Journal of Henan Agricultural Sciences

CONTENTS

2024, 53(9): 0.

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Identification and Evaluation of Resistance to Stripe Rust of 480 Wheat Germplasms

DAI Ziju, LI Wenxu, YANG Huimin, ZHU Xiuhua, WANG Yahuan, XU Fuxin, LIU Dongyang, HOU Jinna, QIN Maomao, WU Zhengqing, ZHOU Zhengfu, LEI Zhensheng

2024, 53(9): 1-15. DOI: 10.15933/j.cnki.1004-3268.2024.09.001

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The main purpose of the present study is to understand the resistance to stripe rust and the utilization of stripe rust resistant genes in current bred wheat varieties,and find the varieties containing novel stripe rust resistant loci.Totally,480 domestic and foreign wheat germplasms were collected from various planting regions.The varieties were inoculated with mixed races of CYR32,CYR33 and CYR34 for testing the adult plant resistance.Simultaneously,the resistance to leaf rust and powdery mildew was investigated in infected field.The KASP markers closely linked or co‑segregated with stripe rust,leaf rust and powdery mildew genes,Yr18/Lr34/Sr57/Pm38(7DS),Yr29/Lr46/Pm39/Sr58(1BL)and Yr30/Lr27/Sr2/Sb3(3BS),were selected for identifying the previous reported pleiotropic resistant genes.The results showed that there were 35(accounting for 7.29%)germplasms showed immunization to CYR32,CYR33 and CYR34,while the numbers of high‑resistant and medium‑resistant germplasms were 69(14.38%)and 79(16.46%),respectively.And the other 297(61.88%)germplasms demonstrated medium‑ or high‑ susceptible to the inoculated races.There were 13,10 and 7 germplasms containing the Yr18,and Yr29 and Yr30 genes,respectively;only 3 germplasms contained two of the resistant genes;no germplasm was detected with all the three resistant genes.In conclusion,183(accounting for 38.12%)germplasms showed resistant to stripe rust at adult plant stage,and 27 germplasms were detected with the above resistant genes.Yr18 or Yr30 gene showed resistant to the current races of stripe rust,however,the resistance level of Yr29 gene was partly lost at adult plant stage.

Effect of Nitrogen and Zinc Application on Filling Characteristics of Grains at Different Positions of Maize Ear

ZHANG Panpan, LI Chuan, ZHANG Meiwei, YU Xiaohong, MU Weilin, GUO Hanxiao, QIAO Jiangfang, ZHANG Guitang

2024, 53(9): 16-27. DOI: 10.15933/j.cnki.1004-3268.2024.09.002

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The high Zn efficiency variety Zhengdan 958(ZD958)and Zn no‑sensitive variety Longping 638(LP638)were used as materials.The field plot experiment was conducted with three N rates[225 kg/ha(HN),180 kg/ha(MN)and 90 kg/ha(LN)]and two foliar Zn treatments［no Zn(F0),spraying Zn of 1∶1 at jointing stage and flare opening stage(F1)],and the effects of N and Zn application on the filling characteristics of grains at different positions and their relationship with grain yield of different Zn efficiency varieties were studied,so as to provide reference for N and Zn application in the maize production.The results showed that HN treatment had the highest yield,followed by MN treatment,and there was no significant difference between HN treatment and MN treatment.HN treatment increased grain weight at different positions.MN treatment delayed the time with the maximum grain‑filling rate(T_Gmax)at different positions,and increased the maximum grain‑filling rate(Gmax)at upper position and active grain‑filling period(D)at basal position of ZD958 ear,while HN treatment delayed T_Gmax,prolonged D of different positions,and enhanced the grain weight with the maximum grain‑filling rate(W_Gmax)at upper and basal positions of LP638 ear,thus increasing grain yield.Compared with F0 treatment,F1 treatment significantly decreased the bare tip length by 9.9%,significantly increased the yield by 2.0%,increased the grain weight,extended D and postponed TGmax at upper and basal positions of ZD958 ear,and increased W_Gmax.The grain yield was significantly positively correlated with TGmax at basal position of ear,and the grain weight at middle position of ear was extremely significantly positively correlated with the grain weight at basal position of ear.To sum up,applying 180 kg/ha N and spraying 4.5 kg/ha Zn of 1∶1 at jointing stage and flare opening stage can delay T_Gmax at upper and basal positions of ear，extend D,and increase W_Gmax and grain weight of high Zn efficiency maize genotype,thus promoting the increase of yield.

Effects of One‑off Basal Application of Different Slow Controlled Release Urea on Yield and Nitrogen Utilization of Sorghum

LIANG Xiaohong, CAO Xiong, ZHANG Ruidong, HUANG Minjia, LIU Jing, NAN Huailin, WANG Songyu

2024, 53(9): 28-36. DOI: 10.15933/j.cnki.1004-3268.2024.09.003

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Taking mechanized sorghum variety Jinza 34 as material，under the condition of an equal nitrogen application rate of 150 kg/ha，six treatments were set up，which were no application of nitrogen fertilizer(CK),application of common urea(C1),application of slow release urea(C2),application of sulphur coated urea(C3),application of Junengwang urea(C4),and application of humic acid urea(C5).The effects of the one‑off basal application of different slow controlled release urea on agronomic traits,yield,nitrogen utilization efficiency and soil nitrate nitrogen accumulation were studied to provide a theoretical basis for improving nitrogen use efficiency of the one‑off basal nitrogen application and reducing soil environmental pollution.The results showed that compared with CK,the ear length,grain weight per ear,1 000‑grain weight and grain yield of sorghum of urea application treatments significantly increased by 8.32%—13.71%,21.05%—42.44%,5.24%—10.32%,16.08%—24.74%,and effective ear number increased by 5.09%—18.38%,respectively.The increases of sorghum yield of urea application treatments were largely due to the increases of grain weight per ear by path analysis.Compared to the C1 treatment,C3 and C4 treatments significantly increased grain weight per ear by 17.25% and 6.25%,1 000‑grain weight by 4.34% and 4.83%,grain yield by 7.46% and 5.84%,and aboveground nitrogen accumulation by 17.99% and 10.00%,respectively;increased nitrogen fertilizer use efficiency(NUE)by 88.03% and 48.89%,agronomic efficiency of nitrogen fertilizer(NAE)by 53.93% and 42.15%,partial productivity of nitrogen fertilizer(NPFP) by 7.47% and 5.84%,and nitrogen uptake efficiency(NUPE)by 18.00% and 10.00%,respectively.The grain weight per ear,nitrogen accumulation in grain,aboveground nitrogen accumulation,NUE and NUPE of C3 treatment significantly increased by 10.35%,7.24%,7.27%,26.88% and 7.27% compared with C4 treatment,respectively.Meanwhile,the nitrate nitrogen accumulation in the 0—60 cm soil layer of C3 treatment significantly decreased by 8.65%—31.30% compared with the other urea application treatments,and the residual nitrate nitrogen was mainly distributed in 0—40 cm soil layer.To sum up,under the conditions of this experiment,one‑off basal application of sulphur coated urea can significantly promote the nitrogen absorption of grains and aboveground part of sorghum,increase nitrogen fertilizer utilization efficiency and yield of sorghum,effectively decrease the accumulation of nitrate nitrogen in soil,reduce the risk of nitrate nitrogen washing to the deep soil layer,and is a suitable nitrogen fertilizer application method for promotion.

Isolation，Identification，and Functional Analysis of Culturable Microorganisms in Cigar Tobacco Leaves from Various Regions

SI Hongyang, YANG Weili, CUI Bing, LAI Miao, ZHAO Mingqin

2024, 53(9): 37-45. DOI: 10.15933/j.cnki.1004-3268.2024.09.004

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To clarify the species and functionality of culturable microorganisms in domestic cigar tobacco leaves,cured cigar tobacco leaves from Wuzhishan,Hainan Province(variety Haiyan 103)and and from Dazhou，Sichuan Province(variety Chuanxue No.1) were used as experimental materials.The culture‑dependent method were employed to isolate microorganisms,which were subsequently identified through molecular sequencing.The enzyme and aromatic substances production abilities of the isolated microorganisms were investigated.The results showed the isolation of 52 different bacterial strains from Haiyan 103 and Chuanxue No.1 cigar leaves,belonging to five different families:Bacillaceae,Planococcaceae,Erwiniaceae,Enterobacteriales,and Moraxellaceae.Bacillus subtilis E‑11,isolated from Hainan tobacco,exhibited the highest protease and pectinase activities of 85.02 U/mL and 256.71 U/mL,respectively.Additionally,four yeast strains were isolated from Hainan tobacco leaves,namely one strain from Pichia,one strain from Rhodotorula,and two strains from Debaryomyces.These yeast strains did not produce extracellular protease and pectinase,and Pichia sp.J‑2 demonstrated the ability to metabolize 2‑phenylethanol.The highest yield of 2‑phenylethanol,333.81 mg/L,was observed after 72 h of fermentation.In conclusion,cigar tobacco contains a rich variety of culturable microbial resources.Bacillus subtilis E‑11 and Pichia sp.J‑2,isolated from Haiyan 103,show promising potential for degrading macromolecular substances and enhancing the aroma content in tobacco leaves.

Identification of HSF Gene Family in Eucommia ulmoides and Its Expression Analysis under Adverse Stresses

WU Shujuan, XU Feng, WANG Guangan, WANG Chen

2024, 53(9): 46-56. DOI: 10.15933/j.cnki.1004-3268.2024.09.005

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Heat shock transcription factors（HSFs）play important roles in plant growth and development and in response to abiotic stresses.In order to understand the information of HSF gene family members in Eucommia ulmoides and reveal the structural characteristics and expression patterns of EuHSFs,this study analyzed the physicochemical properties,protein structure,phylogeny,gene structure,conserved domains,promoter cis‑acting elements,and the expression patterns of EuHSF genes in different developmental periods and abiotic stresses by bioinformatics methods and qRT‑PCR.The results showed that a total of 21 EuHSF genes were identified from Eucommia ulmoides,which differed in protein properties,with amino acid numbers,protein theoretical molecular masses,isoelectric points and instability coefficients ranging from 68—369,7.72—42.06 ku,4.31—9.22,and 7.05—67.28,respectively,and mainly consisted of acidic,hydrophilic,and unstable nuclear proteins.Phylogeneticanalysis showed that EuHSFs were divided into three subgroups,including ClassⅠ(1 EuHSF),ClassⅡ(7 EuHSFs)and ClassⅢ(13 EuHSFs),and promoter cis‑acting element analysis revealed that a large number of light‑responsive and hormone‑responsive elements in EuHSF genes.In addition,EuHSF genes were expressed at different developmental periods in Eucommia ulmoides leaves,but there were significant differences in the expression patterns. qRT‑PCR revealed that all EuHSFs responded to different abiotic stresses(high temperature,low temperature,high salinity,and drought),e.g.,most of the EuHSFs showed elevated expression under high‑temperature stress with the prolongation of the treatment time，and they were strongly responsive to both high and low temperatures.In summary,the EuHSF gene family has the function of regulating plant response to abiotic stresses.

Effects of Cake Fertilizer and Microbial Agents on the Yield and Quality of Flue‑Cured Tobacco and Soil Microbial Characteristics

CHENG Peijun, HAO Haohao, LI Xiang, LI Liang, ZHANG Xiang, TANG Peipei , WANG Xun, SI Xianzong, SUO Yanyan, WU Junlin, XU Fengdan, LI Qian, MA Xinjun, YAN Meng

2024, 53(9): 57-65. DOI: 10.15933/j.cnki.1004-3268.2024.09.006

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In order to improve the soil in tobacco‑growing areas and increase the yield and quality of flue‑cured tobacco，seven treatments[no cake fertilizer and microbial agent(CK),peanut cake(P)，sesame cake(S),compound cake(C),peanut cake+microbial agent(PM),sesame cake+microbial agent(SM),compound cake+microbial agent(CM)]were set up to explore the effects of combined application of cake fertilizer and microbial agent on photosynthetic parameters，antioxidant enzyme activities，physical properties，chemical components of flue‑cured tobacco leaves and soil microbial characteristics.The results showed that compared with CK treatment，the net photosynthetic rate(Pn),superoxide dismutase(SOD),peroxidase(POD),ascorbate peroxidase(APX),catalase(CAT)activities and total sugar，total protein,starch,reducing sugar,total nitrogen,potassium content of tobacco leaves were increased by single cake fertilizer application.The yield of flue‑cured tobacco was increased by 3.6%—14.1%,and the number of soil bacteria was significantly increased by 9.6%—25.9%.When the same cake fertilizer was applied,the application of microbial agents significantly increased the Pn of tobacco leaves,and the activities of SOD,POD and CAT,the output value of flue‑cured tobacco significantly increased by 7.8%—11.7%,and the contents of microbial biomass carbon and microbial biomass nitrogen in soil significantly increased by 11.6%—18.4% and 37.4%—48.6%.Therefore,single application of cake fertilizer or combined application of microbial agents could increase the number of soil microorganisms,enhance plant stress resistance,improve the net photosynthetic rate of flue‑cured tobacco,increase the yield of tobacco leaves and optimize the chemical composition of flue‑cured tobacco.Among them,the combined application of compound cake and microbial agents is better.

Screening of Cellulose and Lignin Degrading Bacteria and Their Effect on Composting of Mushroom Residue

WANG Xianxin, QI Lihua, SONG Jiangfu, ZHANG Qingge, LIU Congcong, ZHANG Qi, SONG Feifei, XU Ailing

2024, 53(9): 66-79. DOI: 10.15933/j.cnki.1004-3268.2024.09.007

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In order to screen high‑efficiency composite bacteria that promote the degradation of cellulose and lignin in the composting process of mushroom residue,sodium carboxymethyl cellulose plate method,Congo red staining method and lignin plate method were used for preliminary screening，combined with filter paper strip disintegration test,aniline blue decolorization rate determination and cellulose and lignin enzyme activity determination test for rescreening from the mushroom residue waste,and then they were used for aerobic composting test to study the physical and chemical parameters and microbial community changes in the process,and to evaluate their degradation ability of cellulose and lignin in mushroom residue.The results showed that the enriched cellulose‑degrading bacteria could completely degrade the filter paper within 6 days,and the activities of carboxymethyl cellulase(CMCase),filter paper enzyme(FPA),exoglucanase(C1)and β‑glucosidase(β‑Gase)reached 2.50,7.30,3.43 and 2.90 U/mL,respectively.The maximum decolorization rate of aniline blue reached 92.10% within 5 days by enriched lignin degrading bacteria，and the activities of lignin peroxidase(Lac),laccase(Lip)and manganese peroxidase(Mnp)reached 0.80,2.42 and 1.35 U/mL,respectively.The results of composting test showed that compared with the natural composting treatment(CK),the highest temperature of the pile body was 58.77℃,and the high temperature(>55℃)lasted for 5 days.The relative decomposition rate of cellulose and lignin increased by 35.83% and 54.33% respectively.The addition of microbial agents increased the richness of bacterial community in the middle stage of composting,and the relative abundance of Pseudomonas,Idiomarina,lutamicibacter,Thiopseudomonas,Globicatella,Pusillimonas,Lysobacter and Brumimicrobium increased. Redundancy analysis(RDA)results showed that temperature,pH value and electrical conductivity(EC value)were the significant factors affecting the composition of bacterial community in the middle stage of composting.In summary,the composite degrading bacteria screened in this study can effectively degrade cellulose and lignin,and prolong the high temperature period of composting

Establishment of RT‑RAA‑CRISPR/Cas12a Based Visual Detection Method for Pepper Mild Mottle Virus

ZHAO Zhenxing, FAN Qixuan, WANG Siyuan, DONG Zheng, HU Zhongze, ZHANG Yongjiang

2024, 53(9): 80-87. DOI: 10.15933/j.cnki.1004-3268.2024.09.008

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Pepper is an important horticultural crop.Pepper mild mottle virus(PMMoV)threatens pepper and other solanaceous crops’production.In order to improve the prevention and control efficiency of PMMoV,this study designed specific RAA primers based on the conserved sequence of its gene encoding coat protein(CP) to achieve rapid isothermal amplification of PMMoV based on recombinase‑aided amplification(RAA).And crRNA was designed to target the RT‑RAA amplification products based on the CRISPR/Cas12a system.The RT‑RAA‑CRISPR/Cas12a system was optimized,and the results showed that the strongest signal was observed in the total system at a final concentration of 400 nmol/L for reporter gene FQ,and a ratio of 1∶5 for Cas12a∶crRNA at the final concentrations of 200 nmol/L and 1 000 nmol/L.The final RT‑RAA amplification reaction and CRISPR chromogenic reaction each only took 15 min,and the positive signal could be directly observed under portable blue light irradiation equipment.This method could specifically detect PMMoV,and the detection limit of RNA of pepper samples carrying PMMoV could reach 1.34 pg/μL,which was 1 000 and 10 times more sensitive than that of conventional RT‑PCR and real‑time RT‑PCR.The detection results of 30 samples showed that the RT‑RAA‑CRISPR/Cas12a assay established in this study could detect PMMoV in leaves,fruits and soils of infected pepper and tomato plants.The established detection system could be used for rapid and sensitive visual detection of pepper mild mottle virus.

Susceptibility to Tebuconazole and Suitability of Resistant Strains of Magnaporthe oryzae in Hainan Province

WANG Yanxia, ZHAO Yuhan, GU Xinyi, FU Zhongju, SUN Yujia, WU Chenglong, WU Weihuai, ZHANG Yaling

2024, 53(9): 88-96. DOI: 10.15933/j.cnki.1004-3268.2024.09.009

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By detecting the susceptibility of Magnaporthe oryzae to tebuconazole in Hainan Province and studying the fitness of resistance strains,the resistance risk of M.oryzae to tebuconazole was understood,which would provide theoretical reference for the scientific use of fungicides and resistance management.The mycelial growth rate method was used to determine the susceptibility to tebuconazole of 144 strains of M.oryzae collected and isolated from Hainan Province in 2022,and the susceptibility baseline of M.oryzae in Hainan Province to tebuconazole was established based on the susceptibility frequency distribution of the strains.Fifteen susceptible strains with low effective inhibitory mid‑concentration(EC₅₀)measured during the establishment of the susceptibility baseline were selected for resistance domestication under continuous selection pressure，and the domesticated strains were classified into resistance levels,and the genetic stability and suitability of the resistant strains as well as the cross‑resistance to the fungicides pyraclostrobin,flucycloxazole,imidacloprid,fenpyroximate，and aliconazole were determined.The EC₅₀ values of the 144 strains tested ranged from 0.176 9 to 0.998 0 μg/mL.The EC50 value of the least sensitive strain was 5.64 times that of the most sensitive strain.The average EC₅₀ value was(0.476 3±0.174 5)μg/mL.The susceptibility frequencies of the strains were distributed in a continuous single‑peak curve,and the susceptibility baseline to tebuconazole of M.oryzae in Hainan Province was 0.476 3 μg/mL.Through indoor domestication of 15 sensitive strains,a total of two resistant strains were obtained.EC₅₀ values were 1.289 5 μg/mL and 1.868 4 μg/mL respectively,and resistance folds were 5.38 times and 6.65 times respectively,both of which were low resistance level.And after eight consecutive generations of transfer,the two resistant strains always maintained low‑level resistance.The EC₅₀ values of pyraclostrobin,epoxiconazole,prochloraz,difenoconazole,and diniconazole were measured and compared between the strains resistant to tebuconazole and their parental strains.The results showed that there was no cross‑resistance in M.oryzae in Hainan Province between tebuconazole and the above chemicals.Through the in vitro fitness test of the tebuconazole‑resistant strains,it was found that compared with the parental strains,the mycelial growth rate of the two resistant strains was significantly reduced,and the conidia production was reduced to varying degrees.In addition,they were more sensitive to strong acid and alkali environment，and their pathogenicity was greatly reduced.Therefore,the frequency of resistance mutations to tebuconazole in rice blast strains in Hainan Province is low,and the environmental suitability of their resistant strains is significantly reduced,resulting in a low risk of resistance.In addition,there is no cross‑resistance between tebuconazole and commonly used fungicides in Hainan Province,and the development of resistance can be delayed by strategies such as cross‑rotation or mixing with multi‑site‑of‑action protectants in production.

Effects of Formula Fertilization and Fertilization Methods on Growth and Woodland Soil of Carya illinoensis

LIU Fukui, DU Yangwen, DU Shiping, ZHANG Rongyang

2024, 53(9): 97-107. DOI: 10.15933/j.cnki.1004-3268.2024.09.010

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The aim is to explore the influence of different formula fertilization and fertilization techniques on leaf mineral nutrients,soil enzyme activity,soil effective nutrient content and fruit quality of Carya illinoensis.Five treatments for different formula fertilization tests,namely OPT(Nutrient experts systematically recommend fertilization),OPT‑N(No nitrogen fertilizer),OPT‑P(No phosphorus fertilizer),OPT‑K(No potassium fertilizer)and CK(No fertilizer),respectively;five treatments were designed for different fertilization techniques,according to time,frequency and proportion and fertilization method. Mineral nutrient,soil enzyme activity,effective nutrient content,oil content and fatty acid content were determined in different periods.Different formula fertilization significantly changed the soil enzyme activity,soil effective nutrient content,plant leaf mineral nutrient content and fruit quality,among which the OPT‑P and OPT‑K treatments had higher soil nutrient absorption and utilization efficiency. Soil phosphatase and urease activities were higher in OPT‑P,sucrase was higher in OPT,and catalase was higher in OPT‑N.The oil content of OPT‑P fruit kernels was significantly increased by 8.23 percentage points over the CK treatment,and there was no significant difference in unsaturated fatty acid content among the treatments.The soil effective N,P,K content and organic matter content with different fertilization techniques were significantly lower than the control treatment,and fertilizing 70% in early March and 30% in early June could improve the leaf nutrient content of N,and K,soil sucrase and catalase activities,the oil rate of fruit kernels was 7.38 percentage points higher than the control.In summary,OPT‑P formula fertilization and twice from the shallow to the deep radial ditch can promote the soil nutrients and increase the oil content of fruit kernels.

Growth Response and Tolerance Evaluation of 32 Spinach Varieties to Cadmium Stress

MIAO Mingjun, LI Jiajia, LEI Xiaokui, LIN Yang, LIAO Jichao, ZHONG Jian, LI Ju, LI Yuejian, CHANG Wei

2024, 53(9): 108-116. DOI: 10.15933/j.cnki.1004-3268.2024.09.011

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Cadmium(Cd)is a highly toxic heavy metal element.To identify Cd‑tolerant spinach varieties and screen related identification indexes,and provide a theoretical basis for the selection and breeding of Cd‑tolerant varieties,in the present study,we conducted a hydroponic experiment using 32 spinach varieties in China.The spinach was exposed to Cd stress(Cd concentration of 50 μmol/L)for 20 days,while the other group of plants was treated as a control(Cd concentration of 0 μmol/L).After the plants reached maturity,we measured the agronomic traits and Cd content.The integrated Cd tolerance of each spinach variety was evaluated by principal component analysis,affiliation function analysis,and correlation analysis.The results showed that plant height,root length,underground fresh weight,aboveground fresh weight,aboveground Cd content,underground Cd content and translocation coefficient of the participating varieties under Cd stress conditions were significantly different from the control.Under Cd stress,the growth and biomass accumulation of spinach were inhibited,and the indicators of each trait decreased significantly.The order of average reduction of each agronomic index was aboveground fresh weight>plant height>underground fresh weight>root length.The principal component analysis of Cd stress tolerance index of the seven indexes was performed,and three principal components were selected，with a cumulative variance contribution of 81.552%;The composite evaluation value(D value)showed highly significant positive correlations(P<0.01)with plant height,underground fresh weight,aboveground fresh weight,aboveground Cd content,and translocation coefficient,which can be used as the preferred indexes for the evaluation of Cd tolerance in spinach.Based on the principal component analysis,we classified the 32 spinach varieties into three categories:Cd‑tolerant,moderate,and Cd‑sensitive varieties.Among the varieties tested,Jin’ge and Micro orchid exhibited strong Cd tolerance,while Desai 176 was found to be Cd‑sensitive.

Preparation of Universal Hapten Immunochromatographic Strip and Its Application in Imidocarb

XING Yunrui, SUN Yaning, HU Xiaofei, FAN Lu, CHEN Linlin, ZHANG Gaiping

2024, 53(9): 117-125. DOI: 10.15933/j.cnki.1004-3268.2024.09.012

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In order to improve the selection efficiency of monoclonal antibodies and lower the preparation threshold of immunochromatographic strip,a universal hapten immunochromatographic strip was established by using biotin‑labeled artificial antigen as a probe and avidin in a test line,and applied for selecting monoclonal antibodies against imidocarb(IM)and detecting IM residues.The results showed that IM artificial antigen and biotin probe were successfully prepared by 1‑(3‑dimethylaminopropyl)‑3‑ethylcarbodiimide(EDC)method.Subsequently,monoclonal antibodies were prepared by immunizing mice and hybridoma cells technology,and the universal immunochromatographic strip was established.Using the strip,a monoclonal antibody cell line against IM with a titer of 1:32 000 was successfully selected,and its sensitivity was up to 6.25 ng/mL.Based on the excellent monoclonal antibody and the universal strip,an immunochromatographic test strip for the detection of IM was established,and its sensitivity was 7 ng/mL in the milk.The immunochromatographic strip and its probe preparation method can be used in general hapten antibody screening,and a hapten immunochromatographic strip test can be established without professional research and development platform,which provides a more efficient method for antibody screening and a new technology for the development of immunochromatographic strip.

Isolation，Identification and Susceptibility Analysis of One Morganella morganii Strain from Swine

PENG Zhifeng, ZHANG Han, HUANG Huimin, SONG Xinghui, ZHU Yabo, LI Ziyi, QIAO Hongxing, BIAN Chuanzhou

2024, 53(9): 126-131. DOI: 10.15933/j.cnki.1004-3268.2024.09.013

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Morganella morganii is an opportunistic pathogen that can cause a variety of diseases and has become an important emerging pathogen worldwide.In order to explore the drug sensitivity and pathogenicity of Morganella morganii from swine,a suspected pathogenic strain was isolated from the intestines of dead pigs in Henan Province.Gram staining,matrix‑assisted laser desorption/ionization time of flight mass spectrometry and 16S rRNA gene homology analysis were performed.Then the minimum inhibitory concentration of 14 drugs against the strain was determined,and the infection experiment wa conducted in mice.The results showed that the bacterium was Gram‑negative brevibacterium，which was identified as Morganella morgenii and named A185.The results of drug susceptibility test showed that the strain was resistant to amoxicillin,meropenem,igacycline,polymyxin,doxycycline and cefoxitin,intermediate to ciprofloxacin and gentamicin,and sensitive to fosfomycin and ceftazidime.The results of infection experiment showed that the Kunming mice inoculated with Morganella morganii exhibited hemorrhage in liver,intestine and lung to varying degrees.The above results showed that the porcine Morganella morganii A185 had multi‑drug resistance,and exhibited strong pathogenicity.

Effects of Mixed Ensiling of Rice with Sorghum‑Sudangrass Hybrid on Forage Nutritional Quality and Microbial Community

LI Lingzi, JI Changli, LIU Ye, FU Mingfang, LI Zongrun, TONG Jundan, CHEN Qiuyuan, ZHANG Jing, ZHAO Yafan, WANG Daoli, PENG Ting

2024, 53(9): 132-140. DOI: 10.15933/j.cnki.1004-3268.2024.09.014

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To investigate the effects of co‑ensiling with rice and sorghum‑sudangrass hybrid on silage nutritional quality,feeding value,and microbial community,three treatments were established:100% rice silage(R),100% sorghum‑sudangrass hybrid silage(SS),and 50% rice mixed with 50% sorghum‑sudangrass hybrid silage(R+SS).After 60 days of ensiling,the nutrient content,feeding value and bacterial community of the silages were analyzed.The results indicated that compared to R treatment，the R+SS treatment significantly reduced the contents of neutral detergent fiber(NDF）and acid detergent fiber(ADF)by 3.10 and 5.35 percentage points,respectively,while increased total digestible nutrients(TDN),net energy for lactation(NEL),digestible dry matter(DDM),and relative feeding value(RFV) by4.23 percentage points,0.44%,4.10 percentage points and 12.46%,respectively.The analysis results of microbial taxonomy composition and diversity revealed that the R+SS treatment significantly reduced the alpha diversity and increased the relative abundance of Lactobacillus.Redundancy analysis indicated that NDF,CP,and NEL were the primary environmental quality indicators influencing the microbial community composition of mixed rice and sorghum‑sudangrass hybrid silage.In conclusion,co‑ensiling with rice and sorghum‑sudangrass hybrid can enhance the abundance of fermentation‑related lactic acid bacteria,thereby effectively improving the nutritional quality and feeding value of rice silage.

Risk Assessment of Citrus Huanglongbing with the Support of Remote Sensing Technology

XIE Guoxue, HUANG Qiting, QIN Zelin, LAN Zongbao, LIANG Yongjian, WEN Guoquan, YANG Shaoe, NING Xia

2024, 53(9): 141-149. DOI: 10.15933/j.cnki.1004-3268.2024.09.015

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Predicting the risk of citrus Huanglongbing(HLB)is an effective measure to control the spread of the disease.In order to achieve precise monitoring and prevention of HLB in a large area,a comprehensive multi‑indicator HLB risk assessment at the plot scale was carried out.Taking the citrus plot in Xinxu Town,Mengshan County,Guangxi as the object,based on multi‑temporal NDVI(normalized difference vegetation index),LSWI(land surface water index)and MTCI(MERIS terrestrial chlorophyll index)remote sensing indicators,a citrus HLB monitoring model was established to obtain HLB‑affected plots.Comprehensive migration distance,slope,citrus growth,elevation and average temperature indicators were used to calculate the weight of indicators using the analytic hierarchy process method,and the comprehensive model for evaluating the risk of citrus HLB infection was constructed to assess the risk level of citrus plots infected with HLB.The results showed that the accuracy rate of the citrus HLB monitoring model was 80.31%,and the leakage rate was 10.53%,which effectively reduced the field investigation of citrus HLB.The weights of migration distance,slope,citrus growth,elevation and average temperature were 0.404,0.289,0.186,0.120 and 0.086,respectively.The ability of migration and diffusion of psyllid was the dominant factor in the spread of the disease. The high‑risk area of HLB infection in Xinxu Town was 475.93 ha,the medium‑risk area was 552.00 ha,and the low‑risk area was 1 188.87 ha,accounting for 21.47%,24.90%,and 53.63% of the citrus area,respectively.The risk of further spread of HLB infection is high,and it is suggested that the relevant departments should strengthen the prevention and control of high‑risk areas to prevent the spread of the disease.

Lightweight Grape Object Detection Fusion Algorithm Based on Improved YOLOv5s

HU Junfeng, LI Songqing, HUANG Xiaowen, LIU Dayang, LI Baicong

2024, 53(9): 150-158. DOI: 10.15933/j.cnki.1004-3268.2024.09.016

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A lightweight grape target detection network YM‑GDM(YOLOv5s‑MobileNetV3 grape detection model) was proposed to meet the requirements of accuracy,real‑time performance,and lightweight of target detection models for agricultural automatic harvesting machinery.MobileNetV3 was employed as the backbone network instead of CSPDarknet53 in YOLOv5s to achieve model lightweighting.The introduction of Res2Net_C2f module and BiFPN(Bi‑directional feature pyramid network)structure aimed to enhance the model’s multi‑scale feature fusion capability.Additionally,the VariFocalLoss loss function was adopted to train the model,mitigating the impact of imbalanced positive and negative samples.The self‑made data set containing five types of table grapes and the open data set(WGISD)containing five types of wine grapes were used as test data sets.The experimental results showed that the YM‑GDM network achieved a mAP50 of 90. 8% for the detection of 10 grape classes.Compared to YOLOv3‑tiny,and YOLOv5s,it improved by 6.2,and 2.2 percentage points respectively.The model size was 9.73 MB,which was reduced by 44.4% and 32.8% compared to YOLOv3‑tiny and YOLOv5s,respectively.Furthermore,by further reducing the number of parameters,a lightweight specialized model,YM‑GDM‑tiny,was obtained with a model size of 4.73 MB and a mAP50 of 86.8%,suitable for deployment on mobile devices with lower computing power.

Parameter Sensitivity Analysis and Optimization of Sesame Phenology Simulation Model

GUAN Haoyue, LI Mengyao, LI Guoqiang, ZHANG Jiantao, GAO Tongmei, CHEN Xianguan, ZHANG Wenyu, WU Jinzhi

2024, 53(9): 159-170. DOI: 10.15933/j.cnki.1004-3268.2024.09.017

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Phenological development is critical for crop field management.In order to accurately predict the development process of sesame by quantifying the response of sesame physiological processes to temperature and photoperiod,a sesame phenology simulation model(SPSM)based on clock model theory was constructed.The sensitivity of parameters was analyzed by extened fourier amplitude sensitivity test(EFAST)to determine the high sensitivity parameters affecting the development process.The parameters of four sesame varieties were determined by the Monte Carlo optimization method and verified in the main sesame producing areas of Henan,Hubei,Anhui and Jiangxi provinces in China.The results showed that the average global sensitivity indexes of optimum temperature(T_opt),basic development coefficient(k),and critical day length(D_c)were the highest among the seven cultivar parameters of SPSM.The R² of the simulated and observed values of Yuzhi 4,Zhongzhi 22,Wanzhi 10 and Ganzhi 7 were0.999,RMSEwas0.50 to 1.32 d,NRMSE was1.06%—3.02%,and d value was 0.999.By independent verification,the standard deviations of the development period simulations and observations of the above four sesame varieties were0—7.65 d,R²were 0.978,0.988,0.992,0.989,RMSEwere5.07,4.43,3.07,4.24 d,NRMSE and Willmott consistency(d)were5.87%—9.44% and0.996—0.997,respectively.The optimized SPSM can well simulate the development dynamics of sesame in the main producing areas of sesame in China.

Effect of Adding Sucrose Fermentation on the Quality of Indonesian BESUKI Cigar Tobacco Leaves

LI Linlin, WANG Ronghao, WU Zhao, ZHAO Zeyu, DU Fu, LIU Jiaqi, TIAN Lijun, SONG Hao, SHAN Yujing, PENG Jian

2024, 53(9): 171-180. DOI: 10.15933/j.cnki.1004-3268.2024.09.018

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To investigate the effect of adding sucrose fermentation on the quality of Indonesian BESUKI cigar tobacco leaves,we studied the changes in conventional chemical composition,volatile components,and sensory quality of cigar leaves with 1.0%(T1),2.0%(T2)sucrose added(calculated by weight of tobacco leaf)during high‑temperature and high humidity fermentation.The results showed that:Adding sucrose fermentation could significantly increase the total sugar content and sugar alkali ratio of tobacco leaves,while potassium content and nitrogen alkali ratio decreased,which was beneficial for balancing the acidity and alkalinity of smoke,reducing irritation,and improving smoke comfort.Adding sucrose fermentation could increase the types of volatile components in tobacco leaves,and the total amount of volatile components first increased and then decreased with fermentation time.The highest content was observed after 16—24 days of fermentation,and adding sucrose was beneficial to the accumulation of ketones,phenolic hydrocarbons,and the total amount of volatile components.The aroma quality,aroma content,and impurity scores of cigar leaves fermented with sucrose were significantly improved.According to the regression model,considering both chemical composition and sensory quality,the best effect on the quality of Indonesian BESUKI tobacco was obtained by fermentation with T1 for 19—20 days.

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