Acta Agriculturae Zhejiangensis

Phenotypic identification and diversity analysis of potato germplasm resources in Zhejiang Province, China

SHEN Shengfa, XIANG Chao, LI Bing, LUO Zhigao, WU Liehong

2022, 34(11): 2319-2328. DOI: 10.3969/j.issn.1004-1524.2022.11.01

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To understand the characteristics of local potato germplasm resources and screen out excellent materials in Zhejiang Province, 13 qualitative traits and 6 quantitative traits of 95 potato germplasm resources collected in the Third National Agricultural Germplasm Census and Collection Action of Zhejiang Province were investigated and analyzed in 2019-2020. The results showed that the phenotypic genetic diversity of the local germplasm resources was relatively rich. Shannon-Wiener diversity index of quality traits ranged from 0.380 to 1.686, of which the diversity index of viral diseases was the highest, indicating that these germplasm resources were with different degrees of viral degradation. The diversity index of flavor was the lowest, and there were 88 germplasm resources with good flavor, accounting for 92.63%. The average diversity index of the 10 morphological traits was 1.079, of which three traits of tuber shape, flesh color and corolla color showed the most abundant diversity. The diversity indexes of the six quantitative traits were high, with an average of 2.84, and the tuber yield and number of main stems were particularly prominent. The tuber dry matter contents of the germplasm resources varied from 18.1% to 25.8%, of which 73 resources with dry matter content ≥ 20%, accounting for 76.84%. The 95 germplasm resources could be classified as small potato germplasm (62 resources), medium and large potato germplasm (21 resources) and colored potato (12 resources). All small potato resources were yellow tuber skin and flesh, high dry matter content and good cooking flavor. Nine excellent resources were screened and could be used for production and breeding.

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Functional analysis of plastid phosphate transporter OsPPT family members in rice

WANG Jichun, LI Ruili, WANG Jiaoling, SHAO Junwen, ZHAO Hongyu

2022, 34(11): 2329-2339. DOI: 10.3969/j.issn.1004-1524.2022.11.02

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The phosphoenolpyruvate/phosphate transporter (PPT) is a member of plant plastid phosphate transporter family (pPTs), which transports phosphoenolpyruvate (PEP) in cytoplasm into the plastid and exchanges phosphate to cytoplasm. To comprehensively analyze the OsPPT gene family and explore its potential function in rice, subcellular localization of OsPPT proteins were clarified by transient transformation of rice protoplasts, and the phosphate transport capacity of OsPPT were analyzed by yeast heterologous expression experiments. The tissue specific expression patterns of OsPPT gene family and their responses to abiotic stress were analyzed by setting up hydroponic experimental treatments such as normal phosphorus supply, phosphorus deficiency and abiotic stress. The results showed that all four members of the OsPPT gene family were localized to the chloroplast membrane and OsPPTs could mediate phosphate transport across the membrane in yeast. In addition, dynamic transcriptional changes determined by qRT-PCR revealed differential expression profiles of OsPPTs in response to environmental stress, such as phosphorus starvation and abscisic acid (ABA), salicylic acid (SA), NaCl stress The OsPPT gene family might be involved in the transportation of phosphate between cytoplasm and chloroplasts, and might also participate in the response of plants to stress.

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Identification of the interaction between ZmNLP5 and promoters of ZmSTP1, ZmAAP2 gene in maize

YUAN Chongyuan, ZHU Yuanfei, CHEN Xia, ZHU Chan, WANG Yi, TAO Haiyan, YU Jiaojiao

2022, 34(11): 2340-2347. DOI: 10.3969/j.issn.1004-1524.2022.11.03

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Nitrogen (N) is the major limitation factor for its grain production, and both genes contain a NRE cis element in the promoter region.N use efficiency of maize may be improved by increasing the availability of carbon (C). Transporters of ZmSTP1 and ZmAPP2 play important roles in the transport and unloading of C-N assimilates in maize. In addition, both ZmSTP1 and ZmAAP2 contain NRE cis elements in promoter regions. NIN-like protein (NLP) is a conserved plant-specific transcription factor family and has been shown in several plant species to be a key player in regulating nitrogen response. ZmNLP5 is a central hub in a molecular network for mediating N signalling and metabolism. To understand whether ZmNLP5 is involved in regulating of ZmSTP1 and ZmAAP2, we used maize B73 as experimental material to explore the interaction between ZmNLP5, ZmSTP1 and ZmAAP2 by yeast one-hybrid system. The results showed that the pGADT7-ZmNLP5-1/2 were transformed into the Y1H(pAbAi-ZmSTP1) and Y1H(pAbAi-ZmAAP2) strains, respectively, they all grew normally on control medium (SD/-Leu). On selection medium (SD/-Leu/+200 ng·mL^-1AbA), the growth of Y1H(pAbAi-ZmAAP2/pGADT7-ZmNLP5-1) strain had been in normal. This result suggested that ZmNLP5 can interact with ZmAAP2 gene, ZmNLP5 is a transcription factor that can bind to the NRE element of ZmAAP2. The results of this study not only contribute to the further understanding of the regulatory mechanism of nitrate signal transduction in maize, but also provide a theoretical basis for the future study of nitrate signal transduction in other plants.

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Effects of different density of Cyperus difformis and Ammannia baccifera on rice yield, processing and appearance quality

LIAO Pingqiang, CHEN Guoqi, LIU Guangming, JIANG Yan, ZHAO Can, WANG Weiling, HUO Zhongyang

2022, 34(11): 2348-2357. DOI: 10.3969/j.issn.1004-1524.2022.11.04

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Both Cyperus difformis and Ammannia baccifera were harmful weeds in rice fields worldwide and caused serious impacts on rice production. In this study, the common rice variety Nanjing 46 in Taihu area was selected as the material. The single factor experiment design was used to design five weed densities and four groups of repetitions. To investigate the effects of different densities of C. difformis and A. baccifera on rice yield, rice processing, and appearance quality and seed nitrogen accumulation. The results showed that the effective panicle number, grain number per panicle, 1 000-grain weight and yield of rice gradually decreased with the increase of density of C. difformis and A. baccifera, the seed nitrogen accumulation, processing and appearance quality of rice also decreased. When the density of C. difformis was 65 plants·m^-2, the yield of rice was 3.28 t·hm^-2, and the yield loss rate was 57.29% compared with the blank control. The rice processing and appearance quality were significantly decreased. When the density of A. baccifera was 250 plants·m^-2, the yield of rice was 3.10 t·hm^-2, and the yield loss rate was 59.16% compared with the blank control. Rice processing and appearance quality decreased significantly. Increasing density of C. difformis and A. baccifera could significantly reduce rice yield, processing quality and appearance quality. The results showed that weeds mainly affected rice yield by affecting the effective panicle number, grain number per panicle and 1 000-grain weight of rice, and field microclimate, water and fertilizer competition may be an important reason for affecting rice processing and appearance quality. In order to ensure rice yield, rice processing and appearance quality, attention should be paid to the prevention and control of weeds in the middle and lower layers of rice fields.

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Effects of Magnolia officinalis leaf litter decomposition on growth trait and physiological characteristic of Fritillaria hupehensis

DUAN Yuanyuan, LIU Xiaohong, WU Jiaqi, GUO Xiaoliang, WANG Fanfan, TANG Tao, YOU Jingmao, GUO Jie

2022, 34(11): 2358-2367. DOI: 10.3969/j.issn.1004-1524.2022.11.05

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In order to investigate the allelopathic effect of Magnolia officinalis leaf litter decomposition, a pot experiment was conducted to explore the effects of M. officinalis leaf litter decomposition in soil on growth traits and physiological characteristics of Fritillaria hupehensis. The results showed that compared with control group, at the seedling stage, plant height, leaf length, leaf width, leaf area, bulb length, bulb width, bulb fresh weight and chlorophyll content of F. hupehensis showed no significant difference in low-dose treatment (P>0.05), and were significantly inhibited in the high-dose treatment (P<0.05). At the flowering stage and the beginning of the withering stage, the plant height, leaf length, leaf width, leaf area and chlorophyll content of F. hupehensis showed obvious trends of “promoting effect at low concentration and suppressing effect at high concentration”. The antioxidant enzyme activities, malondialdehyde content and superoxide anion free radical content of F. hupehensis showed different changes in different developmental stages. With the increase of the dose of M. officinalis, comprehensive effect value increased first and then decreased, and the order was T2 (10.50 g·kg^-1) (1.16)>T1 (5.25 g·kg^-1) (0.44)>T3 (15.75 g·kg^-1) (0.02)>T4(21.00 g·kg^-1) (-0.99). Principal component analysis showed that chlorophyll content, malondialdehyde content and superoxide dismutase activity were the main indicators affecting the growth of F. hupehensis at the seedling stage. During the flowering stage, leaf length, leaf width, leaf area and plant height could be used as the main indicators to characterize the growth of F. hupehensis. In the falling seedling stage, superoxide anion free radicals content and peroxidase activity became the main traits affecting the growth of F. hupehensis, underground traits such as bulb length, bulb width, and fresh bulb weight could be used as the main research indicators for the growth of F. hupehensis. In conclusion, the low-dose M. officinalis litter treatment significantly promoted the growth of F. hupehensis, while the high-dose M. officinalis litter significantly inhibited the growth of F. hupehensis. When the dose of M. officinalis was 10.5 g·kg^-1, growth of F. hupehensis was the best.

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Isolation, identification and analyses for genetic features of one bovine viral diarrhea virus strain from yak

WANG Huihui, FENG Xili, WANG Mengzhu, ZHAO Zeyang, LI Yicong, PU Feiyang, MA Peng, LI Yong, GONG Zhenli, MA Zhongren, MA Xiaoxia

2022, 34(11): 2368-2378. DOI: 10.3969/j.issn.1004-1524.2022.11.06

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In this study, one bovine viral diarrhea virus (BVDV) strain was successfully isolated and termed as GSTZ. By electron microscopy, several 50-60 nm viral particles were observed. GSTZ strain displayed the TCID₅₀ (50% tissue culture infection dose) value of 5.0×10^6.5 mL^-1 as revealed by Karber method. Via reverse transcription-PCR (RT-PCR), the whole open reading frame (ORF) of GSTZ strain was sequenced (whole length of 11 691 nt) and classified into the genotype 1 group at either nucleotide level or synonymous codon usage level. The appearance frequency of A+U was higher than that of G+C in GSTZ ORF, and the synonymous codon usage pattern of GSTZ strain displayed the obvious usage bias in synonymous codons with U/A. It was worth noting that, this strain had a tendency of avoiding the selection of the synonymous codons containing CpG dinucleotides, which strongly suggested that the GSTZ strain might weaken the immune responses from host in order to successfully generate its viral particles. The isolated BVDV strain and the above findings could provide biological materials for the related mechanism study of BVDV, and lay foundations for the analysis of antigen relationship of different genotypes of BVDV and vaccine development.

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Association between polymorphism of vasoactive intestinal peptide type Ⅰ receptor gene and carcass traits in meat quails

WANG Xinle, BAI Junyan, LI Jingyun, LEI Ying, LI Gan, PANG Youzhi, DONG Zhihao, CHEN Yu, FAN Hongdeng, WANG Longwei, CHEN Mengke, ZENG Fanlin, AN Xiaokai, BAI Yongzhen

2022, 34(11): 2379-2385. DOI: 10.3969/j.issn.1004-1524.2022.11.07

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In order to investigate the relationship between the polymorphism of type 1 vasoactive intestinal peptide receptor (VIPR-1) gene and carcass traits of meat quail, the polymorphism of VIPR-1 gene was detected by PCR-RFLP method in two populations, including French giant meat quail and Savimaito meat quail, and the correlation between the polymorphism of VIPR-1 gene and carcass traits of meat quail was analyzed. The results showed that GG, GT and TT genotypes were detected at the BsrDⅠ locus of exon 4-5 of VIPR-1 gene in French giant meat quail and Savimaito meat quail, the GG, GT and TT genotype frequencies of French giant meat quail and Savimaito meat quail were 0.408, 0.490, 0.102; 0.222, 0.444, 0.334, respectively. AA, AG and GG genotypes were detected at the HpyCH4Ⅳ locus of exon 6-7 of VIPR-1 gene in French giant meat quail and Savimaito meat quail, the AA, AG and GG genotype frequencies of French giant meat quail and Savimaito meat quail were 0.020, 0.020, 0.960; 0.056,0.111,0.833, respectively. The results of association analysis showed that the BsrDⅠ locus in exon 4-5 of VIPR-1 gene was significantly associated with the liver weight of French giant meat quail (P<0.05). The HpyCH4Ⅳ locus in exon 6-7 of VIPR-1 gene was significantly associated with body weight, slaughter weight and liver weight of French giant meat quail (P<0.05), and with the body weight, slaughter weight, eviscerated weight, heart weight, chest muscle weight, leg muscle weight, liver rate and chest muscle rate of Savimaito meat quail (P<0.05). The VIPR-1 gene can be used as a candidate gene for marker-assisted selection of carcass traits in meat quail.

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Regulation of endoplasmic reticulum molecular chaperone GRP94 on Pseudorabies virus replication

NI Minshu, CHEN Li, BAO Xi, XU Yue, ZHUANG Tenghan, FENG Lei

2022, 34(11): 2386-2394. DOI: 10.3969/j.issn.1004-1524.2022.11.08

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To explore the effect of glucose-regulated protein 94(GRP94) on Pseudorabies virus(PRV) replication and interaction between GRP94 and PRV structural proteins. BHK-21 cells with GRP94 overexpression were constructed by recombinant lentiviruses transfection and BHK-21 cells with GRP94 coding sequences knockout were constructed by CRISPR/Cas9 system. Progeny PRV titers were measured to compare the changes of virus proliferation efficiency in different cell lines. Western blot was performed to detect the expression level of unfolded protein response (UPR) related proteins in different cell lines. The interaction between virus structural proteins and GRP94 was determined by immunoprecipitation analysis. PRV replication was significantly increased in BHK-21-GRP94 cells, while no significant effect on PRV replication was occurred in BHK-21-GRP94-KO cells. The expression level of GRP78 was significantly up-regulated in BHK-21-GRP94-KO cells and interactions between GRP94 and gB, gD, gI were detected by Co-IP. The overexpression of GRP94 was conducive to the proliferation of PRV. By analyzing UPR related proteins, it was of great significance to study the specific role of GRP94 in the process of virus replication, so as to provide a molecular research basis for PRV and ER stress.

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Genetic diversity of Apis cerana in Zhejiang, China based on mitochondrial DNA tRNA^leu-COⅡ sequence

CAO Lianfei, SHI Jinhu, XU Yalan, SU Xiaoling, HU Fuliang, ZHENG Huoqing

2022, 34(11): 2395-2403. DOI: 10.3969/j.issn.1004-1524.2022.11.09

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To further understand the genetic diversity of Apis cerana in Zhejiang Province, the genetic variation and population genetic structure were determined based on the mitochondrial DNA tRNA^leu-COⅡ sequences of 448 samples from ten prefecture-level cities in Zhejiang, China. Forty-six haplotypes were found and the haplotype diversity (H_d) was 0.665 7, the nucleotide diversity (P_i) was 0.002 61, which indicated that A. cerana in Zhejiang Province had relatively high haplotype diversity and low nucleotide diversity. Neutral test and network of haplotypes showed that A. cerana in Zhejiang Province might have experienced recent expansion. Population genetic structure analysis showed that A. cerana populations in Hangzhou and Ningbo had a certain extent of genetic differentiations with other surrounding A. cerana populations, which was possibly related to the survivorship and adaptation of A. cerana in the local areas over a long period of time and a certain geographical isolation. A new haplotype that highly differentiated with other haplotypes of A. cerana in China was found in this study.

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Effects of different diets on growth performance and environmental water quality of Scylla paramamosain cultured in carb apartment

CAI Yilong, ZHANG Libing, HU Gaoyu, XIAO Guoqiang, CAI Jingbo, ZHANG Xiang

2022, 34(11): 2404-2415. DOI: 10.3969/j.issn.1004-1524.2022.11.10

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To study the effects of different baits on growth performance and feeding environment of Scylla paramamosain in crab apartment, the growth and environmental water quality parameters of S. paramamosain during aquaculture were analyzed by feeding different baits (Sinonovacula constricta, fresh fish meat, compound feed for green crab and eel compound feed). The results showed that the specific growth rate of S. paramamosain in S. constricta group was the highest (1.15%·d^-1), followed by the eel feed group (0.56%·d^-1). Besides the S. constricta group, the specific growth rate of other bait groups were significantly correlated with pH, dissolved oxygen and ammonia nitrogen, etc. In terms of survival rate, there was no significant correlation between S. constricta group and water quality indexes, fish group was only significantly correlated with phosphate and nitrite, and significant correlation between two artificial compound diets and active phosphate, suspended particulate matter, dissolved oxygen, pH, nitrite and other indexes. In addition to water temperature and salinity, water quality was mainly affected by feeding. The comprehensive pollution index of water quality showed that the four bait groups were in the state of pollution and environmental eutrophication (NQI≥8.5), but the eutrophication degree of S. constricta group was the lowest. The results showed that S. constricta was the best bait for crab apartment breeding, and the green crab fed by S. constricta had higher growth performance and better breeding water environment. Considering the breeding cost, compound feed was also suitable for green crab breeding in crab apartment, but the water quality of feeding environment needed to be further improved. The result provided a technical basis for the selection of bait and the optimization of management measures in the facility culture of S. paramamosain.

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Evaluation and genetic diversity analysis of common bean germplasm in Zhejiang Province, China

WANG Baogen, DONG Junyang, WANG Ying, LI Sujuan, WANG Jian, LU Zhongfu, WU Xiaohua, LI Guojing, WU Xinyi

2022, 34(11): 2416-2427. DOI: 10.3969/j.issn.1004-1524.2022.11.11

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To identify the genetic diversity of common bean germplasm in Zhejiang Province, 44 common bean accessions collected from Zhejiang Province in multi-years were evaluated on 13 basic agronomic traits in the field, and it is found that large genetic variation presented on the traits including the growth habits, fresh pod commerciality in this collection. In addition, 185 SNPs from the known BARCBean6K_3 BeadChip were randomly selected and converted into KASP markers, finally the high quality genotypes of the 44 accessions were obtained from 88 KASP-SNPs. Based on the genotype data of this population, PCA and phylogenetic tree analysis both showed that Zhejiang common bean germplasms could be divided into two subpopulations, and the subpopulations differentiation was strongly associated with thousand seed weight, and most of Zhejiang common bean germplasm belong to Mesoamerican gene pool. The study results will further enrich the genetic basis of Chinese common bean germplasms and provide a scientific basis for efficient use of local resources and breeding new cultivars.

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Effect of water-fertilizer interaction on yield, quality and soil nutrients of drip irrigated jujube

ZHANG Jianli, WANG Zhenhua, CHEN Rui, WANG Dongwang, LIANG Yonghui, LIU Ruhua

2022, 34(11): 2428-2437. DOI: 10.3969/j.issn.1004-1524.2022.11.12

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In the present study, the mature red jujube was used as experimental subject. Two irrigation levels (W1, 6 000 m³·hm^-2; W2, 6 600 m³·hm^-2), three base fertilization levels (denoted as S1, S2 and S3, with bio-organic fertilizer application rates of 1 200, 1 650, and 2 100 kg·hm^-2, respectively), were set, and the treatments without bio-organic fertilizer were set as controls (W1CK and W2CK) in the field experiment. It was shown that under the same irrigation level, with the increase of bio-organic fertilizer application rates, the branch length and width, leaf chlorophyll content, fruit morphology, available potassium, available phosphorus, and ammonium nitrogen content in 0-150 cm soil increased. With the elevated irrigation level, the contents of available potassium, available phosphorus and ammonium nitrogen in 0-150 cm soil decreased. Compared with the control, the yield per plant increased by 4.65%-19.83% under W1, and increased by 8.17%-19.08% under W2. The application of bio-organic fertilizer increased the contents of total sugar, soluble solids and vitamin C, and reduced the content of total acidity. The fruit morphology, yield per plant and quality indexes under W1S3 treatment were superior than those under W2CK treatment. Based on the climatic characteristics of aridity and low rainfall in Xinjiang, application of 2 100 kg·hm^-2 bio-organic fertilizer with an irrigation level of 6 000 m³·hm^-2 could replace the local conventional irrigation fertilization management, which is more suitable for jujube cultivation in the arid area of southern Xinjiang.

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Analysis of main quality components and selection of excellent resources of early-sprouting tea germplasm resources in southern Zhejiang, China

SHU Zaifa, LIU Yu, SHAO Jingna, ZHENG Shenghong, ZHOU Huijuan, JI Qingyong, HE Weizhong

2022, 34(11): 2438-2450. DOI: 10.3969/j.issn.1004-1524.2022.11.13

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Cluster analysis and principal component analysis (PCA) were used to analyze the genetic diversity of 64 early tea plant germplasm resources in southern Zhejiang. The results showed that abundant diversity and variation existed in 64 tea resources, and the average coefficient of variation reached 16.68%. The principal component analysis showed that the two principal components represented 64.287% information of 64 early germplasm resources. Sixty-four resources were divided into three groups by cluster analysis. The first group had 33 resources, the second group had 18 resources, and the third group had 13 resources. Ten specific germplasm resources (amino acid ≥5.0% or theanine ≥3.0%), 48 good germplasm resources (tea polyphenols ≥20% or amino acids ≥4.0%), 24 potential excellent tea germplasm resources (water extract ≥48.0% or phenol-ammonia ratio ≤4) were preliminarily screened out. The correlation analysis showed that the five main biochemical components of tea, such as polyphenols and amino acids, were significantly correlated.

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Correlation analysis between taste characteristics and taste compounds of five representative green teas in Guizhou, China

RAN Qiansong, LIU Zhongying, FANG Shimao, YIN Jie, DAI Yuqiao, LI Qin, YANG Ting, PAN Ke

2022, 34(11): 2451-2461. DOI: 10.3969/j.issn.1004-1524.2022.11.14

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Guizhou green tea is of excellent quality, but the current evaluation of its taste characteristics is still lacking data support. Because of this, five representative green tea samples from Guizhou in the 2020 Spring Doucha Competition were used as materials, through the use of artificial sensory evaluation, chemical analysis and intelligent sensory evaluation, combined with mathematical analysis methods such as chemometric analysis and multi-factor analysis, the correlation between the flavor characteristics of five representative green teas and their flavor compounds was explored. The artificial sensory evaluation results showed that the taste characteristics of the five tea samples were slightly different, and the taste characteristics were mainly mellow and refreshing. The fresh mellowness of GZ1 tea sample was better than other tea samples. The results of chemical analysis showed that the caffeine content and total amino acid content of the five tea samples were higher between 37.43-42.66 mg·g^-1 and 38.80-70.64 mg·g^-1, respectively. The content of tea polyphenols and total catechins were lower in the range of 193.47-219.63 mg·g^-1 and 165.42-187.30 mg·g^-1, and the content of savory amino acids such as theanine and glutamic acid was higher. The intelligent sensory evaluation results showed that the five samples detected by the electronic tongue had high umami value and richness, which was more consistent with the sensory evaluation results of “mellow, fresh and refreshing”. The correlation showed that bitterness was highly correlated with the total amount of caffeine and caffeine/catechin. Astringency was highly correlated with epicatechin gallate, water extract, epigallocatechin gallate/total catechin content. Richness had a high correlation with epigallocatechin gallate, tea polyphenols, and total catechins, and freshness had a strong correlation with glutamic acid, aspartic acid, proline and total amino acids. High water extract, high amino acids, high caffeine, low tea polyphenols, and low catechin components formed the mellow and refreshing taste characteristics of Guizhou green tea. The evaluation results of the electronic tongue were consistent with the sensory evaluation results, and the umami taste level determined by the electronic tongue could reflect the quality characteristics of tea to a certain extent.

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Population genetic structure of different resistance levels of Chilo suppressalis based on mitochondrial COⅠ and Cytb sequences in Zhejiang, China

LU Yanhui, GUO Jiawen, TIAN Junce, XUE Zhaohong, ZHENG Xusong, LYU Zhongxian

2022, 34(11): 2462-2470. DOI: 10.3969/j.issn.1004-1524.2022.11.15

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In this study, genetic diversity and genetic structure of eight geographical populations with different resistance levels of Chilo suppressalis in Zhejiang Province were analyzed by using genetic methods based on mitochondrial cytochrome oxidase subunit Ⅰ (COⅠ) and cytochrome b (Cytb) genes. The analysis results of population genetic diversity showed that the COⅠ gene fragment with length of 627 bp and the Cytb gene fragment with length of 455 bp were obtained by PCR amplification and sequencing. Among them, 355 homologous COⅠ sequences detected 68 polymorphic sites, including 22 single mutation sites and 46 reduced mutation sites. A total of 85 haplotypes were defined, with an average of 18.25 haplotypes in each population, of which the Ruian (RA) population had the most haplotypes, with 27 haplotypes. Meanwhile, 326 Cytb homologous sequences detected 45 polymorphic sites, including 19 single mutation sites and 26 simple mutation sites. A total of 64 haplotypes were defined. The average haplotype of each population was 14.375, among which the most haplotypes were 25 in Yueqing (YQ) population. The highest haplotype diversity h of the population was 0.896 3 and 0.934 4, respectively. The analysis of population genetic structure showed that the majority of genetic variation existed among different geographical populations of C. suppressalis from individuals within the population, accounting for 80.30% (COⅠ gene) and 78.16% (Cytb gene). Only 19.43% (COⅠ gene) and 21.22% (Cytb gene) of the genetic differences were found among groups. In addition, no clear geographical differentiation was revealed in the haplotype network and on the neighbor-joining tree. Mantel correlation test showed that there was no significant correlation between genetic distance and geographical distance. These results provided basic data for different resistance level population exchange and control of C. suppressalis in Zhejiang Province.

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Prokaryotic expression of Konjac mosaic virus capsid protein and preparation of polyclonal antibody

TANG Guoliang, ZHANG Yubao, WANG Ruoyu, WANG Yajun, ZHAO Xia, SU Xuesi, JIN Weijie

2022, 34(11): 2471-2481. DOI: 10.3969/j.issn.1004-1524.2022.11.16

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The samples of Angelica sinensis(Oliv.)Diels suspected to be infected with virus disease in Yuzhong County, Gansu Province were sequenced and identified by small RNA sequencing. It was found that Konjac mosaic virus(KoMV) was contained in the sample, and the cp gene of KoMV capsid protein was amplified by RT-PCR. The cloned KoMV cp gene connected to the prokaryotic expression vector pET-28a (+), and introduced into E.coli Rosetta^TM (DE3) to induce expression protein. The CP fusion protein was purified by Ni-NTA gravity column chromatography and immunized New Zealand white-eared rabbits with this protein as antigen to prepare polyclonal antibodies. Sequence analysis showed that the size of KoMV cp gene fragment was 840 bp, encoding a coat protein of 280 amino acids. Compared with KoMV isolates registered in GenBank, the nucleotide sequence similarity was 85.58%-99.41%, and the amino acid sequence similarity was 89.32%-99.29%. There were obvious regional and host differences in the distribution of KoMV virus populations. SDS-PAGE showed that the fusion protein of KoMV CP was expressed in large quantities in the form of inclusion bodies in E.coli Rosetta^TM (DE3) induced by different temperatures, and the molecular weight of the fusion proteins was 36 ku. The results of indirect ELISA and Western blot showed that the prepared polyclonal antibody had a titer of 32 000, which could specifically bind to the leaves of Angelica infected with KoMV and purified protein, showing good specificity. In this study, the polyclonal antibody IgG of KoMV CP fusion protein was successfully prepared, which laid a foundation for the development of serological detection technology of KoMV and the functional research of CP protein.

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Reducing nitrogen loss from paddy filed and ensuring yield of direct seeding rice by adjusting nitrogen base-topdressing ratio

XIA Sujing, QIAO Yue, ZHU Jianqiang

2022, 34(11): 2482-2490. DOI: 10.3969/j.issn.1004-1524.2022.11.17

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Field drainage for waterlogging prevention at seedling stage of direct seeding rice increases the risk of nitrogen loss from paddy field. Appropriate adjustment of nitrogen base-topdressing ratio can provide scientific basis for risk reduction during this process. In the present study, N, P₂O₅, and K₂O application amount were set as 180, 75 and 105 kg·hm^-2, respectively. A field experiment was conducted with different nitrogen base-topdressing ratios. The conventional fertilization method in Jianghan Plain (the proportion of N in basal fertilizer, tillering fertilizer and panicle fertilizer was 6∶4∶0 ) was taken as the control (CK), and treatments were set according to the proportion of N in basal fertilizer, tillering fertilizer and panicle fertilizer, specifically, 4∶6∶0 (T1), 4∶4∶2 (T2), 4∶2∶4 (T3), 2∶6∶2 (T4), 2∶4∶4 (T5), 0∶8∶2 (T6), 0∶6∶4 (T7). The ammonia volatilization loss and nitrogen runoff loss during rice growth and rice yield under different treatments were measured in the field. It was shown that the nitrogen runoff loss was the smallest under T7 treatment (8.79 kg·hm^-2). The ammonia volatilization loss was the smallest under T3 treatment (11.90 kg·hm^-2). The total nitrogen loss was the smallest under T7 treatment (22.89 kg·hm^-2). The highest yield was found under T4 treatment (9 270.0 kg·hm^-2), which was followed by T5 treatment (9 150.0 kg·hm^-2). Given the nitrogen loss and rice yield, the recommended proportions of N in basal fertilizer, tillering fertilizer and panicle fertilizer in the experimental conditions were 2∶6∶2 and 2∶4∶4.

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Crops classification based on GF-3 satellite data and Η/Α/α- decomposition characteristic parameters

ZHAO Lixian, ZHANG Wangfei, LI Yun, ZHANG Tingwei, HUANG Guoran

2022, 34(11): 2491-2503. DOI: 10.3969/j.issn.1004-1524.2022.11.18

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Based on the civil C-band synthetic aperture radar data of GF-3 satellite, Shannon entropy (SE) and its intensity components (SE_I) and polarization components (SE_P), single bounce eigenvalue relative difference (SERD), double bounce eigenvalue relative difference (DERD), polarization fraction (PF), pedestal height (PH), polarimetric asymmetry (PA) and radar vegetation index (RVI) were extracted from GF-3 full polarimetric data by Η/Α/ $α -$ polarization decomposition method. With application of support vector machine (SVM) and random forest (RF), the potential of these parameters extracted by Η/Α/ $α -$ decomposition from GF-3 data in crops classification was preliminarily explored. The results showed that when SERD, PH, PF, RVI and SE_P parameters were used alone, the classification accuracies were relatively high (82%-92%). When PA, DERD, SE and SE_I parameters were used alone, the classification accuracy was less than 80%. However, the combination of PA, DERD, SE and SE_I parameters improved the classification accuracies to 93.02% and 92.05%, respectively, under SVM and RF, and the Kappa coefficients were all greater than 0.8. Therefore, the nine characteristic parameters extracted by Η/Α/ $α -$ polarization decomposition from GF-3 data could well characterize the scattering characteristics of crops and could be used in crops classification.

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Preliminary investigation about structure and probiotic properties of polysaccharides from Dendrobium officinale leaves

LIN Yuqing, LU Shengmin, ZHOU Wanyi, XING Jianrong, YANG Ying

2022, 34(11): 2504-2511. DOI: 10.3969/j.issn.1004-1524.2022.11.19

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To investigate the preliminary structure and probiotic properties of three Dendrobium officinale leaf polysaccharides in the Yandang Mountain area, and provide basis for high-value utilization of D. officinale leaf resources, leaves of D. officinale Yanhu No.1, D. officinale Yanhu No.3 and D. officinale Shenglan No.8 were used as raw materials, and pure polysaccharides were obtained, which were named as DOP1, DOP2 and DOP3. The relative molecular weight of DOP1, DOP2, and DOP3 were 2.56×10⁵, 1.35×10⁵ and 3.26×10⁵, respectively. All of them were composed of arabinose, galactose, glucose and mannose, but with different molar ratios. DOP1 had the tightest organizational structure in aqueous solution, followed by DOP3, and DOP2 was the loosest. The proliferation of Lactobacillus plantarum, Lactobacillus pentosus and Lactobacillus rhamnosus were promoted by all three polysaccharides, among which DOP3 had the most significant effect. DOP2 and DOP3 significantly inhibited the growth of Escherichia coli and Listeria monocytogenes, while DOP1 only inhibited the growth of L. monocytogenes. The relative abundance of Parabacteroides and Koalabacteria were increased and the relative abundance of conditional pathogenic bacteria such as Klebsiella and Bliophila were decreased under the treatment of three polysaccharides in human colon flora. In addition, DOP3 also significantly inhibited the growth of Morganella and Pseudomonas. All three polysaccharides could promote the production of acetic acid, propionic acid and butyric acid, among which DOP2 and DOP3 had better effects than DOP1. The results showed all of these three polysaccharides had probiotic functions, among which DOP3 had a better comprehensive effect and had the potential to be used as a new probiotic factor.

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Primary risk assessment of several fungicides to typical vineyard terrestrial organisms

LYU Lu, WU Shenggan, WANG Qiang, ZHAO Xueping, XU Mingfei

2022, 34(11): 2512-2521. DOI: 10.3969/j.issn.1004-1524.2022.11.20

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Grape is easy to be infected with fungal diseases in the process of planting. Even though there are large number of registered fungicide products, unregistered fungicides have still being used or recommended to control grape fungal diseases. And whether their application would cause harm to the vineyard environmental organisms should be paid enough attention. In this study, the fungicides, i.e., triadimefon, diethofencarb, tetraconazole, isopyrazam and vinclozolin, which have not registered but detected or proposed to use on grape, were selected to conduct primary risk assessment to environmental organisms including birds, bees, non-target arthropods and soil organisms. The results showed that the long-term risk quotient (RQ) values of triadimefon and tetraconazole for birds were both greater than 1, which meant the risks to birds were unacceptable. The RQ values of diethofencarb for predatory non-target arthropods were greater than 5, revealing the unacceptable risk of diethofencarb to predatory non-target arthropods. The risks of isopyrazam and vinclozolin to the several terrestrial organisms were both acceptable. This study could provide reference for the scientific application of the selected fungicides on grapes and the subsequent pesticide registration.

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Real-time detection of orchard cherry based on YOLOV4 model

ZHOU Pinzhi, PEI Yuekun, WEI Ran, ZHANG Yongfei, GU Yu

2022, 34(11): 2522-2532. DOI: 10.3969/j.issn.1004-1524.2022.11.21

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In order to solve the problem of difficulty in target recognition and low detection accuracy in the state monitoring of cherries in different growth periods under natural environment, this paper proposed an improved convolutional neural network cherry classification and detection model based on CSPDarknet53. The feature extraction network used in the classic YOLOV4 had a deeper number of layers and could extract more advanced abstract features, but had a weak local perception of the target. By integrating the CBAM attention mechanism on the CSPDarknet53 network structure, the perception of local features of the target was enhanced. Furthermore, the accuracy of target detection was improved, its feature extraction and target detection capabilities were better than the original algorithm, the feature layer output of the feature extraction network was adjusted, and the third layer output to the second layer output was changed to increase the acquisition of small target semantic information, k-means algorithm was used to optimize the size of the prior frame to adapt to the size of the cherry target, and ablation experiment analysis was conducted. The results showed that the improved YOLOV4 cherry detection model had an average accuracy of 92.31% and an F1 score of 87.3%, which was better than Faster RCNN, YOLOV3 and the original YOLOV4 algorithm. The detection speed was 40.23 frames·s^-1, which was suitable for natural environments. It provided a theoretical and technical basis for realizing automatic monitoring of fruit growth status in orchards.

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Fruit variety recognition based on parallel convolutional neural network

LI Chao, LI Feng, HUANG Weijia

2022, 34(11): 2533-2541. DOI: 10.3969/j.issn.1004-1524.2022.11.22

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In order to solve the defects of traditional fruit image recognition algorithms in feature extraction and the low recognition accuracy of traditional convolutional neural networks, a parallel convolutional neural network was proposed to extract fruit features. ELU activation function was introduced instead of ReLU activation function in the proposed model. Besides, a combination of maximum class spacing loss function and the traditional SoftmaxWithLoss loss function was designed to improve the recognition accuracy of similar varieties. The data of 8 fruit varieties in Fruit-360 data set was selected in the present study, and enhanced by the boundary equilibrium generative adversarial network (BEGAN) combined with the traditional data augmentation to generate a large number of high-quality data for model training. It was shown that the average recognition accuracy of 8 fruit varieties reached 98.85% and exhibited good recognition effect.

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Simulation and optimization analysis and experiment of split plough based on discrete element method

LIU Mingyong, HU Chenglong, XIE Bolin

2022, 34(11): 2542-2552. DOI: 10.3969/j.issn.1004-1524.2022.11.23

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To reduce the resistance of the plough body and improve the efficiency of farming, based on EDEM, the study was carried out on the basis of the parabolic curved plough body. The plough body cultivation process was simulated and analyzed, and the disturbance behavior of the soil particles was observed. The influence of the blade angle, the installation angle and the working speed on the plough body’s tillage resistance was analyzed, and the ideal plough body parameters were obtained. The results showed that compared with the parabolic surface, the resistance of the elliptical surface was 1 591.551 N in the relatively stable stage, reduced by 4.4% compared with the parabolic surface, and had a better effect on soil uplift. Single factor analysis was carried out on the forward speed, blade angle and installation angle of the plough body. When the installation angle was 25°, the resistance of the plough body was the smallest. As the blade angle increased, the resistance of the plough body also increased. Therefore, a lower forward speed and blade angle should be selected within a suitable range to reduce the resistance of the plough body.

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Research on driving factors and combination path configuration of rural cultural tourism entrepreneurship of “new farmers”

LU Jingyi, XIANG Keheng

2022, 34(11): 2553-2566. DOI: 10.3969/j.issn.1004-1524.2022.11.24

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In the present study, Jiangjia Village in Fuyang District, Hangzhou City, was selected as a case, and the actor network theory was used as a framework. An in-depth research on the antecedent factors, implementation process and effectiveness of cultural tourism entrepreneurship of “new farmers” was conducted through the fuzzy set comparison qualitative method on the basis of empirical research, and a case study of 10 “new farmers”. The results showed that the role perception and resource capital were the principle factors promoting the entrepreneurship. Resource capital and government policies were the external guarantee factors, while role perception and internal emotions and beliefs were the internal drivers that drove the innovation and entrepreneurship of “new farmers”. When location conditions and surrounding support became the main push factors, the driving factors influencing the cultural tourism entrepreneurship of “new farmers” tended to be transportation location and resource capital-led type. The driving factors and their combination of cultural tourism entrepreneurship of “new farmers” could vary with the change of policies, the cycle of business model bonus period, the tracking of network hotspots, etc. Finally, a theoretical framework model of actor network was proposed based on qualitative comparative analysis and empirical research in the context of the driving factors and combination paths of the cultural tourism entrepreneurship of “new farmers”.

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Land-shareholding double-layer multi-party interest game cooperation mechanism under background of integration of urban and rural factors

LI Juan, ZHUANG Jincai, ZHENG Jingjing

2022, 34(11): 2567-2582. DOI: 10.3969/j.issn.1004-1524.2022.11.25

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Under the background of urban-rural factors integration, in order to explore the multi-party cooperation mechanism of land-shareholding, a two-layer multi-party evolutionary game model was constructed in the present assay from the theoretical level, and the interest game relationship of the four parties involved in the cooperation of local governments, capital enterprise, farmers, and collective economic organizations was analyzed, and a stable strategy for cooperation within all parties was obtained. Based on the survey data of potato specialty cooperatives in Guyang County, Inner Mongolia, a confirmatory analysis of the stable strategy selection of the multi-party cooperative evolutionary game model was conducted, and the corresponding cooperation mechanism was revealed to promote the continuous and stable progress of land equity cooperation and promote the effective symbiosis of urban and rural factors. It was found that the rational distribution of land value-added income, the government’s policy incentives, the psychological expected income of the participants in the land-share participation cooperation, and the punishment of collusion behavior had great impact on the stability of land-shareholding cooperation. In order to build the cooperation mechanism of the four parties of land investment, suggestions were put forward as follows: expand the cooperation of land investment and increase the excess returns, increase the liquidated damages to reduce the probability of default, reduce additional benefits to improve the stability of cooperation, determine the coefficient of interest distribution according to the proportion of shares to improve the equilibrium of interest distribution, increase government incentives and other income obtained by farmers to mobilize the enthusiasm of all parties for cooperation, increase the income earned by collective economic organizations and enhance their enthusiasm for cooperation, and increase the punishment of collective economic organization collusion to curb the occurrence of collusion behavior.

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