Journal of Henan Agricultural Sciences

CONTENTS

2025, 54(4): 0.

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The timely and accurate identification of cucumber diseases using intelligent identification technology is crucial for the proactive control and rational application of pesticides，which is of great significance for ensuring high‐quality cucumber production and ecological environmental safety.The intelligent identification of cucumber diseases is mainly realized by expert knowledge based on traditional expert systems and knowledge graphs，visible light image processing based on traditional machine learning and deep learning，spectral analysis such as chlorophyll fluorescence and hyperspectrum，and multi‐modal data fusion.The research progress of intelligent identification of cucumber diseases based on the above technologies was reviewed，the existing problems and deficiencies in current research were also summarized，and the development trends of cucumber diseases intelligent identification technology in the future was prospected，in order to provide reference for the application research of cucumber diseases intelligent identification.

Effect of Nitrogen and Zinc Application on Accumulation and Distribution of Mineral Elements in Grain at Different Positions of Ear of Maize with Different Zn Efficiency

ZHANG Panpan, SHAO Yunhui, LI Chunhua, MU Weilin, GUO Hanxiao, LI Chuan, ZHANG Meiwei, ZHAO Shuangsuo, HU Yanqi, HAN Linlin, QIAO Jiangfang

2025, 54(4): 11-26. DOI: 10.15933/j.cnki.1004-3268.2025.04.002

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With zinc（Zn） efficiency（Zhengdan 958，ZD958）and insensitivity types（Longping 638，LP638）as materials，the effects of nitrogen（N）rates［225 kg/ha（HN），180 kg/ha（MN）and 90 kg/ha（LN）］and foliar Zn［no Zn（Zn0），spraying Zn of 1∶1 at jointing stage and flare opening stage（Zn1）］on the yield and content，accumulation and distribution of mineral elements in grain at different positions of maize ear，so as to provide scientific basis for the N and Zn fertilization in summer maize production.The result indicated that in general，the kernel number，1 000‐grain weight，yield and mineral element accumulation showed upper grain<middle grain<basal grain.MN had the highest upper grain yield，and HN had the highest middle and basal grain yield.Foliar Zn improved the yield of upper，basal and whole grain；Compared with LP638，the upper and basal grain yields of ZD958 were higher.Overall，MNZn1ZD958 treatment had high upper，basal and whole grain yield，Zn content in whole grain，and potassium（K）and manganese（Mn）accumulation in upper grain. HNZn1ZD958 treatment had higher yield，N and Mn contents in whole grain，copper（Cu）and Zn contents in upper grain and Cu content in middle grain，and increased the proportions of Mn and Zn in upper grain and Cu in middle grain.There was significantly positive correlation between whole grain yield and basal grain yield；N content in whole grain was significantly positively correlated with middle grain yield，and significantly negatively correlated with Mn and Zn contents.By comprehensive consideration，spraying Zn of 1∶1 at jointing stage and flare opening stage can improve yield under 180—225 kg/ha N condition；Spraying Zn of 1∶1 at jointing stage and flare opening stage can promote N，Zn and Mn accumulation under 180 kg/ha N condition；Spraying Zn of 1∶1 at jointing stage and flare opening stage is conducive to the transport of Cu，Mn and Zn to basal grain under 90 kg/ha N condition；Spraying Zn of 1∶1 at jointing stage and flare opening stage is conducive to the transport of N，Mn and Zn to upper grain under 225 kg/ha N condition.

Effects of Tillage and Straw Returning Methods on Yield and N₂O Emission of Winter Wheat

ZHAO Lishang 1, YANG Tiancong 2, ZHANG Xiaomei2, SONG Quanhao 1, MA Zhiyao 2, FENG Wei 2, DUAN Jianzhao2

2025, 54(4): 27-36. DOI: 10.15933/j.cnki.1004-3268.2025.04.003

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In order to seek technical measures for reducing greenhouse gas emissions and increasing yield in winter wheat production，winter wheat in Huang‐Huai‐Hai Plain was taken as the research object，the effects of different tillage methods［shallow rotating tillage（S）and deep tillage（D）］and straw returning methods［no straw returning（N），direct straw returning（R）and charred straw returning to field（C）］on dry matter accumulation，nitrogen absorption and utilization，yield and N₂O emission of winter wheat were studied.The results showed that deep tillage and straw returning promoted plant growth，increased dry matter accumulation，enhanced nitrogen absorption and utilization，and synergistically increased yield components，thus increasing yield.Compared with SN treatment，the nitrogen use efficiency of SR and SC treatments increased by 16.08%—18.34% and 12.27%—14.03%，and that of DN，DR and DC treatments increased by 19.20%—28.70%，37.60%—40.90% and 24.80%—34.50%，respectively.DR and DC treatments were more beneficial to nitrogen absorption，and improve utilization efficiency.Ear number of DR and DC treatments increased by 7.27%—8.31% and 12.97%—15.03%，grain number per ear increased by 10.34%—9.25% and 6.90%—8.03%，and yield increased by 8.00%—15.15% and 7.00%—9.09%，respectively.Compared with S treatment，D treatment reduced the peak of N₂O emission flux，R treatment increased the peak of N₂O emission flux compared with N treatment，and C treatment reduced the peak of N₂O emission flux compared with N treatment. Compared with SN treatment，N₂O emission accumulation of DR and DC treatments decreased by 1.15%—2.04% and 5.53%—15.14%，respectively.To sum up，deep tillage and straw returning are beneficial to the growth and development of winter wheat，increase yield，save energy and reduce emission，among which DC treatment can be used as the best tillage measure to increase yield and reduce N₂O emission in winter wheat production in Huang‐Huai‐Hai Plain.

Identification of Differentially Expressed ERF Genes under Low Nitrogen Stress in Sesame（Sesamum indicum L.）and Expression Analysis of SiERF08 Gene

TIAN Yuan, ZHANG Pengyu, LI Feng, WANG Dongyong, FU Jinzhou, RONG Yasi, GAO Tongmei

2025, 54(4): 37-46. DOI: 10.15933/j.cnki.1004-3268.2025.04.004

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The AP2/ERF is one of the largest families of transcription factors in plants，which is involved in plant metabolic biosynthesis，organ development，stress response and hormone signal transduction.To explore the expression characteristics of ERF genes under low nitrogen stress in sesame，24 differentially expressed SiERF genes were identified from the former transcriptome result of sesame roots under low nitrogen stress，and their expression characteristics，co‐expression network and the expression profiles of the key gene SiERF08 under different treatments were analyzed.The results showed that the open reading frame length of 24 differentially expressed SiERF genes was 564—1 254 bp，protein molecular weight ranged from 20.92—44.97 ku，and isoelectric point ranged from 4. 39 to 10. 92. And 24 differentially expressed SiERF genes were unevenly distributed on 13 chromosomes.The promoter sequences contained a number of plant hormone response elements（ABRE，AuxRE and so on），stress response elements（such as ARE，DRE core and LTR），growth and development elements（O2‐site，Gap‐box and CAT‐box）and light response cis‐acting elements（G‐box，MRE，I‐box and so on）.The results of gene expression and Venn diagram showed that 24 and 14 differentially expressed SiERF genes were detected in Burmese prolific and Zhengzhi HL05，respectively.Among them，3 differentially expressed genes such as SiERF06 were identified in two varieties at 3，9 days after low nitrogen stress.Five key candidate genes in response to low nitrogen stress，such as SiERF08 were screened by co‐expression network analysis.The expression characteristics of key candidate gene SiERF08 under different stresses were analyzed.The results showed that，SiERF08 gene was induced under low nitrogen，drought，high salinity，GA and SA stresses，and under different treatments SiERF08 gene exhibited different expression patterns.The expression of SiERF08 gene was the highest in drought，high salt，SA，high temperature and low temperature treatment for 6 h，and reached the peak at 12 h under low nitrogen treatment.Under GA treatment，the expression of SiERF08 gene was only induced at 6 h and 72 h，and the difference was significant.Overall，24 differentially expressed SiERF genes under low nitrogen stress were analyzed by bioinformatics，and 5 key candidate genes such as SiERF08 were screened，among which SiERF08 gene was induced to express under low nitrogen，drought，high salinity and SA stress.This research will provide theoretical basis and genetic resources for further exploring SiERF genes function and revealing the molecular mechanism of low nitrogen tolerance in sesame.

Characterisation of the NtHPL Genes and Their Expression Pattern under TMV Stress in Tobacco

WANG Mingxin, LIU Lei, DENG Zhaolong, WANG Haoyang, LIU Pengfei, JIN Weihuan, LI Junying, GUO Hongxiang

2025, 54(4): 47-56. DOI: 10.15933/j.cnki.1004-3268.2025.04.005

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Hydroperoxide lyase（HPL）is a key enzyme in the catalytic fatty acid oxidation pathway and belongs to the cytochrome P450（CytP450）superfamily. HPL plays an essential role in plant responses to both biotic and abiotic stresses，contributing significantly to the regulation of stress tolerance.In this study，the NtHPL gene family in tobacco was identified using genomic data and comprehensively analyzed for their physicochemical properties，protein structures，conserved motifs，and response patterns to tobacco mosaic virus（TMV）.The results indicated that nine NtHPL family members were identified，distributed across eight chromosomes，and classified into three subclasses：B，C，and D.The amino acid lengths of these members ranged from 437 to 527 residues，with theoretical isoelectric points（pI）varying between 5.84 and 9.15.Subcellular localization predictions suggested that these proteins were primarily localized in chloroplasts and cytoplasm.Expression analysis revealed that upon TMV infection，five NtHPL genes were upregulated，one was downregulated，and three exhibited no significant differential expression in tobacco leaves. Overexpression of NtHPL1 enhanced tobacco resistance to TMV，as demonstrated by experimental evidence.These findings collectively suggest that NtHPL may be involved in the regulatory mechanisms underlying tobacco defense against TMV.

Identification of the WOX Gene Family in Paeonia lactiflora Based on Transcriptome Data and Functional Analysis of PlWOX5 Under Salt Stress

PENG Bo, MA Mengmei, ZHAO Ping

2025, 54(4): 57-65. DOI: 10.15933/j.cnki.1004-3268.2025.04.006

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In order to clarify the function，evolutionary relationship，and role in biological processes of the Paeonia lactiflora WOX gene family，members of Paeonia lactiflora WOX family（PlWOXs）were identified based on transcriptome data，followed by a comprehensive bioinformatics analysis of their protein physicochemical properties，structures，evolutionary relationships，and expression patterns.The results showed that five PlWOX genes were identified based on the transcriptome data of Paeonia lactiflora，and their encoded amino acids were mainly hydrophobic and unstable alkaline nucleoproteins.The results of the system evolution analysis showed that the PlWOXs gene was distributed in both WUS/modern lineages and ancient lineages，with no PlWOXs gene distribution in the intermediate lineages.Theresults of protein structure analysis indicated that there were differences in the distribution and quantity of Motif on different PlWOXs genes，and they had significant tissue specificity.PlWOX2 and PlWOX5 were mainly highly expressed in Paeonia lactiflora leaves.The qRT‐PCR detection results showed that PlWOXs genes were responsive to salt stress，but PlWOX5 showed a more significant response to salt stress.Overexpression of PlWOX5 gene could enhance salt tolerance in Arabidopsis by enhancing antioxidant enzyme activity（superoxide dismutase，peroxidase，catalase）and increasing the content of osmoregulatory substances（soluble proteins）to promote reactive oxygen species（ROS）clearance.In conclusion，PlWOX5 gene can improve the salt tolerance of Paeonia lactiflora by increasing the activity of
antioxidant enzymes and the content of osmotic regulators.

Effects of Flue‐cured Tobacco and Wheat Intercropping on Soil Microorganisms and Fertility in Xuchang Tobacco Growing Area

GE Menglong, HE Bing, BAI Feng, HUANG Peiyuan, HE Mengying, LI Jianhua, LI Yuyang, SHEN Hongtao, GAO Jiakai, WANG Yanfang, LIU Ling

2025, 54(4): 66-79. DOI: 10.15933/j.cnki.1004-3268.2025.04.007

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To investigate the effect of wheat/tobacco intercropping on soil quality in Xuchang tobacco growing area，soil samples were collected through multi‐point sampling in the field.High throughput sequencing technology was used to study the differences in soil microbial diversity and community composition between flue‐cured tobacco（CK）and wheat tobacco intercropping（I）planting modes.The relationship between soil environmental factors and soil microbial diversity and community composition was further analyzed by combining soil physical and chemical properties and enzyme activity.The results showed that the wheat tobacco intercropping planting mode significantly increased the Chao1 index of soil bacterial communities and significantly reduced the Simpson index compared to the flue‐cured tobacco planting mode；Significantly increased soil fungal ACE index and Chao1 index.NMDS analysis（Non‐metric multidimensional scaling analysis）showed that there were certain differences in community structure between the two planting modes of tobacco. The wheat tobacco intercropping planting model significantly increased the relative abundance of Acidobacteria and Myxococcata at the bacterial phylum level compared to the flue‐cured tobacco planting model，while significantly reduced the relative abundance of Firmicutes；At the fungal phylum level，the relative abundance of Mortierellomycota significantly increased，while the relative abundance of Ascomycota and Chytridiomycota significantly decreased.LEfSe analysis results showed that the wheat tobacco intercropping planting mode（11 bacterial groups and 35 fungal groups）increased the number of differential indicator fungal groups and decreased the number of differential indicator bacterial groups compared to the flue‐cured tobacco planting mode（16 bacterial groups and 17 fungal groups）. Compared with the monoculture planting mode of tobacco，the intercropping planting mode of wheat and tobacco significantly increased the soil total potassium，alkali hydrolyzable nitrogen content，and sucrase activity，while significantly reduced the soil pH value and available potassium content.The Pearson correlation analysis results indicated that soil total nitrogen，total potassium，alkali hydrolyzed nitrogen，available potassium content，urease and sucrase activity were the main factors affecting soil microbial community diversity.The redundancy analysis results indicated that alkaline hydrolyzable nitrogen content，available potassium content，and sucrase activity were the main environmental factors affecting the structure of soil bacterial and fungal communities.In summary，adopting the wheat tobacco intercropping planting mode could improve the soil microbial community structure，increase soil nutrient content and enzyme activity，and is beneficial for improving soil fertility.

Effects of Submerged Stress on Inter‐root Soil Nitrogen Transformation of Vetiveria zizanoides and Structural Diversity of Bacterial Communities

WEI Jiating, HU Tian, ZHAO Ruibo, LIU Ying, JI Mingfei, ZHANG Jun

2025, 54(4): 80-90. DOI: 10.15933/j.cnki.1004-3268.2025.04.008

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In order to study the effect of inter‐root bacterial community diversity on soil nitrogen transformation in the vegetation of Danjiangkou Reservoir fallout zone，Vetiveria zizanoides was selected as experimental material，and control（CK），semi‐submerged and fully submerged groups were set up to systematically study the effect of submerged stress on inter‐root nitrogen transformation and bacterial community through soil physicochemical analyses，enzyme activity determination combined with high‐throughput sequencing of 16S rRNA and function prediction of PICRUSt2.The results showed that the content of inter‐root soil ammonium nitrogen（NH4+‐N）was significantly elevated under submerged stress，and the content of nitrate nitrogen（NO3-‐N） and nitrite nitrogen（NO2-‐N） were significantly reduced by 66.83%—74.27% and 36.84%—42.11%，respectively，compared with CK.Compared with CK，soil hydroxylamine reductase（HR）activity in the fully submerged group was significantly reduced by 34.48%，and nitrate reductase（NR）and nitrite reductase（NiR）activities were significantly increased.The results of Alpha diversity analysis showed that flooding stress significantly increased the diversity of inter‐root bacterial communities，and the order was fully submerged group>semi‐submerged group >CK.The non‐metric multidimensional scaling analysis and principal component analysis showed that the bacterial community structure showed gradient differences with the increase of submerged depth.Community structure analysis showed that the relative abundance of Firmicutes in the semi‐submerged and fully submerged groups increased significantly by 45.14% and 87.99%，respectively，while the relative abundance of Cyanobacteria in the fully submerged group decreased significantly by 84.98%.Functional prediction revealed that the abundance of genes for the key enzymes of biological nitrogen fixation pathway（Nitrogenase 1.18.6.1）was significantly increased by 198.43% to 198.97% under submerged stress，but it suppressed the abundance of genes for nitrous oxide reductase（1.7.2.4）.Redundancy analysis indicated that soil NO₃^-‐N and NR activities were the main factors affecting the soil bacterial community. In conclusion，submerged stress affects the nitrogen transformation process in inter‐root soils and alters bacterial community structure，which may lead to ecological risks of N₂O greenhouse gas accumulation.

Effect of Heterologous Expression of the vgb Gene Mediated by Different Promoters on Novonestmycin Production

ZHANG Yifan, LIU Xiaoyan, WAN Zhongyi, FANG Wei, ZHU Lei, CHEN Ling, CAI Jun, ZHOU Ronghua, WANG Changgao, MIN Yong

2025, 54(4): 91-100. DOI: 10.15933/j.cnki.1004-3268.2025.04.009

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To enhance the production of the novel agricultural antibiotic novonestmycin，Streptomyces sp.HBERC‐20821 was used as the host strain.Five engineered strains were constructed using different promoters：ermEp，kasOp，kasOp₃，kasOp*，and SP44‑SR12.The expression of the Vitreoscilla hemoglobin gene(vgb)and its differential expression under various promoters were validated by quantitative real‐time PCR(qRT‐PCR)，and the strain with the highest novonestmycin yield was identified through parallel shake flask fermentation experiments.The bioactivity of the functional VHb protein in the selected strain was confirmed using carbon monoxide difference spectroscopy.Furthermore，parallel bioreactor fermentations were conducted to investigate the impact of heterologous expression of the vgb gene，on novonestmycin production.The results showed that five engineered strains containing different promoters were successfully constructed.Under the mediation of the kasOp series promoters，vgb expression was significantly higher than that achieved by the traditional ermEp promoter，with increases ranging from 9‐to 35‐fold；notably，the kasOp₃ promoter mediated vgb expression was 35‐fold that of ermEp.In shake flask fermentations，the parental strain HBERC‐20821 produced 545.01 mg/L of novonestmycin，while the engineered strain 20821‐kasOp‐vgb(20821/PKV)achieved the highest novonestmycin yield with an increase of 46.30% over the parental strain，reaching 797.30 mg/L.Carbon monoxide difference spectroscopy confirmed the bioactivity of the VHb protein in 20821/PKV，as evidenced by the characteristic absorption peak at 420 nm of its CO‐bound complex.In bioreactor fermentations，the parental strain produced 495.67 mg/L of novonestmycin，whereas the yield from 20821/PKV was 803.18 mg/L，corresponding to a 62.04% increase.In summary，the kasOp promoter effectively enhanced vgb expression，thus significantly improving novonestmycin production in the engineered strain 20821/PKV.

Resistance Evaluation and Cross‐resistance Analysis of Mung Bean to Imidazolinone Herbicides

DUAN Yun, SU Wangcang, XUE Fei, WANG Chunyu, XU Hongle, SUN Lanlan, WU Renhai

2025, 54(4): 101-108. DOI: 10.15933/j.cnki.1004-3268.2025.04.010

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To precisely determine the resistance levels of Vigna radiata to imazethapyr and imazapic，explore the underlying molecular mechanisms，and assess potential cross‐resistance and multi‐resistance to other herbicides，the resistance levels of 27 V.radiata varieties to imazethapyr and imazapic were measured using the agar solid medium method.Additionally，the sensitive and resistant varieties were selected to evaluate their cross‐resistance and multi‐resistance to other herbicides. Finally，the mechanism through which V. radiata developed resistance to imidazolinone herbicides was elucidated via acetolactate synthase gene amplification and sequence alignment analysis.The results indicated that the preliminary screening results of resistance of 27 V.radiata varieties to imazethapyr revealed that seven varieties were sensitive，while five varieties were resistant，and one variety was suspected resistant.Both imazethapyr and imazapic could inhibit the growth and development of V.radiata.The resistance results of 13 V.radiata varieties to imazethapyr showed that seven varieties were identified as sensitive，accounting for 53.85% of the total，while six varieties exhibited low‐level resistance，accounting for 46.15%.Specifically，Jilü H1004 was the most sensitive variety，with a GR50 value of 18.80 mg/kg.Baolü 2016 22‐2 demonstrated the highest resistance，having a resistance index（RI）of 3.63.The resistance results of 13 V.radiata varieties to imazapic showed that seven varieties（53.85%）showed high resistance and five（38.46%）displayed extremely high resistance.Pinlü 2019‐26‐8‐17 had the highest resistance level，with a RI value of 237.85.The findings from the ALS gene sequence analysis indicated
that there were no disparities in the amino acid sequences of the two ALS genes between the relatively resistant and relatively sensitive V.radiata varieties.This suggested that the variation in the resistance of V.radiata to imidazolinone herbicides was not attributable to mutations at the ALS gene site.The results of the cross‐resistance analysis revealed that the relatively resistant cultivar Pinlü 2019‐26‐8‐17 exhibited cross‐resistance to choransulam‐methyl and halosulfuron‐methyl，with the RI values of 500.71 and 170.47，respectively.The results of the multi‐resistance analysis indicated that this relatively resistant cultivar demonstrated multi‐resistance to fluroxypyr，with a RI value of 4.38.However，it did not show multi‐resistance to bentazone，bixlozone and lactofen.In conclusion，among the tested varieties，there were variations in the resistance levels of different V.radiata varieties to imidazolinone herbicides.Moreover，the relative resistant cultivar exhibited multi‐resistance to fluroxypyr.These findings provide a
theoretical reference for effectively controlling weeds in V.radiata fields.

A Comparative Study on the Developmental Anatomy of Embryos from Gabengcui Jujube and Golden‐silk Jujube

CHEN Zizhu, QING Jun, BAO Wenhui, WANG Yaping, GE Genzhula, BAI Yu’e

2025, 54(4): 109-116. DOI: 10.15933/j.cnki.1004-3268.2025.04.011

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To investigate the period of embryo failure in Golden‐silk jujube and discuss the causes of such failure，in this study，external morphological and anatomical observations were made on two varieties of jujube，Gabengcui jujube and Golden‐silk jujube，during different developmental periods.The results showed that the development trend of young fruits of Golden‐silk jujube from 3 d after flowering to 35 d after flowering was smooth，and the development trend of Gabengcui jujube was slow‐fast，and the fruit development of Gabengcui jujube was faster than that of Golden‐silk jujube.And 30 percent of Gabengcui jujube had tridentate split stigmas.Anatomical observation found that Golden‐silk jujube and Gabengcui jujube ovary had two compartments，each bearing an anatropous ovule，but Gabengcui jujube had phenomenon of double embryonic compartments and three ovules with a 30% incidence.In most of the fruits an ovule shrivelled，forming a single ovule.At the beginning of the embryo development process，the endosperm development of Golden‐silk jujube was obviously slower than that of Gabengcui jujube，while the embryo development of Golden‐silk jujube was faster than that of Gabengcui jujube；endosperm disintegration，ovule atrophy and embryo failure phenomenon appeared in Golden‐silk jujube at 30 d after flowering，and the endosperm，ovule and embryo of Golden‐silk jujube disappeared at 35 d after flowering.The embryo，endosperm and bead centre of Gabengcui jujube developed normally.On the whole，the fruit development of Gabengcui jujube was faster than that of Golden‐silk jujube.The embryo abortion time of Golden‐silk jujube was 30 d after flowering，and both endosperm and embryo were aborted.

Study on Mechanism of Feizhu Powder for Prevention and Treatment of Deficiency of Qi and Blood in Sows Based on Network Pharmacology and Animal Experiments

GAO Tiantian, LIU Xuesong, LIU Yongti, QIU Zongsheng, LIU Yishan, LI Shuxin, QI Jingjing, LI Ya’nan, WEI Chengwei, MA Tianwen

2025, 54(4): 117-128. DOI: 10.15933/j.cnki.1004-3268.2025.04.012

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In order to screen the action targets of the traditional Chinese veterinary medicine Feizhu powder for preventing and treating deficiency of Qi and blood in sows and explore its molecular mechanism，the active ingredients of the formula of Feizhu powder，their action targets，and the action targets of Qi and blood deficiency were studied，their intersection targets were obtained，and a network of the intersection targets of active ingredients of Feizhu powder and deficiency of Qi and blood was constructed. GO and KEGG pathway enrichment analysis were carried out on the intersection targets，and a network of the core ingredients and core targets and signaling pathways of Feizhu powder was constructed.Molecular docking analysis was performed on the core ingredients and core targets to screen the key targets and signaling pathways of Feizhu powder.Male KM mice were selected，and randomly divided into control group，Qi and blood deficiency model group，ferrous lactate group，and Feizhu powder group.A Qi and blood deficiency model was established for the latter three groups.The control group and Qi and blood deficiency model group were respectively intragastrically administered with 10 mL/kg normal saline，the ferrous lactate group was intragastrically administered with 10 mg/kg ferrous lactate syrup，and the Feizhu powder group was intragastrically administered with 3 g/kg Feizhu powder.The key targets and pathways of Feizhu powder for preventing and treating Qi and blood deficiency were verified through the weight‐bearing test of mice，detection of blood indexes，organ indexes，and candidate proteins.The results showed that 29 active ingredients of Feizhu powder were obtained，and 112 intersection targets of active ingredients of Feizhu powder and Qi and blood deficiency were identified.Eleven core ingredients such as daidzein，kaempferol，luteolin and β‐sitosterol were screened out，and eleven core targets such as PTGS2，PTGS1，HSP90AA1，PRKACA，and TNF were identified.The results of GO and KEGG pathway enrichment analysis showed that the active ingredients of Feizhu powder were mainly enriched in items such as binding of protein，binding of enzyme，cytoplasm，nuclear plasm，inflammatory response and response to exogenous stimuli. A total of 26 signaling pathways were enriched，including the IL‐17 pathway，the regulation of adipocyte lipolysis and the ovarian steroidogenesis pathway，etc.Molecular docking analysis results showed that the binding energies of the core ingredients and core targets were all less than -8 kcal/mol，indicating that the core ingredients and core targets had strong binding activity. The results of the mouse experiment showed that，compared with the Qi and blood deficiency model group，the weight‐bearing swimming time of mice of ferrous lactate group and Feizhu powder group significantly increased，the numbers of white blood cells and red blood cells and the hemoglobin content all significantly increased；the spleen index and thymus index all significantly increased，and the protein expression levels of IL‐17 and TNF‐α in the spleen tissues all significantly decreased.In conclusion，the core ingredients of the traditional Chinese veterinary medicine Feizhu powder are daidzein，kaempferol，luteolin，β‐sitosterol，etc.，and the core targets are PTGS2，PTGS1，HSP90AA1，PRKACA，TNF，etc.，which play important roles in preventing and treating Qi and blood deficiency in sows through signaling pathways such as IL‐17.

Polymorphism of Structural Variation DKK2‐I1‐sv89 in the DKK2 Gene of Five Pig Breeds

LONG Juyan, LU Shengfei, TIAN Jiao, NIU Xi, CHEN Xia, HUANG Shihui, RAN Xueqin, WANG Jiafu

2025, 54(4): 129-135. DOI: 10.15933/j.cnki.1004-3268.2025.04.013

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A structural variation measuring 89 bp，designated as DKK2‐I1‐sv89，was identified in intron 1 of the DKK2 gene of pigs through preliminary genomic analysis.To investigate the polymorphism distribution of this structural variation（SV）and its potential impact on gene expression，specific primers targeting the SV sequence were designed for this study.The population distribution of DKK2‐I1‐sv89 was subsequently evaluated using polymerase chain reaction（PCR）across four Guizhou indigenous pig breeds（Xiang pig，Qianbeihei pig，Kele pig，and Jiangkouluobo pig）and one European pig breed（Large White pig）.The gene expression difference of DKK2 among the various genotypes was determined by using real time quantitative PCR（RT‐qPCR）.Additionally，bioinformatics analyses were employed to assess the functional elements within the structural variation region of DKK2‐I1‐sv89. The findings demonstrated that in the Xiang pig and Qianbeihei pig，three genotypes（insertional genotype II，heterozygous genotype DI，and wild‐type genotype DD）were found，while only one genotype（DD）was found in Large White pig，however in Kele pig and Jiangkouluobo pig，two genotypes（DI and DD）were found.Comparative analysis across the five breeds demonstrated that the four Guizhou local pig breeds（including Xiang pig，Qianbeihei pig，Kele pig，and Jiangkouluobo pig） had a higher frequency of the insertional allele（I allele） compared to the Large White pig. Bioinformatics analysis revealed that the DKK2‐I1‐sv89 contained one intronic splicing enhancer（ISE） and 13 RNA binding protein（RBP） binding sites.Quantitative trait locus（QTL）analysis demonstrated that the QTL at this SV locus was associated with reproduction，meat quality，and health‐related traits. Further analysis of DKK2 gene expression in ovarian tissues of Xiang pigs with different genotypes（II，DI，and DD）revealed that II genotype pigs exhibited significantly higher expression levels compared to both the DI and DD genotypes.The results indicated a notable difference in the distribution frequency of this polymorphism between Large White pig and Guizhou local pigs.Additionally，the SV was found to be beneficial for the expression of DKK2 gene in the ovary and could serve as an early molecular marker for distinguishing between Large White pig and Guizhou local pigs.

Effect of Phosphatidylinositol‐3 Kinase Inhibitors on the Proliferation of Vesicular Stomatitis Virus in Vitro

WANG Mengdi, FAN Wenjie, ZHANG Zhen, ZHAO Ruihong, LI Huawei, LIU Kun, ZHENG Ming, WANG Yakai

2025, 54(4): 136-143. DOI: 10.15933/j.cnki.1004-3268.2025.04.014

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The aims are to study the effect of phosphatidylinositol‐3 kinase inhibitor（PI3K‐IN‐1）on the proliferation of vesicular stomatitis virus（VSV）in vitro on PK‐15 cell model，and to reveal the antiviral effect of PI3K‐IN‐1 on VSV in vitro，which will lay a foundation for seeking and developing drugs to prevent and control VSV infection.The safe concentration of PI3K‐IN‐1 at the cellular level was detected by CCK‐8 method；flow cytometry was used to detect the effect of PI3K‐IN‐1 on the percentage of each period in the cell cycle and apoptosis；and flow cytometry and fluorescence microscopy were used to analyze the difference of virus proliferation in PK‐15 cells infected with VSV after PI3K‐IN‐1 treatment；the mRNA expression level of VSV N gene was analyzed by RT‐qPCR；the expression level of GFP protein in VSV‐GFP was analyzed by immunoblotting；the effect of PI3K‐IN‐1 on the titer of VSV progeny virus was detected by virus titer assay；and the inhibitory effect of PI3K‐IN‐1 on the fluorescence intensity of GFP protein in VSV‐GFP was analyzed by confocal microscopy.The results showed that 0—10 μmol/L PI3K‐IN‐1 had no effect on PK‐15 cell viability，percentage of each period in the cell cycle and apoptosis rate. But with the increase of PI3K‐IN‐1 concentration，the expression level of GFP protein in VSV‐GFP decreased. When the concentration of PI3K‐IN‐1 was 10 μmol/L，the inhibitory effect on VSV proliferation was the best. The relative expression level of VSV N gene mRNA in PI3K‐IN‐1 treatment group was significantly or extremely significantly lower than that in DMSO（CK）group and showed a time‐dependent trend.At 12 h and 24 h after VSV‐GFP infection，the proportion of GFP positive fluorescent cells in PI3K‐IN‐1 treatment group was extremely significantly lower than that in CK group，and the progeny virus titer of PI3K‐IN‐1 treatment group was significantly lower than that of CK at 12 h and 24 h after infection，and was extremely significantly lower at 36 h after infection. In summary，PI3K‐IN‐1 can effectively inhibit the proliferation of VSV in PK‐15 cells，which provides a new strategy and method for the research and development of anti‐VSV drugs.

Identification of Rice in Southern Mountainous Area Based on Object‐Oriented and Machine Learning Methods

WANG Yingying, DUAN Liangxia, ZHAO Yining, SUN Guangrui, YANG Lihong, ZHOU Qing, XIE Hongxia

2025, 54(4): 144-154. DOI: 10.15933/j.cnki.1004-3268.2025.04.015

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The phenology information of rice varies with terrain.In view of the cloudy and rainy conditions and complex terrain in the southern mountainous areas，it is of certain research value to find a remote sensing identification method for rice. With Yongshun County，a typical mountainous terrain in Hunan Province，as the research area，combined with Sentinel‐1 SAR and Sentinel‐2 MSI data，the time series curve was generated by the images of four key phenology periods of rice to grasp the growth trend of rice.Firstly，the object‐oriented method was used to segment the images of transplanting period and harvesting period；secondly，the feature variables optimized by the feature space optimization algorithm（FSO）were input into four models for classification such as random forest（RF）；finally，the results were compared. Otherwise，according to the transplanting order，the rice samples were divided into early moving and late moving samples，the images of transplanting period and harvesting period were reclassified to explore the influence of rice transplanting time on the classification of the two images.The results showed that，compared to imagery from the transplanting stage，imagery from the harvesting stage offered better classification accuracy and was more suitable for rice mapping.The object‐oriented FSO‐RF model achieved the best classification results for imagery during the harvest period，with an overall accuracy of 93.19% and a kappa coefficient of 0. 901. The time of rice transplanting had little effect on the image classification at harvest stage，but had a great influence on the image classification at transplanting stage. The reason was that the earliest transplanted rice was easy to be similar to the spectral characteristics and texture characteristics of some dry land crops and woodlands，and there was a phenomenon of confusion and mis‐classification. In order to improve the recognition accuracy of the image during the transplanting period，it is necessary to improve the time resolution of the image，add more texture features or mask the ground objects.

Detection and Counting of Sesame Capsules per Plant Based on Improved YOLOv8‐Track Model

LI Chenhao, WANG Chuan, LI Guoqiang, ZHAO Qiaoli, YANG Ping, WANG Kai, CHANG Shenglong, ZHENG Guoqing

2025, 54(4): 155-166. DOI: 10.15933/j.cnki.1004-3268.2025.04.016

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Sesame capsules are an essential factor in the composition of sesame yield. In order to realize the accurate detection and counting of sesame capsules per plant，using object detection，multiple targets tracking and other technologies for dynamic tracking of capsules per plant is helpful to improve the efficiency of sesame breeding and cultivation management. Aiming at the phenomena of sesame capsules，such as small target，dense growth and overlapping occlusion，this study taked YOLOv8‐Track as the benchmark model，introduced small target detection head and Shuffle attention mechanism into the feature fusion network，and introduced MPDIOU loss function in the post‐processing stage of the model to construct SD‐YOLOv8‐Track model.In addition，this study utilized the ID counting method of model ByteTrack multi‐target tracking algorithm to track and count sesame capsules using a single rotating video of sesame as the model input. The results showed that when taking a single picture as input，the accuracy，recall，and mean average precision of the SD‐YOLOv8‐Track model for detecting capsules were 92.25%，92.4%，and 92.58%，respectively，indicating 5.94，6.6，and 6.31 percentage points higher than those of the original model YOLOv8‐Track.For the rotating video as input，the multiple object tracking accuracy and multiple object tracking precision of SD‐YOLOv8‐Track model were 89. 42% and 88.23%，respectively，which were 4.23 and 4.60 percentage points higher than the original model.The accuracy，missed detection rate，and error detection rate of the SD‐YOLOv8‐Track model were 93.27%，3.85%，and 2.88%，respectively.The accuracy rate was 5.61 percentage points higher than that of the original model，and the missed detection rate and false detection rate were 3.84 and 1.77 percentage points lower than that of the original model.The improved SD‐YOLOv8‐Track model performs better in detecting sesame capsules and is suitable for dynamic complete counting of sesame capsules in a plant.

Analysis of Yunnan Black Tea Aroma Characteristics Based on HS‐SPME‐GC‐MS and OAV

DING Qihuan, DENG Min, SHI Dekang, SU Jianmei, GU Chonglin, LI Sijin

2025, 54(4): 167-180. DOI: 10.15933/j.cnki.1004-3268.2025.04.017

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In order to investigate the aroma characteristics of Yunnan black tea from different production areas in Yunnan,12 tea samples from four production areas,namely,Simao in Puer,Changning in Baoshan,Menghai in Xishuangbanna and Fengqing in Lincang，were selected as materials.The headspace‐solid phase microextraction(HS‐SPME)combined with gas chromatography‐mass spectrometry(GC‐MS)was used to separate and identify the volatile components of Yunnan black tea from different production areas on a DB‐WAX column，and the volatile components of Yunnan black tea were analysed by using aroma activity value(OAV)combined with chemometrics to determine the volatiles composition and aroma characteristics of Yunnan black tea from different production areas.The results showed that a total of 143 volatile substances，mainly alcohols，aldehydes，esters and olefins，were identified in Yunnan black tea from different production areas，and 50 substances(OAV≥1)contributing to the flavor of Yunnan black tea were screened out with the help of OAV，among which linalool and β‐ionone had a prominent position in OAV，and were important for the composition of the flavour of Yunnan black tea from different production areas.By using orthogonal partial least squares‐discriminant analysis(OPLS‐DA),combined with variable importance projection(VIP),43 differential markers were screened(VIP≥1).Based on the results of OAV and OPLS‐DA，12 characteristic differential substances of Yunnan black tea were screened：2‐methyl butyraldehyde， benzaldehyde， heptaldehyde，Z‐hex‐3‐en‐1‐ol，methyl 2‐(methylamino)benzoate，dihydroactinidolide，D‐limonene,(E)‐3，7‐dimethylocta‐1,3,6‐triene，β‐ionone，α -lonone，dihydro‐β‐ionone，and P‐cymene.Z‐hex‐3‐en‐1‐ol contributed prominently to the clear aroma of black tea from Changning production area；dihydroactinidolide and dihydro‐β‐ionone contributed prominently to the floral and fruity aroma of Menghai production area，and heptaldehyde contributed prominently to the fruity aroma of Fengqing production area.Further analysis showed that the Yunnan black tea aroma types of the four production areas were mainly floral and fruity.Combined with the cluster analysis，Changning and Fengqing production areas had floral and sweet aroma，and the fruity aroma was stronger，absence of honey notes；and the differentiated compounds of Simao and Menghai production areas were more diverse，and the composite aroma was obvious，which involved the sweet aroma，honey aroma，grassy aroma，clear aroma and fat aroma.

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