Acta Agriculturae Zhejiangensis

Identification on disease resistance and molecular markers of F₂ hybrids from Ningchun No.4 and Hedong black wheat

WANG Zhangjun, LIU Yan, ZHANG Shuangxi, LIU Fenglou, LI Qingfeng, ZHANG Xiaogang, LIU Shengxiang, JIA Biao

2019, 31(5): 677-687. DOI: 10.3969/j.issn.1004-1524.2019.05.01

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To provide good resources for Ningxia wheat breeding in disease resistance, Ningchun No.4, Hedong black wheat and 331 individual plants of F₂ generation of Ningchun No.4 × Hedong black wheat were used as materials, disease resistance and molecular markers of the populations were identificated. The results showed that Ningchun No. 4 was moderate susceptible to powdery mildew, stripe rust, leaf rust, and Hedong black wheat was moderate to highly resistant to powdery mildew, moderate resistant to stripe rust and leaf rust. There were 68, 54, 52 individual plants resistant to powdery mildew, stripe rust, leaf rust in hybrids, respectively, and the rate was 20.54%, 16.32% and 15.71% in turn. In them, 50 individual plants were resistant to powdery mildew and stripe rust, 44 plants were resistant to powdery mildew and leaf rust, 32 plants were resistant to stripe rust and leaf rust, and 29 plants were resistant to powdery mildew, leaf rust and leaf rust simultaneously. There were 239 individual plants carried gene Pm16, 22.59% plants carried this gene were resistant to powdery mildew in field, 16.83% of 202 plants carried gene Yr2 were resistant to stripe rust, 17.07% of 246 plants carried gene Lr24 were resistant to leaf rust, 148 plants carried genes Pm16 and Yr2 simultaneously, 185 plants carried genes Pm16 and Lr24, 155 plants carried genes Yr2 and Lr24, 119 plants carried genes Pm16, Yr2 and Lr24 simultaneously. Ten disease resistance quantitative trait loci (QTL) were detected by six SSR markers, which involved in chromosomes 2A, 5A, 3B and 5D, their additive effects were from -0.15 to 0.08, the contribution rates to phenotype were from 2% to 4%, maximum logarithm of odds (LOD) score was 10.40, and QTL enrichment regions of disease resistance existed in 5A, 3B and 5D chromosome.

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Investigation on agronomic characters of dwarf mutant in Panicum miliaceuml and analysis of its sensitivity to GA

ZHANG Bo, JIA Xiaoping, YANG Dezhi, ZHAO Yuan, DAI Lingfeng, KOU Shujun, ZHANG Xiaomei, HOU Dianyun, ZHU Xuehai

2019, 31(5): 688-694. DOI: 10.3969/j.issn.1004-1524.2019.05.02

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In order to investigate the difference between agronomic traits of dwarf mutant and original material of Panicum miliaceuml and whether the cause of dwarf was related to gibberellin (GA) synthesis or signaling pathway, the dwarf mutant 778 and its original material high straw 260 were used as experimental materials. Morphological observation and determination of agronomic traits in plant height, ear length, internode length, internode number, seed length, seed width and number of seeds were determined. Then, the plants were treated by different concentrations of GA in different growth periods, and the changes of plant height, root length, stem width, leaf length and leaf width were measured. The results showed that the plant height of the dwarf mutant material was significantly different from that of the original high straw material, which was mainly caused by the difference between the base of the ground and the length of the first and second elongation joints. Comparing and analyzing the differences of traits between dwarf mutant material 778 and original high straw material 260, it was found that the plant height, ear length, internode number, grain number per ear and internode length of dwarf mutant 778 were significantly lower than that of high straw 260 (P<0.01), the seed length of dwarf mutant 778 was significantly higher than that of high straw 260 (P<0.05). Different concentrations of gibberellin (0, 50, 100, 200 mg·L^-1) had no significant effect on plant height and root length of scorpion dwarf mutant 778 (P>0.05), but different concentrations of gibberellin had significant effects on plant height, root length and GRI value of high straw 260 (P<0.05). And the plant morphology and growth potential of high straw 260 treated with different concentrations of gibberellin were superior to the control group. Under the condition of 100 and 200 mg·L^-1 GA, the difference in plant height between the dwarf mutant and the high straw material decreased with time, and there was no difference at the end. There was no difference in GRI value between different concentrations of gibberellin treatments, indicating that the external gibberellin could restore the scorpion dwarf mutant to the original high straw, and the gene that caused the mutation may be related with the gibberellin synthesis pathway.

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Water footprint of maize production in middle Jilin under different rainfall years

ZHENG Xiaoxue, QIN Lijie

2019, 31(5): 695-703. DOI: 10.3969/j.issn.1004-1524.2019.05.03

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The middle of Jilin province is an important production base of commodity grain in China, and is famous for its high and steady maize yield. In the present study, the characteristic of water consumption and the spatial variations of the water footprint of maize growth were analyzed, which was beneficial for the rational utilization of agricultural water resources and the sustainable development of agriculture. It was shown that the water footprint of maize production was higher in the north of study area, and was lower in the south. The footprint in the drought year was the largest, yet the footprint in the humid year was the smallest. The major proportion of water footprint was the green water footprint, which was almost 75%. The green water footprint was dominated by the effective precipitation footprint, of which the largest proportion was recorded in the humid year, and the smallest proportion was recorded in the drought year. The consumption of soil water was the largest in the drought year, and the smallest in the humid year.

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Molecular cloning and bioinformatics analysis of white king pigeon (Columba livia) ovalbumin-related protein Y gene

XU Xiaoqin, DU Xue, LI Guoqin, ZHANG Long, TAO Zhengrong, LU Lizhi

2019, 31(5): 704-708. DOI: 10.3969/j.issn.1004-1524.2019.05.04

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The experiment was conducted to clone the whole cDNA of OVALY gene in Columba liviaand analyze the bioinformatic information of the cDNA sequence.The nucleotide sequence of a full-length 2 068 bp cDNA for OVALY was obtained, containing 5'UTR (180 bp), CDS (1 164 bp) and 3'UTR (720 bp), and coding 388 amino acids (44.19 ku, GenBank accession number: KX230792). The OVALY cDNA sequence of white king pigeon was most similar to Nipponianippon, and the homology was 81%. This sequence was also similar to Haliaeetusalbicilla and Pteroclesgutturalis, and the homologies were 80% and 79%, respectively. 4 N-glycosylated sites and 22 phosphorylation sites were detected in these amino acids. Conserved domain analysis of OVALY protein showed this protein had a conserved domain for serpin superfamily.

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Isolation, identification and phylogenetic analysis on gp85 gene of subgroup J avian leukosis virus

WANG Yuanhong, LU Zhimin, ZHANG Xueqi, LIU Zimin, HU Zihui, YANG Kankan, LIU Hongmei, PAN Ling, PENG Kaisong, LI Yongdong, WANG Yong

2019, 31(5): 709-715. DOI: 10.3969/j.issn.1004-1524.2019.05.05

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To identify the avian leukosis virus (ALV) in chicken farm in Chaohu, Anhui Province, the virus was isolated from the tumor tissue by using DF-1 cells. The results were confirmed by RT-PCR and transmission electron microscopy. The diseased chicken organs such as liver, heart, spleen and glandular stomach were analyzed by clinical observation, pathological anatomy and histopathological examination. Finally, the gene of envelope protein gp85 were cloned and sequenced by designing specific primers for gp85 motif from ALV reference strains in GenBank, and their nucleotide homologies were compared. The results showed that a virus strain, AHCH-2018, was confirmed by sequencing the 928 bp PCR amplification product. The virus about 100 nm in diameter appeared under the transmission electron microscope, which was enveloped and spherical in shape. Histopathological observation showed that larger areas of the pericardial fat, myocardium, spleen, liver and glandular stomach were infiltrated by lymphocytes and lymphoid cells. The gp85 gene nucleotide sequence analysis showed that the identity of AHCH-2018 strain and other ALV J reference strains ranged from 94.1%-99.5%, amongst which the highest identity (99.5%) was shown with GD1401J strain, when compared with ALV A, B, C, D and E, the nucleotide identity ranged from 46.5%-49.7%. The phylogenetic analysis showed that AHCH-2018 and J subgroup prototype strain were present in the same branch. This study provided a data reference for understanding the molecular epidemiological characteristics of avian leukemia in Anhui Province, China.

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Proliferative characteristics of novel duck reovirus strain JDM10 in DF-1 cells

YUN Tao, HUA Jionggang, YE Weicheng, NI Zheng, CHEN Liu, ZHANG Cun

2019, 31(5): 716-721. DOI: 10.3969/j.issn.1004-1524.2019.05.06

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To explore the proliferative charactistics of novel duck reovirus (NDRV) in chicken embryo fibroblast cells (DF-1). NDRV JDM10 strain was inoculated into DF-1 cells, and after continuous passage. The pathological effect (CPE) of virus on cells, half of tissue infection (TCID₅₀), RT-PCR, indirect immunofluorescence (IFA) and western-blot immunology were studied. The results showed that NDRV JDM10 could effectivly proliferate in the DF-1 cells and produce typical CPE such as cell shedding, cell body enlargement and cell disruption. The σC gene (1 001 bp) was successfully amplified from the infected cells by RT-PCR. NDRV σC was expressed well in DF-1 cells and could specifically reacted with anti-NDRV σC monoclonal antibody. The one-step growth curve showed that NDRV JDM10 strains experienced three periods of incubation, rapid growth and stable phase after infection with DF-1 cells, and the virus titer reached peak at 72 h, and was TCID₅₀ of 1×10^-7.90·(0.1 mL)^-1. These results established foundation for further study of the pathogenesis of NDRV and the development of cell vaccines.

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Correlations between adipokine, biochemical indicators in early lactation cows with fatty liver

ZHU Yingkun, XIAO Jinbang, QIAN Bolin, JIANG Sixun, YOU Liuchao, ZHANG Yue, LIU Hong, MA Li, CAO Suizhong, YU Shumin, SHEN Liuhong

2019, 31(5): 722-729. DOI: 10.3969/j.issn.1004-1524.2019.05.07

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To explore correlations between adipokine, biochemical indicators in early lactation cows with fatty liver, 64 perinatal Chinese Holstein cows from a farm in Sichuan province were used in the present study. Tail vein blood was collected before morning feeding on 7 and 14 days after delivery, serum AST, Glu and NEFA levels of 14 days after delivery were tested. According to fatty liver determination formula, cows were grouped into fatty liver group and control group. Some adipokine and biochemical indicators were tested and their correlations with liver function indexes and energy metabolism indexes were analyzed. The results showed that leptin, adiponectin level of fatty liver early lactation cows were significantly lower than the normal group(P<0.05). TNF-α level of fatty liver early lactation cows was significantly higher than the normal group(P<0.05), IL-6 level appeared no significant difference between fatty liver cows and normal cows(P>0.05). Leptin had significant positive correlations with Glu and ALB levels(P<0.05), significant negative correlations with AST, TBIL, DBIL, TP, NEFA and BHBA levels(P<0.05), indicated that negative energy balance was an important factor of fatty liver. Adiponectin level had significant negative correlations with NEFA, BHBA and AST(P<0.05), significant positive correlations between TP and ALB(P<0.05), speculating liver function and energy metabolism disorders were occurred in fatty liver early lactation cows. TNF-α level had significant positive correlations with TBIL,DBIL and ALT levels(P<0.05), significant negative correlations with Glu and ALB(P<0.05). IL-6 level had a significant positive correlation with ALT level(P<0.05), significant negative correlations with TP and ALB levels(P<0.05), showed that obvious inflammatory stress was appeared in fatty liver cows. Leptin levels showed a significant positive correlation with adiponectin level(P<0.05), significant negative correlation with IL-6 levels, speculating metabolism regulation disorder in fatty liver cows. In summary, the negative energy balance of early lactation cows induced body fat mobilization and downregulated adiponectin and leptin levels, causing lipids accumulate in liver and damaged liver function and triggered inflammatory stress, rising IL-6 and TNF-α levels and induced fatty liver, adiponectin. Thus, leptin and TNF-α levels could be able to be a predicative index of fatty liver in early lactation cows.

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Clinical efficacy and mechanism prediction of Difutongrusan in treatment of bovine mastitis

BAI Dongdong, LI Xinpu, YANG Feng, LUO Jinyin, WANG Xurong, LI Hongsheng

2019, 31(5): 730-736. DOI: 10.3969/j.issn.1004-1524.2019.05.08

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To study the antibacterial activity, clinical effect and mechanism of Difutongrusan in the treatment of bovine mastitis. The antibacterial activities of Difutongrusan against the main pathogens of bovine mastitis were determined by the method of micro broth first, then the cows with mastitis were treated to observe the cure rate and the effective rate. Finally, BATMAN, STRING, Cytoscape databases and softwares were used to analyze the mechanism of treatment of bovine mastitis by Difutongrusan. The results showed that the MIC/MBC of Difutongrusan on Staphylococcus agalactiae was 1.52 mg·mL^-1, the MIC on Escherichia coli and Staphylococcus aureus was 25 mg·mL^-1, and the MBC was twice as high as the MIC; The clinical cure rate of bovine mastitis was 63.16% and the effective rate was 89.47%. A total of 21 active compounds were selected from Difutongrusan, with 173 corresponding targets and 5 pivotal targets. The results indicated that Difutongrusan could have high antibacterial activity in vitro against the main pathogenic bacteria of bovine mastitis, and the clinical efficacy could be definite, which may be mainly through the pivotal target NT5E, IL8, ESR1, PIK3CA and PIK3CB to treat bovine mastitis.

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Effects of different red and blue LED light intensity and irrigation and their interactions on growth of tomato seedlings

YANG Yana, FAN Xiaoxue, XU Gang, ZHANG Yu, LI Yaling, WEN Xiangzhen

2019, 31(5): 737-745. DOI: 10.3969/j.issn.1004-1524.2019.05.09

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In this study, the growth and photosynthesis of tomato seedlings under different light intensity and irrigation were investigated by using red and blue light as the basic light source. The results showed that different light intensity and irrigation had significant effects on the growth of tomato seedlings. Plant height was decreased,stem diameter became smaller, internode was shortened, leaf area was decreased, fresh weight of the plant and dry weight of the shoot was decreased, and photosynthesis of tomato seedlings was decreased with the reduction of irrigation. The main factors affecting chlorophyll content were light conditions, and water was a secondary influence factor. When the light intensity exceeded 3 540 lx, the increase of light intensity had no significant effect on biomass of the plant. The maximal photochemical efficiency F_v/F_m of all treatment combinations in the experiment were stable at around 0.8, indicating that plant growth was in a healthy state. Among all the treatment, tomato plants with 3 540 lx light density and 150 mL·plant^-1 irrigation (L₂W₁) presented the most strong growth vigor.

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Comprehensive evaluation of salt tolerance of 22 alfalfa germplasms at germination stage

LI Mingyu, WANG Yan, LIANG Danni, YAO Yani, LAN Jian

2019, 31(5): 746-755. DOI: 10.3969/j.issn.1004-1524.2019.05.10

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In order to screen salt-tolerance alfalfa germplasms and determine the appropriate concentration for salt-tolerance identification, the relative germination rate, relative germination energy, germination index, radicle dry weight and plumule dry weight were measured with conventional paper germination trial under different NaCl stress (0, 0.4%, 0.8%, 1.2%, 1.6%). Meanwhile, cluster analysis and comprehensive evaluation of salt-tolerance of 22 alfalfa germplasms were made. The results showed that relative germination rate, relative germination energy, germination index, radicle dry weight and plumule dry weight of alfalfa germplasms were all significantly lower than those of the control when the salt concentration increased. The semi-lethal salt concentration was 0.85% (variation coefficient was 26.38%), and 1.2% NaCl was the appropriate concentration to identify the salt tolerance of alfalfa. The salt-tolerant alfalfa germplasms were Herd King, Affinity, German Derby and Golden Empress, and the salt-sensitive alfalfa germplasms were Aohan, A/S Complex, Vernal and Ramble11. These results could provide references for the breeding of new salt-tolerant varieties.

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Application of formula fertilizer based on sunflower stem ammonium sulfate bio-based fertilizer in different growth stages of tomato

LIN Wei, ZHOU Haixia, LAN Zhiqian, ZHANG Kaige, LIU Jiqing, ZHANG Xueyan

2019, 31(5): 756-765. DOI: 10.3969/j.issn.1004-1524.2019.05.11

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Organic water soluble fertilizer can effectively promote crop growth and improve crop characteristics. In view of the different fertilizer characteristics of tomato in different growth stages, the bio-based sulfonate was used and added with formula fertilizer to design the seedling, flowering and fruiting period in this experiment. The total amount of N₂O+P₂O₅+K₂O in seedling stage was 22, and two N-P-K levels were set as 12-4-6(E1) and 9-4-9(E2); the total amount of N₂O+P₂O₅+K₂O at the flowering stage was 50, and two N-P-K levels were set as 20-8-22(M1) and 19-6-25(M2); the total amount of N₂O+P₂O₅+K₂O was 58, and two N-P-K levels were set as 15-9-34(L1) and 13-7-38(L2), and 8 treatments in total were carried out to systematically study the growth, fruit quality and yield of tomato, in order to screen out formula fertilizers that can effectively improve tomato quality, yield and soil nutrient. The results showed that there was no significant difference in plant growth between two N-P-K levels in the early growth stage of tomato. The plant height, stem diameter and number of leaves in the N-P-K treatment of 20-8-22 were superior to that in the treatment of 19-6-25. In the late growth stage, the treatment of 20-8-22(M1)13-7-38(L2) was beneficial to stem diameter, number of leaves and chlorophyll content. The soluble sugar content of E1M2L1 and E2M1L2 was 5.71% and 5.70%, E1M1L1 was significantly lower than E1M2L1. The soluble solid content of E2M1L2 was the highest, 9.1% higher than E2M2L2. The V_C content of E1M2L2 was the highest, E2M1L1, E2M1L2 and E2M2L1 were the second. E1M1L2 significantly increased tomato yield, 88.2% higher than the lowest yield in E2M2L2, followed by E1M2L1 and E2M1L2, and the yield was 3 744.60 and 3 347.81 kg·667m^-2. The available nutrients, EC and pH value of E2M1L2 was all in the middle. In terms of yield, E1M1L2 can significantly increase tomato yield and economic benefits. The principal component analysis showed that E2M1L2 was the optimal fertilizer at seedling stage with the total amount of 22 and N∶P∶K=9∶4∶9, the optimal fertilizer at flowering stage was the total amount of 50 and N∶P∶K=20∶8∶22, the optimal fertilizer in the fruiting period was the total amount of 58 and N∶P∶K=13∶7∶38, which were more conducive to tomato growth, fruit quality and soil quality.

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Study on endophytic bacteria isolated from Eucommia ulmoides with antimicrobial activity against Fusarium graminearum

CHEN Xiaojie, WANG Qi, ZHANG Xinyue, DING Ting

2019, 31(5): 766-776. DOI: 10.3969/j.issn.1004-1524.2019.05.12

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This study was carried out to assess the inhibitory activities of endophytic bacteria from Eucommia ulmoides against Fusarium graminearum. The endophytic bacteria from E. ulmoides were isolated and purified, and the antimicrobial activities of endophytic bacteria against F. graminearum were examined using confront antibiotic culture experiment, disease-control pot experiment. Then, the antimicrobial strain was identified at molecular level and its resistance mechanism against F. graminearum was analyzed. The results indicated 104 endophytic bacteria were isolated from the leaves, stems and roots of E. ulmoides, and DZSG09, DZSJ16 and DZSG23 had strong inhibition against F. graminearum, inhibitory rate of DZSG23 on F. graminearum was 69.04%. Moreover, control efficacy of DZGS23 against F. graminearum was 40.89% by pot experiment after inoculation for 10 d. 16S rDNA sequence analysis showed that DZGS23 belonged to Bacillus. DZGS23 could cause the abnormity and tortuosity of mycelial of phytopathogen and so on. Lastly, bacterial suspension and secondary metabolite of DZSG23 had strong teratogenic effect on conidia of F. graminearum conidia. Deformity rates of conidia were 57.70% and 36.67% after treated with suspension of DZSG23 (1×10⁶ CFU·mL^-1) and its secondary metabolites (10 mg·mL^-1) for 36 h, and they could cause abnormality of conidia and made tubes swollen and contort. However, the bacterial suspension and secondary metabolites of DZSG23 weakly affected germination of F. graminearum conidia. The results afforded scientific basis for DZSG23 application.

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Identification and analysis on pathogenicity-related domain of P3 protein of wheat yellow mosaic virus

ZHANG Yan, QI Yuhua, LU Yanhua, YANG Qiankun, HE Yujuan, LI Junmin, CHEN Jianping

2019, 31(5): 777-783. DOI: 10.3969/j.issn.1004-1524.2019.05.13

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Wheat yellow mosaic virus (WYMV) belongs to the genus Bymovirus, family Potyviridae. Its genome consists of two sense single-stranded RNAs encoding a total of 10 proteins. Previous studies had shown that the conserved protein P3 encoded by potyviruses had multiple functions and played an important role in viral replication, pathogenicity, overcoming host resistance, and cell-to-cell movement. However, whether P3 had similar functions in bymovirus had never been reported yet. In this study, tobacco rattle virus(TRV)-mediated gene silencing system in Nicotiana benthamiana was used to study the function of WYMV P3. Our results indicated that C terminal of WYMV P3 (P3-C), but not the whole P3, had a pathogenic-related functional domain in N. benthamiana. Further frame shift mutation confirmed that, rather than the viral small interfering RNA-mediated host gene silencing, the pathogenicity was caused by the polypeptide of P3-C. In addition, the pathogenicity on N. benthamiana disappeared when any of the two predicted transmembrane domain in P3-C were mutated, indicating the important roles of the transmembrane domains in P3-C pathogenicity.

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Identification of pathotype of Verticillium dahlida isolates on eggplant and their biological characteristics in Shaoxing City, Zhejiang Province

LI Xuqing, ZHANG Jingze, ZHANG Ya, WU Genliang

2019, 31(5): 784-789. DOI: 10.3969/j.issn.1004-1524.2019.05.14

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Based on the method of polymerase chain reaction (PCR) amplification using the specific primers of defoliation and nondefoliation pathotypes of Verticillium dahliae, the pathotype of the isolates causing the verticillium wilt disease of eggplant in Shaoxing City of Zhejiang Province were identified. And the biological characteristics of V. dahliae were also studied. Results of PCR amplification showed that the isolates (VD-sy1, VD-sy2, VD-sy3, VD-sy4 and VD-sy5) obtained all belonged to the defoliating pathotype. The results of biological characteristics showed that VD-sy1 was able to grow at pH between 4.0 to 9.0, and 25 ℃ under 24 h dark condition was beneficial to colony growth; sucrose and yeast extract were the best carbon and nitrogen source for mycelia with linear growth rates of 4.25 and 4.06 mm·d^-1, respectively, while galactose and ammonium chloride showed the least liner growth rates of 2.24 and 2.42 mm·d^-1, respectively.

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Investigation into emission characteristics of odor pollutants in typical livestock and poultry farms in Shanghai

ZHOU Zhongqiang, SHEN Genxiang, XU Chang, SONG Mengjie, CHEN Xiaoting, WANG Zhenqi, FU Kan, QIAN Xiaoyong

2019, 31(5): 790-797. DOI: 10.3969/j.issn.1004-1524.2019.05.15

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Ten typical livestock (layer, dairy, swine) farms in Shanghai were selected as the research object. The characteristics of odor pollutants emissions of typical livestock and poultry farms and different emission stages were investigated by using on-site sampling and laboratory analysis. It was shown that the odor concentration of each farm boundary was between<10 and 128. The odor concentrations of main odor emission stages in layer farms, dairy farms and swine farms were 10-412, 10-511 and 10-538, respectively. Manure composting of the layer farms, waste water storage of the dairy farms and housing of swine farms had greater odor emission than the other odor emission stages of the according livestock and poultry farms, to which more attentions should be paid. There were significant (P<0.01) correlations within NH₃, H₂S contents and odor concentration in swine farms, yet these correlations were not significant in dairy farms. Organic fertilizer production could reduce the odor concentration effectively as compared with simple composting in the manure treatment stage.

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Optimization of straw decomposition with inoculants by using response surface method

SU Yao, JIA Shengqiang, HE Zhenchao, YANG Yanhua, YU Man, CHEN Xijing, SHEN Alin

2019, 31(5): 798-805. DOI: 10.3969/j.issn.1004-1524.2019.05.16

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The rice straw was selected as typical straw to illustrate the influence of 4 straw-decomposing inoculants on the decomposition of straws. Based on the decomposition rate and ratio of carbon to nitrogen of rice straw, one inoculant with better straw decomposing capacity was selected for optimization of straw decomposing conditions by Box-Behnken design. The result demonstrated that among the four factors, namely, straw-decomposing inoculant content, temperature, water content and extra-nitrogen (biogas slurry) content, the effect of temperature, water content and straw-decomposing inoculant content on decomposition rate reached the significant level of P<0.01. The predicted optimum conditions for straw decomposing were temperature of 29.6 ℃, water contents of 90%, straw-decomposing inoculant content of 2.0% of the straw weight, and extra-nitrogen content of 0.17% of straw weight. According to the mathematical model, the maximum decomposition rate on 25th day was 63.25%, and it was consistent with the decomposition rate revealed by verification experiment under the above conditions.

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Natural community species diversity of green manure Vulpia myuros and its correlation with environment factors

HE Qinghai, FANG Ru, HUANG Xubo, QIN Yuchuan, WANG Yanbin, LIU Bentong, YANG Shaozong

2019, 31(5): 806-815. DOI: 10.3969/j.issn.1004-1524.2019.05.17

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To explore the natural community characteristics and species composition of the green manure plant Vulpia myuros, the habitats of 14 wild V. myuros sample plots in Zhejiang provenance were investigated. Analysis variance (ANOVA), correlation analysis and curve estimation were used to analyze the species composition, and the relationship between diversity index or herbage coverage and geographical climatic factors or soil factors. The results showed that there were 49 species of plants belonging to 29 families in the natural community. The major associated species were Eremochloa ophiuroides, Solidago decurrens, Erigeron annuus and Kummerowia striata. There were significant differences in the Shannon-Wiener diversity index, Simpson diversity index, Pielou diversity index and species richness in 14 natural sample plots. There was no significant correlation between diversity index, herbage coverage and species important value of V. myuros with geographical climate factors. There were significant correlations between diversity index with many soil factors, among which the most influential factors were soil temperature, soil salinity and soil organic matter content. The result of curve fitting suggested several models, including reciprocal model, quadratic curve model and linear model. There was a significant positive correlation between herbage coverage and content of available phosphorus, available potassium and organic matter in the soil. This research had clarified some problems of the community composition and community ecological equilibrium of V. myuros. Thus, it might provide the foundation of popularization of V. myuros as a green manure plant.

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Optimization of drying process of blueberry-apple reconstituted fruit cake and its influence on quality

LI Lu, HAN Qiang, WU Weijie, FANG Xiangjun, GAO Haiyan

2019, 31(5): 816-822. DOI: 10.3969/j.issn.1004-1524.2019.05.18

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The cold and hot air temperature drying technology was adopted to optimize the drying process of blueberry and apple pulp to produce high quality reconstituted fruit cake. The parameters of the variable temperature drying process were determined by single factor and orthogonal experiments, and the sensory evaluation, ultrastructure and texture characteristics of the finished product were analyzed. The results showed that the best drying process of blueberry-apple reconstituted fruit cake was 6 mm thickness, 70 ℃, 7 h in the early hot air baking, and 30 ℃ in the late cold air to the final water content of the product was less than 15%. The blueberry-apple reconstituted fruit cake obtained under the dry process condition was sweet and sour, fruity and rich, with higher toughness and not sticky to teeth. The ultrastructure showed that the tissue was smooth and compact, the sensory score was high. The physical and chemical indicators and microbial indicators were in line with the national standards.

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Detection of Vibrio parahaemolyticus in seafood products by real-time quantitative PCR assay

CHEN Lin, ZHOU Qingqing, GU Qing, LI Ping

2019, 31(5): 823-828. DOI: 10.3969/j.issn.1004-1524.2019.05.19

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Vibrio parahaemolyticus is a food-borne pathogen, which can contaminate food and cause serious and acute gastroenteritis to human beings. In this study, a real-time polymerase chain reaction (PCR) assay was developed for the quantitative detection of Vibrio parahaemolyticus in aquatic products. Randomly amplified polymorphic DNA (RAPD) method was introduced to characterize species-specific DNA fragment of V. parahaemolyticus, and specific primers were designed. The proposed method was used to detect V. parahaemolyticus in 20 aquatic products on sale. It was shown that 6 samples out of 20 were carrying V. parahaemolyticus. The minimum detection sensitivity was 50 fg DNA. Compared with the methods recommended in national standard, the proposed method was swifter and more convenient to detect V. parahaemolyticus in auqatic products.

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Optimization of ground wheel components of base fertilizer fixed-point deep applicator

SONG Zhujun, WU Xuemei, LI Guochang, WANG Fang, LIU Hongyun, ZHANG Fugui, ZHANG Peikun

2019, 31(5): 829-836. DOI: 10.3969/j.issn.1004-1524.2019.05.20

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In order to solve the large deviation of fertilization point and marking point caused by the lack of driving force of the ground wheel assembly of base fertilizer fixed-point deep fertilization machine, an optimal parameter experiment is carried out by using a regression orthogonal design. The main factors affecting the deviation between the fertilization point and the marking point were selected as the advance speed of the apparatus, the blade depth, the hub radius and the blade area. The four-factor four-level orthogonal experiment was carried out with the driving torque of the ground wheel assembly as the evaluation index. Simulation experiments showed that the influence of factors on deviation between the fertilization point and the marking point decreased as the blade area>hub radius>blade depth>advance speed of the apparatus. The optimal combination of parameters were as follows: the advance speed of the apparatus was 2.3 m·s^-1, the blade depth was 8 cm, the hub radius was 17.50 cm, and the blade area was 52 cm². Under this condition, the driving torque provided by the soil for the ground wheel assembly was 101.96 N·m, which was increased by 11.76%. Field experiment results indicated that the position deviation between the marking point and the fertilization point was reduced from 4.23 cm to 1.99 cm after optimization, which realized the accurate marking function. The research could provide reference for the optimization of the structure and operation parameters of the base fertilizer fixed-point deep applicator.

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Research on development and revitalization of beekeeping in Zhejiang Province

FU Linlin, MAO Xiaohong, MAO Xiaobao

2019, 31(5): 837-845. DOI: 10.3969/j.issn.1004-1524.2019.05.21

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Zhejiang Province, a national honey province, was taken as an example, and the production and sales of beekeeping in Zhejiang was investigated using bee product market research data and beekeeper survey data from a macro perspective. Subsequently, the production and management behaviors and existing problems of beekeepers were analyzed from a microscopic perspective. On this basis, an empirical analysis of the factors that affected the income of the beekeepers was carried out. The results showed that the cultural level, breeding scale, breeding methods, technology adoption and main marketing channels of beekeepers had a significant impact on the income of beekeepers. Finally, some countermeasures and suggestions were put forward to promote the sustainable development of beekeeping in Zhejiang.

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Recent advances of validation methods in passive microwave remote sensing of soil moisture products

WANG Chunmei, GU Xingfa, YU Tao, ZHOU Xiang, ZHAN Yulin, HAN Leran, XIE Qiuxia

2019, 31(5): 846-854. DOI: 10.3969/j.issn.1004-1524.2019.05.22

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The validation of large-scale soil moisture products in passive microwave remote sensing takes a key role for global climate change and data assimilation studies. However, it is difficult to obtain the real soil moisture values that can represent the soil moisture values and reflect spatial heterogeneity of large-scale passive microwave remote sensing products. It has become a key issue for the validation of passive microwave soil moisture products. At present, two technical approaches were used to build soil moisture observation network: the scale-up methods and the multi-source information fusion methods. Both two approaches involved the ground observation, simultaneous observation, and scale conversion, etc. The progress made in validation methods of large-scale soil moisture products was reviewed in this paper, and five typical methods were summarized, including the measured sample test, image data cross-test, model simulation test, influencing factor test and traditional geostatistical test. The first two methods belongs to the “spot generation” scale conversion test, and they attach importance to the spatial representation of the ground sample. Theoretical basis of the model simulation test and influencing factor test is the relationship between soil moisture and prior knowledge. However, these two methods neglect the importance of sample data. Traditional geostatistical test takes into account both sample points and prior knowledge, but it is insufficient in the comprehensive utilization of prior knowledge information of multi-source data types. Currently, the Bayesian maximum entropy method is widely used in spatial prediction. In some studies, this method has been applied to multi-source data fusion. The advantage of Bayesian maximum entropy test provides a flexible way of data utilization. This method allows multiple sources and types of data sets to be used for spatial analysis of satellite observation scale at the same time, generates high resolution reference map of soil moisture observation field, completes the validation of soil moisture products, and provides a basis for the authenticity test of large-scale soil moisture products. Thus, it would provide a new way for the validation of large-scale soil moisture products.

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