Acta Agriculturae Zhejiangensis

Identification and analysis of HP gene family in wheat

LOU Yuangen, LI Chuang, LI Jingjing, XING Guozhen, LU Yanan, ZHENG Wenming

2023, 35(9): 2023-2032. DOI: 10.3969/j.issn.1004-1524.20221279

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HP (histidine phosphotransfer proteins) plays an important role in regulating growth and development and responding to stress in plants. In order to understand the evolution and function of HP genes in wheat genome, this study identified the HP gene family for Triticum aestivum by the bioinformatics method, and analyzed the physicochemical properties, evolutionary characteristics, gene structure, cis-acting elements, and expression patterns under stress. The results showed that 31 HP genes were identified. The length of amid acids of HP proteins ranged from 116 to 200. Comparison with HP proteins of other plants and analysis of protein structure, gene structure and motif showed that sequences of HP gene family were conserved in wheat. Analysis of cis-acting elements revealed that the promoter regions of HP genes contained regulatory elements involved in abiotic stress, such as light, phytohormone, drought and low temperature. Heatmap analysis showed that HP genes significantly differed in the expression levels under abiotic stress. qRT-PCR analysis indicated that TaHP5-6B was strongly induced under low phosphorus stress. In conclusion, HP family genes identified at the genome-wide level are important gene resources associated with abiotic stress response in wheat.

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Evaluation and analysis of agronomic and quality traits of 115 peanut germplasm resources

ZHANG Xiaoli, ZHU Linglong, LI Fuzhen, TANG Xiumei, XIA Youlin, YOU Yu, ZHONG Ruichun

2023, 35(9): 2033-2044. DOI: 10.3969/j.issn.1004-1524.20221204

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In order to understand the genetic diversity of peanut germplasm resources and improve the utilization efficiency of peanut germplasm resources, 115 peanut germplasm resources were used as materials. Nineteen agronomic and quality traits were evaluated by coefficient of variation, Simpson diversity index, correlation analysis, principal component analysis and cluster analysis. The results showed that the variation coefficients of 19 descriptive traits ranged from 4.21% to 43.42%, in which height of main stem, total branching number, number of branches with pods, pod number per plant and pod yield per plant were rich in variation. The days from sowing to flowering, seed width, shelling percentage, protein content and oil content were relatively stable. Meanwhile, Simpson’s diversity index of the above traits ranged from 1.78 to 2.09. Among them, the linoleic acid content was the smallest, and the seed width was the largest. Correlation analysis showed that the yield per plant was significantly positively correlated with pod number per plant, 100-pod weight, 100-seed weight, pod width and seed width. The 19 agronomic and quality traits were classified into 5 principal components, and the cumulative contribution rate reached 77.142%, which could comprehensively summarize the information of the 19 traits. Cluster analysis divided the 115 peanut germplasm resources into 3 groups. The Group Ⅰ was large-fruit peanut resources with excellent comprehensive character performance, which could be used as high-quality parents in breeding. Sixty-eight accessions with high oil content, 27 accessions with high protein content and 6 accessions with high sucrose content were selected, which could be used as high-quality parents for peanut quality breeding.

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Effects of different cultivation modes on agronomic characters and accumulation of active components of Ganoderma lucidum strains

LI Xuelong, LI Chao, LI Yue, LIU Guoli, ZHANG Peng, ZHANG Min

2023, 35(9): 2045-2055. DOI: 10.3969/j.issn.1004-1524.20221254

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The aim of the study was to explore the effects of wood log cultivation mode and substrate substitute cultivation mode on main agronomic characters and accumulation of active components of different Ganoderma lucidum strains, and to determine the high-quality and efficient cultivation mode of G. lucidum fruiting body. The wood log and substrate (saw dust, wheat bran, gypsum) were used as the cultivation material, and the average mycelial growth rate, mycelial growth vigor, mycelial growth of full bag time and mycelial characteristics of different G. lucidum strains under the two cultivation modes were compared. The main agronomic traits, such as single fruiting body weight, stalk length, stalk diameter, cap diameter, cap thickness and biological efficiency were compared, too. The contents of triterpenoids, sterols, polysaccharides, proteins and 16 kinds of amino acids in fruiting bodies were detected by pharmacopoeia and national standard methods, and the effects of different cultivation modes on the accumulation of main active components in G. lucidum fruiting bodies were compared. There were some differences in average mycelial growth rate, mycelial growth vigor, mycelial growth full time and mycelial characteristics among the six G. lucidum strains tested under the two cultivation modes. The main agronomic traits of G. lucidum strains under different cultivation modes were significantly different, such as single fruiting body weight, stalk length, stalk diameter, cap diameter, cap thickness and biological efficiency. G. lucidum strain CZ22 got the largest fruiting body weight (117.15 g) and the highest biological efficiency (7.54%) under wood log cultivation mode. Strain CZ22 also got the largest fruiting body weight (40.58 g) and the highest biological efficiency (9.02%) under the substrate substitute cultivation mode. There were some differences in the contents of triterpenoids, sterols and polysaccharides in fruiting bodies of the same G. lucidum strain under different cultivation modes. The highest contents of triterpenes and sterols in fruiting bodies was observed in strain CZ27 (1.14%) under the substrate substitute cultivation mode, and the highest content of polysaccharide in fruiting bodies were achieved by strain CZ25 (1.36%) under the substrate substitute cultivation mode. However, the content of protein, total content of 16 amino acids and each amino acid were significantly different. Compared with the wood log cultivation mode, the G. lucidum cultivated by substitute cultivation mode had higher biological efficiency, larger fruiting body yield and higher content of bioactive components, which provides a theoretical basis for efficient cultivation of G. lucidum.

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Identification and analysis of alternative splicing events related to sheep hair follicle development based on RNA sequencing technology

YUAN Xiaochun, WANG Yifan, WANG Yayan, SUN Haoran, MENG Ke, LI Xinhai

2023, 35(9): 2056-2067. DOI: 10.3969/j.issn.1004-1524.20221409

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Alternative splicing is an important molecular mechanism that regulates protein diversity and genetic diversity in animals and plants. In order to identify and analyze the variable splicing during hair follicle development during depilation of Dorper sheep, the Illumina HiSeqTM 4000 sequencing platform was used to sequence the transcriptome of the skin tissue samples in the middle of the side of Dorper adult ewes in September 2019 (peak growth period, S1), January 2020 (telogen, wool growth stops, S2), and March 2020 (early growth period, new wool growth causes old wool to fall off, S3). Using the rMATS software to identify the variable splicing events and differential splicing genes in the samples, 128 033 variable splicing events were identified in three group samples, of which the proportion of skipped exons was the highest, accounting for 75.69%, and the proportion of retained introns was the lowest, accounting for 2.03%. A total of 3 448 significantly (P<0.05) different splicing genes were screened in S1 vs S2, S1 vs S3 and S2 vs S3. The functional enrichment analysis of GO (gene ontology) and KEGG (Kyoto Encyclopedia of Genes and Genomes) showed that differential splicing genes were significantly enriched in VEGF (vascular endothelial growth factor) signal pathway, MAPK (mitogen-activated protein kinase) signal pathway, PI3K Akt signal pathway, Rap1 (RAS-related protein 1) signal pathway, Hippo signal pathway, neurotrophic factor signal pathway and other pathways closely related to hair follicle development, indicating that differential splicing genes played an important role in hair follicle development during depilation in Dorper sheep, The key genes (SMAD2, FGFR2, BMP4, BCL2, etc.) that may be involved in the hair follicle development of sheep depilation traits were further screened out. By analyzing the AS events in the hair follicle development process of Dorper sheep during depilation, this paper reveals the relevant genetic regulation mechanism in the process of hair follicle development and the periodic cycle of wool, which provides a theoretical basis for the subsequent analysis and excavation of the function of development related genes in the process of hair follicle development and the periodic change of wool growth during sheep depilation.

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Genome-wide analysis of pathogenicity and drug resistance of Micropterus salmoides-derived Aeromonas veronii strain AV040

ZOU Wenteng, LIU Zhenhong, YANG Jingxuan, LU Yangyang, PENG Kaisong

2023, 35(9): 2068-2078. DOI: 10.3969/j.issn.1004-1524.20221113

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An outbreak of fry mortality occurred during the ponding period of largemouth bass (Micropterus salmoides) from a farm in Anhui, China in July 2021. After excluding parasitic and viral infections, multiple strains of bacteria were isolated from the spleen and kidney of the affected fish, and a highly pathogenic strain of Aeromonas veronii(A. veronii) strain AV040 was screened by identification, single-colony β-hemolysis test of blood plate and regression infection. Whole genome sequencing of AV040 (GenBank ID: CP095841) revealed 253 coding sequences for virulence genes, of which 156 encoded for adherence molecules, 85 for secretion system and 12 for toxin. The median lethal dose (LD₅₀) of AV040 was 6.53×10⁶ CFU·g^-1 and 2.97×10⁷ CFU·g^-1 for largemouth bass (about 10 g) and Kunming white mice (about 20 g), respectively. The infected largemouth bass exhibited hemorrhage in pectoral and ventral fins, anus, hepatopancreas, spleen and pyloric pendulum, and the infected mice exhibited curling, squinting and trembling, and hemorrhage in the liver and intestine. Five resistance genes in the whole genome of AV040 and K-B drug sensitivity test were performed for phenotypic validation. Among them, OXA-912 and cphA3 were associated with carbapenem resistance and imipenem resistance to carbapenem; rsmA and adeF were associated with aminoglycosides, quinolones and tetracyclines, with sensitivity to gentamicin, kanamycin, neomycin and butamikana of the aminoglycosides, norfloxacin, enrofloxacin, ciprofloxacin and ofloxacin of the quinolones, and doxycycline and tetracycline of the tetracyclines. EF-Tu mutants were associated with resistance to the rifamycins of the rifamycin class. No resistance genes were detected in the genome of strain AV040, but antibiotics phenotypical resistance of antibiotics including clindamycin and lincomycin of the lincomycin class, metronidazole of the nitroimidazole class, meperidine of the methicillin class, penicillin G and ampicillin of the penicillins class were shown. Based on whole-genome sequencing, this study resolved the complex relationship between phenotypes and genotypes of pathogenicity and drug resistance of AV040 strain, and deepened the understanding of the pathogenesis of A. veronii of largemouth black bass origin.

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Comparative analysis of chromosome karyotype between male and female red claw crayfish (Cherax quadricarinatus)

LIU Fangfang, CHEN Honglin, LIU Feng, XU Xiaojun, HUANG Fuyong, LOU Bao, QIAN Haojie

2023, 35(9): 2079-2089. DOI: 10.3969/j.issn.1004-1524.20221534

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In order to obtain accurate chromosome number, arm length and other data to assist chromosome level genome assembly of red claw crayfish (Cherax quadricarinatus), as well as to understand its cytogenetic characteristics, the karyotype differences between male and female red claw crayfish chromosomes were analyzed. The hepatopancreas and testis of red claw crayfish were used as materials to screen the best duration of colchicine. The chromosome specimens were prepared by low permeability and air drying methods. The mitotic phases were obtained from male and female hepatopancreas, and the meiosis phases were obtained from testis. The number of chromosomes of 36 female and male mitotic phases and the number of bivalents of 36 male meiotic phases were counted. The results showed that the highest frequency of the number of chromosomes was 200, which accounted for 39.3% and 50.0% in female and male red claw crayfish, respectively. And the highest frequency of the number of bivalents was 100, which accounted for 50%. The mitotic metaphase with clear chromosome morphology and identifiable centromere was selected for karyotype analysis. It was found that the results of karyotype grouping of female and male red claw crayfish chromosomes were consistent, and the karyotype formula was n=100=36(m)+33(sm)+14(st)+17(t). No heterotypic chromosome was found. The results of independent sample t-test showed that there was no significant difference in chromosome length and chromosome arm ratio of each type between male and female red claw cravfish (P>0.05). The present study compared the chromosome karyotype of female and male red claw crayfish for the first time, which enriched the basic research of red claw crayfish cytogenetics and the chromosome karyotype information of decapods, provided basic data for the genetic breeding of red claw crayfish and reliable reference data for its genome assembly at the chromosome level.

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Identification and functional analysis of the BrLCYB gene of lycopene β-cyclase from Chinese cabbage

LI Biyuan, YUE Zhichen, ZHAO Yanting, LEI Juanli, HU Qizan, TAO Peng

2023, 35(9): 2090-2096. DOI: 10.3969/j.issn.1004-1524.20221285

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In this study, the only lycopene β-cyclase was cloned from Chinese cabbage (BrLCYB) and had 1 482 bp coding sequence. BrLCYB contains the only one intron in its gene sequence, corresponding to 5' UTR. The intron resulted in BrLCYB producing two types of transcripts (BrLCYB-1 and BrLCYB-2) after transcription. BrLCYB-1 accounted for 98.26% of the total BrLCYB transcripts. BrLCYB was widely expressed in a variety of organs and the whole embryonic development stage, and has a higher expression in flowers and mature embryos. The phylogenetic tree of LCYBs showed that the LCYBs of higher plants originated from cyanobacteria. During the evolution of LCYB genes, intron gain and intron loss, occurred successively in LCYBs, and finally no intron existed in coding sequences of LCYBs in higher plants. BrLCYB contained a lycopene β-cyclase domain. In order to further verify the function of BrLCYB, the pET-BrLCYB prokaryotic expression vector was constructed. The co-transformation of pACCRT-EIB and pET-BrLCYB into E. coli can make E. coli enriching orange-yellow β-carotene, proving that BrLCYB of Chinese cabbage had the function of lycopene β-cyclase.

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Effect of glycine betaine on physiological characteristics of eggplant seedlings under high temperature stress

GAO Xiaoping, ZHANG Jing, NIU Tianhang, LIU Yang, CHANG Youlin, LIU Sitian, XIE Jianming

2023, 35(9): 2097-2108. DOI: 10.3969/j.issn.1004-1524.20221258

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To investigate the mitigating effect of exogenous glycine betaine on the injury of eggplant under high temperature stress, eggplant variety ‘Bride’ was used as the test material. Seedlings were foliar sprayed with 0 (CK), 20 (T1), 40 (T2) and 60 mmol·L^-1 (T3) glycine betaine at four leaves and one heart stage for 3 d. The changes in biomass accumulation, photosynthetic characteristics, Calvin cycle key enzymes activity and antioxidant enzymes activity were measured in a climatic chamber at 40 ℃ after high temperature stress for 48 h. The results showed that the growth of eggplant seedlings under high temperature stress could be alleviated to different degrees by spraying different concentrations of glycine betaine, with the most obvious effect of 40 mmol·L^-1 treatment. Compared with CK, the height, fresh weight, root length, root surface area, root volume and root tip number of T2 treated seedlings were significantly (P<0.05) increased by 32.97%, 34.17%, 19.47%, 16.55%, 32.20% and 25.29%, respectively; T2 treatment significantly increased chlorophyll a, chlorophyll b and total chlorophyll content; meanwhile, it significantly increased leaf photosynthetic parameters net photosynthesis rate (P_n), stomatal conductance (G_s) and transpiration rate (T_r) by 51.26%, 40.24% and 49.86%, respectively, and significantly increased the activities of Calvin cycle related enzymes activity such as fructose 1,6 diphosphatase (FBPase), fructose 1,6-bisphosphate aldolase (FBA), 3-phosphoglyceraldehyde dehydrogenase (GAPDH) and transketolase (TK), and the actual photochemical efficiency of photosystem Ⅱ. T2 treatment significantly increased leaf superoxide dismutase (SOD) activity, peroxidase (POD) activity, catalase (CAT) activity and proline (Pro) content by 35.62%, 109.65%, 72.99% and 58.13%, respectively. Glycine betaine at 40 mmol·L^-1 reduced the degradation of chlorophyll accumulation, maintained photosynthetic carbon cycle enzyme activity, promoted photosynthesis, improved leaf antioxidant enzymes activity, and reduced cell membrane injury, thus promoting the growth of eggplant seedlings under high temperature stress

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Numerical estimation of chlorophyll in pepper leaves based on optimized vegetation index combination

WANG Yu, WANG Hong, XIAO Jiujun, LI Kexiang, XING Dan, ZHANG Yongliang, CHEN Yang, ZHANG Lanyue

2023, 35(9): 2109-2120. DOI: 10.3969/j.issn.1004-1524.20221456

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Leaf chlorophyll is closely related to vegetation growth, and it is an important parameter that directly affects vegetation growth and health status. SPAD value can reflect the chlorophyll content of crop leaves. The larger the SPAD value, the higher the chlorophyll content, and the smaller the SPAD value, the lower the chlorophyll content. The shape and physiological and ecological parameters of different varieties of pepper are significantly different, so it is of great significance to accurately and quickly estimate the SPAD value. The SPAD values of 4 different varieties of peppers, Qianjiao 8, Hongla 18, Layan 101, and Red Globe, were measured. The reciprocal, logarithm, reciprocal logarithm, first-order differential and second-order differential transformation of the original spectrum were performed. The vegetation index was optimized by replacing the original spectrum with the transformed spectrum, and the difference between the optimized vegetation index and the classical vegetation index was compared. Finally, the relationship between SPAD values and hyperspectral of different varieties of peppers was obtained, and the optimal inversion model of SPAD values was found. The results showed that there were differences in canopy spectral characteristics of different pepper varieties. SPAD values of modeling set, validation set and full sample of pepper leaves showed that the change trend of line pepper was greater than that of Chao tian pepper. The correlation coefficients of vegetation indices based on reciprocal logarithmic spectral optimization were higher than classical vegetation indices except CI_{red edge}. The random forest model based on lg1/R-VI had good accuracy in both the modeling set and the validation set, which was suitable for the estimation of SPAD values of different varieties of pepper leaves. The coefficient of determination (R²) of full sample model test set was 0.83, mean absolute deviation (MAD) was 1.90, and R²of validation set was 0.45, MAD was 1.26.

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Analysis of the registration of compound preparations of triazole fungicides and strobilurin fungicides

WANG Xiangyun, WANG Luyan, ZHANG Changpeng, LI Yanjie, ZHAO Xueping, JIANG Jinhua

2023, 35(9): 2121-2129. DOI: 10.3969/j.issn.1004-1524.20220917

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In order to clarify the registration of compound preparation of triazole and strobilurin fungicides, the registered information of triazole and strobilurin fungicides obtained from China Pesticide Information Website were analyzed. Based on formulations, toxicity, application methods, crops and targets, the registration of 13 compound preparations were selected for further analysis. Among them, there were 10 types of formulations, and the suspension concentrate, water dispersible granule, wettable powder and emulsifiable concentrate was the top four formulations. It was found more than 92.0% of the compound preparations were low toxic to environmental organisms, and spraying was the main method during their application. The compound preparations of difenoconazole·azoxystrobin was the most widely registered on crops, while the tebuconazole·azoxystrobin compound preparations have the most types of control targets. Results summarized the characteristics of compound preparations of triazole and strobilurin fungicides, and provided scientific data for the development of their environment-friendly preparations.

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pH value sensibility evaluation of pesticides hydrolysis half-life

LI Hui, HU Wenlan, ZHANG Zhiheng

2023, 35(9): 2130-2148. DOI: 10.3969/j.issn.1004-1524.20221205

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In order to quantify the influence of environmental pH value changes on pesticide half-life, a pH value sensibility index evaluation method was established based on pesticides half-life data under different environmental pH values. The sensibility was divided into seven grades including 3A, 2A, 1A, 0, 1B, 2B and 3B according to the index. The results of the evaluation of the hydrolysis half-life of 305 pesticides showed that the pH value sensibility index ranged from -1.67 to 3.31, the average was 0.50, and the average of absolute value for index was 0.60. The pH value sensibility index of the hydrolysis half-life of most pesticides was positive, which will promote hydrolysis when the pH value increased in the environment. Carbamate, diphenyl ethers and aryloxyphenoxypropionate herbicides, carbamate and pyrethroid insecticides, dicarboximidies and benzimidazole fungicides were particularly significant. However, triazine and sulfonylurea herbicides were unique and had a tendency to accelerate hydrolysis when the pH value decreased. The pH value sensibility index of pesticide half-life can directly reflect the influence degree of environmental pH value change on pesticide half-life.

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Antifungal activity and mechanism of wood vinegar against several plant pathogenic fungi

LI Yanxiang, DING Dedong, HE Jing, ZHANG Jinhua, ZHAO Jitao, ZHAO Qian, HOU Caixia, ZHU Zhu

2023, 35(9): 2149-2159. DOI: 10.3969/j.issn.1004-1524.20221348

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To explore the practical application of wood vinegar in the biological control of plant diseases and its mechanism of action against pathogenic fungi, experiments were conducted to evaluate the antimicrobial activity of wood vinegar against several tested plant pathogenic fungi by measuring the colony growth diameter and half effective concentration (EC₅₀). The impact of wood vinegar on spore germination, mycelial dry weight, cell membrane permeability, cell wall integrity, soluble protein content, soluble sugar content, reducing sugar content, and adenosine triphosphate (ATP) content of sensitive fungal strains was assessed, and the ultrastructural changes of the fungal bodies were observed through electron microscopy to reveal its mechanism of action. The results showed that wood vinegar had certain inhibitory effects on all 7 tested plant pathogenic fungi, with the most significant inhibitory effect observed against Fusarium culmorum, with an EC₅₀ value of 4.98 μL·mL^-1, and the inhibition of spore germination and biomass at a volume fraction of 5.5 μL·mL^-1 reached 90.09% and 95.22%, respectively. After treatment with wood vinegar, the cell structure of F. culmorum was damaged, the cell walls of the mycelium became thinner and incomplete, organelle structure was disrupted, and cell membrane permeability increased. Alkaline phosphatase activity, soluble sugar content, reducing sugar content, soluble protein content and ATP content of the mycelium were significantly reduced. Additionally, the fluorescence intensity of iodine-propidium iodide stained spores increased significantly. Wood vinegar mainly achieves its antimicrobial effect by damaging the fungal cell structure and interfering with fungal substance metabolism and energy metabolism.

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Acute toxicity and risk assessment of pesticides aginst pear fire blight on Italian honeybees

BAN Xue, MA Xiangyu, ZHANG Feipeng, ZHANG Wangbin

2023, 35(9): 2160-2168. DOI: 10.3969/j.issn.1004-1524.20221158

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In order to clarify the ecological risk of pesticides against pear fire blight on Italian honeybees (Apis mellifera ligustica), the acute toxicity of seven pesticides was determined indoors through continuous intaking and dripping experiments under the guidance of national standard GB/T 31270.10—2014 “Test guidelines on environmental safety assessment for chemical pesticides part 10: honeybee acute toxicity test”. The risk assessment was carried out according to the introduced hazard quotients (HQ). The results of the acute toxicity test showed that, the 48 h median lethal concentration (LC₅₀, recorded by active ingredients, the same as below) and median lethal dose (LD₅₀, recorded by single bee and active ingredients) of 45% zhongshengmycin TC (technical material), 85% kasugamycin TC and 4% kasugamycin AS (aqueous solution) were 208, 253, 247 mg·L^-1 and 412, 2 434, 223 μg, respectively, which indicated low toxicity. The 48 h LC₅₀ and LD₅₀ of 0.3% ehydroxide AS were 45 mg·L^-1 and 2.86 μg, respectively, which indicated moderate toxicity. In the acute oral toxicity test, the 48 h LC₅₀ of 3% zhongshengmycin SL (soluble concentrate), 3% benziothiazolinone ME (microemulsion) and 47% kasugamycin copper oxychloride WP (water powder) was 110, 65, 107 mg·L^-1, which indicated moderate toxicity; while in the acute contact toxicy test, the 48 h LD₅₀ of these pesticides was 193, 22, 480 μg, respectively, which indicated low toxicity. The HQ of 3% benziothiazolinone ME and 0.3% ehydroxide AS was 75.0 and 340.9, respectively, which indicated moderate risk, while the HQ of 3% zhongshengmycin SL, 4% kasugamycin AS and 47% kasugamycin copper oxychloride WP was 8.5, 5.3 and 3.1, respectively, which indicated low risk. Therefore, in the production of Korla pear, zhongshengmycin, kasugamycin and kasugamycin copper oxychloride were preferred to control pear fire blight. For application of ehydroxide and benziothiazolinone, relevant measures should be taken to reduce the risk of toxicity upon honeybees.

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Transcriptome sequencing and analysis of potential pathogenicity-related genes in Isaria fumosorosea

XIE Meiqiong, WANG Longjiang, HE Yurong, LYU Lihua

2023, 35(9): 2169-2180. DOI: 10.3969/j.issn.1004-1524.20221707

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In order to explore the expression patterns of transcriptome genes of Isaria fumosorosea and to provide data for the functional study of I. fumosorosea, the transcriptom of I. fumosorosea IFCF01 was sequenced and analyzed using high-throughput non-parametric sequencing technology. A total of 540 896 444 clean reads were obtained by transcriptome sequencing. A total of 36 466 unigenes with the total length of 34 551 468 bp and an average length of 947 bp were obtained after assembly. The functional annotation showed that a total of 22 741 unigenes were annotated in 6 major databases,such as Nr, GO, COG and KEGG, accounting for 62.36% of the total unigenes. A total of 21 737 unigenes were annotated in the Nr database, with the highest number of anotation to Beauveria bassiana. 10 888 unigenes were annotated to 25 functional categories in COG database, and the number of genes involved in general function predicted only was the largest, matching on 4 060 unigenes. The GO annotation shoewed that a total of 9 617 unigenes were annotated, distributed in 49 subcategories of three functional modules: biological process function, molecular function and cell composition. A total of 14 364 unigenes were annotated in the KEGG database, involving 108 KEGG metabolic pathways. With the help of pathogen-host interaction gene database, a total of 86 unigenes associated with the potential pathogenicity of I. fumosorosea were identified. A total of 8 572 simple sequence repeats (SSR) loci were detected in transcriptome data of I. fumosorosea, with a frequency of 18.18%. The results of this study could lay the foundation for gene resources mining, functional gene research and SSR molecular markers for I. fumosorosea.

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A soil respiration monitoring equipment based on open chamber method

GU Kechen, JIANG Junjie, PAN Chenxin, SUN Qisong, YIN Wenjie, HU Junguo

2023, 35(9): 2181-2191. DOI: 10.3969/j.issn.1004-1524.20221470

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A soil respiration monitoring equipment was designed to overcome the inconvenience of conventional open chambers which required gas sampling in an infrared gas analyzer to measure CO₂ concentration, and a correction calculation method was proposed for unstable CO₂ concentration. A compact and lightweight low-cost non-dispersive infrared sensor module was installed in a suitable position inside the chamber as an alternative to the infrared gas analyzer through computational fluid dynamics simulation analysis. Under the controlled gas flow rate of 3, 4, 5 L·min^-1, the CO₂ concentration field inside the chamber at soil respiration rates of 0.5, 2, 5, 10 μmol·m^-2·s^-1 was obtained according to the simulation software. The center point of the bottom surface of the chamber was used as the origin to establish Cartesian coordinate system. Combined with the analysis of sensor module dimensions, it was found that for the chamber described in this research, the average error in measuring CO₂ volume fraction to calculate soil respiration rate did not exceed 15% in the spherical area determined by the point (-0.046 5, 0.071 7, 0.209 0) (unit: m) as the center and the diameter of 0.05 m. After the deployment of sensors, the performance of the equipment was compared with the LI-8100 automated soil CO₂ flux system and the self-made closed chamber in field, and the mean error was 10.8%, which was consistent with the results of the simulation analysis.

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Effects of chemical fertilizer combined with organic fertilizer on soil properties, cherry fruit quality and yield

YUE Zongwei, LI Jiaxiao, SUN Xiangyang, LIU Guoliang, LI Suyan, WANG Chenchen, ZHA Guichao, WEI Ningxian

2023, 35(9): 2192-2201. DOI: 10.3969/j.issn.1004-1524.20221052

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To study the effects of chemical fertilizer combined with organic fertilizer on soil physical and chemical properties, soil enzymes activities, and fruit quality and yield of cherry, a field experiment was conducted in Tongzhou District, Beijing, China. A total of 6 treatments were set including control (CK, no fertilizer) and 5 treatments with different proprotion of organic fertilizer. Except CK, the N input in the other treatments were equal. The differences of soil physical and chemical properties, soil enzymes activities, fruit quality indexes and fruit yiled were monitored among treatments. Compared with CK and the treatment with pure chemical ferilizers, the treatment with 50% (based on N input) chemical fertilizer+50% organic fertilizer could significantly (P<0.05) improve the contens of soil organic matter, total nitrogen, available nitrogen, available phosphorus, available potassium, and activities of soil urease, protease, alkaline phosphatase, dehydrogenase. Besides, this treatment could significantly increase viatamin C content and sugar-acid ratio in fruit without significant yield reduction. Thus, for cherry production in the test area, 50% chemical fertilizer+50% organic fertilizer was the recommended fertilization method.

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Activation of soybean shell biochar and its adsorption performance for carbaryl

HAN Jing, ZHU Yiting, ZHENG Chi, MA Lihong, ZHANG Yanan, ZENG Qiuyan, LIU Shuliang, CHEN Shujuan

2023, 35(9): 2202-2211. DOI: 10.3969/j.issn.1004-1524.20220998

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In order to provide theoretical basis and technical support for the utilization of soybean shell and the control of environmental pollution of carbaryl, KOH was used as activator to optimize the preparation conditions of soybean shell activated biochar (A-SBC). The effects of different systems on the adsorption capacity of A-SBC for carbaryl were studied. The adsorption kinetics and adsorption thermodynamics of carbaryl by A-SBC were analyzed. The results showed that the A-SBC obtained by pre-carbonizing the soybean shell at 700 ℃ for 1 h, mixed with KOH at a mass ratio of 1∶2.0, and activation at 750 ℃ for 1.5 h exhibited the best adsorption performance for carbaryl. The adsorption rate and adsorption capacity were 89.63% and 113.28 mg·g^-1, respectively. After activation, the surface of A-SBC was concave, the pores were dense, and the organic functional groups wre reduced. When the pH of the system was 2.0-6.5, the adsorption performance of A-SBC was good, and the maximum adsorption capacity was recorded at the pH value of 5.5. The adsorption capaticy increased with the elevated temperature. The maximium adsorption capacity was found when the ionic strength (NaCl) was 0.01 mol·L^-1. The adsorption of carbaryl by A-SBC was more consistent with the quasi second-order dynamic model. The isothermal adsorption curve was more suitable to be fitted by Langmuir equation, which indicated that this process was mainly chemical adsorption. Thermodynamics test results showed that the adsorption of carbaryl by A-SBC was dominated by hydrophobic interaction and was a spontaneous endothermic reaction. The adsorption performance of A-SBC for carbaryl was excellent, which could provide new ways and new materials for removing carbaryl in water environment.

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Preparation of beta@chitosan fresh-keeping coating and its effect on the quality of Taigu pear jujube

WANG Shiyao, XIN Zhenzhong, WANG Lian, WANG Yu

2023, 35(9): 2212-2221. DOI: 10.3969/j.issn.1004-1524.20221442

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Taigu pear jujube often appears unfavorable storage phenomena, such as cracking, moldiness and decline of quality after harvest because its mature period is rainy season. As a result, the shelf life of fresh fruit is shortened and the commodity quality has decreased. In order to extend its shelf life, a kind of core-shell structure fresh-keeping coating with beta zeolite as the core phase and chitosan as the shell phase was prepared, and the effects of different ratios of the beta zeolite and chitosan on the post-harvest quality of Taigu pear jujube were further explored. The best ratio of beta zeolite and chitosan in fresh-keeping coating was explored through testing the hardness, weight loss rate, cell membrane permeability and soluble solid content. The results showed that after 70 days, the respiratory intensity of Taigu pear jujube treated with the fresh-keeping coating with the quality ratio of beta zeolite and chitosan of 1∶1.5 was 43.99 mg·kg^-1·h^-1, its hardness was 4.99 N. Compared to the initial situation, hardness remained 42%, its weight loss rate was 2.45%, its permeability of cell membrane increased by 48.67%, its soluble solid content decreased. These results suggested that beta @chitosan fresh-keeping coating could delay the increase of weight loss rate and cell membrane permeability of Taigu pear jujube to a certain extent, and inhibit the decrease of hardness and soluble solid content, thus improving the fruit storage quality.

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Chemical composition, antioxidant capacity and protective activity against DNA damage of organic selenium-enriched matcha

LI Xiao, YAN Tingyu, NIU Lili, ZHAI Panpan, ZHANG Huafeng, YANG Xiaohua, PENG Yuanjin

2023, 35(9): 2222-2232. DOI: 10.3969/j.issn.1004-1524.20221253

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In order to elucidate nutritional value, health benefits and their material basis of the novel organic selenium-enriched matcha (OSM), contents of some nutrients (e.g., essential amino acids and trace elements) and bioactive constituents [e.g., epigallocatechin gallate (EGCG)] in 3 kinds of matcha [i.e. organic selenium-enriched matcha, domestic matcha (DM) and imported matcha (IM)] were determined using several instruments such as inductively coupled plasma-optical emission spectrometer. The sensory characteristics of 3 kinds of matcha were analyzed using electronic nose and other methods. Heavy metals and pesticide residues were quantified. In vitro antioxidant capacity as well as protective activity against DNA damage were also studied. The results indicated that content of organic selenium in OSM reached the standard of organic selenium-enrich foods. In particular, selenium concentrations in selenium polysaccharide and selenium protein of OSM were higher than those of DM and IM. In addition, OSM was rich in trace elements such as iron, zinc and strontium. Contents of 3 essential amino acids (i.e. lysine, valine and threonine) and EGCG in OSM were significantly (P<0.05) higher than those in the other 2 kinds of matcha. Lead, arsenic, mercury and cadmium were not detected in OSM and its protein and polysaccharide, and its relative pesticide residues were significantly (P<0.05) lower than those in DM and IM. The sensory characteristics of OSM were better than those of the other 2 kinds of matcha. Moreover, total reducing power, ABTS and DPPH radical-scavenging ability and protective activity against DNA damage of OSM were stronger than those of DM and IM. In conclusion, organic selenium-enriched matcha meets the national food safety standards, possesses high nutritional value and superior sensory characteristics, and exhibits antioxidant capacity and protective activity against DNA damage.

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Design and experiment on portable self-propelled tea picker

YU Guohong, ZHENG Hang, YE Yunxiang, XUE Xianglei, FU Tong

2023, 35(9): 2233-2239. DOI: 10.3969/j.issn.1004-1524.20221118

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In order to solve the problems that the current double hand-lifted tea picker need to be equipped with a large number of operators and the use comfort was poor, a portable self-propelled tea picker suitable for hilly and gentle slope terraced tea gardens was investigated. Its basic structure and working principle of the whole machine were explained, and its key components such as crawler chassis, tea-picking lifting mechanism, hydraulic transmission system and so on were designed and analyzed. Field tea picking experiments were carried out to test the performance of the prototype. The results showed that the fresh leaves under the integrity was more than 81.1%, processable tea rate was more than 93.7%, measured tea machine loss rate was less than 0.82%, measured tea machine leakage rate was less than 0.77%. The portable self-propelled tea picker had a good performance, which meet the requirements of tea farmers for picking.

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Detection of piggery cleaning target under low-light condition

LI Qi, LI Yuzhe

2023, 35(9): 2240-2249. DOI: 10.3969/j.issn.1004-1524.20221447

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The cleaning of piggery without manual intervention can effectively prevent pigs from contracting diseases. Aiming at the problems of weak light, inconspicuous features, and mutual occlusion in the piggery scene, the low detection accuracy of cleaning targets, an improved YOLOv5 algorithm was proposed. In the preconditioning stage, the Retinex algorithm based on bilateral filtering was used to improve the detection ability of the piggery cleaning target under low light conditions; In the cleaning target detection process, the CBAM (convolutional block attention module) attention mechanism was introduced into the backbone of the YOLOv5 network to enable the network to learn more effective feature information of the cleaning target in the piggery; The frame regression loss function in the network was improved, and the DIoU-NMS algorithm was used to filter out the candidate frame of the cleaning target in the piggery, which improved the detection accuracy of the cleaning target in the case of partial occlusion. The experimental results showed that the improved YOLOv5 target detection algorithm improved the accuracy rate by 7.3 percentage point, the recall rate by 7.6 percentage point, and the average accuracy by 7.1 percentage point compared with the benchmark algorithm on the test set of piggery cleaning targets. The robustness was higher in the case of target occlusion. The result provided a basis for the research and development of animal husbandry intelligent cleaning equipment.

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A light-weight model for plant disease identification based on model pruning and knowledge distillation

LIU Yuanyuan, WANG Dingkun, WU Lei, HUANG Dechang, ZHU Lu

2023, 35(9): 2250-2264. DOI: 10.3969/j.issn.1004-1524.20221193

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The emergence of deep learning has provided a new method for plant disease identification, but the current deep learning models have many parameters, which are difficult to use on edge devices such as smartphones or embedded sensor nodes with limited storage and computing resources. In the present study, plant leaves are taken as the research objects, and the methods based on knowledge distillation and model pruning are used to construct a light-weight model for plant disease identification. By improving the ResNet model, one or more teaching assistant network training models are introduced into the knowledge distillation. After sparse training, a light-weight student network model is obtained by model pruning; and the student network is retrained by using the teaching assistant network and learning rate rewinding, which can reduce the size of the model and effectively ensure the performance of the model. The experimental results show that, on a dataset including 38 categories of 14 plants, after pruning the model by 90%, the accuracy of the model is 97.78%, with an increase of 1.49 percentage points over the original model. On the dataset including 5 categories of apple leaves, after pruning the model by 70%, the accuracy of the model is 91.94%, which is 4.85 percentage points higher than the original model. The proposed light-weight model can be transplanted on Android platform and run effectively, which provides a new solution for the embedded terminal to accurately identify plant diseases.

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Law and economics analysis on supervision model of agricultural natural resource assets

ZHANG Wenfei

2023, 35(9): 2265-2274. DOI: 10.3969/j.issn.1004-1524.20221379

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Based on the background of the super ministry system reform, different supervision models of agricultural natural resource assets were compared with analytical paradigm of law and economics, by taking the consistency of supply and demand and cost-benefit as analysis tool. Supervision model of agricultural natural resource assets should be aimed at achieving the balance between supply and demand, to ensure the cost is proportional to the gain. Results show that, from the point of participants, compared with the unilateral supervision model, the matching degree of supply and demand in multiple supervision is higher in consistency of legal systems and feasibility of legal norms. From the point of supervision form, compared with the decentralized supervision model, the comprehensive supervision could obtain greater benefits at lower costs. Realization of multiple comprehensive supervision of agricultural natural resource assets should endow multiple subjects such as government, market and society with the right (power) to govern together, build mechanisms of public participation, in order to maintain social security and maximize the social utility.

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