Journal of Henan Agricultural Sciences

CONTENTS

2024, 53(12): 0.

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Research Progress of Intercropping Promoting Phosphorus Absorption and Utilization

WU Xin, BI Jiayu, GE Yingtong, HE Yangfa, WANG Yuyun

2024, 53(12): 1-9. DOI: 10.15933/j.cnki.1004-3268.2024.12.001

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The application of phosphate fertilizer and mineral weathering are the main sources of soil phosphorus，but the low utilization rate of phosphorus and the limited ability of crops to absorb and utilize phosphorus lead to the accumulation of soil phosphorus，which limits the growth of crops.Intercropping system can improve soil phosphorus availability.The effects of intercropping on phosphorus utilization rate were summarized.The mechanism of intercropping to improve soil phosphorus availability was reviewed from three aspects：soil microorganisms，crop roots，and changes in endogenous hormones.It is suggested that in the future，we should focus on the in‑situ characterization of plant root morphology，auxin‑related gene expression，soil microorganisms and related functional genes，in order to provide reference for improving phosphorus utilization rate through intercropping.

Effects of Chitosan Oligosaccharides with Different Molecular Weight on Growth and Physiological Characteristics of Maize Seedlings under Drought Stress

ZHANG Le, ZHANG Fuguo, JIN Yazhong, ZHANG Yitao, ZHANG Youli

2024, 53(12): 10-22. DOI: 10.15933/j.cnki.1004-3268.2024.12.002

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Under drought stress，50 mg/L chitosan oligosaccharides with different molecular weight （1 197，1 428，1 990 u）were sprayed to explore the effects of chitosan oligosaccharide with different molecular weight on the growth and physiological characteristics of maize seedlings，so as to provide a theoretical basis for the application of chitosan oligosaccharide with different molecular weight in production under drought stress.The results showed that under normal water condition，spraying chitosan oligosaccharide could promote the growth of maize seedlings. Under drought stress condition，spraying chitosan oligosaccharide could alleviate the impact of drought stress on the growth of maize seedlings.Among them，the plant height of treatments with chitosan oligosaccharides with small，medium and large molecular weight increased by 8.55%，9.37% and 5.80%，the stem diameter increased by 10.99%，5.23% and 1.31%，the dry weight of the aboveground part increased by 28.27%，38.82% and 19.83%，the dry weight of the underground part increased by 44.44%，39.58% and 29.17%，the leaf area increased by 43.45%，65.79% and 24.51%，the number of root branches increased by 114.57%，104.84% and 102.33%，the chlorophyll content increased by 43.07%，49.42% and 36.49%，the net photosynthetic rate（Pn）increased by 12.70%，22.49% and 6.80%，the content of superoxide anion decreased by 25.88%，35.74% and 21.83%，the activity of ascorbate peroxidase（APX）increased by 18.77%，29.92% and 11.43%，and the activity of glutathion peroxidase（GPX）increased by 23.30%，11.17% and 16.51%，respectively.Overall，under drought stress condition，spraying chitosan oligosaccharide with small molecular weight had the best effect on promoting the growth of the underground part，while spraying chitosan oligosaccharide with medium molecular weight had the best effect on promoting the growth of the aboveground part.The comprehensive evaluation results showed that the treatment with chitosan oligosaccharide with low molecular weight had the highest comprehensive score，followed by the treatment with chitosan oligosaccharide with medium molecular weight.

Breeding of New Wheat Germplasms with Resistance to Fusarium Head Blight by Backcross Transformation Combined with Molecular Marker‑Assisted Selection

LI Zhengling, HAN Liupeng, ZHAO Mingzhong, QI Xueli, ZHANG Yu

2024, 53(12): 23-30. DOI: 10.15933/j.cnki.1004-3268.2024.12.003

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In order to determine the effects of resistance genotypes and backcross times on Fusarium head blight（Fhb）resistance and agronomic traits of progeny materials，the main resistance genes Fhb1，Fhb2 and Fhb7 were pyramided in Zhengmai 1860，which was a widely planted variety in southern Huanghuai，by backcross transformation and molecular marker‑assisted selection.The Fhb resistance and main agronomic traits of different genotypes and backcross generations were analyzed.The results showed that Fhb1，Fhb2 and Fhb7 could increase the Fhb resistance by 38.9%—76.9%，and there was no significant correlation between these three resistance genes and the bad agronomic traits such as high culm，low density spikelet and low thousand‑grain weight. The percentage of scabbed spikelets of BC₁F₂，BC₂F₂，BC₃F₂populations in two years were 37.8%，31.4%，37.8% and 43.4%，45.8% 43.3%，respectively.Fhb resistance did not decrease with the increase of backcross times. With the increase of backcross times，the plant height of the progeny population materials decreased from 84.95 cm to 70.49 cm，the number of fruiting spikelets per panicle increased from 20.48 to 21.24，the panicle length increased from 10.23 cm to 10.49 cm，and the thousand‑grain weight increased from 37.36 g to 44.80 g.The main agronomic traits of the progeny population materials after three backcrosses were similar to those of the resistance gene receptor parent variety.The new germplasms Zhengzi2021CH182‑33 and Zhengzi2021CH182‑41 bred in this study with excellent comprehensive agronomic properties and Fhb resistance can be further used as disease‑resistance parents.

Identification of Rice B12D Gene Family and Expression Analysis under Submergence Stress

ZHANG Zhaoran, ZHU Boyi, GAO Song, LI Zhaowei

2024, 53(12): 31-42. DOI: 10.15933/j.cnki.1004-3268.2024.12.004

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In order to better understand the function of rice B12D gene，the genome‑wide identification and bioinformatics analysis of rice B12D gene family members were done，and the expression difference of B12D gene in rice（Nipponbare）and upland rice（Kunhan 1）under submergence stress was analyzed by real‑time fluorescence quantitative PCR（qRT‑PCR）.Finally，the possible interaction proteins of B12D were found by Alphafold analysis. The results showed that there were six B12D genes in rice genome，which were distributed on three chromosomes.The number of amino acids of B12D protein ranged from 89 aa to 98 aa，and the isoelectric point ranged from 9.65 to 10.34，which was alkaline.Except that OsB12D3 was located in the cell membrane，the other five B12D proteins were located in mitochondria.The B12D gene family had only one pairs of collinearity genes between Arabidopsis thaliana and rice，and three pairs of collinearity genes between rice and maize.In addition，OsB12D1 gene and OsB12D2 genes had a collinearity relationship with OsB12D3 gene.The protein sequence of rice B12D family had high homology and strong conservation.The promoter of rice B12D gene family contained a large number of light response elements，hormone response elements，stress response elements and development related elements.The expression levels of six B12D genes were different in different tissues，OsB12D1 and OsB12D2 genes were mainly expressed in roots；OsB12D3 gene was mainly expressed in ovary，embryo and endosperm；OsB12D4 gene was highly expressed in most tissues；OsB12D5 and OsB12D6 genes were significantly expressed in stamens，the expression level of OsB12D5 gene was lower，the expression level of OsB12D6 gene was higher，and there were different expression levels in other tissues for OsB12D6 gene.The expression levels of OsB12D1 and OsB12D2 genes in Nipponbare were significantly higher than that in Kunhan 1，the increase of OsB12D1and OsB12D2 gene expression levels after submergence stress was higher than that in Kunhan 1，and the decrease of ATP content was lower than that in Kunhan 1.Rice B12D interacted with heme C and cytochrome C1.In summary，the rice B12D gene family may play an important role in rice growth and development，response to submergence stress and so on.

Basic Study on Material Metabolism of Flue‑cured Tobacco Related to Maturity Based on Non‑targeted Metabolomics

LIU Tengfei, LI Qiujian, ZHANG Li, SHI Qiuhuan, LI Yuanbo, ZHANG Xue, XU Jiayang, JIA Wei, HAN Dan, XU Zicheng, SHI Jiuchang

2024, 53(12): 43-53. DOI: 10.15933/j.cnki.1004-3268.2024.12.005

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To clarify the quantitative indicators of fresh tobacco leaf maturity，Zhongyan 100 was used as the experimental material. Mid‑stem leaves with SPAD values of 10—15（S10~15），15—20（S15~20），and 20—25（S20~25） were harvested to explore the metabolomics characteristics，as well as the appearance evaluation，physical properties，chemical composition，aroma content，and sensory quality of their post‑cured leaves.The results showed that the post‑cured leaves of the S15~20 treatment obtained the optimal quality，including better appearance quality，good physical properties，harmonious chemical components，and the highest total amount of neutral aromatic substances，showing high maturity.According to the partial least squares discriminant analysis（PLS‑DA），32 mutual differential metabolites were identified between two comparison groups，including L‑tryptophan，gamma aminobutyric acid，geranic acid，pyrrole‑2‑carboxylic acid，trehalose，kojibiose，nicotinate D‑ribonucleoside，1‑methyladenosine，sphinganine，melibiitol，indole，scopoletin，18‑hydroxycorticosterone，25‑hydroxycholesterol，and kaempferide，et al.These differential metabolites might be key differential metabolites during the maturation process of tobacco.Then，a total of 48 significantly differential metabolites were identified across the two groups，including 11 amino acids，7 steroids，6 organic acids，6 sugars，3 alcohols，3 nucleotides，2 flavonoids，1 vitamin，and 9 other metabolites.These significantly differential metabolites were primarily enriched in pathways such as amino acid biosynthesis，ABC transporters，arachidonic acid metabolism，lysine degradation，biosynthesis of phenylpropanoids，and butanoate metabolism，which might play important regulatory roles during the maturation of tobacco leaves.

Effects of Different Mulching Methods on Soil Physicochemical Properties，Yield and Quality of Cigar Tobacco Leaves

HE Yuan, LI Longke, GAO Huajun, WU Zongyi, CHEN Guoyu, LIU Chunkui, LI Meng, WU Xiaozong, XU Chunping

2024, 53(12): 54-61. DOI: 10.15933/j.cnki.1004-3268.2024.12.006

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To investigate the effects of different mulching models on the physicochemical properties of tobacco‑planting soil，and the yield and quality of cigar tobacco leaves in Hainan，a field experiment using Haiyan 101 as the test material with five treatments including no mulch（CK），dual color film mulching and removed at 30—35 days after post‑transplant（G1），dual color film covering throughout the whole growth period（G2），biodegradable film mulching within the whole growth period（G3），and crop straw incorporation（G4）was conducted.The change of physical and chemical properties of tobacco‑planting soil under different mulching models was studied，and the regulation on agronomic traits of cigar tobacco during the main growth period，the physical characteristics，chemical properties and combustibility of leaves and the economic benefits were also explored.The results showed that compared with CK，the soil temperature under G3 significantly increased by 7.12% during the vigorous growth period，and that of G4 significantly increased by 10. 01% at maturity. During root spreading period and cluster period，G3 improved the available potassium content by 10.27% and 8.13%，espectively.Compared to CK，the SPAD value in cluster period treated with G3 significantly increased by 8.49%，and the plant height significantly increased by 13.10% in vigorous growth period. At maturity，the stem circumference，maximum leaf length and width under G3 significantly increased by 6.39%，8.09% and 6.17% compared with CK，respectively.After fermentation，compared with CK，G3 reduced the leaf thickness by 5.88%.Among all treatments，G3 had the highest tobacco yield，output value，and economic benefits，with a significant increase of 46.70% in economic benefits compared to CK.In summary，biodegradable film mulching during whole growing period could improve the soil physicochemical properties，maintain well physical characteristic，coordinated chemical composition and good combustibility，and ultimately generate better economic benefits in Hainan tobacco‑planting areas.

Identification of TCP Transcription Factors in Artemisia annua and Analysis of Their Expression Patterns under MeJA Treatment

MA Qiyang, SONG Ge

2024, 53(12): 62-74. DOI: 10.15933/j.cnki.1004-3268.2024.12.007

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Based on the genome data of Artemisia annua，the TCP gene family members were identified，and their protein physicochemical properties，evolutionary relationships，and cis‑acting elements were analyzed. Transcriptome data were used to analyze the expression pattern of TCP transcription factors in Artemisia annua under different light treatments，and qRT‑PCR was used to detect their expression under methyl jasmonate（MeJA）treatment.The results showed that a total of 29 TCP genes（AaTCPs）were identified. AaTCPs differed greatly in protein physicochemical properties，in which the number of amino acids and protein molecular mass ranged from 132 to 443 and 14.60 to 47.86 ku，respectively，and the isoelectric points，instability indices，and fat coefficients were in the ranges of 5.67 to 9.99，33.28 to 69.77，and 51.70 to 77.40，respectively，and all of them were nuclear proteins.Phylogenetic relationships showed that AaTCPs could be classified into class Ⅰ（11）and class Ⅱ（18），unevenly distributed on eight chromosomes.The Motif number of AaTCPs ranged from 1 to 7，all of them contained one conserved TCP2 structural domain，and a large number of light‑，hormone‑，and stress‑responsive elements were present in their promoters.Protein interaction predictions revealed tight interactions among the protein members of AaTCPs except for AaTCP12 and AaTCP8.Transcriptome data showed that the expression levels of AaTCPs varied under different light treatments，and the expression of most genes was induced by light. qRT‑PCR analysis revealed that the expression of AaTCPs all increased to different degrees under MeJA treatment，showing dynamic changes with the extension of treatment time.In summary，the AaTCP gene family plays a significant role in light regulation and hormone response processes.

The Complete Gene Sequencing of Bacilus subtilis Isolated from Rehmannia glutinosa and Study of Genes Related to Growth‑promoting Effects

LA Guixiao, WANG Linlin, GUO Junqi, ZHAO Yulong, GUO Hongxia, JIA Hui, YANG Tiegang

2024, 53(12): 75-85. DOI: 10.15933/j.cnki.1004-3268.2024.12.008

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To investigate the genes associated with the growth‑promoting effects of Bacillus subtilis in Rehmannia glutinosa，whole genome sequencing was performed in this study and the genes associated with the growth‑promoting effects were discovered through functional annotation.The results showed that the genome size of Bacillus subtilis，an endophytic growth‑promoting bacterium of Rehmannia glutinosa，was 4 310 699 bp，with a GC content of 43.44%，and a total of 4 521 predicted coding genes.A total of 3 013，3 354，4 395，206，52，and 212 functional genes were annotated in the clusters of orthologous groups（COG），gene ontology（GO），kyoto encyclopedia of genes and genomes （KEGG），carbohydrate‑active enzymes（CAZyme），comprehensive antibiotic resistance database（CARD），and virulence factor database（VFDB），respectively.The annotation results indicated that there were 46 genes related to growth‑promoting function，with the majority of these genes involved in phosphorus solubilization，nitrogen fixation，and iron carrier processes.In the meantime，anti‑SMASH predicted that a total of 409 genes were enriched into 12 secondary metabolite synthesis gene clusters，with the majority encoding eight repressors，including fengycin，1‑carbapen‑2‑em‑3‑carboxylic acid，bacillaene，bacilysin，subtilosin A，surfactin，pulcherriminic acid，and bacillibactin.Among these，pulcherriminic acid and bacillibactin had been observed to facilitate the chelation of ferric ions in the surrounding environment.

Analysis of Utilization Potential of Major Crop Straws and Livestock Manure in Jiangxi Province

CHE Qifeng, CHEN Xingyue, HU Tingting, ZHANG Jie, ZHANG Jing, HUANG Wenjie, TU Qihong

2024, 53(12): 86-98. DOI: 10.15933/j.cnki.1004-3268.2024.12.009

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This paper analyzed the spatial distribution characteristics of major crop straws and livestock manure in Jiangxi Province and their resource utilization potential，in order to promote the effective use of agricultural waste. Based on the spatial statistical analysis method of GIS，this study estimated the amount of major crop straws and livestock manure generated and their theoretical values of resource utilization in each county（city，district）of Jiangxi Province in 2021，and analyzed their spatial distribution patterns and cold hotspot areas.The results showed that in 2021，the total amount of major crop straw which could be collected in Jiangxi Province was 23.868 2 million tons，and the straw type was mainly rice straw.It generally showed the characteristics of rich in the north and poor in the south，mainly concentrated in the northern part of Poyang Lake Plain，and its theoretical value was equivalent to 11.337 4 million tons of organic carbon，227 000 tons of N，31 600 tons of P，420 800 tons of K，6.613 6 million tons of standard coal or 3.05×10⁹ m³methane gas.The annual production of livestock manure was 70.652 3 million tons，of which poultry manure produced the most，accounting for 40.27% of the total amount of livestock manure.It generally presented a north‑south poor，central rich characteristics，theoretically equivalent to 9.746 3 million tons of organic carbon，487 000 tons of N，175 300 tons of P，315 900 tons of K，11.581 million tons of standard coal，or 8.26×10⁹ m³ methane gas.There were 14 hotspots for crop straw，11 hotspots for livestock manure，and 5 double hotspots for straw‑livestock manure at 99% confidence level.Crop straw hotspots should realistically choose direct return to the field，basal material utilization，fertilizer utilization，energy utilization，feed utilization，raw material utilization，mushroom farming and other suitable utilization direction，livestock manure hotspots should comprehensively consider the development of biogas or way of on‑site return to the field recycling，straw‑livestock manure double hotspots should consider the multi‑level，diversified utilization pattern，giving priority to the integration of agriculture and animal husbandry，and the development of eco‑agriculture.

Detoxication Effects of Eight Kinds of Alleviators on Phytotoxicity of Nicosulfuron to Tobacco

YANG Nan, WANG Mingxin, JIANG Shengfei, LI Junying, FAN Liangxin, CHANG Dong, YIN Guangyu, XU Cuilian

2024, 53(12): 99-109. DOI: 10.15933/j.cnki.1004-3268.2024.12.010

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To address the phytotoxicity of residual nicosulfuron in maize field to following tobacco crop，the detoxification effects of commercially available safeners such as naphtha anhydride，cyclopropane sulfonamide，benoxacor，dichlormid，and plant growth regulators such as gibberellin，brassinolide，DA‑6，and Leu‑Ile‑Val mixture on phytotoxicity of nicosulfuron to tobacco were studied by indor pot experiment，field experiment，and biochemical analysis. The results showed that different amount of nicosulfuron demonstrated great inhibitory effect on the growth of tobacco，and the inhibitory effect increased with the increasing of nicosulfuron.The detoxication was the best when the concentrations of cyclopropane sulfonamide，dichlormid，gibberellin，brassinolide，and Leu‑Ile‑Val mixture were 10，20，40，0.1，30 mg/L，while the detoxication of other three alleviators was not significant.The results of field tests indicated that the detoxication of cyclopropane sulfonamide and dichlormid was better than other three chemicals，and dichlormid showed the best performance.After adding dichlormid for 63 days，the leaf length，leaf width，plant height，leaf number，and stem girth of tobacco was increased by 26.4%，21.6%，13.2%，75.2% and 14.3% compared to the control group.Compared with the control treatment with nicosulfuron，the treatment with cyclopropane sulfonamide and dichlormid by root irrigation showed 51.91%－61.81% and 60.29%－91.95% increase for superoxide dismutase（SOD）activity，catalase（CAT）activity，nitrate reductase（NR）activity and glutathione S‑transferase（GST）activity，respectively.Of note，the dichlormid treatment was better than cyclopropane sulfonamide. In summary，when nicosulfuron was used as herbicide in the previous maize field，the nicosulfuron‑caused phytotoxicity to tobacco can be effectively alleviated by cyclopropane sulfonamide and dichlormid，and 20 mg/L（50 mL/plant）of dichlormid should be used preferentially.

Study on the Diversity and Vertical Distribution of True Bugs in Taihang Mountains，Shanxi Province

CHEN Jing, ZHANG Zhenhua, WEI Jiufeng, ZHAO Qing, ZHANG Hufang

2024, 53(12): 110-118. DOI: 10.15933/j.cnki.1004-3268.2024.12.011

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In order to clarify the species diversity and distribution characteristics of true bugs in Taihang Mountains，Shanxi Province，a survey of true bugs diversity was conducted from June to September in 2022 and 2023，and vertical distribution at five different altitudes was analyzed in this area.The results of biodiversity analysis showed that a total of 6 103 true bugs were collected from June to September in 2022 and 2023，belonging to 25 families，41 subfamilies，110 genera，a total of 189 species.The dominant groups under different altitude gradients were different.The dominant group in altitude Ⅱ（770—1 140 m），altitude Ⅲ（1 140—1 510 m），and altitude Ⅴ（1 880—2 250 m）was Miridae. The dominant group at altitude Ⅰ（400—770 m）was Pentatomidae，and the dominant group at altitude Ⅳ（1 510—1 880 m）was Plataspidae.The altitude had significant effects on the individual number，species number，Shannon‑Wiener diversity index and Pielou evenness index，but had no significant effects on Margalef richness index and Simpson dominance index.The species composition of true bugs community at different altitudes was less similar，and the species similarity coefficient was ranging from 0.040 8 to 0.147 5，the true bug community at altitude Ⅱ and altitude Ⅲ was more similar.The environmental factor that had the greatest influence on the species number of true bugs in Taihang Mountains of Shanxi Province was the precipitation in the hottest quarter of the collection site（BIO18），and the correlation coefficient was 0.238（P<0.05）.The environmental factor that had the greatest influence on the individual number of true bugs in Taihang Mountains of Shanxi Province was the annual precipitation（BIO12），and the correlation coefficient was 0.263（P<0.05）.In summary，the ecological environment of Taihang Mountains in Shanxi Province is diverse，and the resources of true bugs species are abundant.In this area，the species richness of true bugs in low and high altitudes is relatively smaller，while the species in middle altitudes are abundant，and the vertical distribution pattern is single‑peak pattern.

Study on Genetic Diversity and DNA Fingerprint Map of Pumpkin Based on ISSR Markers

WANG Jialong, MA Kun, ZHOU Jing, ZHU Jinfang, GAO Peng, HUANG Fengjiao

2024, 53(12): 119-130. DOI: 10.15933/j.cnki.1004-3268.2024.12.012

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In order to provide theoretical basis for pumpkin variety identification，germplasm conservation and innovation，31 pumpkin germplasm resources were used as materials，and 7 inter‑simple sequence repeat（ISSR）primers with rich polymorphic information were selected to analyze the genetic diversity of the materials and establish DNA fingerprint map.The results showed that a total of 73 bands were amplified by 7 ISSR primers，including 72 polymorphic bands（polymorphic band rate：98.63%）.The average number of observed alleles，the average number of effective alleles，the average Nei’s gene diversity index，and the average Shannon’s diversity information index were 1.986 3，1.570 4，0.329 9and 0.494 4，respectively.The interspecific genetic similarity coefficients ranged from 0.12 to 0.89，with an average value of 0.46.Unweighted pair‑group method with arithmetic means（UPGMA）clustering analysis showed that 31 pumpkin materials were classified into Cucurbita moschata D.，Cucurbita pepo L.and Cucurbita maxima D.at the genetic similarity coefficient of 0. 46.Furthermore，the DNA fingerprint map for 31 pumpkin materials was constructed using the 14 polymorphic sites amplified by primer UBC811 and primer UBC843.All varieties in the DNA fingerprint map were unique，which could be used for molecular identification of varieties.In conclusion，the genetic diversity of 31 pumpkin materials was rich，Cucurbita moschata D.was closer to Cucurbita pepo L.，and the two were more distantly related to Cucurbita maxima D.，which could provide scientific basis for the conservation of pumpkin germplasm resources and variety innovation.The DNA fingerprint map of pumpkin based on the ISSR molecular marker technology could provide a new way for the identification of pumpkin varieties.

Effect of Two Seaweed Fermentation Solutions on Seed Germination and Seedling Growth of Cabbage

PANG Xiaowen, XIE Qinming

2024, 53(12): 131-138. DOI: 10.15933/j.cnki.1004-3268.2024.12.013

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In this study，we investigated the effects of different dilutions of Laminaria japonica and Enteromorpha prolifera fermentation solutions on seed germination and seedling growth of cabbage to provide theoretical basis for the development and application of seaweed resources in crop production.Using cabbage seeds‘Sundance 103’as test material，three different mass fraction groups（1%，5% and 20% groups） of two kinds of seaweed fermentation solution were used to carry out cabbage seed germination test and seedling foliar spraying test.The physiological regulation effects of fermentation solution on cabbage crop was analyzed by measuring the germination indexes of the seeds as well as the growth and physiological indexes of the seedlings.The results showed that the germination indexes of cabbage seeds were significantly different between the two seaweed fermentation solutions in different mass fraction groups：5% Enteromorpha prolifera fermentation solution had the highest germination potential，which increased by 9.32% compared to CK.20% Enteromorpha prolifera fermentation solution had the highest seed germination rate of 94.67%，which increased by 10. 94% compared to CK. 20% Laminaria japonica fermentation solution had the best effect in promoting embryo development.The embryo radicle length and germ length were significantly increased by 22.03% and 28.10%，respectively，compared to CK. The different mass fraction group treatments of the two seaweed fermentation solutions had significant differences in the growth indexes of cabbage seedlings：1% Laminaria japonica fermentation solution had the greatest growth promotion effect on seedlings，with the stem length increased by 17.46% compared to CK，and the fresh weight increased significantly by 34.85% compared to CK. The different mass fraction treatments of two kinds of seaweed fermentation solutions had significant differences in the physiological resistance indexes of cabbage seedlings：as the mass fraction of seaweed fermentation solution increased，the MDA content showed a rising trend，and the 1% treatment group was the lowest，which was smaller than CK.The SOD and CAT content showed a decreasing trend，and the 1% treatment group was the highest，which was significantly higher than CK.In summary，the two seaweed fermentation solutions could not only promote the seed germination and seedling growth，but also reduce the accumulation of MDA，increase the SOD and CAT content，and effectively enhance the level of plant resistance，with the best overall effect from 1% mass fraction Laminaria japonica fermentation solution.

Effect of Different Concentrations of Exogenous ZnONPs on the Heat Tolerance of Snapdragon

LIU Dan, MAO Fengling, BAI Jinxin

2024, 53(12): 139-148. DOI: 10.15933/j.cnki.1004-3268.2024.12.014

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Using snapdragon seedlings as test materials，this study investigated the effects of foliar spraying of different concentrations（0，50，100，200 mg/L）of zinc oxide nanoparticles（ZnONPs）on the growth，chlorophyll content，osmotic regulation substances，and enzyme activity and related substances in the AsA‑GSH cycle of snapdragon seedlings under day/night temperatures of 25℃/20℃，35℃/30℃ ，and 40℃/35℃ stress，in order to explore the role of ZnONPs in alleviating heat stress in snapdragon seedlings. The results showed that under high temperature stress，the height，stem thickness，leaf length，leaf width，and biomass of snapdragon significantly decreased.Applying different concentrations of ZnONPs could alleviate the growth inhibition of snapdragon. Under 40℃/35℃ stress，compared with the treatment of spraying 0 mg/L ZnONPs，the treatment of spraying 50—200 mg/L ZnONPs increased the total chlorophyll content and carotenoid content of snapdragon by 3.8%—7.7% and 4.9%—11.7%，respectively.The soluble sugar content and proline content of osmotic regulating substances increased by 12.7%—44.8% and 18. 2%—32.9%，respectively.Under 40 ℃/35 ℃ stress，the activities of ascorbate peroxidase（APX）， dehydroascorbate reductase （DHAR），monodehydroascorbate reductase（MDHAR），and glutathione reductase（GR）in the AsA‑GSH cycle of snapdragon continuously decreased. After spraying different concentrations of ZnONPs，the activities of APX，DHAR，MDHAR，and GR in snapdragon increased by 35.3%—86.3%，13.5%—24.6%，58.5%—81.5%，and 8.6%—19.7%，respectively. The content of ascorbic acid（AsA），glutathion（GSH），ascorbic acid/dehydroascorbate（AsA/DHA），and glutathione/reduced glutathione（GSH/GSSG） increased by 5.9%—11.7%，9.9%—16.5%，14.4%—31.1%，and 19.5%—37.7%，respectively，while the malondialdehyde（MDA）content，relative conductivity，O₂·^- production rate，and hydrogen peroxide（H₂O₂）content of snapdragon seedlings continued to decrease.The conclusion is that different treatments with ZnONPs can improve the heat tolerance of snapdragon，among which spraying 100 mg/L ZnONPs treatment has the best effect on improving the heat tolerance of snapdragon seedlings.

Analysis of Inhibitory Activity of Bacillus mojavensis against Gram‑Positive Bacteria and Its Antibacterial Secondary Metabolites

LI Haili, ZHANG Wanqi, XU Yindi, WANG Zhifang, ZHU Wenhao, YAN Xiangzhou, FENG Lili, YANG Fan, MA Chunjiang

2024, 53(12): 149-158. DOI: 10.15933/j.cnki.1004-3268.2024.12.015

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In order to screen endophytes of medicinal plants with good antibacterial effect against gram‑positive bacteria，surface disinfection method and plate confrontation method were used to isolate and screen endophytes of rosemary with antagonistic effect against Erysipelothrix rhusiopathiae，Streptococcus and Staphylococcus.Morphological and molecular biological identification of these endophytes were conducted，and their antibacterial active substances were explored through genome analysis.The results showed that seven strains of endophytic bacteria were isolated from different tissues such as roots，stems and leaves of rosemary.A gram‑positive endophyte（HNYJJ2211C）with good antibacterial effect was obtained from seven endophyte strains by plate confrontation assay.It was identified as Bacillus mojavensis（GCF000245335. 1）by morphology and molecular identification，which could effectively inhibit Erysipelothrix rhusiopathiae，Streptococcus and Staphylococcus. Whole genome sequencing analysis showed that the genome size of HNYJJ2211C was 4 147 805 bp，the content of G+Cwas 43.8%，and the number of its coding genes was predicted to be 4 075.3 311，1 453，2 503，440 and 158 genes were annotated by COG，GO，KEGG，VFDB and CAZy respectively.AntiSMASH software predicted that there were six gene clusters related to the synthesis of secondary metabolites. Surfactin，bacillaene，fengycin，bacillibactin，subtilosin A and bacilysin were the main secondary metabolites.In the HNYJJ2211C genome，a gene called TasA was discovered，located at 24 807 14—24 814 999 bp，having a total length of 786 bp（gene2481）and encoding 261 amino acids.From the gene clusters associated with antibacterial active substances contained in its genome，it can be predicted that this strain has the
potential to develop new drugs and serve as a feedable microorganism.

Effects of Adding Different Rations of Qianjinqiao San on Feeding Performance of Finishing Pigs

ZHANG Tao, TANG Yuanjiang, TAO Xiaoyan, LU Yuxi, ZHOU Sixuan, DU Chunlin, ZHANG Xiong, ZHANG Jing, TAN Ya, YANG Yueqian

2024, 53(12): 159-166. DOI: 10.15933/j.cnki.1004-3268.2024.12.016

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This study aimed to investigate the effects of Fagopyrum dibotrys Qian Zhong extract（Qianjinqiao San） on growth performance，carcass traits，meat quality，blood biochemical indexes，antioxidant indexes and immune indexes of finishing pigs，and to explore its application effect on finishing pigs. A total of 24 fattening pigs with similar body weights（Duroc×Landrace×Large White）were randomly divided into 4 groups with 3 replicates per group and 2 pigs per replicate.The control group（R1）was fed a farage‑based diet，and pigs in the experimental groups（R2，R3 and R4）were fed the basal diet supplemented with 0.3%，0.6% and 1.2% Qianjinqiao San.The experiment lasted for 70 days.The results indicated that compared to the R1 group，there were no significant differences in average daily gain（ADG），average daily feed intake（ADFI），and feed‑to‑gain ratio（F/G）among finishing pigs in groups R2，R3，and R4（P>0.05）.Furthermore，the contents of umami amino acids，saturated fatty acids，monounsaturated fatty acids in muscle tissues across different groups did not show significant variation either（P>0.05）；the amount of total polyunsaturated fatty acids in R3 group were significantly reduced（P<0.05）.There were no significant difference in the slaughter rate，backfat thickness，pH_1h，pH_24h，eye muscle area，cooked meat yield，and intramuscular fat content among the different proportion groups of Qianjinqiao San（P>0. 05）.In comparison to the R1 group，blood urea nitrogen levels（BUN）in the R2 group was significantly lower（P<0.05）.Moreover，serum activities of catalase（CAT）and total antioxidant capacity（T‑AOC）were significantly increased in both the R3 and R4 groups（P<0. 05）；additionally superoxide dismutase（SOD）activity levels increased by approximately 8. 04%，2. 66% and 9. 91% respectively across groups R2，R3，and R4（P>0.05）. The contents of immunoglobulin A（IgA），immunoglobulin G（IgG），immunoglobulin M（IgM）in the serum of R3 and R4 groups were significantly increased（P<0. 05）；the levels of interferon‑γ（IFN‑γ）in all test groups were higher than those in the control group，with R2 group showing the most significant increase of 37.43%（P<0. 05）.In conclusion，the supplementation of Qianjinqiao San at concentrations ranging from 0.3% to 1.2% during the late fattening period does not significantly impact growth performance，carcass characteristics，or meat quality. However，it can enhance serum antioxidant capacity and improve body immunity to a certain extent.

Preparation and Identification of Monoclonal Antibody against gE Protein of Porcine Pseudorabies Virus

WANG Ruining, WANG Xun, LI Ge, LI Qingmei, LI Cunfa, YANG Suzhen, GUO Junqing

2024, 53(12): 167-173. DOI: 10.15933/j.cnki.1004-3268.2024.12.017

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In order to develop an immunodiagnostic reagent for porcine pseudorabies virus（PRV），BALB/c mice were immunized with the recombinant gE protein expressed in HEK293F cells.The spleen cells of immunized mice and SP2/0 myeloma cells were fused to generate hybridoma cells.Indirect ELISA and immunoperoxidase monolayer assay（IPMA）were used to screen positive hybridoma cells so as to prepare and identify monoclonal antibodies against the gE protein of PRV.The outcomes demonstrated that two hybridoma cell strains，named 2B5 and 8F7，respectively，which steadily secreted monoclonal antibody against the gE protein were developed.The ELISA titers of ascites were all 1∶1 000 000.IgG1 was the heavy chain and Kappa was the light chain，according to the monoclonal antibody isotype assay.The specificity assay revealed that the two monoclonal antibodies only reacted with PRV and not with other viruses. SDS‑PAGE results showed that the purified monoclonal antibodies had high‑purity specific bands at about 50 ku and 25 ku.The two monoclonal antibody strains could react specifically with 293T cells transfected with gE plasmid according to indirect immunofluorescence assay（IFA）detection.Western blot showed that the specific protein band appearing in the culture supernatant of two hybridoma cells was about 120 ku，indicating that the two monoclonal antibodies could specifically recognize PRV. To summarize，this study successfully prepared two strains of monoclonal antibodies against gE protein，exhibiting excellent specificity and high titer，which provided crucial biological materials required for the subsequent development of diagnostic kits.

Dynamics of Fruit Development and Modeling of Ruanzhidahong Dragon Fruit

SONG Liang, XU Tingting, LU Xuehe, ZHANG Haidong

2024, 53(12): 174-180. DOI: 10.15933/j.cnki.1004-3268.2024.12.018

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The study investigated the dynamic changes in four growth parameters during fruit development of Ruanzhidahong dragon fruit grown in the Yangtze River Delta region.The growth parameters were horizontal and vertical diameter，fresh and dry weight.Four growth equations（Logistic，Gompertz，Richards，cubic polynomial） were chosen to establish fruit growth models using physiological development time（PDT）as the scale. The appropriate mathematical models for each parameter were determined based on the fitting results and validated using independent data.The results indicated that cubic polynomial was appropriate for the dynamic changes of horizontal and vertical diameters，while Logistic equation was suitable for determining the fresh and dry weights of Ruanzhidahong fruits.The R^² values for all fitted models exceeded 0.98.The model’s test results indicated that the relative root mean square error（RRMSE）for horizontal and vertical diameter，fresh and dry weight was less than 15%，demonstrating a successful simulation. Therefore，the models obtained in this study demonstrate high predictive accuracy for growth parameters of Ruanzhidahong dragon fruit.They can be served as a theoretical foundation for cultivation management of this variety in the Yangtze River Delta region.

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