyrroline and p-xylene. The flavor profile analysis showed that the odor intensity of wax was the highest in brown rice, followed by those of grass and nut, while those of floral and fruity odor were the lowest. After extrusion, the nutty, grassy and fruity odor intensities of brown rice were increased significantly, in contrast, the waxy odor intensity decreased significantly. After germination, the waxy, nutty, grassy and floral odor intensities of brown rice were decreased, however, the fruit odor intensity did not change. After extrusion, the nutty, waxy, grassy and fruity odor intensities of germinated brown rice were increased significantly. 【Conclusion】Germination could improve the production of alcohols in brown rice, while it could decrease the contents of other volatile compounds to some extent, resulting in the decline of overall flavor intensity. However, during the process of germination, brown rice could produce volatile flavor precursors through biochemical action, which could promote the formation of more volatile flavor compounds by extrusion in germinated brown rice. Extrusion could have a positive effect on the formation of aldehydes, terpenes, esters, heterocycles and arenes in brown rice as well as germinated brown rice. Moreover, the volatile flavor compounds content in extruded germinated brown rice were significantly higher than those in extruded brown rice. The overall flavor intensities of both raw brown rice and germinated brown rice were increased after extrusion, with the most significant increase in the nutty and fruity odor intensities.

【Objective】The effects of dietary energy level on the production performance, blood index, slaughter performance and meat quality of Holstein steers were studied in this study, so as to provide a theory basis for fattening of Holstein steers.【Method】Thirty-nine healthy Holstein steers weighing about 650 kg were selected and randomly divided into 3 groups(n=13). The trial period was 255 d (the preparation period for 10 days and formal experiment period for 245 days). In the early stage of trial (from day 1 to 87), the three groups were low energy group (LE), medium energy group (ME), and high energy group (HE). The NE_mf were 6.10, 6.30, and 6.50 MJ·kg^-1, respectively, and the CP levels were 11.6%; the late stage of the test (from day 88 to 245) was divided into three stages, I (from day 88 to 130), II (from day131 to 188) and III (from day189 to 245). The dietary energy and CP level of each experimental group was the same in different stages, NE_mf was 6.7, 7.20, and 7.29 MJ·kg^-1, respectively, and CP level was 11.5%.The production performance and other indicators were measured at the end of the early and later period of the feeding experiment. Six steers in each group were randomly selected to collect blood for blood biochemical indicators measurement and to slaughter for slaughtering performance and meat quality measurement.【Result】Compared with LE group and ME group, the average daily gain(ADG) of HE group were increased by 21.50% (P<0.05) and 17.12% (P<0.05) , respectively, but there was no significant effect of dietary energy level on the ADG and feed to weight ratio of Holstein steer in the late and whole period (P>0.05).In the early stage of trial, the serum TG content increased significantly, the high and low order was HE group >ME group>LE group, the LPL content in HE group was the highest, which was 35.17% (P<0.05) and 9.92% higher than that in LE group and ME group, respectively; the NEFA content in HE group was the highest, which was 13.16% (P<0.05) higher than that in LE group, and there was no significant difference with that in ME group. At the later stage of trial, the LPL content of LE group was significantly increased by 18.47% (P<0.05) and 26.17% (P<0.05) compared with ME group and HE group.The carcass weight of Holstein steers was significantly increased by different dietary energy levels, the highest in HE group, which was 6.26% and 5.26% higher than that in LE group and ME group; the slaughter rate in HE group was 4.86% (P<0.05) and 4.10% (P<0.05) higher than that in LE group and ME group, but there was no significant difference in eye muscle area and marbling grade (P>0.05). The dietary energy level had no effect on pH, drop loss, shear force, dehydration rate and colorimetric values (P>0.05); the content of fat in the longissimus dorsi muscle of HE group was the highest, which was significantly increased by 52.54% (P<0.05) and 57.45% (P<0.05) compared with LE group and ME group, respectively.【Conclusion】Under the conditions of this experiment, it was better to adopt the straight-line high-energy fattening mode for Holstein steer. In the stage of body weight 650-770 kg, the level of NE_mf was 6.50 MJ·kg^-1, and the level of CP was 11.6%.

【Objective】 The aim of this study was to to investigate the effect of the heterozygous sex-linked dwarf gene on fat deposition of chickens, so as to provide a theory base for producing quality chicken and local chichen by using dwarf gene. 【Method】 In this study, the normal size Gushi chicken and Guangxi Yao chicken rooster (Z^DWZ^DW) were chosen to cross with the normal type hens (Z^DWW) and the dwarf hens (Z^dwW), respectively, and their offspring were raised under the same condition. 100 chickens (half male and half female) were randomly selected from each hybrid at the ages of 60 days, 90 days and 120 days for body measurement index, respectively. 10 chickens (half male and half female) were randomly selected from the hybrid populations of Gushi chickens to determine the dynamic change of body fat at 60 days and 90 days, respectively. In addition, 10 chickens (half male and half female) were selected from the hybrid populations of Gushi chickens and Guangxi Yao chickens at 120 days to determine the body fat deposition of different populations. The serum biochemical indexes were measured by using automatic biochemical analyzer, the content of intramuscular fat in breast muscle and leg muscle was determined by the Soxhlet extraction method, and the diameter and density of muscle fibers were measured by the paraffin section. 【Result】 The results showed that the offspring were normal in body size, and the male and female offspring from the normal type of female parent presented different characteristics of fat deposition with age. The female body fat indexes including abdominal fat weight, percent of abdominal fat, subcutaneous fat thickness and intermuscular fat width, which were all kept at a low level at 60 days, and increased significantly at the age of 120 days, while the male body fat maintained at low levels. In the meantime, the male and female offspring the dwarf type of female parent presented a similar dynamic change of fat deposition with age, and the body fat levels of the male and the female offspring at 120 days were significantly higher than that at 60 days and 90 days. However, the body fat change of the male offspring from the dwarf type of female parent (the heterozygote of dw) was different from that of the normal type of female parent (the homozygous of DW), whose abdominal fat weight, percent of abdominal fat, subcutaneous fat thickness and intermuscular fat width were significantly higher than that of the normal type of female parent at 90 and 120 days. Combining the data of body fat between Gushi chickens and Guangxi Yao chickens for 120 days, the result showed that population factor was significant for abdominal fat weight, percent of abdominal fat, intermuscular fat width and subcutaneous fat thickness. The abdominal fat weight, percent of abdominal fat, intermuscular fat width and subcutaneous fat thickness of the males from the dwarf type of female parent were significantly higher than that of the normal type of female parent (P<0.01). There was no significant difference in total cholesterol, triglyceride, low-density lipoprotein, and high-density lipoprotein between the population (from the normal type of female parent and the dwarf type of female parent). There was no significant difference in the IMF content of the breast muscle in the female offspring from the normal type of female parent and the dwarf type of female parent. The breast muscle IMF content of male from the dwarf type of female parent was significantly higher than that from the normal type of female parent (P<0.01). There was no significant difference in muscle fiber characteristics, including muscle fiber density, muscle fiber area and muscle fiber diameter between the offspring of the normal type of female parent and the dwarf type of female parent. 【Conclusion】The results showed that the heterozygous dwarf gene changed the characteristics of fat deposition in the males, and it could significantly increase the deposition of abdominal fat, subcutaneous fat, intermuscular fat and breast IMF content in the male. The heterozygous dwarf gene had no significant effect on blood lipid index and the characteristics of muscle fibers.

【Objective】Long non-coding RNAs (lncRNAs), a kind of RNAs with highly conserved secondary and tertiary structures and a length of more than 200 nt, have no protein-coding potential. LncRNAs are widely involved in transcriptional and post-transcriptional regulations, such as dosage compensation, cell differentiation and growing development. The study aimed to perform analyses of cis regulation, antisense lncRNA regulation and competing endogenous RNA (ceRNA) regulation as well as related discussion based on previously obtained high-quality lncRNA omics dataset from mixed samples of Ascosphaera apis mycelia and spores, and further reveal the putative regulatory role of lncRNAs in A. apis.【Method】Protein-coding genes located at 100 kb upstream and downstream of A. apis lncRNAs were predicted on basis of the position of lncRNA genes; these upstream and downstream genes were annotated to GO and KEGG databases using Blast to gain function and pathway annotations. Target mRNAs of lncRNAs were identified using LncTar software and then annotated to KEGG and eggNOG databases. Target miRNAs of lncRNAs and target mRNAs of miRNAs were predicted using TargetFinder, followed by visualization of regulatory networks using Cytoscape v3.7.1. The expressions of partial lncRNAs, target miRNAs and target mRNAs within regulatory networks were validated using RT-PCR.【Result】A total of 5 852 upstream and downstream genes of A. apis lncRNAs were predicted. These upstream and downstream genes could be annotated to 48 functional terms such as cellular process, metabolic process and stress response, as well as 121 pathways including metabolic pathways, biosynthesis of secondary metabolites and biosynthesis of antibiotics, which indicated that part of A. apis lncRNAs could regulate the expression levels of upstream and downstream genes via cis-acting function, thus participating in regulation of basic cell activities including material and energy metabolisms. Seven lncRNAs were found to have sequence complementary pairing relationship with seven target mRNAs. Among them, five targets were annotated to hypothetical proteins in eggNOG database, while gene3444 could be annotated to nuclear pore complex protein An-Nup120 and hypothetical protein in KEGG database, suggesting that the aforementioned one antisense lncRNA might be involved in regulating biological processes including biosynthesis of nuclear pore complex protein. Additionally, 227 lncRNAs were predicted to have target binding relationship with 73 miRNAs; majority of these lncRNAs (79.02%) could only link to 1-2 miRNAs and partial miRNAs could be targeted by several lncRNAs. Furthermore, lncRNA-miRNA-mRNA regulatory networks associated with oxidative phosphorylation pathway and MAPK signaling pathway were constructed and analyzed, the results showed that 222 lncRNAs related to oxidative phosphorylation pathway could target 78 miRNAs and 50 mRNAs; as for MAPK signaling pathway, 222 associated lncRNAs could target 76 miRNAs and 46 mRNAs; the results suggested that part of A. apis lncRNAs were likely to regulate these two pathways via ceRNA mechanism, hence affecting energy synthesis, environmental adaptation as well as growth and development.【Conclusion】Partial lncRNAs may regulate a series of biological processes such as growth and development, material and energy mechanisms as well as environmental adaptation of A. apis; MSTRG.5393.1 as an antisense lncRNA may regulate biosynthesis of nuclear pore complex protein in A. apis.

【Objective】When rice is occasionally stressed by dry-hot wind (DHW) or drought at flowering stage, the yield greatly decreases due to the rapid water loss in developing gamete cells. During this process, calcium as a universal second messenger mediates cellular signal transduction in response to drought or other stresses. However, the underlying molecular mechanism remains elusive. This study attempts to dissect the physiological and molecular function of Oryza sativa Calcium-permeable Stress-responsive Cation channels (OsCSCs) which will provide a new strategy for studying the stress responsive mechanism to DHW in crops.【Method】Based on the methods of genetics, electrophysiology and bioinformatics, a typical receptor-like-channel named OsCSC11 was identified. The expression pattern of OsCSC11 were analyzed by qRT-PCR and GUS staining. Further, subcellular localization of OsCSC11-GFP was observed in Arabidopsis protoplasts and onion epidermal cells. Meanwhile, the oscsc11 mutants were generated by CRISPR/Cas9 gene editing tool. Finally, the phenotype and physiological functions were analyzed by cytological method. 【Result】Sequence alignment results in DUF221 family revealed that OsCSC11 includes a typical conserved domain and some unique motifs, which belongs to an independent subfamily. OsCSC11 is mainly expressed in anthers and leaves in rice. Further analysis showed that the activity of full-length protein of OsCSC11 which is in a resting state can be activated by the hypertonic solution. However, OsCSC11^ΔTM1-3(TM1-3 was truncated in OsCSC11) has constitutive channel activity that specifically mediates divalent cations of calcium and magnesium. Thus, we speculated that TM1-3 is a receptor domain in CSCs/OSCAs channel for sensing DHW stresses, and the rest part of OsCSC11 (OsCSC11^ΔTM1-3) generates calcium signal. OsCSC11 and OsCSC11^ΔTM1-3 are both localized in the plasma membrane which may be related to the receptor function. In loss of function mutants oscsc11-1 and oscsc11-2, anthers become smaller and wrinkler compared to wild type, and pollen sterility rate reached to 60%-70% and water content dramatically reduced.【Conclusion】OsCSC11 functions in regulating anther water status and pollen development through mediating calcium influx and possibly involves in the primary sensing step under DHW stresses.

【Objective】Grain traits are important factors affecting wheat yield. the significant locus of controlling wheat grain traits was explored by genome-wide association analysis of wheat grain traits, which provided a theoretical reference for the study of genetic improvement of wheat grain traits. 【Method】The genome-wide association analysis (GWAS) based on mixed linear model MLM (Q+K) was performed on 121 wheat grown in Xinjiang using wheat 50 K SNP chips for 6 traits which including grain length, grain width, grain length-width ratio, grain area, grain perimeter and 1000-grain weight.【Result】Six grain traits showed wide phenotypic variation in different environments, in which the maximum coefficient of variation of 1000-grain weight was 13.91-17.79 and the heritability of each grain trait was between 0.85-0.90. The polymorphism information content PIC value was 0.09-0.38, and the minimum allele frequency MAF value was 0.05-0.50. Group structure analysis shows that the natural groups used in the experiment can be divided into 4 subgroups. GWAS results showed that a total of 592 significant association sites (P<0.001) were detected in 6 traits, of which 29 SNPs were repeatedly detected in 2 or more environments, distributed in 1A(5), 1B(2), 1D, 2A(5), 3B, 5A, 5D, 6B(4), 6D, 7B and 7D(7) chromosomes, can explain 9.3% to 22.7% of the phenotypic variation. Six markers associated with stable grain length were detected, which distributed on 1A, 2A, and 7D chromosomes to explain the phenotypic variation of 9.9%-22.7%. Two markers associated with stable grain width were detected, which distributed on 3 B and 5 D chromosomes to explain the phenotypic variation of 9.6%-12.2%. Six markers associated with stable grain length-width ratio were detected, which distributed on 2A(2), 5A, 7B, and 7D(2) chromosomes to explain the phenotypic variation of 10.1%-19.4%. Three markers associated with stable grain area were detected, which distributed on 1A, 1B and 1D chromosome to explain the phenotypic variation of 9.9%-18.2%. Six markers with stable correlation with grain perimeter were detected, which distributed on 1A(2), 2A, 6D and 7D(2) chromosomes to explain the phenotypic variation of 9.3%-22.6%. Six markers associated with stable 1000-grain weight were detected, which distributed on 1B, 2A and 6B chromosomes to explain the phenotypic variation 9.7%-12.9%. Five dominant loci of pleiotropism with were found to control wheat grain traits, which distributed 1A, 2A(2) and 7D(2) chromosomes, explaining the phenotypic variation of 9.9%-22.7%.【Conclusion】In this study, the genetic diversity of the materials was abundant, a total of 29 multi-environment stability loci were found in natural population with 2 or more environmental associated with 6 grain traits.

【Objective】Fragrance is an important trait for quality improvement of crops. The 2-acetyl-1-pyrroline (2-AP) is the major component of the aroma flavor. BADH2 controls fragrance in plants, and its null or weak alleles lead to 2-AP accumulation. In this study, the fragrance related genes were modified in JING724, a maize elite inbred line invented by Beijing Academy of Agriculture and Forestry Sciences, using CRISPR/Cas9 to improve its trait of fragrance. 【Method】To find BADH gene family of target species, OsBADH2 protein sequence was used to search against protein databases of Arabidopsis, rice and maize with the Ensembl online BLAST tool. All BADH family members were verified by protein domain information in UniProt database. Furthermore, phylogenetic analysis conducted in MEGA software was used to search for maize BADH2 homologs as gene-editing targets. Based on principles of CRISPR/Cas9, we designed highly specific sgRNA to target the candidate genes. The CRISPR/Cas9 vector containing this sgRNA was introduced into the maize variety JING724 by Agrobacterium-mediated transformation. We obtained transgenic maize plants with PMI resistance. Sanger sequencing was used to confirm the CRISPR/Cas9-mediated mutations. Finally, we used gas chromatography-mass spectrometry (GC-MS) to measure the 2-AP content in T1 seeds of the gene-editing lines. 【Result】Phylogenetic analysis showed that there were two BADH2 homologs in the maize gnome, subsequently they were named ZmBADH2-1 and ZmBADH2-2. ZmBADH2-1 is located in chromosome 4, whereas ZmBADH2-2 was in chromosome 1. Both genes have 15 exons and 14 introns. The 4th exon of ZmBADH2-1 shares high nucleotide identity with that of ZmBADH2-2. A specific target sequence, which is located in the 4th exons of both genes, was designed and introduced into a CRISPR/Cas9 vector. Using this vector, 10 gene-editing lines were acquired after maize transformation. PCR amplification and sanger sequencing revealed that, in each of the 10 gene-editing lines, different type of insertions or deletions were introduced into the target sites of both ZmBADH2 genes successfully. Genotypes of mutations included biallelic and multi-allelic mutations. Mess spectra analysis showed that Zmbadh2-1/Zmbadh2-2 double mutants had 2-AP, which is the same substance of flavor with that in fragrant rice. Using GC-MS, we found that 2-AP contents in grains gathered from four randomly selected T₀ gene-editing lines were 438.29, 404.63, 348.65 and 161.82 μg·kg^-1, respectively. On the contrary, no 2-AP was detected in JING724 wild type. 【Conclusion】With site specific mutations introduced into ZmBADH2-1 and ZmBADH2-2 simultaneously using CRISPR/Cas9, new maize variety with fragrant rice like flavor was created successfully.

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

【Objective】The shadows in UAV multi-spectral remote sensing images were removed to improve the accuracy of the nitrogen inversion model for apple canopy. 【Method】Using the UAV multi-spectral images collected in June 2019 at the apple orchard of Qixia city in Shandong province, as the experimental area, normalized shaded vegetation index (NSVI) and normalized canopy shadow index (NDCSI) were respectively used to remove shadow and to extract the spectral information of the canopy in non shadow area. The correlation analysis method was used to analyze the correlation between the spectral data, including the data obtained based on the original spectral images and the images after removing the shadow based on NSVI and NDCSI, and the measured leaf nitrogen content data, respectively, and then the sensitive wavelength of nitrogen content were screened and spectral parameters were constructed. Partial least squares (PLS) and support vector machine (SVM) methods were used to build the inversion model of nitrogen content and to carry out the precision inspection in the fruit tree canopy. 【Result】The results showed that the green band and red band were sensitive bands for the inversion of nitrogen content in fruit tree canopy based on UAV multi-spectral images. The spectral information of fruit tree canopy was weakened by shadow, and the spectral difference of canopy multispectral bands before and after shadow removal was significant, especially in red-edge band and near-infrared band. The accuracy of nitrogen inversion model based on two shadow indexes after shadow removal was improved, and the optimal model was the support vector machine nitrogen content inversion model based on NDCSI, the modeling set of this model R² and RPD was 0.774 and 1.828, the validation set R² and RPD were 0.723 and 1.819 respectively. 【Conclusion】NDCSI could effectively remove the shadow in the multi-spectral fruit tree canopy image of the UAV to improve remote sensing inversion accuracy of nitrogen content in apple canopy, so as to provide a useful reference for precise nitrogen management in orchard.

【Objective】Cotton leaf curl Multan virus (CLCuMuV) was found to encode three proteins with different sizes near the open reading frame (ORF) of C4, and the size of C4 protein encoded by each isolate of CLCuMuV registered in NCBI database was not the same. The objective of this study is to analyze the effect of proteins encoded by three hypothesized “C4 ORFs” on viral pathogenicity, and to determine the exact location of C4 ORF of CLCuMuV.【Method】According to the conserved protein of geminiviruses, three ORFs with different sizes were found near C4 ORF of CLCuMuV, which were labeled as C4-L (567 nt), C4-M (546 nt) and C4-S (303 nt). The three ORFs were constructed into PVX heterologous expression vector by homologous recombination, respectively. Using an agroinoculation method, the effect of PVX mediated expression of the three proteins on the symptoms of Nicotiana benthamiana were analyzed. Using a homologous recombination method, the infectious clones of CLCuMuV wild type and C4-L or C4-S deletion mutant were constructed, and then together with infectious clone of the associated DNA-β, they were inoculated into N. benthamiana respectively by agroinoculation. The effect of C4-L or C4-S deletion on CLCuMuV pathogenicity was analyzed. Meanwhile, Southern blot and Western blot were used to analyze the accumulation of viral genome and the expression level of viral C4 protein. At the same time, the subcellular localization of C4-L and C4-S proteins in the epidermal cells of N. benthamiana leaves was analyzed by a Gateway system vector.【Result】In the PVX heterologous expression assay, the results showed that the leaf curling and petiole elongation were observed in PVX-C4-L and PVX-C4-S inoculated N. benthamiana plants, and the symptoms of PVX-C4-S inoculated plants were more severe, while those of PVX-C4-M inoculated plants were mild or almost asymptomatic. The expression of C4 protein was also detected by Western blot in PVX-C4-L and PVX-C4-S inoculated plants. The results revealed that the protein encoded by C4-S ORF had the greatest effect on the pathogenicity of PVX heterologous virus. In the infectious clone assay, the results showed that the wild type (CLCuMuV) and C4-L mutant (CLCuMuV-ΔL) inoculated N. benthamiana plants had leaf wrinkled, petiole and stem distorted, while those inoculated with C4-S mutant (CLCuMuV-ΔS) and control (Mock) did not show any symptoms. Furthermore, the height of the CLCuMuV and CLCuMuV-ΔL inoculated plants was significantly shorter than that of CLCuMuV-ΔS and Mock inoculated plants. At the same time, the accumulation of viral genome was detected by Southern blot in both CLCuMuV and CLCuMuV-ΔL inoculated plants, and the expression of C4 protein was also detected by Western blot. The results revealed that the protein encoded by C4-S ORF played a key role in the process of CLCuMuV induced symptoms. The subcellular localization showed that YFP-C4-L was mainly located in the chloroplast of epidermal cells of N. benthamiana leaves, while YFP-C4-S was located in the cell membrane or the cytoplasmic periphery, and formed a point like aggregate structure. 【Conclusion】The protein encoded by C4-S ORF is essential for the infection of CLCuMuV, while the proteins encoded by C4-L and C4-M ORF are not necessary.

【Objective】In order to provide a scientific basis for the selection of novel target for pest control, molecular characterization and expression patterns of chitin deacetylase 2 genes (CpCDA2) of the codling moth (Cydia pomonella) were studied. 【Method】The cDNA sequences of chitin deacetylase 2 genes were identified from the transcriptome data of C. pomonella by bioinformatic methods, and the conserved domain was analyzed, the homologous sequences from other insects were selected to construct phylogenetic tree by using neighbor-joining (NJ) method of MEGA7 software. The protein properties including molecular weight (Mw), theoretical isoelectric point (pI), glycosylation site and hydrophilic/hydrophobic properties were analyzed by online software. RT-qPCR was performed to detect the relative expression levels of CpCDA2a and CpCDA2b in different tissues and developmental stages. To explore the functions of these genes in the process of larval molting, the ecdysone (20E) was injected to the 5th instar larvae, and the expression dynamics of them were detected by RT-qPCR.【Result】The full length cDNA sequences of CpCDA2a and CpCDA2b, alternative splicing variants of C. pomonella CDA2, were identified. The amino acid analysis showed that both variants of CpCDA2 possess the signal peptide, and the open reading frames contain three conserved domains, including chitin binding domain (ChBD), low-density lipoprotein receptor class A domain (LDLa) and chitin deacetylase catalytic domain (CDA). The multi-sequence alignment analysis revealed that the CpCDA2a and CpCDA2b are similar to other insect, and the phylogenetic tree showed that the sequences of insect in the same order are clustered into a clade with high confidence. Three-dimensional structure simulation and comparison revealed that the ChBD functional domains of the two splicing variants are different in structure. Physical and chemical properties analysis showed that there are also differences in hydrophilic/hydrophobic properties and glycosylation site of the two splicing variants. Developmental expression patterns showed that the CpCDA2a expression was high in larval stage, CpCDA2b in early and middle stages of pupae, and the expression of CpCDA2a and CpCDA2b was significantly up-regulated before and after larval molting. Tissue-specific expression analysis indicated that the expression of CpCDA2a and CpCDA2b was the highest in integument, and the expression of CpCDA2b in head was also higher. Expression dynamics of the larvae after injection of ecdysone (20E) indicated that both CpCDA2a and CpCDA2b were up-regulated, but with very different levels.【Conclusion】The CpCDA2 has two alternatively spliced variants, namely CpCDA2a and CpCDA2b, which plays a key role in the development of C. pomonella. While, according to the comprehensive analysis of molecular characterization, developmental stages and tissue expression profiles, and expression trends after ecdysone injection, it is deduced that the differences in protein structure and physical and chemical properties of CpCDA2a and CpCDA2b caused by alternative splicing, resulted in the differentiation of function between them.

【Objective】The objective of this study is to clone the target of rapamycin gene (TOR) of Apolygus lucorum (AlTOR), analyze the expression profile of AlTOR as well as its expression patterns after injection of 20-hydroxyecdysone (20E) and its inhibitor U73122, obtain the recombinant protein and antibody against AlTOR protein, and to lay the foundation for the further study of AlTOR protein function.【Method】The full-length of AlTOR was cloned by RACE method. Using qRT-PCR technique, the expression pattern of AlTOR was determined at different developmental stages and different tissues in A. lucorum, as well as its expression patterns after injection of 20E and U73122. To construct its prokaryotic expression pCzn1-AlTOR, T vector containing the target gene was digested with the restriction enzyme EcoR I and Xho I and subcloned. The recombinant plasmid containing target gene was specifically expressed after induction by IPTG. The target recombinant protein has over expressed and has been purified by using Ni-NTA agarose. The polyclonal antibody was obtained by rabbit immunity, cell fusion and ascites preparation, and Western blot was used to detect the specificity of antibody.【Result】The open reading frame of AlTOR encodes 435 amino acids. AlTOR protein has the typical domains including the SIN1 domain, highly conserved CRIM domain and PH domain. qRT-PCR results showed that AlTOR was expressed in 1-day-old to 16-day-old of A. lucorum, in which the expression level was the highest at 1-day-old. AlTOR was expressed in all tissues of A. lucorum female adults, and the fat body had the highest expression level. 20E could induce the expression of AlTOR at different day-old nymphs, and could also induce the up-regulated expression of AlTOR in the head, wing, ovary and fat body of the female adults, which was increased by 200.00%, 118.89%, 20.53% and 60.82%, respectively, while U73122 significantly suppressed AlTOR expression in the midgut and ovary. The vector containing AlTOR was digested and linked to the vector pCzn1. The recombinant plasmid pCzn1-AlTOR expressed the target recombinant protein of 36 kD after induction by IPTG. The 36 kD target protein from the strain containing AlTOR was obtained by the Ni-NTA agarose and used as the antigen, and one cell line polyclonal antibody could specifically bind to the AlTOR recombinant protein and the total protein of the 3rd instar nymph of A. lucorum, indicating that the prepared AlTOR polyclonal antibody has good specificity.【Conclusion】The expression of AlTOR shows the developmental stage- and tissue-specificity. After the induction of 20E and its inhibitor, AlTOR shows the opposite responses. The obtained polyclonal antibody against AlTOR recombinant protein is highly specific.

【Objective】In order to clarify the latitude characteristics and relationships among soil fertility indices in the typical black soil region of northeast China, clarify the problems of fertility control in the heart of black land and provide strong support for the quality control of black land cultivated land and grain yield stable and increase.【Method】Soil samples were collected following by latitude gradient in the study area after the autumn harvest in 2018. The soil fertility indices, such as soil organic matter, pH, total phosphorus, total potassium, available phosphorus, available potassium, alkali-hydrolyzable nitrogen, bulk density, soil water content, soil temperature the ratio of macro- and micro-aggregates, soil water content and soil temperature, were measured to investigate these indices’ latitudinal variation characteristics and their relationships by means of principal component analysis, redundancy analysis and correlation analysis. 【Result】The soil fertility indices of typical black soil region in Northeast China showed obvious latitudinal variation characteristics. In addition to the weak variation of bulk density, other fertility indices showed medium variation. Soil organic matter was the core influencing index in this region, which was increasing with the increase of latitude, and had a significant positive correlation with the ratio of macro- and micro-aggregates, alkali-hydrolyzable nitrogen, available phosphorus, available potassium, total potassium, total phosphorus and pH. Nevertheless, it was negatively correlated with soil water content, soil temperature and bulk density. The soil fertility indices with latitude between 46.07 ° and 49.18 ° were greatly affected by the change of soil bulk density, while the indices with latitude between 42.99 ° and 45.59 ° were combined action with soil bulk density, organic matter content and pH.【Conclusion】In the latitude range of 46.07 ° to 49.18 °, the soil fertility indices could be regulated by improving soil bulk density, while in the latitude range of 42.99 ° to 45.59 °, the soil nutrient should be comprehensively regulated by soil testing and formulated fertilization. For the whole study area, the regulation of soil organic matte was still an important content of soil fertility.

【Objective】The accumulation and distribution of carbon and nitrogen in sweet potato is the key factor of yield formation, the response relationship between carbon and nitrogen accumulation and distribution in sweetpotato under long-term organic fertilizer addition was studied to provide scientific basis for high-yield and high-efficiency cultivation of sweetpotato in fluvo-aquic soil region. 【Method】Based on the 40-year long-term locating test of fluvo-aquic soil (Xuzhou), the treatments of no fertilizer (CK), nitrogen, phosphorus and potassium fertilizer (NPK), organic fertilizer (M), organic fertilizer + nitrogen, phosphorus and potassium fertilizer (MNPK) were selected as the research objects,soil properties, aboveground / underground biomass and carbon and nitrogen contents of main functional organs of sweet potato under different fertilization measures were measured and analyzed, the effects of different fertilization treatments on the content of carbon and nitrogen in sweet potato and the distribution ratio of carbon and nitrogen in various functional organs, as well as the changes of the ratio of carbon and nitrogen (C/N) in the aboveground and underground parts of sweet potato under different fertilization measures were expounded. The relationship between the distribution of carbon and nitrogen in sweet potato and soil properties was analyzed by principal component analysis.【Result】Compared with single application of organic fertilizer (M) or chemical fertilizer (NPK), long-term application of organic fertilizer combined with nitrogen, phosphorus and potassium fertilizer (MNPK) significantly increased the biomass and dry matter quality of sweet potato root tuber (P<0.05). At the same time, the contents of total nitrogen and available potassium in soil were significantly increased (P<0.01). The correlation analysis between soil properties and C/N fixation and C/N ratio of sweet potato showed that the C/N fixation of sweet potato organs was significantly positively correlated with soil organic carbon (SOC) and total nitrogen (TN) (P<0.01), and significantly positively correlated with soil available potassium (AK) (P<0.05). However, the content of soil available phosphorus (AP) was not correlated with the content of carbon and nitrogen in sweet potato leaves, but positively correlated with the nitrogen fixation of root tuber (P<0.01), with a correlation coefficient of 0.839. The C/N ratio of leaves of sweet potato was positively correlated with soil EC (P<0.01), and the C/N ratio of petioles and stems of sweet potato was negatively correlated with soil EC (P<0.01) . The C/N ratio of underground tubers of sweet potato was negatively correlated with soil available phosphorus (P<0.01); principal component analysis (PCA) was used to analyze the distribution ratio of carbon and nitrogen fixation in aboveground and underground parts and the C/N ratio of aboveground and underground parts. The results showed that the first two axes explained 66.6% of the variation, and the contribution rate of the first principal component axis was 42.8%. The order of CK and NPK treatments on axis 1 was higher, which indicated that the soil pH and EC values were higher under both treatments, and the explanations for the distribution of carbon and nitrogen in the aboveground part and C/N in the underground part of sweet potato were higher.【Conclusion】It can be seen that organic material addition can reasonably allocate C/N of each organ, improve the distribution ratio of carbon and nitrogen in the storage root part of sweetpotato, and promote the formation of sweetpotato yield.

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

【Objective】In this study, the genetic analysis and fine mapping of melon seed coat color gene were performed, and the candidate gene was isolated and the molecular marker developed, so as to provide a foundation for gene function and rational utilization. 【Method】The inbred line HP22 with white seed coat was crossed with the inbred line B8 and B150 with yellow seed coat, and the offspring population was obtained, respectively. The seed coat colors of offspring plants were investigated by visual inspection. The locus of gene was fine mapped through map-based cloning strategy, and the candidate target gene in the mapping region was determined by gene sequencing and function analysis of gene coding region. 【Result】The white seed coat was dominant to yellow, which was controlled by a single dominant gene locus, named CmSC1, showed delayed genetic characteristic. Using 368 yellow seed coat individuals from F₂ population, the target gene CmSC1 was delimited to a 95 kb interval flanked by markers S27 and S28 on chromosome 5 that contained twelve annotated open reading frames (ORFs). Among of them, the MELO3C014406 gene encoded a bHLH transcription factor protein which was homologous to the Arabidopsis AtTT8 gene. Sequence analysis of MELO3C014406 gene in the yellow seed coat inbred lines indicated that two types of sequence variation were identified. The first type was an A insertion at the 47th base pairs position downstream of ATG, and the other was a 14 bp deletion at the 260th base pair position downstream of ATG, which caused premature termination and produced truncated proteins, respectively. Furthermore, the sequencing analysis of 65 yellow seed coat accessions were detected by specific molecular marker YS, and the results showed that they all contained one mutation type of two variation types. These results suggested that the MELO3C014406 gene was an ideal candidate gene for CmSC1 controlling seed coat color.【Conclusion】In this study, the CmSC1 gene regulating seed coat color was finally mapped to a 95 kb interval, and the MELO3C014406 gene was presumed to be the candidate gene. In addition, a specific marker YS was developed based on the 14 bp deletion of MELO3C014406 gene.

【Objective】It is known that the new plant hormone strigolactone is a key factor regulating plant branching development, but the role of strigolactone in the growth and development of strawberry plants is unclear. In this study, the expression characteristics and function of the key gene DWARF27 (D27) for strigolactone biosynthesis in woodland strawberry (Fragaria vesca) was revealed, and the role of FveD27 in strawberry branching growth and development was explored, which would lay a theoretical foundation for studying strawberry plant architecture. 【Method】The coding sequence region of strigolactone synthesis key gene FveD27 was cloned by RT-PCR from woodland strawberry. The phylogenetic relationship between FveD27 and D27 in apple, Arabidopsis and other species was analyzed by using MEGA 6.0. A fusion vector of FveD27 and GFP was constructed to inject tobacco leaves for analyzing the subcellular location of FveD27. The expression level of FveD27 in different organs of woodland strawberry was analyzed by qRT-PCR technology. The FveD27 promoter and GUS fusion expression vector was constructed and transformed into woodland strawberry by Agrobacterium-mediated method, and the expression characteristic of FveD27 was further analyzed by GUS staining. The overexpression vector of FveD27 was constructed and the FveD27 overexpression strawberry lines were obtained by Agrobacterium-mediated transformation with leaf disks of woodland strawberry as explants. 【Result】The coding sequence region of FveD27 with the length of 789 bp was cloned from woodland strawberry. The subcellular localization in tobacco showed that FveD27 was located in the chloroplast. The expression levels of FveD27 in woodland strawberry organs from high to low were followed by young leaves, shoot tips, petiole, mature leaves and roots. The FveD27 promoter sequence with a length of 1670 bp was cloned, and transgenic plants with pFveD27::GUS gene showed that the young leaves and buds had strong GUS activity, the mature leaves and petioles had weak GUS activity, while the roots had little GUS activity. The result of FveD27 expression revealed by GUS analysis was consistent with the result by qRT-PCR. The FveD27 overexpression vector was constructed and the transgenic woodland strawberry lines overexpressing FveD27 were obtained with Agrobacterium-mediated transformation. Overexpressing FveD27 in woodland strawberry inhibited significantly the formation of branch crowns and increased the number of inflorescences. 【Conclusion】FveD27 had the functions of regulating the development of branch crowns and the number of inflorescences in woodland strawberry, and this study provided new ideas for regulating the number of branch crowns and yield of strawberry.

Tenderness has been considered as one of the most important eating quality characteristics of meat, while inconsistent changes of tenderness in postmortem (PM) muscles can significantly reduce the purchasing intention. Therefore, investigating mechanism of postmortem muscle tenderization is becoming more and more important in the past decade. Since apoptosis definition was highlighted in the PM tenderization process, it has been widely concerned. Shortly after slaughter, the reactive oxygen species (ROS) was significantly accumulated, and ATP (adenosine triphosphate) was gradually exhausted within muscle fibers, which inevitably led to skeletal cell death. Both PM cell death and meat tenderization refer to the activation of muscle endogenous enzymes by a series of regulatory factors, followed by the degradation of muscle structural proteins. The two biochemical processes are considered to be highly related. PM muscle cells die mainly through apoptosis. Besides, in the early stage of PM, when a small amount of ROS is produced, the cells start their defense system by autophagy and ATP therefore gradually deplete muscle cells in the later stage, which may change from apoptosis to necrosis. It has been documented that mitochondrial pathway is crucial for the apoptosis activation in PM muscles. The release of apoptotic factors from mitochondria is the master node of the intracellular death cascade reaction. The opening status of mitochondrial outer membrane directly determines the way, in which the muscle fiber dies. In this paper, the release mechanism of apoptotic factors induced by PM mitochondrial damage was discussed from the perspectives of mitochondrial membrane permeability and cristae remodeling, and the regulation of mitochondrial damage on PM muscle tenderization was discussed. Moreover the underlying mechanism behind it was also analyzed to reveal the effect of mitochondria on muscle pH regulation through energy metabolism, the release of apoptotic factors and regulation of apoptosis enzyme activity. At the same time, the interaction between mitochondria and endoplasmic reticulum were discussed, focusing on Ca²⁺ signal transduction and cell apoptosis process. The interaction between mitochondria and lysosome was further investigated, by highlighting the stability of lysosomal membrane and the subsequently released cathepsin to activate Bax and Bid to accelerate mitochondrial membrane permeability. Caspases were activated and involved in the limited degradation of some myofibrils in the early stage of PM tenderization, followed by the inactivation resulting from the decreasing ATP or other factors with the extended PM time. Therefore, caspases maybe only involve the early stage of tenderization. This review could provide a theoretical reference for the perfection of PM muscle tenderization investigations.

【Objective】Fat is an important nutrient element in animal diet, and it is also the main energy supply material, so it plays an important role in animal production. The purpose of this study was to investigate the effects of high fat diet on growth performance, energy metabolism and slaughter performance of early weaned Hu lambs before and after weaning, so as to provide theoretical basis and technical support for healthy breeding of early weaned lambs. 【Method】Thirty pairs of healthy Hu sheep twin lambs with similar birth age, similar weight were randomly divided into two groups, including high fat diet group (HF: 26.89% and 5.07%) and normal fat diet group (NF: 15.15% and 2.80%). The lambs of the two groups were fed with milk replacer and pellet with different fat levels from 7 to 60 days old and weaned milk powder at 60 days old, and then which were fed with the same pellet from 60 to 120 days. Nine pairs of twin lambs were randomly selected at the age of 50-60 and 110-120 days according to the average body weight, and the digestion and metabolism trails were conducted by the method of total feces and urine collection to evaluate the energy metabolism of lambs fed diets with different fat levels before and after weaning. According to the average body weight of lambs, nine pairs of twin lambs were randomly slaughtered at the age of 60 and 120 days to test slaughter performance, organ indexes, and gastrointestinal development. 【Result】Before weaning, the total dry matter intake (DMI), total energy intake (GE), fecal energy (FE), urinary energy (UE), apparent digestibility of total energy, total energy metabolic rate (ME/GE), empty body weight (EBW), slaughter rate, GR value of lambs in the two groups were significantly higher than those in the control group .There was no significant difference in the proportion of stomach and intestines to pre slaughter live weight (P>0.05); The body weight, DE, ME, DE/ME, HCW, head weight, heart weight, hoof weight, the proportion of hoof weight to live weight before slaughter, omasum weight and small intestine weight of 60 day old lambs in HF group were higher than those in NF group (0.05<P<0.1), and the eye muscle area and abomasum weight were significantly higher than those in NF group (P<0.05). After weaning milk powder, all lambs were fed the same pellet to 120 days of age. DMI of lambs fed high fat diet at 61-120 stage and BW, LBW, EBW, HCW, skin + gross weight, heart weight, hoof weight and rumen weight of lambs at 120 days of age were significantly higher than those in NF group (P<0.05), and spleen and kidney weight were also higher than those in NF group (0.05<P<0.1); the feeding high fat diet before weaning did not affect energy metabolism, other organ index and gastrointestinal development of lambs after weaning (P>0.05). 【Conclusion】Feeding high fat diet before weaning could improve the body weight, digestible energy and metabolizable energy, carcass weight and eye muscle area of lambs. Feeding high fat diet during lactation significantly increased feed intake, body weight, live weight before slaughter and carcass weight of lambs after weaning. In conclusion, the increasing dietary fat content before weaning had a positive effect on energy metabolism and slaughter performance of Hu sheep twin male lambs before and after weaning.

【Objective】This study was conducted to explore the effects of early weaning and NDF levels of finishing diets on the growth performance, and nutrient digestion and metabolism of Hu sheep lambs, and to filter out the best combination of feeding methods by calculating the superposition effect between the experimental factors. 【Method】One hundred and twenty Hu lambs with similar body weight [(8.26±2.14) kg] and (20±2 ) days of age were randomly divided into four groups with six replicates in each group and five lambs in each replicate. NDF levels of early finishing diets were 33%(low) and 38%(high), NDF levels of late finishing diets were 28% (low) and 38% (high). Treatments included followings: early weaning (EW) + feeding low NDF diet in early and late finishing stage (EW-LL); early weaning + feeding low NDF diet in early finishing stage and high NDF diet in late finishing stage (EW-LH); early weaning + feeding high NDF diet in early and late finishing stage (EW-HH); ewe reared (ER) + feeding high NDF diet in early and late finishing stage (ER-HH). Early weaned lambs gradually transitioned from breastfeeding to feeding milk replacer at the age of 20 days and supplemented with starter feed. The fattening period was from 61 to 180 days of age. The trial lasted 160 days. The body weight and feed intake of lambs were measured at different stages, and digestibility tests were carried out at 2, 4 and 6 months of age.【Result】There was no significant difference in body weight and average daily gain among four groups (P>0.05) except the average daily gain of lambs from 46 to 65 days of age, which was higher in ER group compared with EW group (P<0.05). The dry matter intake of early-weaned lambs from 21 to 65 days old was significantly higher than that of ewe reared lambs (P<0.05), In the early weaned groups, the pellet intake of lambs fed low-NDF diets during the finishing stage were significantly higher than the other two groups at the age of 121-180 days (P<0.05). The nutrient digestibility, energy and nitrogen utilization of early weaned lambs were lower than those in the ER group at 55 to 65 days of age. But it was inverse at fattening period, the dry matter and organic matter digestibility of lambs fed low NDF diet was higher than those fed high NDF diet in finishing stage (P<0.05). EW-LL group had the highest ADG increase scope by superposition effect analysis. The superimposing effect of the two feeding methods of early weaning and low-NDF diet during the early finishing stage was -50.57%.【Conclusion】According to the whole fattening cycle, early weaning and feeding milk replacer could promote the growth performance of lambs. The fattening effect of the low NDF level diet in the early finishing stage was worse than the high NDF level diet. When lambs have the same diet in the late finishing stage, the weight gain of lambs fed with low NDF diet was better than that of lambs fed with high NDF diet in the early finishing stage. By the comprehensive analysis of the growth performance, digestion and metabolism of lambs, the priority of the combination of feed methods was EW-LL>EW-LH=EW-HH>ER-HH. The combination of early weaning + feeding low NDF diet in early and later finishing stage was recommended in this study.

【Objective】this study was conducted to research effects of addition of cinnamaldehyde in calf starter on growth performance, health status, rumen fermentation and microflora of calves, so as to provide theoretical basis for applying cinnamaldehyde in calf cultivation. 【Method】Twenty-four healthy Holstein male calves at the age of around 15 days, with similar body weight, were selected, and were divided into control group and experiment group, with three replicates in each group and four calve in each replicate. The control group were fed basic starter, while starter of experimental group was added with 0.3% encapsulated cinnamaldehyde which contained 15% effective cinnamaldehyde. The weaning period contained 6 pre-trail period, and 27 days of formal trail period, both groups were fed same amount of milk, the starter and orts grass were given at libtium, and the ruminal fluid was taken by stomach tube at the end of weaning period. After weaned, the animals were kept for another 51 days with same feeding management mode, the ruminal fluid was taken at the end of the experiment. The feed intake, growth performance, body measurement, fence score and ruminal volatile fatty acids concentrations were measured during the two phase of the experiment, and the ruminal bacterial community composition was measured based on 16SrDNA gene high through put sequencing. 【Result】The results showed that: (1) In terms of feed intake and growth performance, there were no difference in average daily gain, dry matter intake and feed efficiency between experiment and control group at pre-weaning, post-weaning stage and whole experiment period (P>0.05), there were no differences in body measurements parameters between experiment and control group as well (P>0.05); 2) In terms of calf health status, the diarrhea rate of the experimental group was lower than that of the control group, but was not significant (P>0.05). 3) For the rumen fermentation parameters, the concentration of volatile fatty acids of rumen fluid in the experimental group on weaning day was tend to lower than that in the control group (P>0.05). At the end of the experiment, the concentration of volatile fatty acids, including acetate, propionate, butyrate and valerate of experimental group was significantly higher, while acetate/propionate ratio lower than that of control group (P<0.01); 4) At the end of experiment, cinnamaldehyde supplementation increased ruminal bacteria Shannon diversity index, decreased Simpson diversity index; addition of cinnamaldehyde increased abundance of Firmicutes and Actinobacteria at phylum and prevotella at genus level (P<0.01). 【Conclusion】The results showed that addition of 0.3% encapsulated cinnamaldehyde which contained 15% effective cinnamaldehyde had no influence on growth performance, but decreased diarrhea rate, increased ruminal volatile fatty acids concentration and decreased acetate/propionate ratio, increased ruminal bacterial community diversity, and abundance of Firmicutes and Prevotella, indicated addition of high amount of cinnamaldehyde had no adverse effects on calf growth performance, but changed ruminal bacterial community composition and ruminal fermentation mode.

【Objective】This study was conducted to investigate the effects of dietary grape seed proanthocyanidins (GSPs) supplementation on growth performance, rumen fermentation, rumen and serum inflammatory factors and antioxidative activity in lambs under the condition of a high-concentrate diet, so as to provide a support for applying GSPs on ruminant. 【Method】A total of forty-eight 1/2 Dorper × 1/2 thin-tailed Han ram lambs with similar body weight (BW = 22.75±1.20 kg, mean ± SD) were randomly divided into four groups, with twelve lambs each group. Lambs were fed a 30:70 forage:concentrate diet with 0 (control), 10 (10GSPs), 20 (20GSPs), and 40 mg·kg^-1 BW (40GSPs) GSPs supplemented, respectively. The feeding experiment lasted for 60 days before sampling, with the first 15 days for diet transition and adaptation. On the first day of the formal experiment period, the BW before morning feeding was weighed as the initial BW. At the end of the trial period, blood samples were collected from the jugular vein, and the serum was separated for the determination of antioxidant and inflammatory indicators and lipopolysaccharide content. At the same time, six lambs in each group were randomly selected to collect rumen fluid by using an oral stomach tube at 1, 3, 4, 6, 8 and 12 hours after feeding, respectively, and the rumen pH were measured immediately. The fluid samples at 3 hours after feeding were used to measured fermentation parameters and lipopolysaccharide content. The other 6 lambs in each group were slaughtered, and the samples of rumen tissue were collected to determine the antioxidation index and inflammatory factors. 【Result】The final weight of lambs in 10GSPs and 20GSPs groups were significant greater than that in control group (P<0.05), with no difference between 40GSPs and 10GSPs or 20GSPs (P>0.05). Lambs in 10GSPs and 20GSPs groups also had higher average daily gain and average daily feed intake than control and 40GSPs (P<0.05), with no significant difference between control and 40GSPs (P>0.05). The dietary GSPs supplementation had a certain regulatory effect on rumen pH. The rumen pH increased linearly with the supplementation of GSPs (P<0.05) at 3, 8 and 12 h after feeding, and tended to increased linearly at 4 h (P=0.057). The concentrations of acetate, butyrate and total volatile fatty acid in rumen fluid tended to decrease after GSPS was added (P<0.1), but there was no significant effect on propionate, isovalerate, valerate, and the ratio of acetic acid to propionic acid (P>0.05). After GSPs was supplemented, the serum lipopolysaccharide concentration was significantly lower than that of the control group (P<0.05), but which did not affect the lipopolysaccharide concentration in the rumen fluid. The activity of GSH-Px in rumen tissue of 20GSPs and 40GSPs groups was significantly higher than that of the control group and 10GSPs group (P<0.05) , however, the content of MDA was significantly lower. The serum SOD activity of 20GSPs and 40GSPs group was significantly higher than that of the control group, and the GSH-Px activity was higher than control and 10GSPs group (P<0.05). There was no significant effect was observed in rumen inflammatory factors when GSPs was supplemented, but there was a trend of decreasing IL-6 and IL-10 (P<0.1). The level of TNF-α in 20GSPs and 40GSPs group were significantly lower than that in the control group and 10 GSPs group (P<0.05). The level of IL-10 in 40GSPs group was significantly lower than control group, with no significant difference between 40GSPs and 10GSPs or 20GSPs (P>0.05). 【Conclusion】Supplementing appropriate amount of GSPs to high-concentrate diet could improve the rumen pH, and the antioxidant capacity of serum and rumen tissue of lambs, suggesting that appropriate amount of GSPs had the potential protective effect on lamb health. The optimal feeding dose was 20 mg·kg^-1 BW under the condition of this experiment.

【Objective】The activity of superoxide dismutase (SOD) in wheat grains has a significant effect on the color and nutritional quality of wheat flour. Identification of associated loci and candidate genes for SOD activity in wheat grains is important for discovering the genetic mechanism of SOD activity in wheat grains and genetic improvement of wheat flour color. 【Method】The SOD activity of 212 common wheat varieties (lines) planted in 3 environments was detected by photoreduction method of nitro-blue tetrazolium (NBT), and the genome-wide association study (GWAS) of SOD activity in wheat grains was carried out by 16 705 high-quality SNP markers of 90K SNP chip, and candidate genes of significantly associated loci of stable inheritance were identified. 【Result】The phenotypic variation of SOD activity among wheat varieties (lines) was significant in different environments, with the coefficient of variation ranging from 4.34% to 5.23%, the correlation coefficient ranging from 0.60 to 0.90 (P<0.001). Polymorphic information content (PIC) ranging from 0.24 to 0.29 and the whole genome linkage disequilibrium (LD) attenuation distance of 7 Mb. The analysis of population structure showed that the tested materials could be divided into 3 subgroups. GWAS analysis showed that 29 loci (P≤0.001) were significantly associated with SOD activity, which were distributed on chromosomes 1A, 1B, 2A, 2B, 2D, 3B, 3D, 4B, 4D, 5A, 5B, 5D, 6A, 6B, 6D and 7B. A single locus can explain the phenotypic variation(R²) between 5.47% and 32.43%, of which 14 loci were detected in 2 or more environments. Nine significant associated loci were detected in three environments, distributed on chromosomes 1B, 2B, 4B, 5A, 5B, 6B and 6D, with a contribution rate of 6.21%-16.62%. SOD genes of TraesCS2B01G567600, TraesCS3D01G069900, TraesCS3D01G070200, TraesCS5B01G525700, TraesCS6A01G021400 and TraesCS6D01G431500, and SOD-activity-related candidate genes of TraesCS5A01G263500 and TraesCS6B01G707800 were used to identify the candidate genes of significantly associated loci of stable inheritance. The functions of the candidate genes were mainly related to the inhibition of cell reactive oxygen species accumulation and the participation in antioxidant regeneration. 【Conclusion】Twenty-nine SNP loci associated with SOD activity in wheat grains were detected, and 7 SOD genes and 2 candidate genes related to SOD activity were screened out.