Journal of Henan Agricultural Sciences

Research Progress of Nematophagous Fungi against Plant Parasitic Nematodes

HAO Xin, WANG Bowen, CHEN Jie, LI Yang, MA Ling

2021, 50(1): 1-9. DOI: 10.15933/j.cnki.1004-3268.2021.01.001

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Plant parasitic nematodes,as a kind of plant pathogens,cause harm to global agriculture and forestry.The loss caused by plant parasitic nematodes was S157 billion per year. Nematode-trapping fungi have attracted wide attention as a safe,efficient and low-toxic biotechnology for the control of nematodes.In this paper,the classification,action mechanism,control effect,influencing factors and application status of nematode-trapping fungi were discussed,and the prospect of interaction between nematode-trapping fungi and plant parasitic nematodes was prospected.

Application Advance of Soft Computing Technology in Agricultural Production

TIAN Ye, SUN Ruizhi

2021, 50(1): 10-15. DOI: 10.15933/j.cnki.1004-3268.2021.01.002

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Soft computing is a collection of methodologies that aim to exploit tolerance for imprecision and uncertainty to achieve tractability,robustness and low solution cost.The problems with uncertain factors in complex systems is solved by combining fuzzy logic with intelligent technology,to access the reality of the objective facts.With the development of Chinese agriculture into the intelligent stage, soft computing technology has been used in agricultural production more and more widely.The application research of soft computing techniques in crop yield prediction,weed detection and monitoring of land use and land cover were elaborated,and the prospects of soft computing techniques in agriculture application research and development was discussed in this paper so as to provide theoretical support for ensure food security in our country and the construction of a world-class level of agriculture.

Cloning and Function Verification of Two Phytochrome C Genes in Zea mays

DING Wusi, CHEN Shizhan, LIU Lei, FAN Xiaocong, DING Mengyue, SUN Guanghua, WANG Lijian, YANG Jianping

2021, 50(1): 16-26. DOI: 10.15933/j.cnki.1004-3268.2021.01.003

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In order to study the similarities and differences of PHYC functions between maize and Arabidopsis and the functions in photomorphogenesis and shade avoidance of two maize PHYC genes,we cloned two maize PHYC genes,analyzed the functional domains of maize PHYC and phylogenetic tree of PHYC proteins maize and common plant species by bioinformatics,and verified the functions of two maize PHYC genes in Arabidopsis by transgenic techniques.The results showed that the coding regions of ZmPHYC1 and ZmPHYC2 from maize inbred line B73 were cloned.The coding region length was 3 405 bp,and encoded a protein with 1 135 amino acid residues with molecular weight of 126.14 ku and 126.07 ku.Similar to the PHYC proteins of Arabidopsis and rice,ZmPHYC1 and ZmPHYC2 contained a PAS_2domain,a GAF domain,a PHY domain,two PAS repeat domains,a histidine kinase A domain and a histidine kinase like-ATPase domain. Amino acid sequence homology analysis showed that ZmPHYC1 and ZmPHYC2 had the highest homology with the sorghum PHYC protein.ZmPHYC1 and ZmPHYC2 could not only complement long hypocotyl phenotype caused by Arabidopsis phyC-2 mutant under red light,but also promote the de-etiolation of seedlings under continuous red and blue light conditions,and inhibit the elongation of hypocotyl under shade condition.For promotion of seedling de-etiolation and inhibition of shade avoidance,ZmPHYC1 play more important role than ZmPHYC2.

Genetic Structure Analysis of Parental Inbred Lines of Main Maize Hybrid Varieties in China

LI Jingjing, ZHANG Wenyang, WANG Lifeng, WANG Hao, LI Huiyong

2021, 50(1): 27-34. DOI: 10.15933/j.cnki.1004-3268.2021.01.004

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Genetic structure of parental inbred lines of the common maize varieties was analyzed using 56K maize gene chip,and heterosis modes of parental inbred lines of the main maize varieties were analyzed,so as to provide guidance and reference for maize breeding.The results showed that a total of 35 261 high-quality single nucleotide polymorphism(SNP) markers were obtained from 56 000 SNPs,the average of minimum allele frequency (MAF) was 0.28,the MAF of 12.56% of the SNP markers were 0.45—0.50,the polymorphism level was high.Polymorphic information content(PIC) was 0.10—0.50,with an average of 0.37.The results of cluster analysis showed that,five groups were divided,of which,group Ⅰ was mainly BSSS germplasm,which was represented by PH6WC;group Ⅱ mainly contained PH4CV,male parents of hybrid DK517 and DH605 and so on; group Ⅲ contained Reid, P group germplasm and European male parents;group Ⅳ contained domestic lines,such as Zong 3 and Zong 31;group Ⅴ contained classic SPT germplasm,Suwan and CIMMYT tropical germplasm.At the same time,the parental SNP differences of 16 maize hybrids were analyzed,and a total of 7 615 polymorphic sites and 7 592 conserved sites were selected.

Effect of Nitrogen and Zinc Application on Contents and Distribution of Nitrogen and Zinc in Wheat Grain

SHAO Yunhui, ZHANG Panpan, MA Geng, LU Hongfang, WANG Lifang, WANG Chenyang, LI Xiangdong, YUE Junqin, QIN Feng

2021, 50(1): 35-43. DOI: 10.15933/j.cnki.1004-3268.2021.01.005

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The field experiment was conducted with a split plot design. Grain was pilled into testa,aleurone layer,subaltern aleurone layer,three transition layers of endosperm and central layer of starch endosperm from out to inner (F1—F7). Main plot treatments were two Zn rates,0,9.60 kg/ha,sub-plot treatments were two N rates,0,240 kg/ha,and the effect of N and Zn application on the contents and distribution of N and Zn in the grain fractions was studied,so as to provide reference for improving zinc content in wheat grain endosperm and optimal management of nitrogen and zinc fertilizer.The results indicated that distribution of N and Zn was nonuniform in the wheat grain.The N content was higher in the F2 and F3 fractions,and the content of Zn decreased in the grain from out to inner.The distribution of N and Zn was the highest in the F7 fraction,which were 40.40% and 26.20%,respectively,and the lowest in F1 and F6 fractions.The grain yield,N contents in the grain,Zn accumulation and distribution in grain’s fractions of the treatment with Zn were not significantly different from the treatment without Zn.Zn application significantly increased grain Zn content by 49.3%,maximum increase amplitude was in the F5 fraction, and Zn accumulation in each fraction increased by 50.1%—63.6%, maximum increase amplitude was in the F7.The Zn distribution in the grain fractions was not obviously affected by Zn application. N application significantly increased the grain yield,N content in grain and its fractions,N accumulation in the grain fractions, maximum increase amplitude was in the F3 fraction, and the distribution of N obviously increased in F3 fraction, but decreased in F1 and F7 fractions under N application treatment.N application significantly increased Zn contents in grain and its fractions,the Zn content in F7 fraction increased by 24.9%,the Zn distribution proportion in F7 fraction increased by 14.3%,but the Zn distribution proportions in other fractions decreased (expect F2 fraction).Correlation analysis was significant between Zn content in the F7 and N contents in the grain,F1 and F3 fractions.Relationship was also significant between Zn content in the F7 fraction and N content in the F1 fraction.Therefore, N and Zn application can significantly improve Zn content in grain, accumulation and distribution in the inner of wheat grain,and is a reasonable and efficiency way to increase the Zn nutrition quality in the endosperm of wheat grain.

Effects of Nitrogen Application Levels and Root Separation on the Root Morphology and Nitrogen Fixation Ability of Alfalfa in the Maize/Alfalfa Intercropping System

WANG Libo, SHAO Zeqiang, LU Wenlong

2021, 50(1): 44-51. DOI: 10.15933/j.cnki.1004-3268.2021.01.006

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In this study,pot experiments were conducted to study the effects of nitrogen application levels and root separation patterns on the root morphology and nitrogen fixation ability of alfalfa in the maize/alfalfa intercropping system using random block design and root separation technology,so as to provide a scientific basis for reduction and efficiency enhancement of chemical fertilizer in the maize/alfalfa intercropping system. The results showed that nitrogen application increased the root length,root surface area and root volume of the alfalfa at different diameter-class by 22.4%, 18.4% and 16.9%, but decreased the number of nodules,dry weight of the nodules,the rate of nitrogen fixation,fixed amount and the leghemoglobin content by 33.8%, 64.5%, 29.4%, 17.0% and 55.4% respectively.Under no nitrogen fertilizer(N0) treatments, compared with plastic sheet separation (PSS) treatment, the root length,root surface area and root volume of alfalfa at different diameter-class of no separation(NS) and nylon net separation (NNS)treatments were reduced by 26.1%,29.1%,36.9% and 13.6%,12.9%,23.5%,and the number of root nodules,dry weight of root nodules,nitrogen fixation rate,fixed amount and the content of leghemoglobin of NS and NNS treatments were increased by 69.5%,76.4%,148.2%,52.9%,73.3% and 49.3%,61.4%,66.0%,41.9%,43.3%,respectively;Compared with the NNS treatment,the root length,root surface area and root volume of alfalfa at different diameter-class of NS treatment were reduced by 14.0%,18.8% and 19.5%,and the number of alfalfa nodule,dry nodule weight,nitrogen fixation rate, fixed amount and the content of leghemoglobin of NS treatment were increased by 13.5%,9.3%,49.5%,7.8% and 20.9%respectively.Under nitrogen fertilizer (N1) application treatment,compared with PSS treatment,the root length,root surface area and root volume of alfalfa at different diameter-class of NS and NNS treatments were reduced by 23.9%,28.3%,32.3% and 12.6%,15.3%,23.5%,and the number of root nodules,dry weight of root nodules,nitrogen fixation rate,fixed amount and the content of leghemoglobin of NS and NNS treatments were increased by 45.5%,97.6%,105.5%,30.0%,42.6% and 42.8%,45.5%,58.6%,26.2%,18.1%,respectively;Compared with the NNS treatment,the root length,root surface area and root volume of alfalfa at different diameterclass of NS treatment were reduced by 12.9%,15.4% and 18.1%,and the number of alfalfa nodule,dry nodule weight,nitrogen fixation rate,fixed amount and the content of leghemoglobin of NS treatment were increased by 1.9%,35.8%,29.6%,2.9% and 20.7%,respectively.Therefore,both nitrogen application and root separation promote the growth of root at different diameter-class in the maize/alfalfa intercropping system,while inhibite the nitrogen-fixing ability of alfalfa.

Effects of Exogenous Gibberellin on Flue-cured Tobacco Leaf Trichome and Aromatic Substances

HE Lingxiao, XUE Gang, SUN Jutao, ZHANG Zhiqiang, DING Yongle, YANG Tiezhao, XU Shixiao, CHENG Changhe

2021, 50(1): 52-59. DOI: 10.15933/j.cnki.1004-3268.2021.01.007

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In order to increase the content of the neutral aroma components of flue-cured tobacco,improve the quantity,and explore the role of gibberellin in the anabolic pathway of cembranes,trichome density and trichome exudate content were detected at 50,60 days old leaves of NC71 and Zhongyan 100,which were sprayed with gibberellin at seedling stage.The results showed that the density of trichome and the content of trichome exudate increased by 30.97% and 37.44%, respectively, and the neutral aroma components of flue-cured tobacco increased by 78.38%.Moreover,the content of endogenous gibberellin increased,and the expression levels of positive regulatory genes CPS and GA20ox in the gibberellin synthetic pathway were up-regulated,while the expression levels of negative regulatory genes GA2ox was down-regulated.In conclusion, spraying gibberellin on the stem tip can increase the density of leaf trichome,the secretion capacity of trichome, the content of trichome exudate and terpene degradation products of neutral aroma components at seedling stage.

Effects of Different Light Transmittance and Growth Regulator Concentrations on Panax notoginseng

LIN Jie, LIANG Zengyan, ZHAO Zhi, LUO Chunli, LUO Fulai, WANG Hualei

2021, 50(1): 60-67. DOI: 10.15933/j.cnki.1004-3268.2021.01.008

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In order to screen out the light transmittance and 24-epibrassinolide (EBR) concentration suitable for triennial Panax notoginseng growth, shading network was used to set five different light transmittance(57.31%,31.61%,18.90%,11.44%,8.04%) and six concentrations of EBR solution[0(water),0.05, 0.20, 0.40, 0.80, 1.60 mg/L] was set up to explore the effect of different light transmittance and different concentrations of EBR solution on agronomic characteristics,quality and yield of triennial Panax notoginseng in field experiment.The results showed that the morphologic indexes of Panax notoginseng were different under different treatments,when the light transmittal rate was 18.90%,the plant height,middle leaf length,middle leaf width and fresh weight per plant of Panax notoginseng were significantly higher than those under other treatments,which were 33.67 cm,14.06 cm,4.67 cm,53.86 g,and total saponins content was high(8.14%).The effects of different transmittance on water content,ash content,extract and total saponins contents of Panax notoginseng were not significant.When the concentration of EBR was 0.05 mg/L,the plant height,root-shoot ratio,water content and ash content of Panax notoginseng were the highest,which were 32.41 cm,2.15,6.50% and 5.22%,respectively.When EBR concentration was 0.20 and 0.40 mg/L,petiole length and relative chlorophyll content of Panax notoginseng were the highest, which were 10.36 cm and 47.21, respectively.When the concentration of EBR was 0.80 mg/L,the yield of ginseng Rg1 and individual plant was the highest (4.00% and 46.98 g,respectively),and the total saponin content was higher(7.30%).In conclusion,when the transmittance of Panax notoginseng was 18.90%,which is more suitable for the accumulation of underground biomass and saponins. When the concentration of EBR was 0.80 mg/L,the quality and yield of Panax notoginseng were the best.

Effects of Pretreatment with Alkaline Non-alkali Solutions on Anaerobic Fermentation of Corn Straw under Mild Hydrothermal Condition

WANG Yuxiaofei, WANG Yu, ZHU Guoqiang, SUN Yunbo, YANG Yi, QU Wei

2021, 50(1): 68-80. DOI: 10.15933/j.cnki.1004-3268.2021.01.009

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To provide an economic and effective preprocessiog method of corn straw,Dry yellow corn straw was used as feedstock.Four alkaline non-alkali solutions(Na₂CO₃,Na₂SO₃,K₂HPO₄,CaO₂,each had 3 gradients of 2%,4%,6%) were taken as pretreatment agents,no pretreatment as control,soaked in condition of TS content of 10% for 24 h,heated in 80℃ water bath kettle.The lignin,hemicellulose and cellulose content of straw,and COD(chemical oxygen demand)value and TVFA( total volatile fatty acid) content of aqueous extracts of straw and final biogas slurry after anaerobic fermentation,as well as the pH values of fermentation brothes, daily biogas yields and methane content of biogas during the anaerobic fermentation were analyzed.The results showed that,lignin,hemicellulose and cellulose content of corn straw decreased significantly after pretreament with alkaline non-aliki solutions under mild hydrothermal condition.For most pretreament groups,the biogas yields of unit solid and the peak methane content of biogas increased,pH values of fermentation brothes finally stabilized at 7.50—9.00.Among all groups,CaO₂ pretreament group could dissolve lignocellulose best,with the content of lignin,hemicellulose and cellulose decreasing by 59.75%, 69.61% and 47.09% respectively at most.For 2% CaO₂
pretreatment group,the best biogas production effect was got,with the biogas yield of unit solid reaching 291.8 mL/g and the peak of methane content of biogas increasing by 39.02%;COD value and TVFA content of the final biogas slurry of this group were also the lowest among all groups,decreasing by 56.25% and 61.45% compared with the control group respectively. Through analysis,it was found that types and concentrations of pretreatment solutions influenced the removal rates of lignocellulose of corn straw significantly,and both removal rates of lignocellulose and pretreament methods had influence on the anaerobic fermentation effect of corn straw,so comprehensive consideration should be conducted in the
actual production.

Soil Microbial Community Structure and Functional Diversity in Vegetable Greenhouse for Different Planting Years in Wuling Area of Guizhou Province

ZHAO Hui, WANG Xiying, XU Shiqiang, TAN Zhiyong

2021, 50(1): 81-91. DOI: 10.15933/j.cnki.1004-3268.2021.01.010

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In order to reveal the quality of greenhouse soil for vegetable cultivation,the relationships between soil microbial community structure,functional diversity,and soil quality of vegetable greenhouse for different planting years (3,5 and 7 a) and uncovered vegetable field (CK)were investigated byphospholipid fatty acids(PLFAs)analysis and Biolog-Eco micro plate method in Wuling area of Guizhou Province,so as to provide a scientific basis for exploring the microbial mechanisms underlying soil quality degradation.Results showed that as the greenhouse vegetable planting years increased,total nitrogen,ammonium nitrogen and nitrate nitrogen contents gradually accumulated,soil acidification was obvious.The soil microbial biomasss carbon and nitrogen contents were the highest for the planting 3 a treatment,which were 74.51% and 4.17% higher than those of CK;the amount of soil fungi increased,and the bacterial/fungal decreased gradually,which led to the decrease of soil stability.The results of hierarchical clustering and principal components analysis(PCA)showed that the planting years had significant impacts on the soil microbial community structure.The soil microbial community structures of the planting 3 a treatment and the control group showed significant differences from those for the planting 5 a and 7 a treatment.The average soil well color development(AWCD)and the microbial functional diversity index(Shannon index,Simpson index,McIntosh index) of the planting 3 a treatment were higher than those of other planting years treatments. The main carbon sources absorbed by the greenhouse soil were amino acids and carbohydrates,and the planting years of 3 a treatment had the highest capacity to use all carbon sources.pH value,organic carbon content,total nitrogen content,and C/N were the main factors affecting soil microbial community structure and microbial metabolism.According to the comprehensive analysis results,the soil microbial community structure and functional diversity changed significantly,and the soil microecological balance was disturbed,impeding the efficient and sustainable use of greenhouse soil after greenhouse vegetable planting 3 a.

Relationship between Incidence of Gastrodia elata and Changes of Fungal Community Structure

BAI Qiuyue, DENG Baiwan, XIE Xiuchao, ZHANG Caini, LIU Lanlan, YUAN Minqian

2021, 50(1): 92-100. DOI: 10.15933/j.cnki.1004-3268.2021.01.011

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The study of the differences in fungal community structure in healthy and diseased Gastrodia elata provides clues for the investigation of the mechanism of Gastrodia disease.Traditional culture method and high-throughput sequencing technology were used to analyze the changes of fungal commu ity structure in healthy and diseased plants tubers.The results of culture analysis showed that the healthy and diseased Gastrodia tubers fungi covered 3 phyla and 14 genera.Among them, there were 8 genera including Trichoderma and Talaromyces in healthy Gastrodia.There were 10 genera including Cladosporium,Alternaria and Fusarium in the Gastrodia elata strain.A total of 1 692 OTUs(operational taxonomic units) were obtained.The classification of species showed that at the phylum level,the main groups of fungi in the healthy and diseased groups of Gastrodia were basically the same,mainly including Ascomycota and Basidiomycota,but there were obvious differences in the relative abundance of fungi in healthy and diseased tubers of Gastrodia elata.At the genus level,the composition and relative abundance of fungi healthy and diseased Gastrodia were very different.Among them, Ilyonectria, Helotiales and Fusarium may be the pathogenic bacteria group that caused diseases of Gastrodia.It can be seen that both the traditional culture method and the high-throughput sequencing technology show that there are significant differences in the fungal community structure between healthy and diseased Gastrodia elata.The occurrence of Gastrodia elata disease is related to the change of the fungal community structure in Gastrodia elata,suggesting that it can be prevented by adding superior probiotics in the early stage.

Analysis of Jasmonic Acid Content Change after Infected by Aphid in Different Tobacco Varieties#br#

SUN Jiping, CHEN Fei, LI Xuejun, SUN Huan, LI Xuhui

2021, 50(1): 101-107. DOI: 10.15933/j.cnki.1004-3268.2021.01.012

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Jasmonic acid(JA) is closely related to plant disease resistance and insect resistance.In order to select aphid resistant tobacco varieties and study its mechanism,leaf test in vitro and pot experiment were used to identify the aphid resistance of 12 tobacco varieties,and the changes of JA content before and after aphids infestation were analyzed.The results showed that four tobacco varieties,such as Qieke,Tailifei 2411, Shubalaomuji 2113 and Wangpoer showed resistance to aphid; the aphid resistance of Wuchuanheiyan,Hongxianxiangdayeyan, MC Nair and Vesta 33 were moderate; Zhongyan 100 and SpeightG-28 were susceptible to aphid; the Luositou 2474 and Yunyan 97 showed high sensitivity to aphid.The content of endogenous JA in aphid resistant tobacco varieties was significantly higher than that in other varieties.The content of JA increased significantly in 12 tobacco varieties after aphid infestation.The increase amount and increase range of JA in aphid resistant tobacco varieties were greater than those in aphid susceptible tobacco varieties.JA content had a good correlation with aphid resistance,which could be used as a physiological index for identification of aphid resistance in tobacco varieties.

Effects of Ethephon and STS on Cucurbita moschata Sex Differentiation and Expression of Ethylene Related Genes

LI Qingfei, ZHANG Mannan, WANG Mengmeng, CHEN Bihua, WANG Guangyin, ZHOU Junguo, LI Xinzheng

2021, 50(1): 108-112. DOI: 10.15933/j.cnki.1004-3268.2021.01.013

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Three inbred lines of Cucurbita moschata were used as materials to investigate the effect of spraying ethephon and ethylene inhibitor STS on shoot meristem during seedling stage on sex differentiation,as well as the response of ethylene synthesis and receptor genes (ACO,ETR) to this treatment.The results showed that,compared with untreated plants,ethephon improved the expression levels of ACO and ETR.Furthermore,through investigating the sex differentiation within 20 nodes,it was found that the first female flowers node decreased and the number of female flowers increased significantly. Its effects on male flowers number varied with different inbred lines.In contrast, STS inhibited the expression of ACO and ETR,delayed the first female flowers emerging,increased the number of male flowers, and decreased the number of female flowers.The results indicate that ethephon and ethylene inhibitor STS can affect both ethylene synthesis and ethylene response genes,the promoting effect of ethephon and the inhibiting effect of STS on female flowers may be related to its induction or inhibition on the expression levels of ethylene synthesis and response gene.

Comprehensive Evaluation of Aluminum Tolerance of 26 Grape Germplasms Based on Inorganic Nutrition

ZHANG Yongfu, XU Shiqin, JING Zange, MO Liling, HUANG Li

2021, 50(1): 113-120. DOI: 10.15933/j.cnki.1004-3268.2021.01.014

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To prevent or reduce the harm of aluminum stress to grape,an effective identification method of aluminum tolerance from inorganic nutrition was searched, and aluminum tolerance genotype was obtained.The contents of 12 elements N,P,K,Ca,Mg,Fe,Mn,Zn,B,Mo,Cu and Al in leaves of 26 grape germplasm resources from different sources were determined under aluminum stress by soilless cultivation.Taking aluminum tolerance coefficient as the index of aluminum tolerance identification,principal component analysis, membership function and cluster analysis were used to evaluate the aluminum tolerance of different grape germplasm resources.The results showed that under aluminum stress,the contents of inorganic nutrient elements in different grape germplasm resources were significantly different.From the correlation analysis, it was found that there was a certain correlation between the aluminum tolerance coefficients of each element, which indicated that the information provided was overlapping,and it was inaccurate to use a single element to evaluate the aluminum tolerance of different grape germplasm resources.After principal component analysis of 12 single indexes,5 comprehensive indexes representing 85.095% of the original data information of grape aluminum tolerance were obtained.Then the contribution rate of these 5 comprehensive indexes was used to calculate their respective membership function values,and the comprehensive evaluation value(D value) of aluminum resistance of various germplasm resources was obtained by weighting. Based on the cluster analysis of D value,26 grape germplasm were divided into 3 categories, 7 cultivars, such as Vitis sinocinerea,Hongdiqiu,Heimeigui and so on,had strong aluminum tolerance;Taishan-1,Heitiwuji,Shuijing and so on,were moderately tolerant to aluminum;Tonghua-3,Gaoshan-2,Beta and other 9 germplasm resources were weakly tolerant to aluminum. Using the aluminum tolerance coefficient of each element as independent variable and D value as dependent variable,stepwise regression was carried out,and the optimal regression equation was obtained.The correlation between D value and predicted value of aluminum tolerance in 26 grape germplasm resources was very significant.The contents of K,Mn,Zn and B were selected as the identification index of aluminum tolerance of grape.This method not only could avoid the randomness and one-sidedness of evaluating aluminum tolerance of grape by using a single index,but also reflect the relationship between different aluminum tolerance related traits and aluminum tolerance in grape,so the evaluation results were more reliable.

Heat Tolerance Threshold and Physiological and Biochemical Responses of Leaves of Different Apple Varieties

TIAN Jia, LI Jia, MENG Qingbo, LI Zhongyong, XU Jizhong

2021, 50(1): 121-128. DOI: 10.15933/j.cnki.1004-3268.2021.01.015

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In order to study the heat tolerance threshold and physiological and biochemical changes of leaves of different apple varieties,the new shoots of Orin,Fuji and Gala were used as the test materials under the artificial simulated high temperature environment.The time of symptom onset,heat injury rate and heat injury index were investigated,and changes in relevant physiological and biochemical indicators were measured.The main results showed that the heat tolerance threshold of the leaves of Orin,Fuji and Gala were 41,43 and 44℃ ,respectively. In the field,Orin leaves had the highest damage rate and damage index of 53.50% and 18.05,and Gala leaves had the lowest damage rate and damage index of 5.36% and 1.57,respectively.With the prolongation of high temperature,the chlorophyll contents of the three apple varieties showed a gradual decrease trend,the soluble protein content and the activities of superoxide dismutase and peroxidase showed a law of increasing first and then decreasing,and reached the peak value at 6 hours after high temperature stress.In summary,with the prolongation of high temperature stress, the photosynthetic pigments of apple leaves are degraded, the membrane peroxidation is aggravated,the antioxidant enzymes activity of apple leaves increase,and the physiological metabolism of
apple leaves changes significantly.

Effects of Different Tomato Rootstocks on Growth and Physiological Characteristics of Eggplant Grafted Seedlings under Heat Shock Treatment

QIN Tao, LIU Xinshe

2021, 50(1): 129-133. DOI: 10.15933/j.cnki.1004-3268.2021.01.016

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To investigate the resistance of eggplant seedlings grafted with different tomato rootstocks to high temperature stress,four different tomato varieties(Jinpeng No.1,Dahongxiaoguo,Fenguozhongguo,Xiaoyingtao,named as T₁,T₂,T₃,T₄,respectively) were selected as rootstocks for Datianlvfeng eggplant.After normal temperature and heat shock, the effect of four different tomato stocks on physiology and heat tolerance of grafted eggplant was studied by determining the height,stem diameter,number of leaves,plant type,heat injury index,SOD activity,MDA and chlorophyll contents,electrolyte leakage rate at seedling stage.The results showed that the height,plant type and leaf number of eggplant grafted with different tomato varieties did not change significantly compared with self-root seedlings; the electrolyte leakage rate and MDA content of eggplant seedlings grafted with different tomato varieties decreased significantly compared with self-root seedlings; the chlorophyll content increased, the T₁,T₂,T₃were significantly higher than that of self-root seedlings; the SOD activity increased,the T₁,T₃were significantly higher than that of self-root seedlings,the T₃ increased by 53.31%;the heat damage index was the lowest after T₃ grafting, only 21.67%.Comprehensive analysis showed that the grafted eggplant seedlings with Fenguozhongguo(T₃) tomato as rootstock showed high heat resistance.

Genetic Evaluation for Breed Conservation Effect of Four Populations of Taihu Goose

LIU Hongxiang, WANG Jian, SONG Weitao, ZHU Chunhong, TAO Zhiyun, XU Wenjuan, ZHANG Shuangjie, LI Huifang

2021, 50(1): 134-141. DOI: 10.15933/j.cnki.1004-3268.2021.01.017

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In order to fully understand the dynamic change process of breed conservation effect of Taihu goose in Jiangsu Province and compare with that in Zhejiang Province,the genetic parameters of Taihu goose populations in Jiangsu (TS,TC,TK) and Zhejiang Province (TZ) were analyzed by simplified genome sequencing.The results showed that compared with the previous two generations,the inbreeding coefficients of TC and TK populations decreased to 0.087 5 and 0.075 3,respectively,due to the using of strict stemma equimultiple random select cross method and appropriate introduction of breeding-use individuals between different populations;the genetic structure analysis and principal component analysis showed that TZ population was independent of the other three populations,and differentiated significantly.The genetic differentiation coefficient(F_st) of TZ and TS population reached 0.051 5,and the inbreeding coefficient was 0.110 7,which was significantly higher than that of populations in Jiangsu Province.The selection signature analysis of TZ and TS population showed that the genes for fatty acid metabolism related pathway were selected.The results showed that the effect of Taihu goose population conservation was better in Jiangsu Province,and the increase of inbreeding coefficient could be avoided by stemma equimultiple random select cross method and appropriate blood exchange.

Establishment and Application of Real-time PCR for Differentiation of Porcine Reproductive and Respiratory Syndrome Virus Strains NADC30-like and JXA1-R

ZHOU Xinyu, CHEN Xinxin, QIAO Songlin, GUO Zhenhua, LI Rui, DENG Ruiguang, ZHANG Gaiping

2021, 50(1): 142-151. DOI: 10.15933/j.cnki.1004-3268.2021.01.018

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To distinguish JXA1-R vaccine strain and NADC30-like HNhx strain of porcine reproductive and respiratory syndrome virus(PRRSV),TaqMan real-time quantitative PCR and SYBR Green Ⅰ realtime quantitative PCR were established by using specific primers based on the difference of the regions of Nsp2 genes between HNhx and JXA1-R.The results indicated that the standard products had a good linear relationship in the range of 10³—10⁸copies/μL,and the R² of the standard curves were over 0.990 in two methods.The detection limit of the TaqMan-based fluorescent quantitative PCR was 10⁴copies/μL(JXA1-R) and 10³copies/μL(HNhx),respectively.The detection limit of SYBR Green Ⅰ fluorescent quantitative PCR was 100 copies/μL.The reproducibility of intra-and inter-assay displayed that the coefficient of variations was less than 1.5%,the repeatability was good.In conclusion,the method for differentiation of HNhx and JXA1-R was successfully established in this study, which exhibited good stability and high specificity, and provided a new technical support for the differential detection of PRRSV.

Epidemiological Investigation of Main Viral Infectious Diseases in Pig in Southwest Henan Province

BAO Yin, ZHAI Hongyue, LI Huimin, YUAN Zhiqiao, MA Yujing, DING Yushan, KAN Yunchao, YAO Lunguang, TIAN Zhijun, LENG Chaoliang

2021, 50(1): 152-157. DOI: 10.15933/j.cnki.1004-3268.2021.01.019

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This study investigated the prevalence and antibody levels of the major viral diseases in largescale pig farms in southwest Henan Province, so as to formulate the scientific and reasonable comprehensive diseases prevention and control measures. 723 tissue samples of sick pigs organization(lymph nodes,lung,spleen,kidney,brain,etc.)and 2 476 blood samples were collected from 57 pig farms(Nanyang、Zhumadian and Xinyang) during 2017—2019,and the antigen and antibody levels of CSFV,PRRSV,PCV2 and PRV were detected by PCR and ELISA,respectively.The results showed that the infection of CSFV,PRRSV,PCV2 and PRV were common,and the average positive rates of pathogens reached 42.20% (192/455),56.85% (328/577), 45.03% (136/302) and 28.85% (73/253),respectively. In addition,the co-infection of PRRSV mixed with other viruses existed in some pigs.The antibody detection results indicated that the average positive rates of CSFV,PRRSV,PCV2,PRV gE and PRV gB were 78.37%(1 841/2 349),75.93%(1 565/2 061),84. 62%(682/806),33.12%(627/1 893) and 94.00% (1 504/1 600),respectively.Interestingly,the pathogen detection rates of CSFV,PCV2 and PRV decreased significantly in 2019.

Identification and Susceptibility of Actinobacillus pleuropneumoniae from Henan Province

PENG Zhifeng, HUANG Hui, ZHANG Xiaozhan, JIANG Zenghai, SHI Hongtao, QIAO Hongxing, BIAN Chuanzhou

2021, 50(1): 158-163. DOI: 10.15933/j.cnki.1004-3268.2021.01.020

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To investigate antibiotic resistence of Actinobacillus pleuropneumoniae(APP),APP isolates were isolated and identified from different regions in Henan Province.Susceptibility of 15 antibiotics for APP was determined with K-B disc method,then tetracyline-resistant genes tetA,tetB,tetC,tetD,tetE,tetH,tetG,tetK,tetL,tetM and tetO were detected and sequenced. The results showed that 5 type Ⅰ APP clinic strains were isolated.Susceptibility test showed that APP strains were sensitive to cefotaxime,ceftriaxone,levofloxacin, ofloxacin and sulfamethoxazole, and resistant to tetracycline (5/5, 100%),penicillin(3/5,60%),ampicillin(3/5,60%) and metronidazole(3/5,60%).The drug resistance genes tetB and tetA were detected(5/5,100%).The above results indicated that tetracyline-resistance of these APP strains was mediated by drug efflux.

Isolation and Identification of Aeromonas veronii from Micropterus salmoides and Pathological Lesions of Its Infection

DENG Longjun

2021, 50(1): 164-171. DOI: 10.15933/j.cnki.1004-3268.2021.01.021

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This study was conducted to investigate the bacterial pathogen that caused the death of Micropterus salmoides farmed in Sichuan Province.Pathogenetic bacterial was isolated from naturally infected Micropterus salmoides by using traditional methods, and the pathogenicity was determined by artificial infection test.The pathogenic isolate was identified by biochemical identification micro reagent tube and 16S rRNA sequence analysis.Its antibiotic susceptibility was tested by Kirby-Bauer discagar diffusion method,and the histopathological characterization of organs of the diseased Micropterus salmoides
was further observed by conventional paraffin section technique.The pathogenic strain MYLY01 was isolated from the diseased Micropterus salmoides and was identified as Aeromonas veronii based on phenotypic characteristics and 16S rRNA sequence analysis.It had an median lethal dose(LD₅₀) of 4.79×10⁸cfu/mL for healthy Micropterus salmoides,and showed the same clinical symptom as the diseased Micropterus salmoides.Drug sensitivity tested results showed that the MYLY01 was sensitive to cefradine,ceftazidime pentahydrate, gentamicin, neomycin and so on, but resistant to doxycycline, tetracycline,amoxicillin and so on.Clinical symptom of the diseased Micropterus salmoides showed ulcer in dorsum and side of the body,congestion and hemorrhage in gill,swelling in liver,spleen and kidney. Histopathological observation showed obvious degeneration,necrosis and inflammation in liver,spleen,kidney and muscles.The findings confirm that A.veronii is the cause of this epidemic which induces the body damages and even death because of the pathological changes in organs,and the neomycin will be used for the control of this disease in the future.

Screening and Application of Thermophilic Bacteria for Degrading Soybean Dregs

CHEN Xiaojia, LI Li, MA Xinxin, SHI Jiping, ZHAO Zhijun

2021, 50(1): 172-179. DOI: 10.15933/j.cnki.1004-3268.2021.01.022

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In order to degrade soybean dregs in high temperature,thermophilic bacteria were screened from soybean dregs, horse manure, pig manure and domestic sludge. The results showed that 7 thermophilic bacteria strain were gained,which could utilize soybean dregs to grow at 70℃ .Through 16S rDNA identification, they belonged to Geobacillus toebii, Geobacillus sp. , Methylobacterium sp.and Thermus thermophilus,respectively. Among them,Geobacillus toebii (G.T1) was the best for degrading soybean dregs(4.80%).The optimal growth temperature for strain G.T1 was 65℃ ,and optimal initial pH value was 7.5,the strain reached stable growth phase within 24 h.The strain G.T1 was inoculated to soybean dregs (including 5% soybean water sludge) and fermented at 70℃ for 5 d,the carbon-nitrogen ratio (C/N) of the experimental group decreased to 14.43 after 5 d of fermentation,which was lower than the blank(CK).Furthermore,the seed germination index of the experimental group was 135.00%,far higher than that of the CK (86.42%).In addition, as important nutritional indicators of organic fertilizers,the contents of nitrogen,phosphorus and potassium were 27.72,7.69 and 17.92 g/kg in the experimental group,respectively,which were 18.46%,25.04% and 22.74% higher than those in the CK.Therefore,strain G.T1 could not only promote the maturity process of soybean dregs,but also improve the contents of nitrogen,phosphorus and potassium nutrients.

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