Journal of Henan Agricultural Sciences

CONTENTS

2025, 54(12): 0.

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Effects of Long⁃Term Application of Subsoiling and Organic Fertilizer on Distribution of Hydrogen and Oxygen Isotopes in Precipitation and Water Utilization of Maize

YANG Yonghui, WU Jiabin, ZHANG Yunhong, ZHANG Tinghao, GAO Cuimin, HAN Weifeng, PAN Xiaoying, HE Fang, Devotha G.NYAMBO, LUO Ning

2025, 54(12): 1-12. DOI: 10.15933/j.cnki.1004-3268.2025.12.001

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In order to elucidate the effects of long⁃term subsoiling tillage and application of organic fertilizers on distribution of rainfall in soil and utilization，based on hydrogen and oxygen isotopes tracing method，the effects of long⁃term subsoiling tillage and application of organic fertilizers on the distribution of stable hydrogen and oxygen isotopes in soil and water use of maize were studied with conventional tillage treatment as control.The results showed that long⁃term subsoiling tillage and application of organic fertilizers improved soil structure and the distribution and utilization of precipitation in the soil.The soil moisture generally gradually decreased with the increase of soil depth at jointing and grain filling stages of maize.At harvest stage，the soil moisture obviously increased，and increased with the increase of soil depth.At jointing and grain filling stages，subsoiling tillage notably increased the 0—80 cm soil moisture.At harvest stage，both the subsoiling tillage and application of organic fertilizers treatments increased the
0—100 cm soil moisture.At jointing stage，the contribution rate of water from the 0—40 cm soil was obviously higher than those in other layers，reaching over 90%，and the contribution rate of water from the 20—40 cm soil was the highest.At grain filling stage，conventional tillage，subsoiling tillage and application of organic fertilizers treatments mainly utilized the water in 20—60 cm soil，and the water contribution rate reached 46.3%，53.4% and 92.7%，respectively.As the growth stage progressed，the soil water storage capacity of all treatments generally gradually increased.Compared with conventional tillage treatment，subsoiling tillage and application of organic fertilizers were more beneficial for increasing soil water storage at jointing and harvest stages，and decreasing water consumption during sowing—jointing and grain filling—harvest period；meanwhile，subsoiling tillage and application of organic fertilizers were more beneficial for increasing the net photosynthetic rate and stomatal conductance at grain filling stage of maize，as well as improving leaf water use efficiency at jointing and grain filling stages. Ultimately，subsoiling tillage and application of organic fertilizers significantly increased maize yield（10.6%，9.4%）and water use efficiency（13.6%，12.2%），although the difference between the two treatments was not significant. Correlation analysis showed that the contribution rate of water in 0—20 cm soil was significantly positively correlated with maize yield and water use efficiency at jointing stage，and significantly negatively correlated with maize yield and water use efficiency at grain filling stage.In conclusion，long⁃term subsoiling tillage and application of organic fertilizers can effectively regulate the rational distribution of precipitation in soil，improve the photosynthetic
characteristics of maize，promote the efficient utilization of rainfall，and increase the yield and water use
efficiency of maize.

Effects of Irrigation Amount and Nitrogen Application Rate on Dry Matter Accumulation and Yield of Maize under Different Densities

ZHANG Meiwei, GUO Hanxiao, ZHAO Shuangsuo, HU Yanqi, MU Weilin, ZHANG Panpan, LI Chuan, ZHENG Fei, QIAO Jiangfang

2025, 54(12): 13-22. DOI: 10.15933/j.cnki.1004-3268.2025.12.002

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In order to definite the response of dry matter accumulation and yield formation of maize to irrigation and nitrogen under different densities，Xundan 509 and Zhengdan 958 were used as materials to study the effects of irrigation amount［reduced irrigation（W1，irrigating 37.5 mm at flare opening and silking stages），full Irrigation（W2，irrigating 75 mm at flare opening and silking stages）］and nitrogen application rate［no nitrogen fertilizer（N0），low nitrogen level（N1，150 kg/ha nitrogen fertilizer），normal nitrogen level（N2，225 kg/hanitrogen fertilizer）］on dry matter accumulation，ear height coefficient，chlorophyll fluorescence parameters（Fv/Fm and Fv/Fo），leaf area index（LAI），yield and its components of maize under different densities［67 500 plants/ha（D1），90 000 plants/ha（D2）］.The results showed that，full irrigation and application of nitrogen both increased dry matter accumulation of population，LAI，Fv/Fm，Fv/Fo，ear height coefficient，yield and its components of two maize cultivars.Simultaneously，the appropriate irrigation amount and nitrogen application rate promoted transfer of dry matter to grain.Increase of density also increased dry matter accumulation of population，LAI and grain yield，while had adverse effects on Fv/Fm and Fv/Fo.Two maize cultivars generally had the highest dry matter accumulation of population，LAI and grain yield under D2W2N2 treatment. Compared with D1W1N0 treatment，the yields of Xundan 509 and Zhengdan 958 increased by 60.76% and 55.38% under D2W2N2 treatment respectively.Overall，D2W2N2 treatment with nitrogen application rate of 225 kg/ha，irrigating amount of 75 mm at the flare opening and tasseling⁃silking stages of maize respectively，and density of 90 000 plants/ha is the optimal measure to obtain high yield under high density.

Effects of GS3 Gene Haplotypes on Japonica Rice Panicle and Grain Traits under Different Alkaline Conditions

DUAN Kairong, DONG Yue, WANG Jingwen, LIU Chang, LIU Songrui, NIAN Hua, TIAN Lei

2025, 54(12): 23-31. DOI: 10.15933/j.cnki.1004-3268.2025.12.003

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Thirty japonica rice germplasm resources were used as experimental materials and planted in normal paddy soil（control，CK）and soils with different alkalization degrees［alkalized saline soil（T1，alkalization degree of 39.7%）and alkaline soil（T2，alkalization degree of 57.3%）］.The full⁃length sequence of the GS3 gene was obtained by Sanger sequencing.Combined with panicle and grain phenotypic traits such as 1 000⁃grain weight，grain length and panicle length，as well as the change rates of panicle and grain traits under different treatments，haplotype analysis was carried out to explore the regulatory effects of different haplotypes of the GS3 gene on panicle and grain traits of japonica rice under alkaline stress condition.Comparative analysis of the full⁃length GS3 gene in 30 japonica rice germplasm resources was studied，and a total of 27 effective variation sites，including 13 missense mutation sites and 14 Indel sites，were detected.A correlation analysis between these sites and the change rate of 1 000⁃grain weight revealed that two Indel sites，SDP（significant differences are present in all）and SOA（significant differences are only under higher alkaline stress），which were located at positions 16730526 and 16732228 on chromosome 3，respectively，were significantly correlated with the change rate of 1 000⁃grain weight，and together formed four main haplotypes，SDP1，SDP2，SOA1 and SOA2.A frameshift mutation in the coding region of SDP2 haplotype led to the premature termination of the protein domain，which weakened the sensitivity of GS3 to changes of alkalinity，mainly reflecting in the non⁃significant difference in 1 000⁃grain weight，grain length，panicle length and their change rates between T1 and T2 treatments.The SOA2 haplotype had a 2 bp insertion mutation in the intron region，which reduced the response ability of GS3 to alkaline stress and weakened its functional effect，mainly
manifesting in that the 1 000⁃grain weight under T2 treatment was extremely significantly higher than that of SOA1，and the change rate of 1 000⁃grain weight under T2 treatment was extremely significantly different from SOA1，but there were no significant difference in grain length，panicle length and their change rates compared with SOA1.In summary，this study is the first to reveal the functional trade⁃off mechanism of the GS3 gene between alkali tolerance and grain weight trait regulation，and clarify the specific regulatory effects of the SDP2 and SOA2 haplotypes on panicle and grain traits under alkaline environments.

Identification and Analysis of Amylase Genes Related to Pre⁃harvest Sprouting Resistance in Wan 1329 Based on Transcriptome Sequencing

WANG Zhen, ZHANG Bin, SHI Lichao, LIU Rui, LI Jinxiu, ZHOU Ran, YANG Ling, ZHAO Qian, LI Jinbang

2025, 54(12): 32-39. DOI: 10.15933/j.cnki.1004-3268.2025.12.004

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Using Jimai 22 with high sensitivity to pre⁃harvest sprouting（PHS）as the control，the PHS resistance and α⁃amylase activities of the new white⁃grain wheat variety Wan 1329 and its parent Lianmai 2 were analyzed. During the critical period when α⁃amylase activity changed，transcriptome sequencing，KEGG and GO enrichment analysis were employed to identify differentially expressed amylase genes，and real⁃time fluorescence quantitative PCR（qRT⁃PCR）was used for verification their expression patterns.In addition，AlphaFold was employed to analyze the effects of coding sequence variations on protein structure. The results showed that both Wan 1329 and Lianmai 2 exhibited moderate resistance to PHS，while Wan 1329 had a lower relative germination index，indicating stronger resistance. At 24 h after seed imbibition，the α⁃amylase activities of Wan 1329 and Lianmai 2 were markedly lower than that of Jimai 22，and α⁃amylase activities at 20 h，24 h，and 72 h were highly positively correlated with the relative
germination index. Transcriptome sequencing，KEGG and GO enrichment analysis revealed that two α⁃amylase genes located on the long arm of chromosome 6D， TraesCS6D02G313300 and TraesCS6D02G313500，were candidate α⁃amylase genes. The third exon of TraesCS6D02G313500 in Wan 1329 was elongated by 138 bp，extending into the 3´UTR region. qRT⁃PCR validation confirmed that the expression patterns of both genes were consistent with transcriptome analysis，and the cumulative relative expression level of TraesCS6D02G313500 in Wan 1329 was significantly lower than that in Lianmai 2. AlphaFold prediction suggested that this variation caused a C⁃terminal extension of α⁃amylase，resulting in decreased structural stability and increased conformational flexibility.Collectively，TraesCS6D02G313300 and TraesCS6D02G313500 are key genes related to PHS resistance
in Wan 1329，among which the structural variation of TraesCS6D02G313500 may further enhance resistance to PHS.

Effects of Shading on Source⁃Sink⁃Translocation Traits of Different Peanut Varieties

ZHANG Jun, ZHANG Man, ZHANG Penglei, ZHANG Zhongxin, ZHANG Xinyou, LIU Juan, HAO Xi, DONG Wenzhao, ZANG Xiuwang

2025, 54(12): 40-48. DOI: 10.15933/j.cnki.1004-3268.2025.12.005

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To evaluate the shade tolerance of different peanut cultivars and explore the impact of shading on source⁃sink⁃translocation characteristics，seven commonly cultivated peanut（Arachis hypogaea L.）varieties were selected to investigate their photosynthetic capacity，stem anatomical structure，reproductive development，and yield components under both normal light and shading conditions.The results showed that shading significantly reduced net photosynthetic rate and SPAD chlorophyll values，indicating a decline in assimilate synthesis capacity. In terms of transport traits，shading decreased the number of vascular bundles and xylem vessels by 9.52% and 30.10%，respectively.Reproductive development was also affected. The maximal flowering accumulation rate of Kaifeng 49，Yuhua 31，and Huayu 20 decreased by over 40%，while Puhua 28，Yuhua 22，and Yuhua 23 experienced declines of around 25%. Theoretical maximum flower number decreased by more than 50% in Kaifeng 49 and Yuhua 31，and by 12.48% and 15.86% in Yuhua 23 and Yuhua 22，respectively.While shading had little effect on the peak pod⁃filling rate，it delayed the time to reach this rate by an average of 5 days and shortened the duration of rapid accumulation by 6 days. Yield performance varied across cultivars：under shading，Yuhua 22 and Yuhua 23 showed relatively higher yields，with Yuhua 23 producing 1.33 times and 1.67 times more than Huayu 20 and Puhua 28，respectively.These findings suggest that Yuhua 22 and Yuhua 23 are suitable for shaded environments，with management strategies focused on enhancing photosynthesis. In contrast，Huayu 20 requires agronomic interventions that promote flowering and regulate assimilate translocation under shade.

Identification of NPF Genes in Sesame and Expression Analysis under Waterlogging Stress

WU Ke, DU Zhenwei, FAN Yadong, JIANG Xiaolin, CUI Chengqi, WANG Jingjing, ZHAO Fengli, ZHANG Ruping, LIU Yanyang, MEI Hongxian

2025, 54(12): 49-61. DOI: 10.15933/j.cnki.1004-3268.2025.12.006

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The nitrate transporter 1/peptide transporter family（NRT1/PTR family，NPF）plays a crucial role in nitrogen uptake and utilization in plants，and is also involved in diverse biotic and abiotic stress responses.To elucidate the function of NPF in the waterlogging stress of sesame（Sesamum indicum L.），a systematic bioinformatics analysis was conducted to identify SiNPF genes. Furthermore，the dynamic expression profiles of selected SiNPFs under submergence conditions were characterized.The results of genome⁃wide analysis indicated that a total of 66 SiNPF members were identified in sesame，distributed across 12 chromosomes（excluding chr13），with chr2（12 genes），chr6（11），chr8（9），and chr9（8）being gene⁃enriched regions.And after constructing the phylogenetic tree（Mega 7，NJ method），it was found that these genes were divided into 10 subfamilies.The analysis of gene structure and conserved motif revealed that the majority of SiNPFs had similar gene structures and high conservation，with 44 SiNPFs containing all conserved motifs.Protein property analysis indicated that SiNPF proteins ranged in molecular weight from 20.96 to 135.01 ku，with the majority being alkaline（48 proteins）and stable（52），all of which were hydrophobic（GRAVY>0）.Transmembrane topology prediction（TMHMM）showed that these proteins contained multiple transmembrane domains（4 to 17）.The prediction results of the cis acting elements of the promoters showed a high frequency of cis⁃acting elements，including light⁃responsive elements，and ABA⁃responsive，anaerobically induced，and methyl jasmonate⁃responsive elements.Transcriptome analysis revealed that under waterlogging stress，15 SiNPFs were significantly downregulated in the roots of sesame varieties C177 and C185，while SiNPF2，SiNPF48，and SiNPF56 were significantly upregulated.The results of cloning and sequencing of these three significantly upregulated SiNPFs from varieties C177 and C185 showed that the amino acid sequence differences between the two varieties were limited，and the overall similarity was high. RT⁃qPCR validation confirmed that the expression patterns of these three genes under waterlogging stress were highly consistent with the transcriptome sequencing results.These findings indicate that specific SiNPF genes are closely associated with waterlogging response in sesame，providing a theoretical basis for further investigation on the biological functions of SiNPF genes and breeding of new waterlogging⁃tolerant sesame varieties.

Establishment of a Bioreactor Culture System for Anoectochilus roxburghii Protocorm⁃Like Bodies

ZHANG Fangling, KONG Xiangjun, ZHOU Xiaojun, WANG Hongzhen, WU Mei, FANG Li

2025, 54(12): 62-69. DOI: 10.15933/j.cnki.1004-3268.2025.12.007

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To screen the optimal hormone combination and germplasm for inducing Anoectochilus roxburghii protocorm⁃like bodies（PLBs），and to investigate the effects of inoculum density and culture duration on PLBs biomass and kinsenoside content for optimizing large⁃scale culture conditions，this study utilized tissue⁃cultured seedlings of Anoectochilus roxburghii.The effects of different concentration combinations of 6⁃benzylaminopurine（6⁃BA），1⁃naphthaleneacetic acid（NAA），zeatin（ZT），and 2，4⁃dichlorophenoxyacetic acid（2，4⁃D） on the induction of PLBs from five different Anoectochilus roxburghii germplasms were tested.The PLB induction rate，kinsenoside content，and proliferation characteristics were compared among the different plant germplasms.Using a plant bioreactor，the influence of inoculum density（20—80 g/L） and culture duration（10—40 days） on biomass and
kinsenoside synthesis was studied.The results indicated that the combination of MS+3 mg/L 6⁃BA+0.5 mg/L NAA+0.8 mg/L ZT+0.2 mg/L 2，4⁃D medium had the best effect on improving the induction rate of adventitious buds of different species of Anoectochilus roxburghii germplasms.Under identical culture conditions，the ZY1（Nanjing，Fujian） germplasm demonstrated superior PLB induction rate（93%），kinsenoside content（5.03%），and radial proliferation capacity compared to other germplasms.In the bioreactor culture system，both the proliferation coefficient（1.46）and kinsenoside content（7.80%）of the PLBs peaked at an inoculum density of 60 g/L after 40 days of culture in darkness.In conclusion，utilizing the superior ZY1 germplasm with the MS+3 mg/L 6⁃BA+0.5 mg/L NAA+0.8 mg/L ZT+0.2 mg/L2，4⁃D medium，combined with bioreactor culture conditions of 60 g/L inoculum density and 40 days of culture in darkness，enables the highly efficient accumulation of both Anoectochilus roxburghii PLB
biomass and kinsenoside content.

Effects of LED Red⁃Blue Light Quality Ratio on the Growth and Physiological Characteristics of Dendrobium fimbriatum Test⁃Tube Seedlings

ZHANG Ningjing, WANG Jiangning, MENG Xinya, FU Qiunuan, HUO Jiaran, LU Jiajing, WANG Yuxin

2025, 54(12): 70-80. DOI: 10.15933/j.cnki.1004-3268.2025.12.008

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To cultivate high⁃quality commercial seedlings of Dendrobium fimbriatum，six different light quality treatments were set up，including 100% red light（R），100% blue light（B），80% red light+20% blue light（8R∶2B），70% red light+30% blue light（7R∶3B），and 50% red light+50% blue light（5R∶5B），with white fluorescent lamp as the control（CK）. After 60 days，growth indicators（plant height，leaf number，leaf length，stem diameter，fresh weight，dry weight），antioxidant indicators［superoxide dismutase（SOD）activity，malondialdehyde（MDA）content，ascorbate peroxidase（APX）activity，peroxidase（POD）activity，catalase（CAT）activity］，photosynthetic indicators（chlorophyll a content，chlorophyll b content，chlorophyll a+b content，chlorophyll a/b ratio，and carotenoid content），chlorophyll fluorescence indicators［potential activity of PS Ⅱ in plants（Fv/Fo），maximum photochemical efficiency（Fv/Fm），actual photochemical efficiency（Φ_PSⅡ），photochemical quenching coefficient（qP），electron transfer rate（ETR Ⅱ），and non⁃photochemical quenching coefficient（qN）］，as well as the content of nutrients（soluble proteins and soluble sugars）and other related indices，totaling 24 indexes were measured in order to screen the LED light quality ratio for the healthy growth of D. fimbriatum test⁃tube seedlings.The results indicated that monochromatic red light was beneficial for the height and elongation of test⁃tube seedlings，while monochromatic blue light was beneficial for increasing soluble protein content. 8R∶2B treatment was
beneficial for leaf elongation，leaf number，and stem thickness increase of test⁃tube seedlings of D.fimbriatum.The overall antioxidant index was best when they were treated with 5R∶5B，and the dry and fresh weight，soluble sugar content，chlorophyll content，and chlorophyll fluorescence of test⁃tube seedlings were also best when they were treated with 5R∶5B.There was a significant correlation between growth indicators，antioxidant indicators，nutritional indicators，chlorophyll fluorescence parameters，and chlorophyll content.The results of principal component analysis indicated that the physiological and biochemical indicators of Dendrobium fimbriatum test⁃tube seedlings under 5R∶5B treatment showed the best performance，with the highest comprehensive score.In summary，the light environment of 50% red light+50% blue light（5R∶5B）LED is the most conducive to the growth and development of the test⁃tube
seedlings of D.fimbriatum，and is the most suitable light environment for cultivating strong seedlings.

Effects of Nitrogen，Phosphorus，Potassium，Magnesium，and Boron Fertilizers on Yield and Nutrient Uptake of Seed Lotus

LI Jun, YANG Liangbo, ZHU Dandan, LIU Dongbi, HOU Jun, TANG Jiping, ZHENG Xingwen, XIE Yibai, YU Yanfeng, NI Chengfan

2025, 54(12): 81-90. DOI: 10.15933/j.cnki.1004-3268.2025.12.009

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A micro⁃plot（2.25 m^²）experiment was conducted throughout the entire growth period of seed lotus（Taikonglian No. 36）to investigate the effects of major nutrients on yield and nutrient uptake.The experiment comprised seven treatments：balanced application of nitrogen，phosphorus，potassium，magnesium，and boron（BF）；nitrogen reduction（BF⁃N）；phosphorus reduction（BF⁃P）；potassium reduction（BF⁃K）；magnesium reduction（BF⁃Mg）；boron reduction（BF⁃B）；and a no⁃fertilizer control（CK）.The results indicated that under the BF treatment，the seed yield and total biological yield were 398.7 g/plant and 3 473.0 g/plant，respectively. The cumulative amounts of nitrogen，phosphorus，potassium，magnesium，and boron were 44.1，8.9，45.5，4.8，and 0. 044 g/plant，respectively. Compared with BF treatment，BF⁃N treatment significantly affected the growth of lotus seeds，reducing the single leaf area，petiole thickness，and leaf SPAD value（relative chlorophyll content）of lotus pods by 20.3% to 37.5%. Both BF⁃N and BF⁃K treatments significantly reduced the yield，biological total yield，and nutrient accumulation of lotus seeds，with BF⁃N treatment reducing 60.3%，70.0%，and 61.1% to73.9%，respectively，and BF⁃K treatment reducing 23.4%，37.7%，and 26.0% to 46.8%，respectively；The effects of BF⁃P，BF⁃Mg，and BF⁃B treatments on lotus yield were not significant，but they significantly reduced the biological total yield of lotus by 10. 0% to 13.9%. The accumulation of nitrogen，phosphorus，and potassium in BF⁃P treatment was significantly reduced by 12.2% to 18.0%，while the accumulation of various nutrients in BF⁃Mg treatment was significantly reduced by 11.9% to 19.7%.The boron accumulation in BF⁃B treatment decreased by as much as 51.5%.The effects of BF⁃P treatment on lotus magnesium and boron，and BF⁃B treatment on lotus nitrogen，phosphorus，potassium，and magnesium accumulation were not significant.The contributions of effective receptacle count，average number of carples，average fruit set percentage and average single seed weight to yield were 66.7%，19.3%，3.1% and 10.9% respectively.Except for fruit setting rate，they all showed significantly positive correlation with lotus yield at the level of 0.05 or above.In summary，under the conditions of this experiment，the comprehensive effects of nitrogen，phosphorus，potassium，magnesium，and boron on the yield and nutrient absorption of lotus are as follows：nitrogen>potassium>magnesium>phosphorus>boron.The seasonal utilization rates of phosphorus and boron are relatively low.In practical production，it is necessary to optimize the application strategies of phosphorus and boron fertilizers.

The Stress Tolerance Characteristics of Plant Growth⁃Promoting Rhizobacteria JHS0317 and XL0404 and Their Growth⁃Promoting and Disease Control Effects on Tobacco

LI Xiaojie, QIU Rui, LIU Chang, BAI Jingke, ZHANG Mengdan, CHEN Yuguo, MIAO Shuyue, LI Shujun

2025, 54(12): 91-100. DOI: 10.15933/j.cnki.1004-3268.2025.12.010

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The aims are to clarify the stress tolerance characteristics，growth⁃promoting and disease control effects，and mechanisms of Pantoea agglomerans JHS0317 and Enterobacter ludwigii XL0404，which were plant growth⁃promoting rhizobacteria.The sensitivity of strains JHS0317 and XL0404 to temperature，pH value，and ultraviolet radiation was determined using the plate counting method.The growth⁃promoting effect on tobacco and the control effect on tobacco root rot caused by Fusarium were determined through indoor pot inoculation experiments.The results showed that the suitable growth temperature for strain JHS0317 was 28—35 ℃，and the suitable growth temperature for strain XL0404 was 28—45 ℃，with a lethal temperature of 65 ℃.Both strains could grow under pH conditions of 5—9，and the optimal pH was 5—7.They were sensitive to ultraviolet radiation.The results of pot experiments showed that the treatment of strains JHS0317 and XL0404 had a significant promoting effect on tobacco plant height，aboveground fresh weight，root fresh weight，total root surface area，and total root volume，which increased by 62.58%，44.76%，55.23%，63. 83%，71.43%，and 82.32%，49.84%，43.02%，43.38%，61.11% respectively compared to the control treated with water.The content of plant hormones such as brassinolide，gibberellin，and auxin，and the activity of resistance enzymes such as phenylalanine ammonia lyase，per oxidase，polyphenol oxidase and superoxide dismutase in tobacco leaves were also increased compared to the control.The treatment of strains JHS0317 and XL0404 could reduce the incidence rate and disease index of tobacco Fusarium root rot，and the control effects were 49.99% and 24.98%，respectively.In summary，the plant growth⁃promoting rhizobacteria strains JHS0317 and XL0404 have good stress resistance to high temperature and acid and alkali，and have significant growth⁃promoting and disease prevention effects on tobacco，with good application prospects.

Isolation and Identification of the Dominant Pathogen of Salvia miltiorrhiza Root Rot and Observation of Its Infection Process in S.miltiorrhiza Roots

2025, 54(12): 101-109. DOI: 10.15933/j.cnki.1004-3268.2025.12.011

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To identify the dominant pathogen causing Salvia miltiorrhiza root rot in Henan Province and clarify
its infection process，diseased root samples were collected from three main producing areas（Yuzhou City，Mianchi County，and Fangcheng County）in Henan Province.The dominant pathogens were identified based on morphological characteristics and molecular biological analyses. Highly pathogenic isolates were screened through pathogenicity assays.The infection process of a selected isolate in S.miltiorrhiza roots was observed using green fluorescent protein（GFP）labeling and fluorescence microscopy.The results showed that Fusarium oxysporum exhibited the highest isolation frequencies（63.16%，48.28%，and 39.63%，respectively）among pathogens isolated from the three areas，indicating it as the dominant pathogen of S. miltiorrhiza root rot in Henan Province.Following inoculation of S.miltiorrhiza seedlings with F.oxysporum spore suspension：spores adhered to the root surface and began to germinate at 12 h post⁃inoculation（hpi）；germinated spores developed into hyphae，which spread along the intercellular spaces of epidermal cells between 24—48 hpi；hyphae expanded into the cortex and extended along parenchyma cells，eventually surrounding entire cells by 3—5 days post⁃inoculation（dpi）；extensive hyphal networks formed randomly within cortical cells by 7—9 dpi；hyphae surrounded the entire root cortex and extended to the root tip by 11 dpi；all root cells were infected by 14 dpi.At this stage（14 dpi），the stem base turned brown，fibrous roots became brittle and easily broken，and root tips of some plants turned yellow.In conclusion，F.oxysporum is the dominant pathogen causing S.miltiorrhiza root rot in Henan Province.This study elucidated the infection process of GFP⁃labeled F.oxysporum in S.miltiorrhiza roots from a histopathological perspective，providing a basis for targeted control strategies against this disease.

Investigation of Weed Community in Soybean Field and Screening of Highly Effective Herbicides

YANG Ruosheng, LI Jiuying, XUE Fei, SUN Lanlan, SU Wangcang, XU Hongle, WU Renhai

2025, 54(12): 110-120. DOI: 10.15933/j.cnki.1004-3268.2025.12.012

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To clarify the occurrence of malignant weeds in soybean fields in Henan Province and identify herbicides for controlling harmful broadleaf weeds therein，field surveys were carried out to statistically analyze the weed community in 19 major soybean⁃producing regions of Henan Province，and laboratory bioassays were performed to evaluate the activity of 11 soil treatment herbicides and 14 foliar treatment herbicides against the malignant weeds in soybean fields，including C.melo，C.argentea，C.communis，and A.retroflexus.The results of weed community survey showed that 8 weed species，including D.sanguinalis，A.australis，T.aestivum，A.retroflexus，E.indica，C.melo，P.oleracea，and E.crus‑galli，had a relative abundance of over 20% in soybean fields，while the relative abundance of A. theophrasti，E. prostrata，C.argentea，V.radiata，S.viridis，and C.ficifolium ranged from 3% to 20%.The results of laboratory bioassays indicated that the soil treatment herbicides metribuzin，prometryn，pyroxasulfone，and foliar treatment herbicides such as glufosinate⁃ammonium，lactofen，fluoroglycofen⁃ethyl，and MCPA⁃Na had high efficacy against C.melo.The soil treatment herbicides metribuzin，prometryn，oxyfluorfen，pyroxasulfone，flumioxazin，and s⁃metolachlor，along with foliar treatment herbicides such as fomesafen，bentazone，glyphosate，imazamox，glufosinate⁃ammonium，and imazethapyr exhibited high efficacy against C.argentea.For C.communis，only acetochlor with different application amounts showed higher efficacy and fluroxypyr had high control efficacy on it among foliar treatment herbicides.The soil treatment herbicides acetochlor，flufenacet⁃methyl，metribuzin，and pyroxasulfone，along with foliar treatment herbicides such as MCPA⁃Na，imazethapyr，and lactofen showed high efficacy against A.retroflexus.In conclusion，eight weeds species including D.sanguinalis，A.australis，A.retroflexus，and C.melo were the dominant noxious weeds in soybean fields of Henan Province. Metribuzin had the best pre⁃emergence control effect on C.melo and C.argentea，while acetochlor exhibited the best pre⁃emergence control effect against C.communis and A.retroflexus.The most effective foliar herbicides for C.melo，C.argentea，C.communis，and A.retroflexus were glufosinate⁃ammonium，fomesafen，fluroxypyr，and MCPA⁃Na，respectively.

Identification of R2R3⁃MYB Gene Family and Screening of Genes Related to Flower Color Formation in Paeonia suffruticosa

YAN Gengyang, GAO Wen, LI Hang, SHANG Ye, ZHANG Juping

2025, 54(12): 121-137. DOI: 10.15933/j.cnki.1004-3268.2025.12.013

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Based on the whole genome data of Paeonia suffruticosa，the R2R3⁃MYB gene family was identified to study its role in the formation of P.suffruticosa flower color and identify key regulatory genes.The physicochemical properties，chromosome location，phylogenetic analysis，gene structure，cis⁃acting elements，expression correlation and protein interaction were analyzed by bioinformatics methods.The expression patterns of genes in P.suffruticosa samples were verified by qRT⁃PCR. A total of 78 R2R3⁃MYB transcription factors of P.suffruticosa were identified，all of which were located in the nucleus and were mostly hydrophilic unstable proteins with abundant cis⁃acting elements and highly conserved R2 and R3 domains at the N⁃terminal.The 78 members were divided into 26 subfamilies（P1—P26）.Among them，Ps2RMYB21，Ps2RMYB38 and Ps2RMYB33 were homologous with S4 and S7，and had conserved motifs specific to the corresponding subfamilies respectively.The expression levels of Ps2RMYB33 and Ps2RMYB38 in normal sepals，colored sepals and normal petals were increased successively，and the expression patterns of structural genes PsCHS，PsCHI3，PsF3H1，PsDFR1，PsANS and PsUFGT4 were the same.There was a significant positive correlation between them，which was consistent with the accumulation trend of anthocyanins.However，Ps2RMYB21 was opposite to the expression pattern of these structural genes，and had a very significant negative correlation with the accumulation trend of anthocyanins.The results of qRT⁃PCR were basically consistent with the transcriptome data.To sum up，Ps2RMYB21，Ps2RMYB33，Ps2RMYB38are three key genes regulating the formation of peony flower color.

Establishment of Fluorescence Quantitative PCR Method for Detection of Sacbrood Virus in Honeybee

WANG Hongkun, ZHANG Xuan, ZONG Deqin, LI Yushi, LÜ Mingkui, ZHAO Hongmu

2025, 54(12): 138-147. DOI: 10.15933/j.cnki.1004-3268.2025.12.014

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To establish a highly sensitive detection method for sacbrood virus（SBV） in bees，the predominant circulating strain（GenBank accession no. MZ821922. 1）of SBV in Apis cerana cerana and Apis mellifera ligustica from Mengzi City，Yunnan Province，was identified by RT⁃PCR sequencing，and specific primers were designed based on the conserved regions of this strain. The total RNA of the samples was extracted and the cDNA was obtained by reverse transcription.The RT⁃qPCR detection method was established by SYBR Green Ⅰ dye method，the specificity，sensitivity，repeatability and practicality were verified.The results demonstrated the high quality of the designed primers.The concentration of SBV positive plasmid standard was 2.62×10^¹⁰ copies/μL，and the linear range was 2.62×10^³—2.62×1010 copies/μL（R^²=0.999 1），and the amplification efficiency was 102.6%.Furthermore，the
assay was proved to be highly sensitive（detection limit：2.62×10^⁰ copies/μL），specific（no cross⁃reactivity
with other common honeybee viruses），and reproducible（intra⁃and inter⁃assay CV<1.19%）.Two⁃step
RT⁃qPCR and conventional RT⁃PCR were performed on bee samples from Mengzi City and Gongshan
County，Yunnan Province. It was found that the detection sensitivity of two⁃step RT⁃PCR was significantly
higher than that of RT⁃PCR：the positive detection rate of total samples（56.1% vs 11.5%），Chinese bee
in Gongshan County（46.2% vs 5.4%），Chinese bee in Mengzi City（80. 0% vs 250%）and Italian bee in Mengzi City（53.3% vs 13.3%）.These findings further demonstrated the clinical applicability of the established method. In conclusion，the developed two⁃step SYBR Green Ⅰ⁃based RT⁃qPCR assay for SBV is sensitive and reliable，and can provide effective technical support for the rapid detection，early diagnosis，and epidemiological surveillance of SBV in China.

Preparation of Different Types of Tobacco Leaf Dry Distilled Flavor and Application in Heated Cigarettes

XU Da, LI Jiaxin, WANG Xuanjing, GUO Lei, LI Lipeng, LIU Wei, TIAN Yaowei, SUN Zhengguang, SUN Qiang, XU Chunping

2025, 54(12): 148-166. DOI: 10.15933/j.cnki.1004-3268.2025.12.015

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To expand the core flavor source of heated cigarettes，flue⁃cured tobacco，sun⁃cured yellow tobacco，burley tobacco and oriental tobacco were selected as raw materials to prepare flavors by dry distillation process. Firstly，the contents of conventional chemical components in four kinds of tobacco leaves were determined.Then，the dry distillation flavors were prepared and the extraction rate was calculated.The composition differences of volatile components were explored by GC⁃MS and principal component analysis.Finally，the aroma⁃enhancing effect was evaluated by sensory evaluation.The results showed that the nicotine and total nitrogen contents of burley tobacco were significantly higher than those of other tobacco leaves，the total sugar and reducing sugar contents of sun⁃cured yellow tobacco were the highest，and the chemical components of flue⁃cured tobacco were more balanced.The extraction rate of dry distillation perfume was 2. 8% for flue⁃cured tobacco，2.7% for oriental tobacco，2.5% for sun⁃cured yellow tobacco and 1.5% for burley tobacco. A total of 205 volatile components were identified by GC⁃MS，and the total contents were 7 024.00 μg/g for flue⁃cured tobacco，1 407.79 μg/g for sun⁃cured
yellow tobacco，3 881. 90 μg/g for burley tobacco and 6 951. 56 μg/g for oriental tobacco，respectively.Phenols，ketones and heterocyclic compounds were the main aroma components. Principal component analysis showed that the aroma components of flue⁃cured tobacco and oriental tobacco were similar，while burley tobacco and sun⁃cured yellow tobacco were significantly different.The sensory evaluation showed that the addition of dry distillation flavors could improve the smoking quality of heated cigarettes.Among them，the effect of flue⁃cured tobacco dry distillation flavor was the best，and the improvement effect was significant in aroma quantity，sweetness and strength.In summary，the aroma components of flue⁃cured tobacco are coordinated，the content of volatile components is high，and the burnt sweet and smoked aroma are prominent，which can effectively make up for the problem of insufficient aroma release of heated cigarettes.

Comprehensive Quality Evaluation of 11 Different Varieties of Pear Paste

ZHANG Sipu, CUI Wei, ZHANG Ke, LU Yunfeng, MIAO Jianyin, GUO Chaofeng, XIE Yuanming, ZHANG Yanmin, NIU Jiajia

2025, 54(12): 167-180. DOI: 10.15933/j.cnki.1004-3268.2025.12.016

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To identify pear varieties suitable for pear paste processing，this study measured the colorimetric indices，total acidity，vitamin C content，total phenolic content，amino acid composition，and volatile components of syrups produced from 11 major pear varieties cultivated in Henan Province.The pastes were assessed using an essential amino acid（EAA）scoring system，and a comprehensive evaluation was conducted via principal component analysis.The results showed that Huangguan pear paste had the highest clarity（87.65%）and the lowest browning degree（0.53）.Yuluxiang pear had the highest essential amino acid/total amino acid（25.04%）and essential amino/non⁃essential amino acid（34.17%），which was close to the requirements of ideal protein. Wonhwang pear paste achieved the highest essential amino acid ratio coefficient（SRC，87.35）.The essential amino acid index（EAAI）values of Hongxiangsu，Zhongli No.1，Yuluxiang，and Wanxiu pear pastes were all near 1.0，indicating high nutritional quality of their essential amino acids.A total of 41 volatile compounds were identified across varieties，with Hwasan pear paste having the highest number of volatile component types（21）.There was a strong correlation between various indicators.By introducing 71 indicators of different varieties of pear paste into the factor analysis calculation system，10 common factors were obtained. The cumulative contribution rate of the first 7 factors was 89.03%，and the model was obtained according to the variance contribution rate：Y=0.244 9F1+0.153 4F2+
0.143 6F3+0.113 6F4+0.100 4F5+0.073 0F6+0.061 5F7.Comprehensive scores ranked the varieties in descending order as follows：Akizuki，Hwasan，Wanxiu，Hwanggeumbae，Yuluxiang，Huangguan，Hongzaosu，Wonhwang，Suli，Hongxiangsu，Zhongli No. 1 pear paste.

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