Journal of Henan Agricultural Sciences

Progress on Application of MYB Transcription Factor in Plant Drought Tolerance Genetic Engineering

LI Junxia, DAI Shutao, CHEN Yuxiang, ZHU Cancan, QIN Na, SONG Yinghui, WANG Chunyi, RUI Zhanxu, LIANG Qiufang, LI Fu, WANG Shengxuan

2020, 49(11): 1-9. DOI: 10.15933/j.cnki.1004-3268.2020.11.001

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Drought seriously affects the growth,yield and quality of plant. Drought tolerance breeding is an important measure to ensure plant production. It is an effective way to improve plant drought tolerance through genetic engineering technology compared with the conventional breeding methods.MYB transcription factors are one of the largest transcription factor families in plant,which play an important role in regulation on plant tolerance to drought stress. This paper systematically and comprehensively elaborated the application of MYB transcription factors in plants(Arabidopsis thaliana, Nicotiana tabacum,Oryza sativa,Zea mays and Glycine max,etc) drought tolerance genetic engineering,so as to provide some references for the utilization of MYB transcription factors and drought tolerance genetic improvement and breeding.

Effects of Benzoylphenylurea Insecticides on Growth,Development and Reproduction of Scatopse sp.

SHEN Dengrong, LI Jiajia, HA Shalading, HE Chao, ZHANG Rui

2020, 49(11): 10-18. DOI: 10.15933/j.cnki.1004-3268.2020.11.002

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To determine the toxicity and toxic effects of benzoylphenylurea insecticides(BPUs) against Scatopse sp.,impregnation method was used to determine the effects of four BPUs,lufenuron,chlorfluazuron,flufenoxuron and hexaflumuron,on the toxicity, growth, development and reproduction of larvae stage.The results showed that the toxicity of lufenuron and chlorfluazuron to the 2nd instar larvae was relatively high,with LC₅₀ of 2.627 mg/L and 3.791 mg/L,respectively.The developmental duration of 2nd instar larvae and pupal stage were prolonged under the sublethal concentration of four BPUs.The 4th larva stage of chlorfluazuron and lufenuron under LC₄₀ treatment were extended by 2.31 d and 2.11 d respectively,compared with the control(use sterile water instead of liquid medicine).Under LC₄₀treatment,chlorfluazuron had the strongest inhibition effect on the pupation rate and eclosion rate of 2nd instar larvae,which were 60.56% and 72.56% respectively,and pupation rate and eclosion rate of lufenuron were 65.21% and 76.45% respectively. Chlorfluazuron and lufenuron had significant inhibition effect on the eggs laid per female,which decreased by 41.78% and 37.53% respectively,compared with the control.These results indicate that chlorfluazuron and lufenuron are ideal pesticides against Scatopse sp., the young larvae stage(before 2nd instar larvae) is the best time to use two kinds of BPUs,the inhibition effect of BPUs on the growth,development and fecundity of Scatopse sp can be gave full play.

Genome-Wide Identification of Maize GATA Gene Family and Expression Analysis under Heat Stress

WANG Yanzhao, ZHOU Bo, HAN Xiaohua, HUANG Bao, LU Xiaomin, CHENG Junling, WANG Shufeng, NIE Lihong

2020, 49(11): 19-25. DOI: 10.15933/j.cnki.1004-3268.2020.11.003

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The members of GATA gene family were identified, their physicochemical characteristics,chromosome distribution, phylogenetic evolution, gene structure and conserved motif were analyzed by bioinformatics,and their expression levels were analyzed under heat stress,so as to provide basis for function study of GATA genes.The results showed that a total of 37 GATA genes were identified,which were unevenly distributed on 10 chromosomes in maize genome, and harbored a conserved motif CX₂CX₁₈CX₂C.Phylogenetic tree analysis showed that the maize GATA gene family could be divided into four groups,A,B,C and D,containing 21,10,4 and 2 genes,respectively,and the genes in the same group had similar gene structure.Nine GATA genes including five group A genes( Zm00001d010785,Zm00001d023539, Zm00001d041883, Zm00001d025953 and Zm00001d031135), two group B genes(Zm00001d011771 and Zm00001d034751)and two group C genes(Zm00001d036494 and Zm00001d01 4656) were up-regulated,and two group B GATA genes(Zm00001d016361 and Zm00001d009193)were down-regulated under heat stress.This study suggested the above up-regulated nine GATA genes may play important roles in response to heat stress.

Effects of Returning Straw to Field through Cow’s Belly and Applying Nitrogen on Annual Yield of Wheat and Maize and Soil Physical and Chemical Properties

ZHAO Lingxiao, JIANG Li’na, MA Jianhui, WEI Jituo, ZHAO Dongyang

2020, 49(11): 26-36. DOI: 10.15933/j.cnki.1004-3268.2020.11.004

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The effects of straw returned to field directly and through cow’s belly combined application of nitrogen fertilizer for 3 years on annual yield of wheat and maize and soil physical and chemical properties were studied, so as to to provide theoretical reference for efficient utilization of straw and rational application of nitrogen fertilizer.The results showed that the annual yield of wheat and maize of the treatment with straw returned to the field through cow’s belly(applying dry cow drug 4 000 kg/ha) was higher than that of the N₁ treatment without nitrogen fertilizer and N8 treatments with 695 kg/ha nitrogen per year when the straw was directly returned to the field,and the N₂ treatment with maize straw returned to the field and 695 kg/ha nitrogen had the highest annual yield of wheat and maize,followed by N₃ treatment with maize straw returned to the field and 653 kg/ha nitrogen and N₄treatment with maize straw returned to the field and 625 kg/ha nitrogen.Under the condition of returning straw to the field through cow’s belly,the N₄ treatment at the mature stage of wheat and the N₇ treatment with wheat straw returned to the field and 625 kg/ha nitrogen at the mature period of maize had significantly increase of organic matter and total nitrogen contents in the 0—20 cm soil compared with N8 treatment.The treatment with straw returned to the field reduced the relative abundance of Actinomycetes and increased the relative abundance of Proteobacteria.Taken together,N₄ treatment had higher annual yield,although it was slightly lower than that of N₂ treatment,but the nitrogen application decreased by 70 kg/ha compared with N₂ treatment,the surface soil organic matter and nitrogen contents of N₄ treatment were the highest,and the improvement effect of N₄ treatment on soil bulk density and porosity was better,which was better than N₂ treatment.Therefore,N4 treatment was the best treatment under the conditions of this study.

Genome-Wide Identification and Expression Pattern of LOX Gene Family of Sorghum bicolor

LIU Meng, WEI Yulei, DING Dong, ZHAO Xunchao, LUO Weiyi, XU Jingyu

2020, 49(11): 37-44. DOI: 10.15933/j.cnki.1004-3268.2020.11.005

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Bioinformatics technology was used to identify lipoxygenase(LOX) family genes in sorghum,and their evolutionary relationship,gene structure,conservative motif and expression patterns in different tissues and under low temperature stress were analyzed,so as to lay a foundation for the functional study of sorghum LOX gene. The results showed that 11 sorghum LOX genes were identified,named SbLOX1—SbLOX11 according to their chromosomal positions,which could be divided into two subfamilies, and the gene structure of the same subgroup was basically similar.The gene expression profiles analysis of sorghum LOX family in various plant tissues revealed that expression patterns of different LOX genes varied. The expression levels of SbLOX10 and SbLOX9 were lower in most tissues,whereas the expression levels of SbLOX2—SbLOX5 and SbLOX8 were higher in almost all different tissues. The transcriptome data of LOX genes were analyzed under low temperature (4℃ ),the expression levels of SbLOX1,SbLOX3,SbLOX5 in 9-LOX subgroup and SbLOX4,SbLOX9 in 13-LOX subgroup of cold tolerant sorghum were upregulated,and which of cold sensitive sorghum were higher too, speculating that LOX gene had important role in response to low temperature stress.

Effects of Intercropping Green Manure on Growth of Flue-Cured Tobacco and Soil Enzyme Activities

LI Jinqiu, HE Guoqiang, WANG Jingying

2020, 49(11): 45-53. DOI: 10.15933/j.cnki.1004-3268.2020.11.006

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In order to explore the suitable fertilization model for the growth of flue-cured in Binxi tobacco region of Heilongjiang,using field experiment,Y1(intercropping rhinoceros+30% reduction in fertilization),Y2(intercropping rhinoceros+no fertilization),Y3(intercropping wheat+30% reduction in fertilization),Y4(intercropping wheat+no fertilization),Y5(not intercropping green manure+conventional fertilization),Y6(not intercropping green manure+30% reduction in fertilization) and CK(not intercropping green manure+no fertilization)treatments were set,and the effect of intercropping green manure and application of chemical fertilizers on agronomic traits,soil enzyme activity,potassium content in different parts of tobacco plant,and economic traits of flue-cured tobacco were studied.The results showed that,during the flattening period of tobacco,intercropping green manure+30% reduction in fertilization treatment had obvious increase effect in the number of leaves and maximum leaf area of flue-cured tobacco,which increased by 1.56%—3.47% and 1.58%—2.57% compared with Y5 treatment ,improved the potassium content in the stem and leaf parts of tobacco,but had no significant effect on the potassium content in the roots.For soil sucrase and catalase activities,intercropping green manure+30% reduction in fertilization was appropriate,but urease and phosphatase activities were better when only intercropping green manure.In addition,intercropping green manure+30% reduction in fertilization treatment could increase the economic traits of flue-cured tobacco,the quality of single leaf,orange smoke ratio,yield and output value,compared with Y5 treatment,the increases were 3.01%—3.28% (P>0.05),44.29%—131.72%(P<0.05),1.36%—4.58%(P>0.05) and 1.45%—9.82%(P>0.05).In summary,from the perspective of sustainable development of green agriculture,intercropping wheat or rhinoceros+30% reduction in fertilization is more suitable in the Binxi area of Heilongjiang.

Comparison of RNA Extraction Methods in Five Tissues of Fennel

WANG Xiaomin, WANG Lin, WANG Caiyan, PAN Bingqing, ZHOU Xuerong

2020, 49(11): 54-60. DOI: 10.15933/j.cnki.1004-3268.2020.11.007

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In order to screen the optimal extraction method of total RNA from each tissue of fennel,in this study,the fennel from Haiyuan in Ningxia was used as the material,modified hexadecyl trimethyl ammonium bromide(CTAB) method,CTAB water saturated phenol method,modified Trizol method and OMEGA plant RNA extraction kit (the kit method for short) were compared in the effects of total RNA extraction from roots,stems,leaves,flowers and seeds of fennel.The concentration,purity and integrity of total RNA from each tissue of fennel were determinated by nucleic acid and protein detector as well as agarose gel electrophoresis.The results showed that for fennel roots,steams,leaves and flowers,the quality of RNA extracted by the kit method was the best,the yield was higher,and the extraction time was short.The quality of RNA extracted by the modified CTAB method was better,the yield was high,but the extraction time was longer.The yield of RNA extracted by the modified Trizol method was higher,but the concentration and integrity were poor.The yield and quality of RNA extracted by the CTAB water saturated phenol method were lower. For fennel seeds,the effect of RNA extraction by the modified CTAB was the best.

Microbes Functional Diversity of Alfalfa Soil in the Rainfed Loess Plateau

CHENG Wenwen, LUO Zhuzhu, NIU Yining, CAI Xia

2020, 49(11): 61-70. DOI: 10.15933/j.cnki.1004-3268.2020.11.008

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To explore the response of soil microbial functional diversity on alfalfa planting years in the Loess Plateau,based on the long-term localization experiment(2002—2016) in the rainfed agricultural area of Loess Plateau,alfalfa plamting soils with three cultivated years(6,13 and 15 a) in Loess Plateau were selected with farmland as control,the influence of different alfalfa planting years on soil organic carbon content,microbial biomass carbon content and microbial functional diversity were studied.The results showed that compared with farmland,planting alfalfa could significantly increase soil microbial biomass carbon content. With the extension of the planting years of alfalfa,soil organic content and microbial biomass carbon contents showed an obvious trend of decline. Biolog eco plate TM analysis of the carbon source utilization ratio of the microbial community showed that the utilization of carbohydrates and amino acids in the alfalfa planting soils decreased with the deepening of the soil layer.At the soil depth of 0—30 cm and 30—60 cm,the average rate of color change of alfalfa planting soils for 6 a was significantly higher than that of farmland,13 a and 15 a after 120 h cultivation.There was no significant difference in soil microbial diversity index (H),evenness index (U) and carbon source richness index (S) at the soil depth of 0—30 cm;the H and S values of 6,13 a of 30—60 cm were significantly different from farmland.While carbohydrates and amino acids are well utilized by soil microorganisms in alfalfa planting soils,carbohydrates and carboxylic acids are mainly used by farmland soil microorganisms.The principal component analysis showed that the utilization characteristics of microbial carbon sources in alfalfa planting and farmland soils were different,and the preferences of different carbon sources of them were different.

Improvement Effect of Feed Rapeseed on Saline-alkali Soil

FAN Qianyu, LI Jin, LIU Zhenhua, HUANG Chunguo, YANG Zhenping, GAO Zhiqiang

2020, 49(11): 71-78. DOI: 10.15933/j.cnki.1004-3268.2020.11.009

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In order to reveal the regularity of salinity degree and nutrient change of soil in saline-alkali land after planting feed rapeseed,an experiment was carried out in saline-alkali land in Yingxian,Shanxi Province.The effects of feed rapeseed on soil saline-alkali degree,soil nutrients and mineral elements contents in different saline-alkali soils were studied under the same conditions. Soil samples were collected before sowing,plant samples were collected at the early flowering stage,and pH value,total salt,nutrient and mineral element contents were determined.The results showed that feed rapeseed could reduce the pH value,total salt and Na⁺contents of saline-alkali soil,and Na⁺was mainly concentrated in the root system of feed rapeseed.The contents of available phosphorus and available potassium in saline-alkali soil could be improved to a certain extent by planting feed rapeseed,but there was no obvious advantage in the increase of content of soil organic matter in the same year after planting feed rapeseed(harvest at early flowering).After planting feed rapeseed,the content of available Fe in soil increased significantly,while the contents of available Ca,Mg,B and Mn decreased significantly.In conclusion,feed rapeseed has certain effects on soil improvement in saline-alkali land,and can improve soil fertility and soil environment.

Distribution of Microfilament in Phaneroplasmodium of Myxomycetes

CHANG Yuting, REN Qiyuan, LI Yanshuang, WANG Xiaoli, LI Yu

2020, 49(11): 79-84. DOI: 10.15933/j.cnki.1004-3268.2020.11.010

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In order to study the state of existence of microfilaments in phaneroplasmodium of Fuligo septica and realize the purpose of studying phaneroplasmodium from the nucleus level,through the cultivation and the fluorescence staining of phaneroplasmodium of Fuligo septica,it was found that there was microfilament existed in plasmodium.In further research,it was found that the microfilament presented the border of gel-like ectoplasm and inner more fluid endoplasm of veins of plasmodium,which provided strong evidence for microfilament participated in the protoplasm movement in myxomycetes.It is concluded that the microfilament may be an important component of the cluster network in the phaneroplasmodium.

Effects of Benzoylphenylurea Insecticides on Growth,Development and Reproduction of Scatopse sp.#br#

SHEN Dengrong, LI Jiajia, HA Shalading, HE Chao, ZHANG Rui

2020, 49(11): 85-90. DOI: 10.15933/j.cnki.1004-3268.2020.11.011

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To determine the toxicity and toxic effects of benzoylphenylurea insecticides(BPUs) against Scatopse sp.,impregnation method was used to determine the effects of four BPUs,lufenuron,chlorfluazuron,flufenoxuron and hexaflumuron,on the toxicity, growth, development and reproduction of larvae stage.The results showed that the toxicity of lufenuron and chlorfluazuron to the 2nd instar larvae was relatively high,with LC₅₀of 2.627 mg/L and 3.791 mg/L,respectively.The developmental duration of 2nd instar larvae and pupal stage were prolonged under the sublethal concentration of four BPUs.The 4th larva stage of chlorfluazuron and lufenuron under LC₄₀ treatment were extended by 2.31 d and 2.11 d respectively,compared with the control(use sterile water instead of liquid medicine). Under LC40 treatment,chlorfluazuron had the strongest inhibition effect on the pupation rate and eclosion rate of 2nd instar larvae,which were 60.56% and 72.56% respectively,and pupation rate and eclosion rate of lufenuron were 65.21% and 76.45% respectively. Chlorfluazuron and lufenuron had significant inhibition effect on the eggs laid per female,which decreased by 41.78% and 37.53% respectively,compared with the control.These results indicate that chlorfluazuron and lufenuron are ideal pesticides against Scatopse sp., the young larvae stage (before 2nd instar larvae) is the best time to use two kinds of BPUs,the inhibition effect of BPUs on the growth,development and fecundity of Scatopse sp can be gave full play.

Effects of Biomaterial Carbon Application on Fusarium Wilt of Watermelon Seedlings and Its Mechanism#br#

SHU Xiuyu

2020, 49(11): 91-97. DOI: 10.15933/j.cnki.1004-3268.2020.11.012

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This paper used watermelon variety Qianli No. 1 as experimental material,set four biomaterial carbon application levels of 0%(CK),0. 5%(T1),1%(T2) and 2%(T3),and studies the effects of biomaterial carbon application rates on Fusarium wilt of watermelon seedlings and its mechanism.The results showed that biomaterial carbon could significantly reduce the disease index of watermelon Fusarium wilt and improve the control effect. Compared with CK,on the 20th day after inoculation,the disease index of watermelon seedlings treated with T1, T2 and T3 significantly decreased by 10.02%, 19.61% and 29.62% respectively,and the control effects increased by 48.83%,62.66% and 77.25% respectively(P<0.05). At the same time,with the increase of biomaterial carbon application rate,the Fusarium wilt disease index,the number of fungi,the number of Fusarium oxysporum and the content of MDA in watermelon seedlings decreased gradually,while the control effect of Fusarium wilt,biomass of watermelon seedlings,total soil microorganisms number,bacteria number,actinomycetes number,bacteria/fungi,actinomycetes/fungi,protective enzymes(superoxide dismutase, peroxidase, catalase and phenylalnine ammonialyase)and the activity of disease resistance related enzymes(chitinase and β-1,3-glucanase) increased gradually.In conclusion,the external application of biomaterial carbon could effectively improve soil microbial community structure,reduce the number of Fusarium oxysporum,promote the growth of watermelon seedlings,increase the activities of protective enzymes,and disease resistance related enzymes,so as to enhance the resistance of watermelon to Fusarium wilt.

Effect of Reductive Soil Disinfestation on Growth,Yield,Diseases and Insect Pests of Three Continuous Cultivation Vegetables in Plastic Greenhouse#br#

GUO Chenxi, ZHOU Guifang, CHEN Bihua, GUO Weili, CAI Zucong, HUANG Xinqi, WANG Guangyin

2020, 49(11): 98-109. DOI: 10.15933/j.cnki.1004-3268.2020.11.013

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In order to explore field application effect of reductive soil disinfestation(RSD)method,a plas tic greenhouse soil of 30 years’ continuous cropping in Xinxiang of Henan Province was selected as the research object,and one RSD treatment plastic greenhouse(adding organic matter 19 500 kg/ha) and a control greenhouse(CK,without adding any treatment),in which the RSD treatment plastic greenhouse was set with two areas of “RSD treatment” and “RSD+968 bio-bacterial fertilizer”,the changes of soil physical and chemical properties and biological properties after RSD treatment were measured,and the
growth,early yield and occurrence of diseases and insect pests of tomato or cucumber were investigated and counted for three consecutive crops.The results showed that compared with CK,Fusarium oxysporum number of RSD treatment decreased 98.95%,and the number of root-knot nematodes per 100 g dry soil decreased from 3 297.33 to 41.00.Compared with CK,RSD treatment could promote the growth and strengthen the first autumn tomato and the second spring cucumber,RSD treatment and RSD+968 treatment increased the first autumn tomato first fruit cluster yield by 73.30% and 109.59% respectively,and increased three fruit yield of second stubble spring cucumber by 7.14% and 21.43% respectively. Compared with CK,the disease rate of stem base rot of RSD and RSD+968 treatments of first stubble autumn tomato reduced by 28.64% and 40.41% in plastic greenhouse respectively,and the control effect on rootknot nematode in first stubble autumn tomato was 91.11% and 100% respectively.Meanwhile,RSD treatment greatly delayed occurrence of Fusarium wilt of second stubble spring cucumber in the plastic greenhouse.The disease rate of cucumber fusarium wilt was 100% in CK greenhouse,whereas the disease rate of cucumber fusarium wilt of RSD and RSD+968 treatments were only 10.61% and 3.82% respectively.In additoin,the disease rate of stem base rot in the third cropping autumn tomato with RSD treatments was 57.84% lower than that of CK,and the control effect of RSD treatments on root-knot nematode was still
87. 10%. It can be seen that RSD treatment has long-term effectiveness and RSD+968 treatments have additive
effect. Therefore,the RSD treatment of solid organic materials can significantly improve soil quality,reduce disease rate and increase yield of continuous cropping system in plastic greenhouse.This is an effective measure to alleviate the continuous cropping obstacle of vegetable cultivation and realize sustainable cropping.

Establishment of Paeonia ostii ‘Fengdan’ Embryo Culture System and Analysis of Methylation Variation in Malformed Embryo Culture Seedlings#br#

LIAN Xiaofang, LI Yuying, ZHANG Wanqing, GUO Lili, ZHANG Youfu, HOU Xiaogai

2020, 49(11): 110-119. DOI: 10.15933/j.cnki.1004-3268.2020.11.014

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In order to improve the application efficiency of oil peony Paeonia ostii ‘Fengdan’ tissue culture technology in production practice,the experiment was carried out with tree peony seed embryo as material.Different light and temperature culture conditions,different sizes of P.ostii ‘Fengdan’ mature seed embryos,different plant growth regulator ratios and different concentrations of additives were set to perfect the P.ostii ‘Fengdan’ mature seed embryo culture system.Methylation-sensitive polymorphism technology(MSAP) was used to analyze the methylation level and pattern variation of malformed embryo culture seedlings that appeared during the cultivation process.The results showed that the optimal culture conditions in the test range were dark culture for 7 d after inoculation and then switched to light culture.The suitable culture temperature was (25±1)℃ .Suitable explants were mature embryos ≥3 mm.The plant growth regulator ratio was 0.8 mg/L NAA combined with 1.0 mg/L 6-BA,and the basic medium was modified MS(double Ca²⁺).Compared with no exogenous additives,the addition of P.ostii ‘Fengdan’endosperm had a significant inhibitory effect on the seedling rate;75 mL/L coconut milk treatment had better effect with seedling rate of 85.66%;1.0 g/L activated carbon treatment significantly had the highest
seedling rate of 90. 46%. Compared with normal primary cultured seedlings,the total amplification sites number of malformed seedlings increased by 21.20%,and the total methylation sites number decreased by 12.97%. The total methylation rate of normal tissue cultured seedlings was 85.38%,and the total methylation rate of malformed seedlings was 74.30%.The variation of malformed seedlings methylation dominated by polymorphisms(82.37%),among which demethylation variation accounted for the largest proportion of 46.45%.The results of comprehensive analysis showed that the optimal culture protocol for aseptic culture of P. ostii ‘Fengdan’ seed embryos were modified MS(double Ca²⁺)+0.8 mg/L NAA +1.0 mg/L 6-BA+1.0 g/L activated carbon(or 75 mL/L coconut milk).The culture conditions was dark culture for 7 d and then switch to light culture,temperature conditions was (25±1)℃ .The total methylation rate of P.ostii ‘Fengdan’ malformed embryo culture seedlings was lower than that of normal seedlings,the variation of malformed seedlings demethylation dominated by polymorphisms.

Effect of Boron on Growth and Resistance Related Indicators of Red Globe Grape Seedlings

YANG Yang, TAN Yuchao, CHANG Ruizhe, REN Fengshan, JIANG Xilong, WU Xinying

2020, 49(11): 120-127. DOI: 10.15933/j.cnki.1004-3268.2020.11.015

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In order to study the effect of boron on the growth and resistance related indicators of grapes,we chosed one-year-old Red Globe grape seedlings as test materials,and treated them by nutrient solution containing 0,0. 2,3. 1,6. 2 mg/L boron respectively.After 121 days of treatment,we analyzed the physiological and biochemical indicators of grape seedlings under different boron concentrations.Those indicators included plant growth,contents of boron,soluble sugar,starch,protein,proline and malondialdehyde,activities of SOD,PAX,POD and CAT and so on.The results showed that there were significant differences in the growth of grape seedlings treated with different concentrations of boron.When the concentration of boron was 3. 1 mg/ L,the new shoot growth and biomass accumulation were maximum.The boron content of grape leaves and roots increased with the increase of exogenous boron concentration.For the treatment of different boron,the boron contents in leaves and roots were 448.0 mg/kg and 75.3 mg/kg,respectively. The results showed that the shoot growth,root soluble sugar content,leaf soluble protein content,leaf proline content,leaf SOD activity and leaf CAT activity were the lowest,root starch content,leaf MDA content and POD activity were the highest,with the increase of exogenous boron concentration,root soluble sugar content,leaf soluble protein content,proline content,and leaf CAT activity increased,starch content in roots and POD activity in leaves decreased,SOD and APX activities increased first and then decreased,and malondialdehyde content decreased first and then increased. High boron(6.2 mg/L) treatment had higher root soluble sugar content,leaf soluble protein content,leaf proline content and CAT activity.There were significant positive correlations between the concentration of exogenous boron and the concentration of boron in grape leaves and roots(r=0.950 1,r=0.989 4),significant negative correlation between exogenous boron concentration and starch content in root system(r=-0.978 2),positive correlation between exogenous boron concentration and soluble protein content in leaves(r=0.923 3),negative correlation between exogenous boron concentration and POD activity in leaves(r=-0. 849 6),while not significant.Appropriate amount of exogenous boron promoted the growth of grape seedlings,no boron(0 mg/L)and high boron(6.2 mg/L) treatments affected the growth and resistance related physiological and biochemical indexes of grape seedlings.

Isolation and Phylogenetic Analysis of Fowl Adenovirus Serotype 4 Isolate ZZ

JIN Qianyue, WANG Yinbiao, CHAI Yongxiao, LU Qingxia, LI Peng, GUO Zhenhua, XING Guangxu, DENG Ruiguang, ZHANG Gaiping

2020, 49(11): 128-133. DOI: 10.15933/j.cnki.1004-3268.2020.11.016

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In order to study the genomic characteristics and molecular evolution of FAdV-4 strains circulated in Henan Province,a FAdV-4 strain,named FAdV-4 ZZ,was isolated from HPS-affected chicken and subject to whole genome sequencing and phylogenetic analysis.It was found that FAdV-4 ZZ could stably propagate on LMH cells(chicken hepatoma cell) and produce obvious cytopathic effects.Compared with the non-pathogenic strains FAdV-4 ON1 and FAdV-4 KR5,ORF19,ORF27 and ORF30 genes were absent in FAdV-4 ZZ,CH/JSXZ/2015,JSJ13 and SDSX1 circulated in different areas of China.Compared with the ORF29 of first isolated JSJ13 strain,the ORF29 sequence of FAdV-4 ZZ had 33 base pairs deletion.The identities of Hexon genes between FAdV-4 ZZ and CH/JSXZ/2015,SDSX at the nucleotide level were all 100%,and were 98. 69% and 98. 90% between FAdV-4 ZZ and strains ON1,KR5,respectively.The current study successfully isolated FAdV-4 ZZ strain,obtained its whole genome sequence and established its phylogenetic relationship with other FAdV strains at home and abroad.

Study on PCV2 Prevalence in a Pig Farm through Detection of Swine Oral Fluid

GUO Zhenhua, RUAN Haiyu, QIAO Songlin, ZHANG Gaiping

2020, 49(11): 134-140. DOI: 10.15933/j.cnki.1004-3268.2020.11.017

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To explore the application of swine oral fluids in porcine circovirus type 2(PCV2) detection,PCV2 infectious status with oral fluids were evaluated in a large-scale pig farm in the north of Henan.The results showed that the sample rate of 28 oral fluid samples collected from 560 pigs was about 20.0%,while the 70 serum samples were only 2.5%.Importantly,the detection rate of PCV2 was 100.0%(28/28) in oral fluid samples and only 2.86%(2/70) in serum samples with PCR tests.To further identify the genotype of PCV2 in this farm,6 PCV2 genomes were successfully sequenced.The analysis of nucleotide identity showed that the PCV2 strains shared about 97.0%,95.4%,93.3%,94.4% and 91.6% identities with representative strains of PCV2a,PCV2b,PCV2c,PCV2d and PCV2e,respectively.The phylogenetic analysis showed that the isolates were clustered into the same branch with PCV2a.These results suggested that the genotype of PCV2 strains in this pig farm was PCV2a.

Detection of Aminoglycoside Antibiotic Resistance Genes in Pig Infectious Porcine Contagious Pleuropneumonia Actinobacillus pleuropneumoniae Isolated from Pigs

LI Haili, FENG Lili, WANG Yinghua, XU Yindi, ZHANG Qingxian, WANG Zhifang, ZHU Wenhao, XU Feng, YOU Yi, WANG Keling

2020, 49(11): 141-146. DOI: 10.15933/j.cnki.1004-3268.2020.11.018

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In order to study the aminoglycoside antibiotic resistance of pig infectious porcine contagious pleuropneumonia Actinobacillus pleuropneumoniae(APP) in Henan Province and neighboring regions in the scale of swine.A drug sensitivity test was carried out in 85 APP strains isolated from lung and trachea pigs in 56 large scale pig farms in Henan Province.The serotype identification results showed that isolates were 1,3,7,8,9,10,12,15.Antimicrobial susceptibility disk method were used for resistant phenotype identification.At the same time,the PCR method for detection of aminoglycoside antibiotic resistance genes(ant2-Ⅰ a,aac3-Ⅱ c and aac3-Ⅳ).The drug resistance rates of APP to aminoglycoside antibiotic drug resistance genes(aac3-Ⅱ c)and(aac3-Ⅳ) were 57.6% (49/85) and 81.2% (69/85),respectively.The ant2-Ⅰ a resistance genes were not detected.

Analysis of SSR Site Information in Transcriptome of Oreochromis aurea Based on RNA-seq

2020, 49(11): 147-152. DOI: 10.15933/j.cnki.1004-3268.2020.11.019

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In order to develop molecular markers related to salt tolerance in Oreochromis aurea,the nextgeneration sequencing RNA-seq method was used to mine microsatellite tags in O.aurea.The results showed that a total of 71 009 unigenes were obtained by splicing and assembly,and a total of 10 875 SSR loci were found in all sequences.The number of sequences contained SSR loci was 8 315,accounting for 11.71% of the total sequences.There were many types of SSR loci in the transcriptome of O.aurea,and 5 different nucleotide repeat types and 126 different repeat motifs were recognized.Among them,the content of the dinucleotide tandem repeat unit type was the most,with 5 779,accounting for 53.14% of the total number of sites.And among dinucleotide repeat types AC/GT the motif type had the highest frequency(3 954 times,36.36%).In addition,SSR loci were dominated by 6 repetitions(3 048),accounting for 27.77% of the total number of loci;SSR loci sequence length was 12—254 bp,of which 3 480 loci were highly polymorphic(29.14%),and 8 462 loci were moderate polymorphic(70.86%).

Effects of Insect Hormone on ALP Activity and Its Gene Expression Level in Bombyx mori

TANG Fenfen, YANG Weike

2020, 49(11): 153-158. DOI: 10.15933/j.cnki.1004-3268.2020.11.020

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In order to explore the effects of insect hormones on the alkaline phosphatase(ALP) activity and its gene expression level in Bombyx mori,using the 3rd day of the fifth instar silkworm(Dazao) as test materials,the ALP gene expression level and enzyme activity were detected in the hemolymph and the midgut at different time after injecting 20E and JH.The results showed that the ALP expression level was significantly up-regulated at 6 h,12 h and 24 h after 20E or JH treatment in the hemolymph of Bombyx mori,but it was up-regulated only at 12 h and 24 h in the midgut. In the 20E treatment group,the ALP relative expression level was the highest at 6 h in the hemolymph,which was 2.3 times of the control group;but it was the highest at 12 h in the midgut,which was 2.8 times of the control group.After JH treatment,the ALP expression level in the hemolymph and midgut were the highest at 24 h,which were 3.7 times and 4.7 times of the control group respectively. In addition,the ALP enzyme activities were significantly activated in the hemolymph and midgut by 20E and JH.The hemolymph ALP enzyme activity was significantly higher than the control group at 6 h,12 h and 24 h after 20E treatment,and the highest activity at 12 h was 33.34 U/mg;While the activity of ALP in the midgut were significantly higher than the control group only at 12 h and 24 h,their activities were 37.68 U/mg and 40.95 U/mg respectively.Meanwhile,the ALP activity were also significantly higher in the hemolymph and midgut at 6 h,12 h and 24 h after JH treatment,among them,the ALP activity in the hemolymph had the highest value of 42.29 U/mg at 6 h,while the enzyme activity was rosed to the highest value of 46. 84 U/mg at 12 h in the midgut.The above results indicated that the ALP expression level was regulated by 20E and JH in Bombyx mori,and the effects of hormones on the gene expression level were consistent with the change rule of enzyme activity.

Detection of Regulatory Genes in Genetically Modified Crops Using TaqMan Probe Based on Real-Time PCR

LUO Jianxing, HAI Xiao, LIU Guoqiang, QILE Muge, GUO Liang

2020, 49(11): 159-165. DOI: 10.15933/j.cnki.1004-3268.2020.11.021

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In order to detect the genetically modified plant components in food,three positive plasmids ppCaMV35S,ptNOS and ptCaMV35S as positive test controls were independently constructed,then two endogenous genes (pUbi,pRice-actin) and three regulatory genes(pCaMV35S,tNOS and tCaMV35S)were selected to detect the genetically modified components in four genetically modified crops (corn,rice,cotton and soybean) and two non-genetically modified crops(corn and wheat) using TaqMan real-time PCR.The result of specific test showed that the constructed positive plasmid could meet the actual detection needs, endogenous genes and other regulatory genes had good specificity.The limit of detection(LOD)of pCaMV35S and tNOS in transgenic cotton samples were 1 and 10 ng,respectively,which could effectively screen and test genetically modified crops and products that were commercialized or in the intermediate test stage.

Numerical Simulation and Experimental Study on Hot Air Drying Process of Wheat

GENG Tie, WANG Shiqi, DUAN Erya, SHENG Jie, YUAN Yutao

2020, 49(11): 166-171. DOI: 10.15933/j.cnki.1004-3268.2020.11.022

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Drying is an important means of grain storage,so optimizing the drying process can make the drying process more scientific and energy-saving.The mathematical model of moisture and heat transfer in the process of wheat drying was established in this paper. Based on the UDF function of fluent,the corresponding self-defined program was compiled to simulate and verify the drying process of wheat in the small vertical mixed flow dryer.The results showed that the calculated results of the average humidity of wheat and the average temperature of wheat region were basically consistent with the experimental results;the humidity of wheat decreased with the increase of the drying wind speed,and the drying quality of wheat was better and the energy consumption was less when the drying wind speed was 8—10 m/s.It was proved that the model and calculation program in this paper were effective.

Winter Wheat Agrometeorological Drought Zoning and Analysis in Henan Province Based on Grid and Fuzzy Clustering Algorithm

LI Shimin, ZHANG Hongli, WANG Laigang, ZHENG Guoqing, GUO Yan, GAO Jianhua

2020, 49(11): 172-180. DOI: 10.15933/j.cnki.1004-3268.2020.11.023

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110 ground observations of the precipitation distance percentage in Henan Province were applied to evaluate the agrometeorological drought zoning and risk probability analysis for winter wheat with fuzzy clustering algorithm based on grid. Indicators of spatial association indices-Moran scatter plots and LISA cluster map were adopted to conduct to the spatial exploratory data analysis.The results indicated that 17 counties(cities) of Jiyuan,Puyang and Shangqiu and so on fell into the first quadrant with “high high”;12 counties(cities) of Xinyang,Zhumadian and Kaifeng and other regions fell into the third quadrant with “ low-low”,where these regions existed strong positive correlations with more precipitation;6 counties(cities) of Xinyang,Zhumadian and Jiaozuo,Qixian,Huaiyang fell into the fourth quadrant with“high-low” and the second quadrant “low-high” respectively, where these regions existed strong negative correlations.It could be concluded that the correlations of spatial variation of precipitation among counties was small.Then,fuzzy k means was used to divide the precipitation distance percentage data into five classes,while the fuzzy performance index(FPI) and normalized entropy(NCE) were the smallest.There existed significance between different zones at the 0.05 level by One-Way ANONA test.According to the distribution maps,the spatial and differential laws of drought could happen in Henan.The south-west of Xinyang and Nanyang belonged to the humid area with lower drought risk probability;Sanmenxia,parts of Puyang,Anyang and Kaifeng counties belonged to the region of severe drought with higher drought risk probability;most areas of Zhoukou and Luohe belonged to the drought-stricken area.The high risk probability was mainly distributed in the western mountainous areas such as Sanmenxia,Nanyang and other western mountainous areas and Xuchang and Zhengzhou.The low risk probability value areas were mainly in the rice growing areas such as northeast and Xinyang.This was consistent with the spatial distribution of the drought, but there were also differences in some regions,such as the high level of drought in parts of
Puyang,but low risk probability value.

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