Journal of Henan Agricultural Sciences

CONTENTS

2025, 54(3): 0.

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Cloning and Function Analysis of Rice Sucrose Transporter OsSUT4 Gene

CAI Yumeng, WANG Ying, HE Fei, LIU Yanqing, LIU Jingyan, GUO Yanli, SUN Yue, SU Jingping, WANG Shengjun, LI Junling

2025, 54(3): 1-7. DOI: 10.15933/j.cnki.1004-3268.2025.03.001

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The rice sucrose transporter gene OsSUT4 was cloned，and sucrose transport capability of OsSUT4 was verified by yeast heterologous expression system.The gene was overexpressed in rice，the phenotype of transgenic plants was observed，and the salt tolerance and drought resistance was identified，so as to explore the role of the OsSUT4 gene in stress resistance in rice and provide new gene resources and theoretical foundations for rice genetic improvement. The results showed that OsSUT4 gene was primarily expressed in the flag leaves at jointing stage；The sucrose transport capability of OsSUT4 was confirmed for the first time using the yeast heterologous expression system；Overexpression of OsSUT4 gene did not significantly alter the grain morphology or the thousand⁃grain weight，but significantly increased plant height and survival rates under salt and drought stresses，thereby enhancing the salt
tolerance and drought resistance of rice.

Effect of Spraying Amino Acid Chelated Zinc after Anthesis on Synthesis and Accumulation of Starch and Protein in Wheat Grains

ZHANG Bowen, JIN Haiyang, XU Haixia, ZHENG Fei, HE Ning, SU Yazhong, ZHAO Guojian, CHENG Hongjian, CHENG Xiyong, LI Xiangdong

2025, 54(3): 8-19. DOI: 10.15933/j.cnki.1004-3268.2025.03.002

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Zinc sulfate，zinc aspartate acid and zinc glycinate of 0.2% and 0.4% were sprayed on wheat after anthesis with water as the control（CK），and the effects of spraying amino acid chelated zinc on the contents of soluble sugar，starch，amino acid and protein，and the activities of enzymes related to starch synthesis，nitrogen metabolism in wheat grains were studied in different periods，so as to identify the most efficient zinc fertilizer，and provide theoretical basis and technical reference for the high⁃yield and quality enhancement of wheat production. The results showed that compared with CK，spraying zinc glycinate and zinc aspartate significantly increased the activities of soluble starch synthase in grains at 10 days after spraying at Longting and Dancheng experimental sites，with increase rates of 16.26%—27.45% and 24.51%—39.13%，respectively；At Dancheng experimental site，spraying zinc sulfate，zinc glycinate and zinc aspartate significantly increased the activities of bound starch synthase in grains at 10 days after spraying，with increase rates of 9.93%—17.65%，while at Longting experimental site，only spraying 0.2% zinc aspartate significantly increased the activity of bound starch synthase in grains by 12.15%；Spraying zinc glycinate and zinc aspartate significantly increased the starch content in grains at 10 days after spraying at Longting and Dancheng experimental sites，with increase rates of 14.49%—20.82% and 13.26%—20.68%，respectively，there was no significant difference between zinc sulfate treatments and CK，and there was no significant difference in the starch content in grains at 20 days after spraying and mature stage between zinc treatments and CK.Compared with CK，spraying zinc glycinate significantly increased the activity of glutamine synthetase in grains at 10 days after spraying at Longting and Dancheng experimental sites，with increase rates of 16.67% and 10.42%—14.58%，respectively；Significantly increased the activity of glutamate synthetase in grains at 10 and 20 days after spraying at Dancheng experimental site，with increase rates of 12.18%—15.53% and 11.84%—12.76%，respectively.Spraying 0.2% zinc aspartate significantly increased the activities of glutamine synthetase and glutamate synthase in grains at 20 days after spraying at Longting experimental site，with increase rates of 23.21% and 10.83%，respectively. Spraying 0.2% zinc aspartate or 0.2% zinc glycinate significantly increased the protein content in grains at mature stage at both experimental sites，spraying 0.4% zinc glycinate only significantly increased the protein content in grains at mature stage at Dancheng experimental site，and 0.2% zinc aspartate treatment had the highest increase rates at both experimental sites，which were 7.90% and 7.25%，respectively.In conclusion，spraying 0.2% zinc aspartate，zinc glycinate after anthesis can significantly increase the protein synthesis and accumulation in wheat grains，improve the protein content in grains at mature stage，and 0.2% zinc aspartate has the best effect.

Effect of Bacillus on Seed Germination and Physiological and Biochemical Characteristics of Maize under Salt⁃Alkali Stress

DENG Cong, MA Lu, WANG Qingsong, FU Jian, WANG Yufeng, YANG Kejun

2025, 54(3): 20-30. DOI: 10.15933/j.cnki.1004-3268.2025.03.003

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In order to study the mitigation effect of Bacillus on salt⁃alkali stress of maize at germination stage，maize with different salt⁃alkali tolerance（Kennian No.1，Xianyu 335，Jiaqiu 105）were used as materials，the germination test was carried out by inoculating Bacillus under salt stress（100 mmol/L NaCl）and alkali stress（60 mmol/L NaHCO₃），and the effects of Bacillus on the length，antioxidant enzyme（POD，SOD，CAT，APX）activity，osmotic adjustment substance（soluble protein，soluble sugar，free proline）content，membrane lipid peroxidation（superoxide anion and malondialdehyde contents）of maize plantule and embryo root under salt⁃alkali stress were studied.The results showed that the growth of maize plantule and embryo root was inhibited under salt⁃alkali stress，and the activities of antioxidant enzymes，the contents of osmotic adjustment substances and the degree of membrane lipid peroxidation all showed an upward trend. Compared with salt⁃alkali stress，Bacillus could increase the length，antioxidant enzymes activities and contents of osmotic adjustment substances，and reduce the degree of
lipid peroxidation of plantule and embryo root of three maize varieties under salt⁃alkali stress.Under salt stress，inoculation of Bacillus decreased the content of superoxide anion of maize plantule by 14.74 % —49.30%，and the content of superoxide anion of maize embryo root by 0.37%—57.55%；Under alkali stress，inoculation of Bacillus decreased the content of superoxide anion of maize plantule by 18.11% —46.31%，and the content of superoxide anion of maize embryo root by 15.36%—46.83%. Overall，Bacillus can scavenge reactive oxygen species and maintain osmotic pressure balance by increasing the activities of antioxidant enzymes and contents of osmotic adjustment substances，reduce the damage caused by salt⁃alkali stress to maize germination，and then improve the
tolerance to salt and alkali.

Effects of Irrigation Methods on Crop Yield and Water⁃Nitrogen Use Efficiency under Wheat⁃Peanut Rotation System

GAO Cuimin, TIAN Yuan, ZHANG Ruiqing, HE Fang, HAN Weifeng, ZHANG Yunhong, PAN Xiaoying, YANG Yonghui

2025, 54(3): 31-39. DOI: 10.15933/j.cnki.1004-3268.2025.03.004

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Based on a four⁃year field positioning experiment，three irrigation methods，including traditional irrigation（TI），sprinkler irrigation（SI）and drip irrigation（DI）were used to explore the effects of irrigation methods on crop photosynthetic characteristics，grain yield and water⁃nitrogen use efficiency under wheat⁃peanut rotation system，with no irrigation treatment as control（CK），so as to provide a theoretical basis for promoting the popularization and application of water⁃saving irrigation technology in wheat⁃peanut rotation system. The results showed that irrigation methods had significant effects on crop SPAD value，net photosynthetic rate，transpiration rate，yield and its components，water and nitrogen use efficiency under wheat⁃peanut rotation system，and DI treatment was the highest among all treatments，followed by SI treatment. Under wheat⁃peanut rotation system，compared with other treatments，net photosynthetic rate，transpiration rate and SPAD value of DI treatment increased by 3.3%—105.4%，5.6%—109.6% and 1.0%—19.4%，respectively；Grain number per spike，1 000⁃grain weight and grain yield of wheat increased by 6.9%—21.1%，4.6%—39.2% and 9.2%—50.6%，respectively；Number of full pods per plant，100⁃pod weight，kernel rate and yield of peanut increased by 8.8%—35.5%，2.8%—16.2%，1.7%—5.6% and 4.1%—30.1%，respectively.The annual grain yield，water use efficiency and partial factor productivity of nitrogen of wheat⁃peanut showed DI>SI> TI .Compared with TI treatment，the total yield，water use efficiency and partial factor productivity of nitrogen of wheat⁃peanut of DI treatment increased by 17.9%，19.5% and 17.9%，respectively，and those of SI
treatment increased by 10.1%，14.9% and 10.1%，respectively. In summary，under the condition of this
study，drip irrigation is the best irrigation method for increasing yield and efficiency of wheat⁃peanut，followed by sprinkler irrigation，both of which can achieve the goal of high yield and water and nitrogen use efficiency.

Effects and Comprehensive Evaluation of Different Plant Regulators on Growth of Sugar Beets

YUAN Tuantuan, LIU Changbing, BEN Qiuping, DING Zhaofei, WANG Kaiyong, FAN Hua, LI Weinong, LI Zhiqiang

2025, 54(3): 40-49. DOI: 10.15933/j.cnki.1004-3268.2025.03.005

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To identify suitable growth regulators for sugar beets under drip irrigation in Xinjiang and improve the sugar beets yield，the effects of different plant growth regulators on agronomic traits，photosynthetic characteristics，yield of sugar beets were investigated. At 50，70，and 90 days after the emergence of sugar beets，four plant growth regulators DPC，CCC，S3307 and ETH，were sprayed three times on the sugar beets，with water as the control（CK），in 2021 and 2022.The results showed that plant growth regulators could effectively reduce plant height，increase photosynthetic pigment content in sugar beet leaves by 7.38%—16.81% and the net photosynthetic rate by 0.54%—21.54%.The four treatments could increase the yield of sugar beet by 5.97%，12.91%，2.84%，14.20% in 2021，and 10.46%，13.08%，8.39%，18.17% in 2022，respectively，compared with CK. The sugar production of the four regulator treatments was increased by 4.56%，10.90%，6.87%，10.35% in 2021，and 8.34%，11.87%，9.44%，0.43% in 2022.By combining PCA（principal component analysis）and entropy weighted TOPSIS comprehensive evaluation，the impact level of the four growth regulators on sugar beets showed CCC>ETH>DPC>S3307>CK.Overall，CCC was the best regulator in regulating the growth，yield，and quality of sugar beets.

Screening and Identification of Plant Growth⁃Promoting Rhizobacteria and Their Growth⁃Promoting Effects on Tobacco

NIU Wenyan, LI Xiaojie, XU Min, QIU Rui, BAI Jingke, LIU Chang, CHEN Yuguo, SI Junke, LI Shujun, SONG Pengyu, SONG Shouye

2025, 54(3): 50-59. DOI: 10.15933/j.cnki.1004-3268.2025.03.006

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To explore multifunctional beneficial microbial strains that promote tobacco growth，bacterial strains with growth⁃promoting potential were isolated from the rhizosphere soil of healthy crops in Jiaxian County，Pingdingshan City，Henan Province.The bacterial strains were identified through morphological，physiological，biochemical analyses，and 16S rDNA sequence analysis.Their growth⁃promoting effects were confirmed through petri dish seed germination tests and pot experiments involving tobacco. The results indicated that strains JLS0502 and JLS0413 had indole⁃3⁃acetic acid（IAA）concentrations of 28.35 mg/L and 22.03 mg/L in their fermentation broths， respectively， with 1⁃aminocyclopropane⁃1⁃carboxylic acid（ACC）deaminase activities of 546.64 U/L and 521.21 U/L.Both strains also exhibited phosphate solubilization，potassium release，and nitrogen fixation capabilities.Identification confirmed that strain JLS0502 was Leclercia adecarboxylata，while strain JLS0413 was Enterobacter ludwigii.Treatment with strains JLS0502 and JLS0413 significantly improved the germination rate of tobacco seeds and promoted root elongation. Pot experiments showed that strains JLS0502 and JLS0413 significantly improved agronomic traits，including plant height，fresh weight，and root morphology of tobacco. Specifically，compared to the uninoculated control，the plant height，total fresh weight，root fresh weight，total root surface area，and total root volume increased by 66.05%，55.82%，59.30%，22.55%，29.37%，and 75.38%，66.29%，105.23%，90.15%，and 140.48%，respectively.Furthermore，the levels of plant hormones，including brassinolide（BR），gibberellin（GA），and indole⁃3⁃acetic acid（IAA），as well as the activities of defense enzymes，such as phenylalanine ammonia lyase（PAL），catalase（CAT），peroxidase（POD），polyphenol oxidase（PPO），and superoxide dismutase（SOD），were significantly higher in the tobacco leaves than in the control group.These findings highlight the significant growth⁃promoting effects of strains JLS0502 and JLS0413 on tobacco.

Effects of 6⁃BA on Physiological Characteristics of Primary Rhizomes of Polygonatum sibiricum Red.

WANG Rongrong, YUE Hewei, ZHANG Bo, HU Peng, LIU Yang, YANG Xue, LI Juan, LI Lianzhen

2025, 54(3): 60-70. DOI: 10.15933/j.cnki.1004-3268.2025.03.007

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To investigate the effects of 6⁃benzylaminopurine（6⁃BA）on the physiological characteristics of the initial root tubers of Polygonatum sibiricum Red.，four mass concentrations of 6⁃BA（0 mg/L，CK；50 mg/L，M1；100 mg/L，M2；200 mg/L，M3）were applied to the initial root tubers.The germination and growth of the initial root tubers were observed，and the activities of peroxidase（POD），catalase（CAT），superoxide dismutase（SOD），lipase，malate dehydrogenase（MDH），6⁃phosphogluconate dehydrogenase（G6PDH），and the contents of malondialdehyde（MDA），lipids，free amino acids，proteins，starch，and soluble sugars were measured.The results showed that the germination rate，seedling leaf length，leaf width，diameter，main root length，lateral root number，and dry weight of the primary rhizomes treated with 6⁃BA were significantly higher than those in the CK group（P<0.05）.On the 61st day，the emergence rates of M1，M2，M3，and CK were 33.10%，60.12%，46.66% and 3.68%；Among them，the M2 treatment showed significant differences compared to other treatments（P<0.05）. On days 1—31，the soluble sugar，starch，protein，free amino acids，and fat content in the primary rhizomes of Polygonatum sibiricum Red.treated with 6⁃BA were significantly lower than those in the CK group（P<0.05），and the
lipase activity was significantly higher than that in the CK group（P<0.05）；The activities of POD，SOD，CAT，MDH，and G6PDH enzymes in the primary rhizomes of Polygonatum sibiricum Red.treated with 6⁃BA were higher than those in the CK group，while the MDA content was lower.The activities of SOD，CAT，and G6PDH reached their highest values on the 41st day after treatment.Among them，the SOD activity in M1，M2，and M3 increased by 39.87%，86.99%，and 51.02%，respectively，compared to the CK group；The CAT activity in M1，M2，and M3 increased by 30.25%，54.06%，and 45.79% respectively compared to CK；The G6PDH activity in M1，M2，and M3 increased by 51.90%，132.42%，and 76.89% respectively compared to CK.The results of correlation and principal component analysis showed that the eight indicators of starch content，soluble sugar content，protein content，free amino acid content，fat content，peroxidase activity，superoxide dismutase activity，and catalase activity could be used as core evaluation indexes to characterize the emergence and growth of primary rhizomes of Polygonatum sibiricum Red. under different concentrations of 6⁃BA（0，50，100，200 mg/L）.In conclusion，6⁃BA can significantly improve the emergence rate，substance metabolism level，and antioxidant enzyme activity of the primary rhizomes of Polygonatum sibiricum Red.，and the optimal concentration is 100 mg/L.

Effect of Piriformospora indica on Growth and Physiological Characteristics of Endangered Tibetan Medicine Codonopsis convolvulacea Kurz.var.vinciflora（Kom.）L.T.Shen

CHAI Guozhu, WANG Meihui, QIU Zejiang, YANG Qifan, YUAN Fang

2025, 54(3): 71-76. DOI: 10.15933/j.cnki.1004-3268.2025.03.008

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In order to improve the yield and stress resistance of cultivated Codonopsis convolvulacea Kurz.var.vinciflora（Kom.）L.T.Shen，the promotion effects of endophytic fungus Piriformospora indica on its growth were explored.The effects of P.indica on the growth and physiological characteristics of C.convolvulacea were studied by outdoor pot experiment with two treatments of control group（CK）and inoculation group（PI）.The results showed that the symbiotic system of P.indica and C.convolvulacea was successfully established by soaking and watering inoculation，and the colonization rate was 70.37%.Compared with CK，the leaf number，stem number，main stem length，aboveground fresh weight and dry weight of C.convolvulacea colonized by P.indica were significantly increased by 101.27%，102.41%，76.07%，278.23% and 265.52%，respectively.The fibrous root number，fibrous root length，underground fresh weight and dry weight were significantly increased by 52.57%，63.39%，23.65% and 32.52%，respectively，and the tuber diameter was increased by 12.14%.The activities of catalase（CAT）and peroxidase（POD） were significantly increased by 9.79% and 19.35%，respectively，and the activity of superoxide dismutase（SOD） was reduced by 28.02% compared to CK.The contents of soluble sugar and proline were significantly increased by 42.84% and 92.19%，respectively.Root activity was significantly increased by 65.57%.In summary，P.indica can promote the growth and enhance stress resistance of C.convolvulacea，and can be used to improve the cultivation quality of C.convolvulacea.

Identification and Functional Characterization of Endophytic Microbes from Mulberry Tree（Morus alba L.）

LI Sha, YANG Jiahu, WANG Yanhui, HUANG Xinqiu, YANG Wen

2025, 54(3): 77-86. DOI: 10.15933/j.cnki.1004-3268.2025.03.009

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This study used methods such as microbial culture，target gene sequence amplification，and homology identification to isolate and identify endophytic microbes in Morus alba L.，and explored the strain types，distribution characteristics，and functional characteristics of endophytic bacteria and fungi in the roots，stems，and leaves of Yunsang No.6 mulberry.The results showed that a total of 43 endophytic bacteria belonging to 9 genera and 22 endophytic fungi belonging to 14 genera were isolated from mulberry trees. Among them，21 bacteria and 11 fungi were isolated from roots，13 bacteria and 5 fungi were isolated from stems，and 9 bacteria and 6 fungi were isolated from leaves.There were significant differences in the distribution of endophytic microbes in the roots，stems，and leaves of mulberry trees，there were Falsibacillus pallidus，Bacillus velezensis and Pseudomonas koreensis shared in the roots and stems of mulberry trees，Pantoea agglomerans，Bacillus subtilis and Pseudomonas koreensis shared in the stems and leaves，and Pseudomonas koreensis shared in the roots，stems，and leaves；Endophytic fungi Colletotrichum siamense was shared in mulberry roots and leaves，but no common fungi were found to exist in the stems and leaves，stems and roots.Further screening of functional microbes showed that 8 strains of nitrogen fixing microbes（7 bacteria，1 fungus），4 strains of phosphorus solubilizing microbes（2 bacteria，2 fungi），and 8 strains of potassium solubilizing microbes（6 bacteria，2 fungi）were obtained from 65 endophytic microbes，respectively.In summary，endophytic bacteria and fungi in mulberry trees exhibit diversity and distribution differences in roots，stems，and leaves.Among endophytic bacteria and fungi in mulberry trees，there are functional strains that can dissolve phosphorus，nitrogen，and potassium.

Effect of PGPR and Melatonin on the Antioxidant Properties and Arsenic Absorption of Coreopsis grandiflora

LIANG Ya’nan, WU Mengxi, XIAO Yongqin, ZENG Jingjin, DUAN Yili

2025, 54(3): 87-98. DOI: 10.15933/j.cnki.1004-3268.2025.03.010

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Using Chaoyang seedlings of Coreopsis grandiflora as test materials，an indoor pot experiment was conducted to investigate the effects of inoculating plant growth promoting bacteria（PGPR）and foliar spraying of melatonin（MT）at concentrations of 5 μmol/L（MT₁），20 μmol/L（MT₂），and 50 μmol/L（MT₃）under 80 mg/kg arsenic（As） stress on the growth，root architecture，chlorophyll content，chlorophyll fluorescence parameters，nutrient elements，As absorption，and antioxidant properties of C.grandiflora.The results showed that compared with the CK treatment without As stress，the overall plant height，leaf area，and total biomass（aboveground and root dry weight）of C.grandiflora were significantly reduced under As stress.Foliar spraying of different concentrations of MT and MT+PGPR treatment could alleviate the damage of As stress to C.grandiflora. Under As stress，compared with the treatment without PGPR inoculation or MT spraying，the root system configuration（total root length，total volume，total surface area，root tip number）of C.grandiflora treated with As+MT2+PGPR was significantly improved.The root activity increased by 166.1%，and the chlorophyll content，maximum photochemical efficiency
（Fv/Fm） of PS Ⅱ ，and photochemical quenching coefficient（qP） increased by 56.7%，15.6%，and 62.8%，respectively，while the non photochemical quenching coefficient（NPQ）decreased by 23.7%.As stress significantly reduced the nitrogen，phosphorus，and potassium content in the aboveground and root systems of C.grandiflora，while As+MT2+PGPR treatment could increase the nitrogen，phosphorus，and potassium content in the aboveground and root systems to varying degrees.At the same time，foliar spraying of different concentrations of MT and MT+PGPR could increase the As content in the aboveground and root parts of C. grandiflora，improve the As transport coefficient and enrichment coefficient，activate the activities of superoxide dismutase（SOD），catalase（CAT），ascorbate peroxidase（APX），and glutathione reductase（GR），and increase the content of ascorbic acid（AsA）and glutathione（GSH）to alleviate oxidative stress on C.grandiflora.In summary，As+MT₂+PGPR treatment has the best effect on improving the resistance of C.grandiflora to As pollution.

Identification，Biological Characterization and Screening of Prevention and Control Agents of the Pathogens Causing the Rot Disease of Malus×robusta（CarriŠre）Rehder

DU Chenyang, SHEN Fengying, WU Junwei, ZHAO Xinrui, WANG Xue, WU Weigang

2025, 54(3): 99-109. DOI: 10.15933/j.cnki.1004-3268.2025.03.011

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By using Malus×robusta（CarriŠre）Rehd as the test material，the pathogenic fungi were isolated using tissue isolation method，and their pathogenicity was determined according to Koch’s postulates.The morphological characteristics of the pathogenic fungi and rDNA⁃ITS sequence analysis were combined to identify the types of pathogenic fungi，their biological characteristics were investigated，and through indoor toxicity tests，the most virulent chemical fungicides were selected against the pathogenic fungi of Malus×robusta rot disease，in order to provide reference basis for the comprehensive prevention and control of Malus×robusta rot disease. The results showed that the two strains of pathogenic fungi HL⁃23 and HL⁃100 isolated from Malus×robusta suffering from canker were both Valsa mali.The biological characteristics of strains HL⁃23 and HL⁃100 were analyzed by measuring the colony diameter via the cross⁃method.The results showed that strains HL⁃23 and HL⁃100 were most suitable for
cultivation on PDA medium.The optimal pH value for mycelial growth was 5.0，and the optimal temperature was 25—30 ℃.They were suitable for growth under 12 hours of light and dark alternation conditions，the optimal carbon source was sucrose，and the most suitable nitrogen source was ammonium dihydrogen phosphate.The toxicity of nine commonly used chemical fungicides，such as difenoconazole and azoxystrobin，against the pathogenic fungi HL⁃23 and HL⁃100 of Malus×robusta rot disease was determined through indoor plate experiments. It was found that the inhibition rates of nine fungicides on the same type of fungal strains HL⁃23 and HL⁃100 were significantly different.The best inhibition effect was difenoconazole，with EC₅₀ values of 0.010 9—0.0110 mg/L；The second was azoxystrobin，with EC₅₀values of 0.015 8—0.018 1 mg/L.

Influence of Intercropping with Different Root Segregation on Soil Nutrients，Extracellular Enzymes and Their Stoichiometric Characteristics of Chinese Cabbage

SUN Xinying, FAN Bowen, ZHAO Liqin, LIU Juncai, DONG Ziming, YANG Fengjun

2025, 54(3): 110-119. DOI: 10.15933/j.cnki.1004-3268.2025.03.012

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This study investigated the soil nutrient content and extracellular enzyme activities related to soil carbon，nitrogen and phosphorus cycles and their stoichiometric change characteristics under Chinese cabbage and garlic intercropping，so as to provide scientific basis and theoretical support for efficient Chinese cabbage production and sustainable land use. Spring and Autumn Imperial cabbage and Zhaozhou white garlic were used as experimental materials，and four treatments were set up including Chinese cabbage monoculture（CK），Chinese cabbage and garlic plastic film root⁃separated intercropping（SM），Chinese cabbage and garlic nylon net root⁃separated intercropping（NW），and Chinese cabbage and garlic non⁃separated intercropping（BF），to determine the nutrient nature of the soil and the four kinds of extracellular enzyme activities and their stoichiometric ratios，and to calculate the lengths and angles of vectors used for the indication of the resource limitation of microorganisms.The results showed that，compared with CK treatment，BF treatment was able to increase soil organic carbon（SOC），total nitrogen（TN），total phosphorus（TP）and quick⁃acting phosphorus（AP）contents by 10. 51% to 21.21%，24.51% to 266.07%，35.71% to 102.86%，and 18. 88% to 154.17%，respectively，and intercropping was able to improve soil nutrient profiles and soil fertility. Compared with CK treatment，NW and BF treatments were able to significantly increase NAG and ALP activities，with NAG enzyme activities increased by 17.54% to 104.58% and 48.04% to 159.05%，respectively，and ALP enzyme activities increased by 9.60% to 31.55% and 20.67% to 48.32%.By calculating enzyme stoichiometric ratios，it
was found that CK and SM treatments significantly increased vector length（VL） and angle（VA）compared with NW and BF，and potting soils were mainly limited by microbial carbon and phosphorus.The degree of carbon and phosphorus limitation was slowed down under NW and BF treatments compared to CK treatment. The carbon and phosphorus limitations faced by soil microorganisms are easily exacerbated by monocropping；Chinese cabbage and garlic non⁃separated intercropping can improve soil nutrient effectiveness，increase soil alkaline phosphatase content，and effectively alleviate microbial resource limitations.

Cloning and Bioinformatics Analysis of PomiR2592bj and Its Target Gene PoSDR1 of Tree Peony

LEI Yang, GAO Jingshan, NAN Qiongqiong, GUO Lili, HOU Xiaogai

2025, 54(3): 120-128. DOI: 10.15933/j.cnki.1004-3268.2025.03.013

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Based on the previous analysis of transciptome，miRNA and degradome of early⁃flowering Fengdan，early⁃flowering Fengdan mutant plants，and late⁃flowering Lianhe，we identified PomiR2592bj and its target gene PoSDR1，which were excavated to regulate the flowering period of tree peony.To further elucidate the biological characteristics and functions of PomiR2592bj and PoSDR1，we cloned a 113 bp sequence for PomiR2592bj and a 1 380 bp sequence for PoSDR1.Secondary structure analysis revealed that the precursor sequence of PomiR2592bj could form a stable stem⁃loop structure with mature sequences located at the 3' arm end；multiple sequence alignment indicated conservation within these mature sequences；phylogenetic tree analysis unveiled that the miR2592 family was subdivided into 10 branches with homology.The protein encoded by PoSDR1 contained 296 amino acids.The protein was hydrophilic with no transmembrane domain and signal peptide. 19 Ser sites of PoSDR1 were phosphorylated，while 11 Thr sites were phosphorylated，and 2 Tyr sites were phosphorylated.PoSDR1 had a NADB_Rossmann superfamily conserved domain. PomiR2592bj was predicted to have a targeting relationship with PoSDR1.

The Regulatory Effect of the SP1/IGF2BP3 Axis on the Proliferation of Porcine Skeletal Muscle Satellite Cells

HUANG Yuxin, WANG Wei, LI Yixing, TANG Zhonglin

2025, 54(3): 129-138. DOI: 10.15933/j.cnki.1004-3268.2025.03.014

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This study investigated the effect of insulin⁃like growth factor 2 mRNA⁃binding protein 3（IGF2BP3） on the growth and development of porcine skeletal muscle and analyzed its biological function and expression regulation mechanism so as to provide a theoretical basis and candidate genes for genetic improvement in pigs.Porcine skeletal muscle satellite cells were utilized as the experimental material.RT⁃qPCR and Western blot were employed to measure the expression of cell proliferation marker genes PCNA、CDK4、Cyclin A2 and MKI67，and EdU and CCK⁃8 kits were used to detect cell
proliferation viability and rate.The dual⁃luciferase assay of SP1 binding site mutations in the promoter region of the IGF2BP3gene and the ChIP assay were employed to investigate the expression regulatory mechanism of the IGF2BP3 gene by the transcription factor SP1.The results showed that knocking down IGF2BP3 in porcine skeletal muscle satellite cells significantly upregulated the expression of cell proliferation marker genes，increased the proportion of EdU positive cells，and significantly increased the cell proliferation rate at 48 and 72 hours.The dual⁃luciferase activity was significantly reduced by mutations in the SP1 binding region，whereas the activity was significantly increased by SP1 overexpression. SP1 could bind to the promoter region of the IGF2BP3 gene.Interference with SP1 decreased IGF2BP3 expression，whereas overexpression of SP1 upregulated IGF2BP3 expression.Overexpression of SP1 partially eliminated the phenotype that knockdown of IGF2BP3 promoted proliferation of porcine skeletal muscle satellite cells. In conclusion，the transcription factor SP1 and the IGF2BP3 gene interact to regulate the proliferation of porcine skeletal muscle satellite cells.

Effects of Feeding Frequency on Growth Characteristics，Digestive Enzymes Activities and Muscle Nutrient Composition of Juvenile Pseudobagrus ussuriensis

CHEN Junping, WU Huihui, SHEN Fangfang, ZHANG Jiaxin, YU Ruomeng, FU Yongjie, HU Jibang, ZHAO Daoquan, XIE Guoqiang, SUN Wenxi

2025, 54(3): 139-146. DOI: 10.15933/j.cnki.1004-3268.2025.03.015

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The effects of feeding frequency［1（F1），2（F2），3（F3）and 4（F4）times per day］on growth characteristics，intestinal digestive enzyme（lipase，α⁃amylase，trypsin）activity，liver aminotransferase（aspartate aminotransferase，alanine aminotransferase）activity and muscle nutrient composition of juvenile Pseudobagrus ussuriensis were studied，so as to determine the optimal feeding frequency and improve the feed utilization and nutritional quality of juvenile Pseudobagrus ussuriensis.The results showed that the final body weight，specific growth rate of body weight，relative increase rate of body weight，daily increase of body weight and specific growth rate of body length of Pseudobagrus ussuriensis of F2 group were significantly higher than those of F1 and F3 groups. The activity of intestinal lipase of F1 group was significantly higher than that of other groups.The intestinal trypsin activity of F2 group was the highest，and significantly higher than that of F3 and F4 groups. The feeding frequency had no significant influence on the activity of aspartate aminotransferase in liver tissue，and the activity of alanine aminotransferase of F1 group was significantly higher than that of other experimental groups.The crude protein contents of F2 and F4 groups were significantly higher than those of F1 and F3 groups.The crude lipid content of F4 group was the lowest，which was significantly lower than that of other experimental groups. There was significant difference in total amino acid content among different groups，the order was F1>F3>F2>F4. In conclusion，it is suggested that the feeding frequency should be 2 times per day，which is more conducive to the growth of Pseudobagrus ussuriensis.

Development of a Climate Suitability Model for the Flowering Period of Chimonanthus praecox in Chongqing and Identification of Potential Planting Zones

JI Li, LI Qiang

2025, 54(3): 147-156. DOI: 10.15933/j.cnki.1004-3268.2025.03.016

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Under the influence of global climate warming，the light，temperature，and water resources required for the growth and development of Chimonanthus praecox have undergone significant changes.Conducting research on the climatic adaptability of its key growth periods can provide a scientific basis for optimizing its cultivation structure.This study established a climate suitability model for wintersweet based on the light，temperature，and water requirements during its flowering period in Beibei，Chongqing，integrating climate suitability theory and fuzzy mathematics.The model was used to analyze climatic suitability characteristics during wintersweet’s flowering stage in Chongqing，identify potential planting areas，and offer insights for adjusting cultivation practices and rationally utilizing climatic resources.Results indicated that Beibei exhibited the highest temperature suitability（0.83—0.99），conducive to
flowering，while precipitation suitability ranged from 0.45 to 1.00，sufficiently meeting water demands.However，sunlight suitability was the lowest（0.28），suggesting that light conditions partially constrained flowering.Comparative analysis of the integrated climate suitability index and actual flowering quantity from 2019 to 2022 revealed consistent upward trends，validating the model’s rationality. Chongqing experienced diverse climatic conditions from December to January，with average temperatures of 3—9 ℃，uneven precipitation distribution，and increasing sunshine duration from west to east.Southeastern Chongqing and northeastern Chongqing exhibited more low⁃temperature days and greater diurnal temperature variations，slightly impacting natural growth.Among climatic factors，temperature suitability outweighed precipitation and sunlight suitability，with overall favorable conditions for wintersweet flowering in Chongqing，though sunlight remained a limiting factor.Overall，the integrated climate
suitability during wintersweet’s flowering period in Chongqing is generally favorable，with western Chongqing offering optimal climatic conditions.

Analysis of Thermogravimetric Characteristics and Stepwise Thermal Release Aroma Components of Different Types of Tobacco

LIANG Miao, LIU Yuxuan, ZHU Haibo, SU Man, ZHENG Yongjie, XIE Yifei, JIANG Xi, ZHANG Junsong, ZHU Yuanyang

2025, 54(3): 157-169. DOI: 10.15933/j.cnki.1004-3268.2025.03.017

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In order to study the weight loss characteristics and component release characteristics of different types of tobacco leaves in different thermal weight loss temperature ranges，flue⁃cured tobacco，cigar tobacco and burley tobacco were used as the research subjects. By thermogravimetric analysis method and self⁃built programmed temperature control heating platform coupled with flue gas capture device，the thermal weight loss characteristics of different types of tobacco leaves and the heat release products under stepwise heating conditions were compared and analyzed.The results showed that the thermal weight loss process of different types of tobacco leaves could be divided into four stages，but the thermal weight loss characteristic parameters of each sample were different.The weight loss rate and maximum weight loss rate of flue⁃cured tobacco in stage Ⅱ were high，the initial decomposition temperature of cigar tobacco and burley tobacco was high，and the final decomposition temperature was low.Flue⁃cured tobacco had lower comprehensive pyrolysis index［9.84×10^⁃5%（/ min·℃²）］and residual mass（28.47%）.The pyrolysis kinetics analysis showed that the activation energy（E_a）and pre⁃exponential factor（A）of flue⁃cured tobacco in stage Ⅱ were higher than those of cigar tobacco and burley tobacco.The Ea value of burley tobacco in stage Ⅲ was higher than that of flue⁃cured tobacco and cigar tobacco，while the E_a value of cigar tobacco in stage Ⅳ was higher than that of flue⁃cured tobacco and burley tobacco. Under the condition of stepwise heating，different types of tobacco leaves showed the highest weight loss rate in the thermal decomposition stage Ⅲ，and the weight loss rate of stage Ⅱ showed flue⁃cured tobacco>cigar tobacco>burley tobacco.The weight loss rate of flue⁃cured tobacco and cigar tobacco in stage Ⅲ was significantly higher than that of burley tobacco（34.60%）.The
heat release aroma products of different types of tobacco leaves in each heating stage were significantly different. The total amount of aroma substances released showed cigar tobacco（13 883.48 μg/g）>burley tobacco（11 970.11 μg/g）>flue⁃cured tobacco（8 753.42 μg/g）.The content of aroma substances released in stage Ⅲ accounted for 42.86%，48.68% and 61.85% of the total content，respectively.In summary，the thermal weight loss characteristics and stepwise heat release aroma components of different types of tobacco leaves are different. The type of tobacco raw materials and the heat release components at the corresponding temperature should be considered in the formulation design of cigarette products.

Effect of Different Enzymes on the Quality Improvement Effect of Discarded Tobacco Extract

LUO Haitao, WANG Shuoying, QI Yating, LI Ruili, CHEN Dong, RAO Chaoqi, ZHANG Junsong, ZOU Enkai

2025, 54(3): 170-180. DOI: 10.15933/j.cnki.1004-3268.2025.03.018

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In order to improve the use value of discarded tobacco leaf，deionized water treatment was used as a control，pectinase，papain，cellulase，α⁃amylase，and laccase were selected to pretreat discarded tobacco leaf，extracts were prepared from discarded tobacco leaf by using ethanol extraction，and the aroma components of each extract were analyzed and cigarette flavoring sensory quality evaluation was carried out.Results showed that a total of 61 volatile components were detected in the extracts of six groups by GC⁃MS analysis，among them，the total volatile components of CK（tobacco extract obtained by deionized water treatment）were 12 335.78 μg/g，and the total volatile components of the extracts prepared by different enzyme treatments were 12 968.96—14 993.75 μg/g，which were increased to varying degrees compared with CK，and the total volatile components of tobacco leaf extract prepared by laccase treatment was the highest. Principal component analysis（PCA）was used to comprehensively evaluate the volatile components of each extract，the results showed that the tobacco extract treated with laccase had the highest comprehensive score，followed by the tobacco extract treated with cellulase and α⁃amylase，and the tobacco extracts treated with papain，pectinase and deionized water had lower comprehensive scores；Compared with CK，the cigarettes flavored by tobacco leaf extract prepared after enzymatic treatment had less impurity and irritation，more comfortable aftertaste，the comprehensive score of cigarettes flavored by tobacco leaf extract prepared by laccase treatment was the highest，which was 87.4 points，7.5 pionts higher than CK.In summary，laccase is the optimal biological enzyme for the preparation of discarded tobacco leaf extracts suitable for cigarette flavoring.

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