Acta Agriculturae Zhejiangensis

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2023, 35(3): 0-0.

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Assessment of genetic diversity and variety identification based on insertion site-based polymorphism (ISBP) markers developed in wild species related to sweet potato

MENG Yusha, WANG Yin, LAI Qixian, LIU Lei, XIANG Chao, WU Yonghua, ZHENG Yanran, GU Xingguo, FANG Hao, MIAO Miao, WU Liehong, TANG Yong

2023, 35(3): 489-498. DOI: 10.3969/j.issn.1004-1524.2023.03.01

Abstract ( 427 )

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Based on genome wide sequencing of wild species related to sweet potato Ipomoea triloba and Ipomoea trifida, 2 360 long terminal repeat retrotransposons (LTR-RT) sequences were identified. One hundred insertion-site-based polymorphisms (ISBP) primer pairs were developed and four sweet potato germplasm resources were used to evaluate the polymorphisms of ISBP primers. Twenty four pairs of ISBP primers were found to be highly polymorphic, which were then applied to genetic diversity analysis and variety identification of 56 sweet potato germplasm resources collected in Zhejiang province. The results showed that 24 pairs of ISBP primers generated 182 polymorphic bands in 56 germplasm resources, and each germplasm resource had unique fingerprint information. The genetic analysis results of NTsys, UPGMA and PCA showed that the genetic distance between different germplasm resources in the population ranged from 0.027 5 to 0.653 8, with an average of 0.373 2, the average of polymorphic information content (PIC) was 0.289 6, and these resources were divided into two populations containing 11 and 45 germplasm resources, respectively. Studies have shown that the genetic variation of this population is small and the genetic background among different germplasm resources is narrow. The ISBP primers developed in this study not only increased the number of transposon molecular markers, filled in the gap of application of ISBP markers in genetic analysis of sweet potato, but also provided a favorable tool for the identification, protection and utilization of sweet potato variety resources.

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Effects of NaCl stress on seed germination and physiological characteristics of winter oilseed rape

XU Yaozhao, ZENG Xiucun, WANG Zhenchao, DANG Shizhuo, LIU Yongjing

2023, 35(3): 499-508. DOI: 10.3969/j.issn.1004-1524.2023.03.02

Abstract ( 491 )

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In this study, we investigated the salt tolerance of winter oilseed rape cultivars by subjecting Brassica rapa cultivars of Longyou 16, Tianyou 2, Tianyou 4, and Brassica napus cultivars of Ningyou 2 to NaCl stress at concentration of 34, 68, 102, 136, 170 mmol·L^-1. We measured the seed germination indexes and physiological characteristics. In addition, salt tolerance index and salt tolerance critical value were determined by correlation analysis and regression analysis, respectively. The results showed that seed germination rate, radicle length, sprout length, seedling dry weight and fresh weight all showed a decreasing trend with the increase of NaCl concentration, and the decrease of Brassica rapa cultivars of Longyou 16 and Tianyou 2 and Tianyou 4 was less than that of Brassica napus cultivars of Ningyou 2, and the inhibition degree of high salt concentration (170 mmol·L^-1) on radicle was significant. With increasing salt stress concentration, proline content, cell membrane permeability, malondialdehyde (MDA) content and peroxidase (POD) activity of seedlings gradually increased. Correlation analysis showed that seed germination rate was significantly (P<0.01) positively correlated with radicle length, sprout length, seedling fresh weight and seedling dry weight, while germination rate was significantly (P<0.01) negatively correlated with MDA content, cell membrane permeability, proline content and POD activity; MDA content in rapeseed seedlings was significantly (P<0.01) positively correlated with cell membrane permeability and proline content. Regression analysis showed that critical value of salt tolerance for seed germination and growth index of winter rapeseed was 164.87-251.99 mmol·L^-1, and the median lethal concentration and lethal concentration were 316.29-494.95 mmol·L^-1 and 558.57-883.67 mmol·L^-1, respectively. The critical value of germination growth index of winter rape showed germination rate>sprout length>fresh weight of seedlings>dry weight of seedlings>radicle length. The salt tolerance of winter oilseed rape showed Longyou 16>Tianyou 2>Tianyou 4>Ningyou 2. The salt injury index of winter oilseed rape showed Ningyou 2>Tianyou 4>Tianyou 2>Longyou 16. In conclusion, seed germination rate, radicle length and MDA content of winter oilseed rape could be used as indicators of salt tolerance. These results could provide references for germplasm innovation of salt tolerance winter oilseed rape.

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Aluminum subcellular distribution and its combining characreristics with cell wall in tea leaves

LI Chunlei, XU Hongmei, LIU Jie, ZHANG Rujun, MA Xingyun, ZHANG Hua

2023, 35(3): 509-514. DOI: 10.3969/j.issn.1004-1524.2023.03.03

Abstract ( 393 )

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Tea tree (Camellia sinensis) is a very typical aluminium (Al) concentrating plant, which is mainly enriched in the leaves. In order to analyze the characteristics of Al content in tea leaves, this paper explored the accumulation characteristics of Al in tea leaves using mature leaves of the high-Al variety Nongkangzao and the low-Al variety Pingyangtezao as materials, and analyzed the subcellular distribution of Al in tea leaves and the possible binding mode with the cell wall. The results showed that Al in the leaves of the two varieties (Nongkangzao and Pingyangtezao) was mainly distributed in the cell wall (46.2%-54.9%), and to a less extent in the nucleus and chloroplasts (24.2%-31.2%). In the cell wall fractions, pectin was the main Al-enriched component of the cell wall, accounting for 60.1%-79.0% of the Al content in the cell wall. Both methyl-modified and esterified treatments significantly reduced the Al content of the cell wall, while the cellulase and pectinase enzymatic treatments had less effect on the Al content of the cell wall, suggesting that the -COOH and -NH₂ in the cell wall play a key role in binding Al, while the length of the sugar molecule chains had no significant effect on the Al binding. Therefore, the cell wall of tea leaves is the main location where Al accumulates, preventing it from entering further into the cell interior and reducing its toxic effects; Al is most probably bound to the carboxyl and amino groups in the cell wall in the form of a hydrogen bond and is immobilised in the cell wall.

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Prediction and bioinformatics analysis of target genes of plant development-related miR828 gene family

HAN Xueyang, LIU Ning, WEN Xin, WEI Jicheng, REN Ruyi, HAO Aiping

2023, 35(3): 515-522. DOI: 10.3969/j.issn.1004-1524.2023.03.04

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MicroRNAs (miRNAs) are a class of small regulatory RNA. miRNAs play an important role in many natural and pathological processes in organisms. In order to study the characteristics and biological functions of plant miR828 family, the secondary structure, target gene prediction, sequence alignment and evolution tree of miR828 members were analyzed using bioinformatics method. The precursors and mature sequence of miR828 family members were obtained from the PmiREN database, and the secondary structure was predicted online through the precursor sequence, and the sequence comparison, target gene prediction and evolution tree were constructed through the mature sequence. A total of 45 mature miR828 members were obtained from the database, which only distributed in angiosperms. Precursor and mature miRNAs are highly conserved in plant evolution. The secondary structure prediction found that all members of the miR828 family could form stable stem ring structures. Target gene prediction analysis found that miR828 family target gene include domain proteins, transcription factors, carrier proteins, ubiquitin ligases, etc. Most of them are MYB transcription factors and MYB domain proteins. The phylogenetic tree analysis showed that the members of miR828 family were close to each other. These results provide a basis for further study on the characteristics and functions of plant miR828 family.

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Effects of intercropping tea with landscape trees on ecosystem of tea garden and tea production

WANG Jinfeng, ZHOU Qi, LYU Yulong, CHEN Zhuomei

2023, 35(3): 523-533. DOI: 10.3969/j.issn.1004-1524.2023.03.05

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In order to find a suitable intercropping mode for landscape trees and tea, three intercropping modes of landscape trees-tea, including Camellia sinensis(L.) O. Ktze.-Yulania denudata (Desr.) D.L.Fu, C. sinensis-Sapindus saponaria L. and C. sinensis-Prunus campanulata (Maxim.) Yü et Li were comprehensively studied and evaluated by measuring the microclimate, soil physical and chemical properties, and tea yield and quality. Tea monoculture was also studied as a control. The results showed that all the three intercropping modes were beneficial to the microclimate of the tea garden by reducing temperature and increasing humidity of tea garden, especially for the C. sinensis-Y. denudata mode and C. sinensis-S. saponaria mode. All three intercropping systems could improve the physical and chemical properties of the soil in the tea garden. In particular, for the tea garden soil intercropped with P. campanulata, the content of organic matter, total nitrogen, total phosphorus and the enzyme activities of urease and sucrase were significantly improved, which meant that the soil nutrients were comprehensively enhanced. While Y. denudata and S. saponaria intercropping with tea would promote the uptake of available potassium and phosphorus by tea trees, which would be beneficial for tea yield and quality. In addition, all three types of intercropping could improve tea yield and quality to some extent. At the time of promoting the growth of tea trees, the three intercropping modes were also beneficial to the accumulation of amino acids and other substances, and a suitable phenol-ammonia ratio was formed, which meant the tea quality was improved. Especially for the tea trees intercropped with S. saponaria, tea quality was significantly improved and tea yield was 51.81% higher than that of the monoculture tea garden. In conclusion, M. denudata, S. mukorossi and C. campanulata were all suitable landscape trees which could be intercropped with tea trees.

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Preliminary analysis on research hotspot of crop root exudates in China

ZHAO Shuhui, ZHANG Zhenhua, OU Zhangdan, TIAN Maoping, CHEN Yumei, ZHAO Ziwei

2023, 35(3): 534-546. DOI: 10.3969/j.issn.1004-1524.2023.03.06

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Based on the experimental studies on crop root exudates published in Chinese from 2000 to 2021 in the databases of CNKI, CQVIP and Wanfang, the hotspot of crop root exudates studies was preliminarily explored. It was shown that 42% of the published studies focused on wheat, soybeans, maize, rice, faba beans. Crops, microorganisms and soil were the main factors involved in the interaction between external environment and root exudates. Interaction studies mainly focused on the influence of crop root exudates on the environment, and the response of root exudates to the environment. Based on the analysis accuracy of crop root exudates, relative studies could be divided into three levels, namely, total detection (39% of the total studies), functional component analysis (50% of the total studies) and total component analysis (11% of the total studies). Although the types of large molecular weight compounds such as lipids, alkanes and alcohols in crop root exudates accounted for a relatively higher proportion of all compounds (41%), the number of studies on low molecular weight and a few types of primary metabolites such as acids, amino acids and sugars accounted for a relatively higher proportion (24%). In general, grain crops are the focus of root exudates research in China’s agricultural system. The full analysis of root exudates is still in the early development stage. The interaction between the functions of acids, amino acids and primary metabolites of sugars in crop root exudates and the external environment is the hot topic at present.

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Effects of Hyriopsis cumingii polysaccharides on growth performance, antioxidant activity and immune function in growing broiler

XIA Lunbin, MA Longlong, QIAO Deliang, HE Yanfei, JIANG Ping

2023, 35(3): 547-555. DOI: 10.3969/j.issn.1004-1524.2023.03.07

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The objective of this study was to investigate the effects of Hyriopsis cumingii polysaccharides (HCP) on growth performance, antioxidant activity and immune function in growing broiler. A total of 240 one-day-old Arbor Acres (AA) broiler chickens were randomly allotted into 4 groups, with 6 replicates in each group and 10 chickens in each replicate. Normal control group (HCP-0) was fed with basal diet, while low dose group (HCP-1), medium dose group (HCP-2) and high dose group (HCP-3) were supplemented with 200, 400 and 800 mg·kg^-1 HCP in basal diet, respectively. The experimental period was 42 days. The results showed as follows:1) Compared with HCP-0 group, the ADG of broilers in HCP-2 group and HCP-3 group was extremely significantly increased (P<0.01), and there was no significant difference in ADFI and F/G among all groups(P>0.05). 2) Compared with the HCP-0 group, the activities of T-AOC, T-SOD and GSH-Px in serum of broilers in HCP-2 group and HCP-3 group were significantly increased(P<0.05 or P<0.01), the content of MDA was significantly decreased(P<0.05), and only T-SOD activity of broilers in HCP-1 group was significantly increased(P<0.01), the activities of CAT showed no significant difference among all groups (P>0.05). 3) The spleen index of broilers in HCP-2 group was significantly increased compared with HCP-0 group (P<0.05), while there was no significant difference among other groups (P>0.05), and there was no significant difference in bursa of fabricius index and thymus index among all groups (P>0.05). 4) On the 21st day of experiment, compared with HCP-0 group, the content of IgG in serum of broilers in HCP-2 group was significantly increased (P<0.05), and the content of IgA in serum of broilers in HCP-1 group, HCP-2 group and HCP-3 group were significantly increased (P<0.05); At the 42nd day of the experiment, the content of IgG in the serum of broilers in HCP-2 group and HCP-3 group was extremely significantly increased compared with HCP-0 group and HCP-1 group (P<0.01), there was no significant difference between HCP-1 group and HCP-0 group(P>0.05); At the 21st and 42nd day of the experiment, the content of IgM in serum of broilers showed no significant difference among all groups (P>0.05). Combined with the above results, dietary HCP supplementation can significantly improve ADG, antioxidant activity and immune function of broilers, and the appropriate amount of HCP added to the feed of broilers was 514-679 mg·kg^-1by regression equation analysis under this experimental conditions.

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Correlation of PRLR gene polymorphisms and reproductive traits in Kele pig

YANG Suan, GUO Xiaojiang, YANG Hongwen, XIONG Li, LI Chen, TAN Yuancheng, WANG Chunyuan, ZHANG Yiyu

2023, 35(3): 556-564. DOI: 10.3969/j.issn.1004-1524.2023.03.08

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The aim of this study was to investigate the polymorphisms of the prolactin receptor gene (PRLR) and their effects on reproductive performance in Kele pigs, with a view to finding molecular markers for improving reproductive performance in Kele pigs. In this study, SNPs in the exon region of the PRLR gene were screened in 204 Kele pigs and identified by direct PCR product sequencing. At the same time, the correlation between different genotypes and diplotypes of SNPs and seven reproductive traits was analysed by single-factor variance method in SPSS software, and the influence of base variation on the secondary structure of mRNA was analysed by online software. The results showed that g.20655220C>T mutation was detected in the 8th exon of PRLR gene, which caused serine to proline change, which was a missense mutation. Two synonymous mutations g.20648859C>T and g.20648931C>T were detected in the fourth intron. There were three genotypes in all three SNPs, g.20648931C>T was low polymorphism and the other two loci were medium polymorphism. Chi-squared test showed that the genotype distribution of g.20655220C>T loci was in Hardy-Weinberg equilibrium (P>0.05). The genotypes of g.20648859C>T and g.20648931C>T deviated significantly(P<0.01) from Hardy-Weinberg equilibrium; g.20655220C>T locus led to changes in the secondary structure of the PRLR gene mRNA sequence, resulting in decreased stability of the secondary structure of the mRNA; the three SNPs consisted of 5 haplotypes and 9 diplotypes. The dominant haplotype H1 (CCC) frequency was 0.610 and the dominant diplotype H1H1 (CCCCCC) frequency was 0.382. Correlation analysis showed that the average birth weight, weaning litter weight and weaning average weight of CC genotype were significantly(P<0.05) higher than those of CT genotype. CT genotype of g.20648931c>T locus had significantly (P<0.05) higher birth weight, birth litter weight and weaning litter weight than those of CC genotype and TT genotype, and CC genotype had significantly (P<0.05) higher birth weight than that of TT genotype. H1H5 (CTCTCT) was the dominant doublet type in terms of litter size and live litter size, whereas H2H3 (CCTTCT) was the superior doublet type in terms of litter weight and average weaning weight. In conclusion, PRLR gene could be used as a candidate gene for molecular marker-assisted selection of breeding traits in Kele pig.

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Association analysis between GnRH-1 gene polymorphism and egg quality in quail

WANG Longwei, BAI Junyan, JIA Xiaoping, LEI Ying, CHEN Mengke, FAN Hongdeng, LU Xiaoning, HE Yuhan, ZENG Fanlin, ZHANG Rongkai

2023, 35(3): 565-574. DOI: 10.3969/j.issn.1004-1524.2023.03.09

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To explore the correlation between GnRH-1 gene of gonadotropin releasing hormone (GnRH) gene subtype and quail egg quality, the single nucleotide polymorphism (SNP) mutation site of GnRH-1 gene in Chinese yellow quails, Beijing white quails and Korean quails was detected by PCR amplification and DNA sequencing, the polymorphism of GnRH-1 gene and its association with quality of quail eggs were analyzed in order to obtain the molecular markers related to the quality of quail eggs. The results showed that 14 SNP mutations of GnRH-1 gene were detected in Chinese yellow quails, Beijing white quails and Korean quails, which were C71T, C108T, C168T, C178T, A184G, C206T, A209C, C215T, A252G, A279T, C281T, C293G, C339T, C458T. Only two genotypes of A209C and C281T were detected in Chinese yellow quail and Beijing white quail, while other 12 SNP loci detected three genotypes in three quail varieties. Correlation analysis showed that there were significant (P<0.05) correlations between C71T, C215T, A209C and A279G loci of GnRH-1 gene and yolk height and yolk index, and between the A209C locus and the yolk width. There was a significant (P<0.05) correlation between A279G locus and eggshell thickness of quail. In conclusion, GnRH-1 gene could be used as a molecular marker or candidate gene for quail egg quality.

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Effects of sublethal doses of dinotefuran on genes expression and enzymes activities related to immune and detoxification in foragers of Apis mellifera ligustica

ZHANG Yuhao, MA Weihua, LIU Jinjia, MA Xiumei, JIANG Yusuo

2023, 35(3): 575-581. DOI: 10.3969/j.issn.1004-1524.2023.03.10

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To determine sublethal doses of dinotefuran to Apis mellifera ligustica and explore the effects of sublethal doses of dinotefuran on the expression of immune detoxification genes and enzyme activities on Apis mellifera ligustica, in this study, the feeding trials was used to determine the virulence curve of dinotefuran to Apis mellifera ligustica. The effects of LC₂₀ and LC₅₀ dinotefuran on the relative expressions of Abaecin, Apidaecins, Defensin-1, Hymenoptaecin, AChE-2, PPO, GST and Cyp450 mRNA were investigated by qRT-PCR. Determination of changes in the activities of PPO, AChE, SOD, GST, CAT in Apis mellifera ligustica. The LC₂₀ and LC₅₀ of dinotefuran on Apis mellifera ligustica determined by feeding trials were 0.458 mg·L^-1 and 0.988 mg·L^-1. Compared with the control group, the expression levels of PPO, Apidaecins, Defensin-1, Hymenoptaecin genes were significantly (P<0.05) up-regulated in the LC₂₀ group. Apidaecins, In LC₅₀ group, the expression of AChE-2, PPO, GST, Cyp450, Apidaecins, Defensin-1 showed a significant up-regulation trend, and the expression of Hymenoptaecin and Abaecin showed a significant down-regulation trend. In additon, the expression levels of Abaecin and Hymenoptaecin in the LC₅₀ group were significantly lower than those in the LC₂₀ group. The expression levels of AChE-2, PPO, GST, Cyp450, Apidaecins in the LC₅₀ group were significantly higher than those in the LC₂₀ group. Compared with the control group, the activities of AChE, CAT and SOD of LC₂₀ group decreased significantly. Compared with the control group, the activities of AChE, PPO, CAT and SOD of LC₅₀ group decreased significantly. In conclusion, dinotefuran inhibits gene expression of Abaecin and Hymenoptaecin, and has an inducing effect on Apidaecins, Defensin-1, AChE-2, PPO, GST, Cyp450 affects the normal immune system of Apis mellifera ligustica. The decline of AChE, PPO, CAT and SOD enzyme activities indicates the damage of dinotefuran to the detoxification system of Apis mellifera ligustica. Our stduy provides a further basis for further research on the effects of sublethal dose dinotefuran on the metabolism and physiology of Apis mellifera ligustica.

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Quality analysis of five kinds of tomato germplasm resources with different fruit colors

LOU Qianqi, LIANG Yan

2023, 35(3): 582-589. DOI: 10.3969/j.issn.1004-1524.2023.03.11

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In order to deeply tap the utilization potential of tomato with different fruit colors in quality traits, 83 tomato germplasm with different fruit colors were used as materials to analyze the variation characteristics, correlation and cluster analysis of 17 main tomato quality traits. The results showed that the coefficient of variation of fruit weight of red tomato was the largest (101.05%), the coefficient of variation of vitamin C (V_C) content of pink tomato was the largest (82.27%), the coefficient of variation of fruit weight of orange tomato was the largest (106.11%), the coefficient of variation of fruit weight of brown tomato was the largest (56.91%), and the coefficient of variation of lycopene of purple tomato was the largest (54.65%). Fruit color was positively correlated with lycopene content, and lycopene content was positively correlated with soluble solids content. The tested materials were divided into eight categories at the European distance of 9.5, of which six red tomatoes and one orange tomato were more durable and were clustered in class Ⅱ; three orange tomatoes were grouped into class Ⅴ, which tasted good; one red tomato and one puroke tomato had excellent nutritional quality and were clustered in class Ⅷ.

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Construction of high-density bin marker genetic map and QTL mapping for fruit length in Luffa spp.

CUI Junjie, LYU Zhen, YANG Tianwen, WANG Jing, HONG Yu, CAO Yi

2023, 35(3): 590-597. DOI: 10.3969/j.issn.1004-1524.2023.03.12

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To explore the construction of a high-density genetic map of Luffa spp. and to identify the genetic loci for fruit length variation, a BC₁ population of 67 individuals was developed by crossing two inbred lines, Luffa acutangula ‘Shengyou’ and L. cylindrica ‘Pingguo’, with significant differences in fruit length. A high-density interspecific genetic map of Luffa spp. was constructed using the bin markers based on whole genome resequencing. The map consisted of 9 299 bin markers distributed in 14 linkage groups spanning 2 956.57 cM (about 80.47% of the Luffa cylindrica genome) with an average genetic distance of 0.32 cM. Combined with the genetic map, the MQM model in MapQTL 6 software was used to detect QTL for fruit length in Luffa spp. A QTL locus qFL1.1 for fruit length in Luffa spp. was mapped between markers scaf1-bin33 (19.76 cM) and scaf1-bin359 (92.84 cM) on linkage group Scaf 1. The LOD of qFL1.1 was 4.21, the contribution rate was 28.90% and the additive effect was 6.04. The results provided a theoretical basis for genetic mining based on the interspecific genetic map in Luffa spp.

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Carboxymethyl chitosan-induced transcriptome WRKY genes analysis and resistance-related gene mining in grapefruit fruit

YANG Qing, LIU Shenghong, HUANG Erbin, DU Rongyu, WANG Fang, DENG Jia

2023, 35(3): 598-614. DOI: 10.3969/j.issn.1004-1524.2023.03.13

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In this experiment, postharvest grapefruit fruits were treated with a 1.5% carboxymethyl chitosan (CMCS) solution and sterile water for 24 hours to study their resistance to green mould caused by Penicillium digitatum and Penicillium expansum. Results showed that the CMCS treatment group significantly reduced the incidence of green mould caused by P. digitatum and P. expansum and inhibited the growth rate of spot diameter.Non-reference transcriptome sequencing was conducted on CMCS and sterile water-treated grapefruit peels to screen for CMCS-induced differentially expressed genes (DEGs). A total of 85 DEGs were obtained, including 67 up-regulated genes and 18 down-regulated genes. GO and KEGG analyses were performed on the DEGs, and their functions were categorized into 37 GO entries in three major categories: biological processes, cellular components, and molecular functions. These DEGs were significantly enriched in 20 KEGG metabolic pathways. Furthermore, WRKY genes were mined, and five WRKY genes were screened as the most likely to be involved in the plant disease resistance process. All five WRKY genes were up-regulated, and the evolutionary tree analysis with Arabidopsis showed that the five WRKY genes were functionally similar to those of Arabidopsis genes. The functions of the involved DEGs were mainly related to secondary metabolism, phytohormone signaling, peroxisome, protein processing in the endoplasmic reticulum, glutathione metabolism, and phenylpropane biosynthesis pathways. qRT-PCR validation of the eight key differentially expressed genes showed that the regulatory trends were consistent with transcriptomic trends. These results may provide a theoretical basis for mining CMCS-induced resistance-related genes in grapefruit and studying postharvest molecular resistance mechanisms in grapefruit.

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Effects of litter leaves of Populus tomentosa Carr. on seed germination and phenotypic plasticity of Aegilops tauschii Coss.

CHEN Hao, ZHANG Yuewei, WANG Ning, WANG Zi, CHANG Qingshan

2023, 35(3): 615-623. DOI: 10.3969/j.issn.1004-1524.2023.03.14

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The aim of this study was to investigate the allelopathic effects of leaf litter extraction and litter decomposition of Populus tomentosa Carr. on seeds and seedings of Aegilops tauschii Coss. Seeds and seedlings of A.tauschi were used as materials in the experiment, and the dish method and pot method were used respectively. The results showed that the regulation of allelopathic substances of P. tomentosa Carr. litters had a concentration effect. The concentration aqueous extract of P. tomentosa Carr. promoted germination at low concentration (5 mg·mL^-1) and significantly (P<0.05) inhibited it at high concentration (40, 60 mg·mL^-1). The biomass accumulation and total leaf area were significantly (P<0.05) increased with low leaf litter addition (30 g) of P. tomentosa Carr. to the soil, while in the treatment of high addition (120 g), biomass accumulation, total leaf area, mean leaf area, leaf area ratio, leaf area root mass ratio and relative growth rate were significantly inhibited. The leaves of P. tomentosa Carr. had an allelopathic effect on A. tauschii Coss, high concentrations of allelochemicals in the fallen leaves of P. tomentosa Carr. significantly inhibit seed germination and plant growth of A. tauschii Coss. On the other hand, A. tauschii Coss. showed high phenotypic plasticity in adapting to the environment of fallen leaves of P. tomentosa Carr, leaf had the highest phenotypic plasticity among all plant components, phenotypic plasticity index of total leaf area, leaf biomass, leaf biomass allocation were 0.81, 0.78 and 0.34, respectively.

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Effects of different temperature on supercoolings point and freezing point of Picromerus lewisi Scott

LENG Meng, YAO Dihui, LONG Kai, ZHAO Haiyan, YANG Maofa, YU Xiaofei

2023, 35(3): 624-629. DOI: 10.3969/j.issn.1004-1524.2023.03.15

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In order to clarify the changes of supercooling point and freezing point of Picromerus lewisi Scott under different temperature, female adult, male adult and third instar of P. lewisi Scott were treated at different temperatures(30, 35, 40 ℃) for 1 h, and the changes of supercooling point and freezing point were measured. It was shown that high temperature changed the supercooling point and freezing point of P. lewisi Scott. After treatments under 30, 35, 40 ℃, the supercooling point and freezing point of the third instar of P. lewisi Scott increased significantly (P<0.05) as compared with those under control (25 ℃). The supercooling point of male and female P. lewisi Scott treated at 30 ℃ did not show significant difference with those under control, yet the supercooling point and freezing point of male and female P. lewisi Scott treated at 35 ℃ and 40 ℃ were significantly higher than those under control. The supercooling point and freezing point of male adult were the lowest under different treatments amomg all the tested insect states of P. lewisi Scott. Thus, high temperature could elevate the supercooling point and freezing point of P. lewisi Scott, reduce the temperature adaptation range. Among all the tested insect state of P. lewisi Scott, male adult could better adapt to the temperature change.

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Development and application of bio-organic fertilizers for prevention and control of bacterial wilt of Curcuma wenyujin

FENG Lin, ZHOU Quan, CHEN Weiliang, MAO Bizeng

2023, 35(3): 630-638. DOI: 10.3969/j.issn.1004-1524.2023.03.16

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Bacterial wilt of Curcuma wenyujin (BWCW) caused by Ralstonia solanaceraum was becoming a common destructive disease in the planting of C. wenyujin at present. In order to control the disease, a bio-organic fertilizer (BOF) was developed by using Paenibacillus jamila ZJU-C612-3 and Bacillus amyloliquefaciens ZJU-C612-4, which were screened early in our laboratory and showed strong antagonistic activities to R. solanaceraum. The incidence rate of C. wenyujin tuber treated with BOF in greenhouse pot culture was reduced by 60 percentage point. After using 200 kg BOF in 667 m² field, the incidence rate of healthy C. wenyujin tuber was reduced by 17.66 percentage point, and the plants grew well. In addition, the BOF could alleviate the monocropping obstacle of C. wenyujin. After applying bio-organic fertilizer in the 5-year continuous cropping field, the incidence rate of continuous cropping C. wenyujin tuber decreased by 9.43 percentage point and 19.75 percentage point in July and August. This study result could provide a basis for the industrialized development of bio-organic fertilizer for the control of bacterial wilt of C. wenyujin in future.

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Effects of different soil disinfection methods on soil fungal diversity and community structure

ZHU Shijun, WANG Lili, JIN Shuquan, ZHOU Jinbo, WANG Feng, LU Xiaohong

2023, 35(3): 639-646. DOI: 10.3969/j.issn.1004-1524.2023.03.17

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In the present study, strawberry greenhouse with severe continuous cropping obstacles was used as the research object, the effects of four soil disinfection methods of high temperature control (T1), quicklime disinfection (T2), calcium cyanamide disinfection (T3), and dazomet disinfection (T4) on soil fungal diversity and soil fungal community structure were compared. It was shown that the soil fungal diversity under different soil disinfection methods were reduced to varying degrees as compared with the control (CK), and various alpha diversity indexes decreased as T4>T3>T2>T1. For soil dominant fungi at genus level (relative abundance>1%), the relative abundance of Fusarium, Thielavia, Mortierella, Verticillium and Acremonium were decreased under four soil disinfection treatments compared with CK, while the relative abundance of Chaetomium and Penicillium were increased. According to the impact of soil disinfection methods on soil fungal diversity and soil fungal community structure, the performance of dazomet disinfection was the best and was followed by calcium cyanamide disinfection. Given the economic cost, calcium cyanamide disinfection was a preferable choice.

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Screening thermophiles to promote co-composting of biogas residue and black soldier fly larval frass

XIAO Xiaolan, ZHANG Hao, FU Chuanhui, LIU Hao, RUAN Wenquan

2023, 35(3): 647-657. DOI: 10.3969/j.issn.1004-1524.2023.03.18

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Thermophiles were screened from the thermophilic phase of co-composting of biogas residue of kitchen organic slurry (BR) and black soldier fly larval frass (LF). The strains those could present high activities of degrading organic matter were selected to be compounded into thermophilic inoculum, and the effects of thermophilic inoculum on co-composting of BR and LR were examined. It was shown that 4 bacterial strains and 4 fungi strains were isolated. Among them, the isolated bacterial strains of Bacillus licheniformis and Bacillus aerius had higher activities of glucoamylase, cellulase, laccase, urease, protease and xylanase, and the isolated fungi strains of Thermomyces lanuginosus and Aspergillus fumigatus had higher activities of cellulase and laccase. After compounding and inoculating the above four strains into the co-composting of BR and LF, the thermophilic phase of co-composting was prolonged for 2 days compared with the blank group, and the degradation rate of organic matter, germination index, humic substance content and the ratio of humic acid to fulvic acid were increased from 12.09%, 85.98%, 107.95 g·kg^-1, 2.67 to 15.08%, 90.77%, 117.40 g·kg^-1, 3.01, respectively. In addition, inoculation of thermophilic inoculum increased the relative abundance of Firmicutes and Actinobacteria to 76.84% and 18.12%, respectively, on the fifth day of composting, yet decreased the relative abundance of Saccharomycetales to 1.70%. In general, inoculation of thermophilic inoculum could prolong the period of thermophilic phase, reduce plant toxicity, promote humification process, optimize microbial community structure and improve the quality and efficiency of composting.

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HPLC fingerprint of cultivated Sanghuangporus and content determination of main component

ZHONG Shi, CAO Yinglong, WANG Wenqiong, HUO Jinxi, SUN Yuqing, XUAN Lijiang, LI Yougui

2023, 35(3): 658-665. DOI: 10.3969/j.issn.1004-1524.2023.03.19

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To establish HPLC fingerprint of cultivated Sanghuangporus(Sanghuang), screen out the signature component of Sanghuangporus through chemical pattern recognition, the content determination method of Sanghuangporus was established based on this index. According to the corresponding methods in the general rules of The Chinese Pharmacopoeia (four parts) in the 2020 edition, the fingerprint of Sanghuangporus was established, and the similarity was evaluated by using the similarity evaluation system of traditional Chinese Medicine Chromatographic Fingerprint 2004 Version A to determine the common peak. The content of Hypholomine B in Sanghuangporus was also determined. HPLC fingerprints of 10 batches of Sanghuangporus were established, with 12 common peaks, and the similarity was greater than 0.92. The results of HCA, PCA and OPLS-DA were consistent, and the samples were divided into two categories. The injection volume of Hypholomine B had a good linear range with its peak area in the range of 1.25-20.00 μg (R²=0.999 9). The RSD values of precision, stability and repeatability were all less than 2.0% (n=6). The recovery rate was 95%-99%, and the RSD was 1.48%. The content of Hypholomine B in 10 batches of mulberry yellow samples ranged from 0.15% to 0.95%. The fingerprint can be combined with HCA, PCA and OPLS-DA to comprehensively evaluate the quality of Sanghuangporus. The method is stable and reliable, providing reference for the quality evaluation of Sanghuangporus.

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Optimization and computational fluid dynamics analysis of fan operation for cascading cage-rearing meat duck house in summer

LIN Yong, DAI Weiwei, BAO Encai, WANG Qiang, BAI Zongchun, XIA Liru, ZHANG Yao, SUN Yulun, OUYANG Lihu

2023, 35(3): 666-675. DOI: 10.3969/j.issn.1004-1524.2023.03.20

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To study and determine the key environmental parameter problem areas in the cascading cage-rearing meat duck house in summer, optimize the number and layout of fan operation, and realize precision environment control, a total of 23 measuring points were set in the 96 m (length)×17 m (width)×5.3 m (height) duck house to monitor the dynamics of temperature, relative humidity, wind speed and CO₂ concentration. Computational fluid dynamics (CFD) model of duck house was conducted to simulate the effects of different fan operation schemes on the distribution and evenness of temperature field and airflow field. The results showed that the indoor temperature ranged from 22.9 ℃ to 30.9 ℃, and the outdoor temperature varied from 17.6 ℃ to 36.6 ℃ during the test period. The relative humidity inside and outside the duck house was 48.1%-92.5% and 31.2%-100.0%, respectively. The variation tendency of indoor temperature and relative humidity were similar to that outside the duck house, while the changes of indoor temperature and relative humidity were smaller than that of outside. The indoor wind speed and CO₂ concentration were 0.79-1.32 m·s^-1 and 1 161-1 685 mg·m^-3, respectively. The distribution of temperature field and airflow field inside the duck house were simulated under the scenario when the highest air temperature outside the duck house was 36.6 ℃. When 10 fans with wet curtain cooling were used, the ventilation rate was 287 518 m³·h^-1, the mean temperature and mean wind speed were 30.9 ℃ and 1.08 m·s^-1, respectively. When additional two or four fans were running in response to the high temperature aera which was near the four column cages of center area inside the duck house, the ventilation rate was raised to 314 140, 367 384 m³·h^-1, respectively, and the temperature was reduced by 0.2, 0.4 ℃, respectively, and the mean wind speed was increased by 0.10, 0.30 m·s^-1, respectively, as compared with the original design. In addition, the area with weak ventilation under the original design was improved and the non-uniformity coefficient of airflow inside the duck house was decreased, which could meet the cooling requirement. Thus, accurate regulation of the number and combination of running fans for the high temperature area inside the cascading cage-rearing meat duck house could improve the airflow distribution and realize better cooling effect in summer.

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Can Climate-Smart Agriculture Project reduce agricultural carbon emission: experience evidence from Climate-Smart Staple Crop Production Project in Anhui, China

WEI Xinyan, LIU Ying, ZHANG Junbiao

2023, 35(3): 676-687. DOI: 10.3969/j.issn.1004-1524.2023.03.21

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Based on the panel data of 23 counties in Anhui, China during 2011-2019, under the guidance of Joseph Schumpeter’s innovation theory and Baud’s innovation-driven theory, the synthetic control method was adopted to evaluate the effects of the Climate-Smart Staple Crop Production Project on agricultural carbon emissions in Huaiyuan County and to explore the corresponding action mechanism. It was shown that the Climate-Smart Staple Crop Production Project had certain time lag effect on agricultural carbon emission, as it did not inhibit the per capita agricultural carbon emissions in Huaiyuan County in the initial stage, but it effectively suppressed per capita agricultural carbon emissions in Huaiyuan County from 2016 to 2019. It was calculated that the average annual decline of per capita agricultural carbon emissions was 1.977 6 kg (or 1.43%) in Huaiyuan County during 2015-2019. The intermediary test results showed that the project had promoted the innovation of agricultural development pattern by using modern technology and ideas, and had produced structural and technological effects through innovation, by promoting the structural transformation and upgrading of agriculture in Huaiyuan County and the development of biochemical technologies. Thus, the agricultural carbon emissions in Huaiyuan County has been curbed.

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Research on promotion mechanism of “standard land” reform to high quality development of modern agriculture

ZHANG Yanbin, GENG Bin, LIANG Ying, XU Tiaojuan, XU Baogen, ZENG Xin, ZHANG Yueyue

2023, 35(3): 688-697. DOI: 10.3969/j.issn.1004-1524.2023.03.22

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In order to achieve modern agriculture of high-quality development, governments implement a series of industrial policies to promote the willingness to modern agricultural industry development, and the “standard land” land supply policy plays an important role in regulating and guiding modern agriculture. Based on the survey data of “standard land” in 21 provincial modern agriculture industrial park of Zhejiang Province, this paper constructed the action mechanism of “standard land” policy on the high-quality development of modern agriculture, empirically analyzed the influencing factors of “standard land” policy on the attract of modern agricultural enterprises, and discussed the mechanism of “standard land” policy promoting the high-quality development of modern agriculture. The research showed that factors passed the independent sample T test. The results showed that there were significant differences among the factors affecting the willingness to modern agricultural enterprises development and the “standard land” policy had a strong attraction for the development of modern agriculture. The regression coefficients of standard land control index refinement cognition, standard land credit supervision construction, government service construction, standard land property rights management and crowding-out effect cognition were 1.43, 1.35, 1.28, 0.98, 0.94, respectively, indicating that these five factors had a significant impact on the willingness to modern agricultural enterprises development. For promoting modern agriculture of high-quality development, it is suggested to refine the reform of “standard land” policy by improving the control index system of “standard land”, establishing a unified and strict regional supervision system, maintaining market development order, simplifying approval procedures, establishing regional classification guidelines.

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Effects of mechanization service degree and farmland operation scale on maize production efficiency

WANG Ningke, ZHANG Rui, ZHANG Shengyong

2023, 35(3): 698-707. DOI: 10.3969/j.issn.1004-1524.2023.03.23

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Based on the panel data of 20 major maize producing provinces (autonomous regions, municipality) in China from 2003 to 2018, combined with the methods such as the super efficiency DEA model, regression model and intermediary effect analysis, the effects of mechanization service degree and farmland operation scale on maize production efficiency were analyzed from the intermediary perspective of farmland operation scale. The results showed that the average maize production efficiency of the sample provinces (autonomous regions, municipality) from 2003 to 2018 was 0.775, and there were significant differences among the sample provinces (autonomous regions, municipality). The improvement of mechanization service degree could significantly enhance maize production efficiency. The farmland operation scale had an obvious mediating effect between mechanization service degree and maize production efficiency, and the mediating effect accounted for 32.26% of the total effect. Among the selected control variables, irrigation rate and unemployment rate had significantly positive effects on maize production efficiency, while aging rate and disaster rate had significantly negative effects on maize production efficiency. The effect of farmland operation scale on maize production efficiency exhibited an inverted U shape with an effect boundary. At present, farmland operation scale did not reach the boundary. Based on these findings, it was suggested to properly expand the farmland operation scale and promote the development of mechanization service, so as to further enhance the maize production efficiency.

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Progress on Nep1-like proteins (NLPs) of phytopathogens

XIANG Jiang, CHENG Jianhui, WEI Lingzhu, WU Jiang

2023, 35(3): 708-716. DOI: 10.3969/j.issn.1004-1524.2023.03.24

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NLPs[necrosis-and ethylene-inducing peptide 1 (Nep1)-like proteins] are a type of apoplast secreted protein, which exist in a variety of phytopathogens, and play an important role during plant-pathogen interactions. According to structural characteristics, NLP proteins are divided into three types: Ⅰ, Ⅱ and Ⅲ, with cytotoxic and noncytotoxic forms. Cytotoxic NLPs bind to NTCD4 protein to facilitate oligomerization of NLP proteins, resulting in cell death and disease susceptibility in eudicots, but not monocots. However, the function of noncytotoxic NLP proteins remains unclear. NLPs can also act as microbe-associated molecular patterns(MAMP)to trigger plant immunity. Many plants have evolved immune systems that specifically recognize NLP proteins, e.g., Arabidopsis thaliana recognizes the conserved peptide nlp20/24 of type Ⅰ NLP proteins via the receptor-like protein RLP23, thereby stimulating immune responses. This article reviewed the research progress on NLPs, such as protein structures, gene expression patterns, mechanisms of pathogenicity and host recognition, to provide a theoretical basis and reference for further research on pathogen-host interactions and disease prevention and control.

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Research progress on harm of mycotoxins in feed to aquatic animals

QIU Mei, DENG Qi, FANG Zhijia, WANG Yaling, SUN Lijun

2023, 35(3): 717-726. DOI: 10.3969/j.issn.1004-1524.2023.03.25

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Mycotoxin contamination in aquatic feed poses a health risk to aquatic animals, and causes significant economic losses in the aquaculture industry. This article focused on the current state of mycotoxin contamination in aquatic feed, the toxic effects of mycotoxins on aquatic animals and their residues in meat and organs. The adverse effects of mycotoxins on aquatic animals mainly include decreased appetite, organ damage, and immunosuppression. Mycotoxin-contaminated aquatic food becomes a threat to human health. Mycotoxin residues in aquatic animals have potential food safety risk. Therefore, global limits and regulations for mycotoxins in aquatic feed and aquatic products need to be further refined.

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