Journal of Henan Agricultural Sciences

Contents

2023, 52(5): 0.

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Research Progress of Enterotoxigenic Escherichia coli Vaccine for Piglets

LÜ Linfen, PANG Shengmei, LI Baoliang, LIANG Yuxuan, DUAN Qiangde

2023, 52(5): 1-8. DOI: 10.15933/j.cnki.1004-3268.2023.05.001

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Enterotoxigenic Escherichia coli（ETEC）is an important pathogen causing diarrhea of piglets in the world. Various strategies such as giving antibiotics in the feed，oral administration of specific antibodies，dietary and diet supplementary treatment，and genetic breeding have been used to prevent and treat ETEC infection.Compared with other preventive measures，vaccination is the most economical and effective mean to prevent diarrhea of piglets caused by ETEC.In this paper，the latest research progress of ETEC vaccine for porcine was reviewed，the challenges in ETEC vaccine research were analyzed，and the research strategies for effective vaccines were proposed，in order to provide the basis for the development of novel，safe，efficient and broad‐spectrum ETEC vaccines.

Effects of Nitrogen Fertilizer Management on Nutrient Uptake and Yield of Direct‐Seeding Rice under Long‐Term Integrated Rice‐Crayfish Model

PENG Chenglin, SI Guohan, ZHAO Shujun, YUAN Jiafu, LÜ Hongjiu, XU Xiangyu, XU Dabing, XIE Yuanyuan, ZHOU Jianxiong

2023, 52(5): 9-16. DOI: 10.15933/j.cnki.1004-3268.2023.05.002

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In order to provide technical support for the high‐yield，high‐quality，high‐efficiency and ecological cultivation of rice under long‐term integrated rice‐crayfish model，a field experiment was carried out to study the effects of different nitrogen managements（base nitrogen accounted for 50%，40%，30%，20%，0 and no nitrogen fertilizer，respectively）on the nutrient uptake and yield of direct‐seedingrice under long‐term integrated rice‐crayfish model with rice monoculture model as control.The results showed that the 20% and 0 treatments under the long‐term integrated rice‐crayfish model，and 30% and 20% treatments under the rice monoculture model，were good for nitrogen，phosphorus and potassium absorption of rice.Under the same nitrogen fertilizer management，the long‐term integrated rice‐crayfish model could promote the absorption of nitrogen，phosphorus and potassium compared with the rice monoculture model.The rice yields of 20% and 0 treatments were significantly higher than the other treatments under the long‐term integrated rice‐crayfish model，and those of 30%，20% and 0 treatments were significantly higher than the other treatments under the rice monoculture model，which were mainly achieved by increasing the effective panicle number and seed setting rate.The rice yield under the long‐term rice‐crayfish co‐cropping mode increased by 11.1% on average compared with that under the rice monoculture model，which was mainly achieved by increasing the effective panicle number.With the decrease of the ratio of base nitrogen，the same trend as rice yield was found in nitrogen fertilizer partial productivity，agronomic efficiency，apparent utilization efficiency and contribution rate under the two models. Compared with the rice monoculture model，the long‐term integrated rice‐crayfish model greatly increased partial productivity and apparent utilization efficiency of nitrogen fertilizer by 9.5% and 21.3% respectively，but decreased the agronomic efficiency and contribution rate of nitrogen fertilizer.In summary，the recommended ratios of base nitrogen were 20% and 0 under the long‐term integrated
rice‐crayfish model.

Effects of Combined Stress of High Temperature and Drought on Photosynthetic Physiology of Maize

WANG Yakun, LI Hongping, XU Zhenzhen, LIU Tianxue

2023, 52(5): 17-23. DOI: 10.15933/j.cnki.1004-3268.2023.05.003

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The effect of high temperature（H），drought（D）and combined stress of high temperature and drought（HD）on photosynthetic physiology of maize seedling was studied with high temperature‐tolerant Zhengdan 958（ZD958）and high temperature‐sensitive Xianyu 335（XY335）by pot experiment，so as to provide theoretical basis for breeding of excellent maize varieties resistant to high temperature and drought.The results indicated that compared with CK without stress，the chlorophyll content of ZD958 leaves significantly decreased under stresses，there was significant difference among treatments，and the HD treatment showed the greatest reduction.The change of chlorophyll content of XY335 was consistent with that of ZD958，and the influence was greater.The ratio of chlorophyll a to b in ZD958 leaves increased under D and HD conditions，while it increased for XY335 only under HD condition.The net photosynthetic rate，stomatal conductance，transpiration rate，electron transport rate in ZD958 and XY335 leaves all（even significantly）decreased，D treatment had the greatest changes，and the decrements in ZD958 were more than that in XY335 under D and HD conditions.The activity of phosphoenol‐pyruvate carboxylase（PEPCase） in ZD958 leaves significantly increased under H condition，while no obvious changes of that were observed under D and HD conditions，but the activity of ribulose‐1，5‐bisphosphate carboxylase/oxygenase（Rubisco） significantly decreased under D and HD conditions.The activity of PEPCase in XY335 leaves decreased compared with CK under stresses，and the decrement of HD treatment was the greatest.The activity of Rubisco in XY335 leaves significantly decreased compared with CK under stresses，and the decrement of D treatment was the greatest. Overall，D and HD stresses have greater influence on photosynthetic physiology of maize seedling，and different varieties have different responses to the above three kinds of stress.

Genetic Diversity and Population Structure Analyses of Maize Inbred Lines Based on DUS Test Traits in Huang‐Huai‐Hai Region

DOU Dandan, SUN Jianjun, GUO Yuxi, WANG Dexin, GUO Xinhai, DING Chaoming

2023, 52(5): 24-32. DOI: 10.15933/j.cnki.1004-3268.2023.05.004

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Based on the 39 basic characters in the DUS test guide of maize，the genetic diversity and population structure of 100 test inbred lines and 9 typical inbred lines were analyzed，so as to provide a theoretical basis for the efficient utilization of maize germplasm resources and the improvement of breeding efficiency in the Huang‐Huai‐Hai region. The results showed that the average allelic variation of all characters in 109 inbred lines was 5.026，with a range of 1—9；The mean effective allelic variation was 2.919，with a range of 1.000—6.053；The average Shannon index was 1.133，with a range of 0.000—1.928.The diversity of supporting root color intensity（Chr.22）was the highest，followed by filament color intensity（Chr.9），and the diversity of top traits of the first leaf（Chr.1）was the lowest.The effective allelic variation of 48.72% characters was greater than 2.919，and the Shannon index of 61.54% characters was greater than 1.133.The clustering analysis results indicated that the genetic background of maize germplasm in the Huang‐Huai‐Hai region was relatively rich.The 109 materials were clustered into 5 groups，except group D which contained 9 inbred lines with the smallest number，the other 4 groups had basically the same number of inbred lines，and group D had a relatively small proportion of genetic similarity coefficient in other groups.The genetic distance among the five groups
varied from 0.200 to 0.665，the genetic distance between group D and group B was the greatest with 0.665，followed by the genetic distance between group D and group C（0.586）；The genetic distance between group C and group B was the smallest with 0.200，followed by the genetic distance between group E and group A（0.212）.

Comprehensive Evaluation of Grain and Its Quality Traits of 150 Maize Inbred Lines

ZHAO Haijun, SHI Jiaqing, WANG Bin, GUO Yiyang, HU Xiaoli, HAN Zanping

2023, 52(5): 33-39. DOI: 10.15933/j.cnki.1004-3268.2023.05.005

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The grain（grain length，grain width，100‐grain weight and bulk density）and its quality traits（grain water content，contents of oil，protein，starch and lysine）of 150 improved maize inbred lines were analyzed，and correlation analysis，cluster analysis and principal component analysis were used to comprehensively evaluate them to screen out excellent inbred lines for high‐quality maize breeding.The correlation analysis results showed that the water content of grains of inbred lines was significantly positively correlated with contents of protein，starch and lysine，and significantly negatively correlated with bulk density；Starch content was significantly negatively correlated with contents of protein，oil and bulk density；100‐grain weight was significantly positively correlated with grain length and grain width.The cluster analysis results showed that inbred lines of maize were divided into six groups.17N0358 and 17N0443 of groups 1 and 2 had obvious specificity in terms of quality traits，the water content，protein content，starch content，lysine content and 100‐grain quality of 17N0358 reached the highest，and 17N0443 had the characteristics of low grain water content，protein content，100‐grain weight，but high oil content.The results of principal component analysis showed that the cumulative contribution rate of the first four principal components（100‐grain weight，water content，protein content and lysine content）reached 73.703%.The comprehensive scores of 8 inbred lines were higher than that of the control line B73，which were 17N0536，17N0560，17N0544，17N0358，17N0373，17N047，17N0556 and 17N0540.A total of 9 inbred lines with better grain and its quality traits were screened out，which were 17N0358，17N0443，17N0536，17N0560，17N0544，17N0373，17N0470，17N0556 and 17N0540.

Comprehensive Evaluation of 176 Waxy Sorghum Germplasm Resources Based on Principal Component，Grey Correlation and DTOPSIS Analyses

WU Guojiang, ZHOU Wei, YU Zhonghao, LI Yan, LÜ Jingbo, WANG Zhenguo, ZHOU Yaxing

2023, 52(5): 40-51. DOI: 10.15933/j.cnki.1004-3268.2023.05.006

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In order to screen out the high‐quality waxy sorghum germplasm resources suitable for popularization and utilization in the Xiliaohe Plain，16 main agronomic traits of 176 waxy sorghum germplasm resources were analyzed，and the comprehensive evaluation and ranking were carried out by combining principal component analysis method，grey correlation degree method and DTOPSIS method.The results showed that the coefficient of variation of the main agronomic traits of 176 waxy sorghum germplasm resources ranged from 6.69% to 47.84%，and there were generally significant or extremely significant correlation between the traits. Through principal component analysis，16 main agronomic traits of 176 waxy sorghum germplasm resources were integrated into 9 independent comprehensive indexes（principal components），and the cumulative contribution rate of these 9 principal components was87.169%.The top 30 germplasm resources were screened based on comprehensive evaluation scores by principal component analysis，weighted correlation degree by grey correlation degree method and relative closeness degree by DTOPSIS method respectively. By the combination of the above three methods for comprehensive evaluation，10 germplasm resources with excellent comprehensive traits were selected from 176 germplasm resources，namely QNR，9705，2011‐30，Xinliang 52，102A，HubocuoP4，Jan‐88，9701，0657 and 4126 ，respectively，which laid a foundation for the breeding of waxy sorghum varieties in the Xiliaohe Plain.

Cloning and Expression Analysis of NtCCX2 Gene in Tobacco

FENG Kang, YANG Yuanyuan, LIU Qiong, CHENG Lan, HAO Haohao, WANG Pingping, JIN Weihuan, GUO Hongxiang

2023, 52(5): 52-60. DOI: 10.15933/j.cnki.1004-3268.2023.05.007

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To investigate the role of NtCCX2 gene in tobacco in response to cadmium and other stresses，it was cloned from tobacco and its expression pattern was analyzed. The results showed that the total length of NtCCX2 gene was 2 464 bp，and the CDS region was 1 926 bp，encoding 641 amino acids.It had significant characteristics of CCX family，and there were two α‐repeat regions in the transmembrane region：α1 pattern motif GNGAPD and α2 pattern motif G（N/D）SxGD. NtCCX2 gene had obvious spatial and temporal expression characteristics.Its expression was the highest in leaves at seedling stage and flourishing stage，while was higher in roots and flowers at mature stage.The promoter of NtCCX2 gene contained ABA and other hormone signals as well as stress‐related response elements.In addition to drought，salt and cadmium stresses，the expression of NtCCX2 gene in roots and leaves also had different responses to ABA，MeJA and ethylene.Among them，after 2 hours of ABA treatment，the relative expression level of NtCCX2 gene in the root was 3.7 times that of 0 hour；After 8 hours of MeJA treatment，the relative expression level of NtCCX2 gene in the leaves was 2.1 times that of 0 hour；After2 hours of ethephon treatment，the relative expression level of NtCCX2 gene in both the root and leaves increased. Cadmium‐reducing agents could down‐regulate the expression of this gene.

Relationship between Meteorological Factors and Yield and Quality of Flue‐cured Tobacco Upper Leaves under Different Transplanting Date in Sanmenxia

SU Yongshi, JIANG Weifeng, YANG Yang, CHANG Jianbo, WANG Jing, LI Zhixin, XU Min, GUAN Weidong, ZHANG Xiaoquan, FU Yunpeng

2023, 52(5): 61-73. DOI: 10.15933/j.cnki.1004-3268.2023.05.008

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In order to clarify the effects of meteorological factors on the yield and quality of upper leaves of flue‐cured tobacco in Sanmenxia，and to establish key meteorological indicators for the formation of high quality upper leaf，the main flue‐cured tobacco cultivar Yunyan 87 was used as the material for field experiments in Lingbao from 2020 to 2021.The changes of meteorological factors，yield and quality of upper leaves of flue‐cured tobacco were studied，and the relationship between meteorological factors during upper leaves growth and yield and quality was analyzed.The results showed that the meteorological factors at each growth stage of the upper leaves of flue‐cured tobacco changed with the delay of transplanting date，especially at the mature stage.With the delay of transplanting date，the yield，output value，proportion of superior tobacco，average price of upper leaves，reducing sugar，the ratio of sugar to nicotine，aroma quality，aroma quantity and the total score of sensory quality increased first and then decreased，while nicotine gradually decreased.The economic characters and quality of the upper leaves of flue‐cured tobacco were the best after transplanted on May 7.The correlation analysis showed that the economic indexes，reducing sugar content，nicotine content，the ratio of sugar to nicotine，aroma quality，aroma quantity and total score of sensory quality of upper leaves were significantly or extremely significantly positively correlated with the average daily temperature at maturity stage.The correlation degree order of each meteorological factor with aroma quality，aroma quantity and total score of sensory quality of the upper leaves was temperature，sunshine hours and rainfall，especially the correlation degree of average daily temperature at mature stage was the highest.When the average daily temperature of the mature stage was 20.20—25.16℃，aroma quality，aroma quantity and total score of sensory quality of the upper leaves were better.

Effects of Graphene Oxide Seed Soaking on Growth and Yield of Brassica napus L.

ZHANG Zishuang, SONG Wei, XU Benbo, XU Jinsong, ZHANG Xuekun, XIE Lingli

2023, 52(5): 74-80. DOI: 10.15933/j.cnki.1004-3268.2023.05.009

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To explore the effects of seed soaking with graphene oxide（GO） on seeds germination，seedlings growth and yield of Brassica napus，seeds of five B.napus varieties were soaked with GO solution of 0（control），5，10，25，50，100 mg/L，and the germination potential and yield in mature stage were determined.After Zhongshuang No. 9 was treated with different concentration of GO solution，cytochrome（chlorophyll a and b，carotenoids，total chlorophyll）content，net photosynthetic rate，stomatal conductance，transpiration rate，intercellular CO2 concentration，ABA and IAA content in plants were detected. The results showed that GO treatments promoted seed germination of B. napus，and 25 mg/L GO treatments significantly increased the yield of B. napus and could be used for seed soaking.Lower than 25 mg/L GO treatments had no significant effect on cytochrome of Zhongshuang No.9 seedlings compared with the control，while the plant cytochrome content significantly reduced after treating with 50 and 100 mg/L GO；Net photosynthetic rate significantly increased after treating with 5 mg/L and 10 mg/L GO；Lower than 25 mg/L GO treatments had no significant effect on the IAA content，but 50 and 100 mg/L GO treatments significantly decreased the IAA content；ABA content increased with the increasing of GO mass concentration on the 10th day of treatment.Low concentration GO treatment promoted seed germination and increased net photosynthetic rate of leaves，but high concentration GO treatment inducedrapid increase in ABA content，decreased cytochrome content，and inhibited the growth and development of B.napus.The results proved that the appropriate concentration of GO seed soaking treatment promoted the early germination and rapid development of B.napus，which was conducive to the improvement of yield.

Effects of Straw Returning and Rotating Tillage on Soil Physical Characteristics and Carbon and Nitrogen Content in Double‐Cropping Paddy Field

XIONG Rui, ZHANG Wei, ZHONG Kangyu, ZHOU Wentao, OU Xi, WANG Hongrui, LONG Pan, XU Ying, FU Zhiqiang

2023, 52(5): 81-90. DOI: 10.15933/j.cnki.1004-3268.2023.05.010

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In order to explore the effects of straw returning and rotating tillage on soil physical properties，carbon and nitrogen in double‐cropping rice area，we set four different tillage modes，including rotary tillage for early rice and turning tillage for late rice with straw returning（RTS），rotary tillage for early rice and no tillage for late rice with straw returning（RNS），rotary tillage for early rice and turning tillage for late rice without straw returning（RT），rotary tillage for early rice and no tillage for late rice without straw returning（RN）in Liuyang City，Hunan Province. The results showed that the particle size distribution and carbon and nitrogen contribution rate of soil aggregates were dominated by 0.25—2 mm in each treatment. Soil bulk density under no‐tillage for late rice was higher than that under rotation tillage for early rice，the rotary tillage of early rice reduced the soil bulk density of no-tillage late rice last year.Compared with no straw returning，straw returning increased the stability of soil aggregates（0—20 cm soil layer），and the concentration of large aggregates（>0. 25 mm），mean weight diameter（MWD），soil organic carbon（SOC），total nitrogen（TN）were increased by 2.49%—12.72%，1.23%—12.34%，8.46%—36.70%，8.96%—81.25%，respectively.After using different tillage methods for late rice，the concentration of >0. 25 mm aggregates was significantly increased. And no tillage could improve the aggregate concentration（>0.25 mm）more than that of turning tillage in 10—20 cm soil layer.Compared with other treatments，the concentration of large aggregates and organic carbon content in 0—20 cm soil layer under RNS increased by 1.87%—21.67% and 5.76%—19.36%，respectively. Therefore，RNS is helpful to maintain soil aggregate stability and increase soil organic carbon content.

Effects of Activated Phosphate Rock Powder on Transformation of Cadmium Fraction in Soil and Cadmium Uptake by Pepper

CHAI Guanqun, YANG Jiaojiao, WANG Li, LIU Guihua, LUO Muxinjian, QIN Song, FAN Chengwu

2023, 52(5): 91-98. DOI: 10.15933/j.cnki.1004-3268.2023.05.011

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In order to explore the effect of activated phosphate rock powder on the transformation of cadmium（Cd）fraction in Cd‐safely‐utilized lime soil and the absorption of Cd by pepper，and to provide scientific basis for the development and utilization of phosphate rock powder，the Cd‐safely‐utilized lime soil and pepper variety‘Qing Hong Yuan Shuai’were selected as test materials. The effects of no phosphate rock powder（CK），0.5% phosphate rock powder（PR‐0.5%），0.5% oxalic acid activated phosphate rock powder（CPR‐0.5%）and 0.5% citric acid activated phosphate rock powder（NPR‐0. 5%）on soil Cd fraction，pepper biomass and fruit Cd content were studied by indoor culture experiment and pot experiment.The results showed that compared with the control（CK），the dry weight of pepper fruits treated with unactivated and activated phosphate rock powder increased significantly by 13.99%—41.79%，and the dry weight of pepper fruits treated with NPR‐0.5% was the largest.With the prolongation of culture time，the content of available Cd in soil decreased. Compared with 0 d，after 90 d of soil culture，the decrease of available Cd in CK，PR‐0.5%，CPR‐0.5% and NPR‐0.5% was 26.67%，26.67%，31.11% and 35.56%，respectively.The application of unactivated and activated phosphate rock powder promoted the transformation of activated Cd fraction in soil to inert fraction，and the content of activated Cd fraction in NPR‐0.5% treatment was the lowest. Compared with CK，the proportion of activated Cd fraction in NPR‐0.5% treatment decreased by 16.45 percentage points.Compared with CK，the Cd content of pepper fruits treated with unactivated and activated phosphate rock powder decreased by 14.04%—31.58%，and the Cd content of pepper fruits treated with NPR‐0.5% decreased the most.In summary，NPR‐0.5% treatment promoted the transformation of activated Cd fraction in soil to inert fraction and inhibited the absorption of Cd by pepper.

Study on the Fertilization Effect of Subsurface Pipe Drainage and Okra Planting in Improving Coastal Saline Soil

TAN Pan, WANG Shichao, XIE Jin, JIANG Gaoqian, HAN Lipu

2023, 52(5): 99-109. DOI: 10.15933/j.cnki.1004-3268.2023.05.012

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To explore the promoting effect of different fertilization treatments on subsurface pipe drainage and subsurface pipe salt drainage and the influence on soil salt content，a simulation experiment was carried out for two consecutive years by using the simulation box method.Three fertilization treatments were designed，including organic fertilizer+chemical fertilizer（NPK+OM），organic fertilizer（OM）and chemical fertilizer（NPK），and no fertilization（CK）was as the control.With the subsurface pipe drainage water samples and soil samples in 0—80 cm soil layer as the research objects，the volume of subsurface pipe drainage，the amount of subsurface pipe salt drainage，the cumulative amount of each salt ion in subsurface pipe drainage，the soil electrical conductivity and the soil salt content were measured.The results showed that under the conditions of subsurface pipe drainage and okra planting，fertilization increased the volume of subsurface pipe drainage，and the cumulative volume of subsurface pipe drainage with different fertilization treatments was 44.50%—74.55% higher than CK. Fertilization increased the amount of subsurface pipe salt drainage，and the cumulative amount of subsurface pipe salt drainage was 33.35%—128.83% higher than CK，with chloride ion（Cl^-）and sodium ion（Na⁺）being the highest.Fertilization significantly reduced the soil electrical conductivity and soil salt content.In 2021，the soil electrical conductivity in 0—80 cm soil layer was 44.82%—74.06% lower than the initial value，and the soil salt content was 24.97%—66.80% lower than the initial value. The NPK+OM treatment had the best effect.In the process of combining subsurface pipe drainage with okra planting to improve coastal saline soil，fertilization can increase the volume of subsurface pipe drainage，the amount of subsurface pipe salt drainage，reduce the soil electrical conductivity and soil salt content，and accelerate the improvement of coastal saline soil.Applying organic fertilizer together with chemical fertilizer has the best effect and is the best fertilization compensation method.

Effect of pH Value on Acid Production Characteristics and Microbial Community in Co‐Fermentation of Pig Manure and Spent Mushroom Substrate

REN Yuansen, ZHANG Haibo, CHENG Hongyan, WANG Yumeng, LUO Yuan, LIU Na

2023, 52(5): 110-120. DOI: 10.15933/j.cnki.1004-3268.2023.05.013

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Aiming at the problems of single product component and low efficiency in the current resource utilization of agricultural waste，this study used pig manure and spent mushroom substrate for anaerobic co‐fermentation. In this paper，the variation rules of acid production and microbial community structure were studied by the anaerobic co‐fermentation of pig manure and spent mushroom substrate，which was regulated by various pH values. A total of 7 treatments（the pH value was not adjusted，and pH values were 4.0，5.5，7.0，8.5，10.0，11.5）were set up to measure and analyze the changes of volatile fatty acids（VFA），ammonia nitrogen，total phosphorus，soluble chemical oxygen demand（SCOD） and microbial community. When pH value was 8.5，VFA concentration reached the maximum of 36 736.29 mg/L on the 11th day of fermentation，which was 1.36 times the control. Acetic and n‐valeric acids were greatly affected by pH，in general，the proportion of acetic acid increased with the increase of pH value，while the proportion of n‐valeric acid decreased with the increase of pH value. Under different pH values control，SCOD showed the same trend as VFA with fermentation time.The results of high‐throughput sequencing analysis showed that pH 8.5 was the optimal acid producing group，and the microbial community richness was the highest.On the maximum acid accumulation day，Firmicutes was the dominant microflora in all pH control groups.In the optimal acid‐producing group，the relative abundance of Bacteroidetes increased while that of Firmicutes decreased with fermentation.At the genus level，the relative abundance of Lactobacillus，Clostridium_sensu_stricto_1 and Terrisporobacter decreased，while the relative abundance of Caldicoprobacter increased with anaerobic fermentation.Therefore，when pH value is 8.5，the VFA yield of anaerobic co‐fermentation of pig manure and spent mushroom substrate can be increased，and its utilization rate can be improved.

Optimization of Fermentation Conditions and Stability Evaluation of Antagonistic Bacteria against Wheat Crown Rot

ZHANG Qiang, WU Limin, LI Pengyan, ZHAO Na, HU Shen’ao, LU Ninghai

2023, 52(5): 121-129. DOI: 10.15933/j.cnki.1004-3268.2023.05.014

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In order to determine the antifungal activity of Bacillus amyloliquefaciens strain HB‐081 against Fusarium pseudograminearum，the fermentation conditions were optimized by single factor test and orthogonal test，the mycelial growth rate method was used to analyze the stability of the cell‐free fermentation broth and the control effect of strain HB‐081 was measured by pot experiment.The optimized medium composition was maltose 10.0 g/L，peptone 15.0 g/L，KCl 5.0 g/L.The optimal fermentation conditions were as follows：initial pH 6，temperature 28—32℃，liquid volume 50—75 mL，inoculation amount 2%—4%，culture time 48 h.The antifungal activity of cell‐free fermentation broth were free from 100℃，ultraviolet light for 180 min，pepsin，trypsin，and proteinase K.However，the cell‐free fermentation broth were not resistant to strong acid and alkali.Two biosynthetic genes of lipopeptide，bacAB and fenB were detected by using PCR.Pot experiment showed that the control efficacy of strain HB‐081 optimized fermentation broth on wheat crown rot was 69.74%，which was higher than the control efficacy（64.31%）of original fermentation broth.

Identification，Evolution and Expression Analysis of WOX Gene Family in Capsicum annuum

ZHANG Yeda, WANG Xing, WANG Liping, TONG Jing, WU Zhanhui, GAO Yankui

2023, 52(5): 130-141. DOI: 10.15933/j.cnki.1004-3268.2023.05.015

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The WUSCHEL（WUS）‐related homeobox（WOX）transcription factor family is a kind of plant specific transcription factors，which maintain the dynamic balance of plant stem cells，shoot tip meristem formation，embryonic development and other important life activities.In this study，10 WOX genes were identified in pepper（Capsicum annuum）variety Zunla‐1 by using bioinformatics methods，named CaWUS to CaWOX13.CaWOXs were unevenly distributed on 5 chromosomes and are divided into 3 branches.All subcellular predictions showed being located in the nucleus.The number of genes varied greatly among different branches，and members of the same branch had similar gene structures and conserved motifs.CaWOX family promoters contained 14 kinds of action elements related to plant growth and development，hormone regulation and stress，and the most widely distributed was methyl jasmonate response element.No tandem duplication and large fragment duplication were found in CaWOX family，but there was a collinear relationship with Arabidopsis thaliana and Solanum lycopersicum.As a key protein，CaWUS played a pivotal role in the protein interaction network.The gene expression of CaWOXs was significantly different，and some genes played an important role in the regulation of root，stem and fruit development.

Isolation and Identification of Oyster Mushroom Growth‐Promoting Bacteria from Fermented Materials

ZHANG Junjie, WANG Jingqi, LIU Qin, LIU Qing, CUI Xiao, ZHANG Yuting, WU Jie, KONG Weili

2023, 52(5): 142-149. DOI: 10.15933/j.cnki.1004-3268.2023.05.016

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The fermentation culture of oyster mushroom contains indoleacetic acid（IAA），which promotes the growth and development of oyster mushroom.In order to find the microbial strains with strong IAA production ability，firstly，samples of oyster mushroom fermentation material at different time were collected，and bacterial strains were separated by dilution coating plate method.Secondly，120 strains were selected for qualitative analysis and quantitative determination of IAA‐producing capacity by chromogenic reaction，and high‐yield IAA strains were used to test the oyster mushroom growth‐promoting effect.Finally，the genomic DNA of the IAA‐producing strains was extracted by GUTC，16S rRNA gene amplification and sequencing analysis were carried out，and phylogenetic analysis was performed based on the constructed 16S rRNA gene phylogenetic tree to determine the taxonomic status of the strains.The results showed that in 120 bacterial strains isolated，116 strains produced color reaction，which were proved to have the ability to produce IAA.Compared with the control group，IAA color development degree of 88 strains was lower，28 strains were relatively higher.28 strains with relatively high IAA color rendering degree after qualitative determination were selected for quantitative determination，among them，AFX‐13‐3，XWW‐4‐3，LTX‐11‐3，AMB‐8‐2，AMB‐10‐2，AMB‐14‐3，ST‐1‐2，ST‐3‐2，ST‐4‐2 could produce IAA more than 20 mg/L.Among the nine strains，AFX‐13‐3 had the strongest ability to produce IAA，with average mass concentration of 85.18 mg/L，followed by AMB‐10‐2，ST‐3‐2，AMB‐8‐2 and XWW‐4‐3，with the average mass concentrations of IAA of 39.35，34.12，30.94 and 30.71 mg/L，respectively. The ability of the remaining four strains ST‐1‐2，ST‐4‐2，LTX‐11‐3 and AMB‐14‐3 to produce IAA was relatively weaker，and the average concentrations of IAA were 24.41，24.09，22.26 and 21.94 mg/L，respectively.After the growth promotion test of oyster mushroom，it was found that AFX‐13‐3，XWW‐4‐3 and LTX‐11‐3 had good growth promotion effect on oyster mushroom.Through 16S rRNA gene sequencing and phylogenetic analysis，it was determined that AFX‐13‐3 belonged to Priestia，and XWW‐4‐3 and LTX‐11‐3 belonged to Glutamicibacter.

Analysis of Porcine PVALB Gene Polymorphism and Differential Expression in Muscle Tissues of Different Pig Breeds

HAN Yuqing, BO Suxue, YAN Feifei, DUAN Mengqi, CHAMBA Yangzom, SHANG Peng

2023, 52(5): 150-155. DOI: 10.15933/j.cnki.1004-3268.2023.05.017

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To investigate the single nucleotide polymorphism（SNP）loci of the small albumin（PVALB）gene in Tibetan and York pigs and the expression pattern in the leg and longest dorsal muscles of different pig breeds，Tibetan and York pigs at 30，90 and 180 days of age were used as test subjects，SNP screening and genotyping of DNA sequences in the 2 000 bp region upstream of the start codon of PVALB gene in Tibetan and Yorke pigs were performed. Detection of PVALB gene differential expression in the leg and longest dorsal muscles was carried out by using real‐time fluorescent quantitative PCR.The results showed that two SNP sites，G1659C and C1269T，were identified by polymorphism detection in the 2 000 bp region upstream of the PVALB gene start codon，and both conformed to the Hardy‐Weinberg law（P>0.05）.The results of the predicted transcription factors showed that G1659C and C1269T mutant sites led to the disappearance of NRF2 and NCOR1 transcription factors and the addition of NR1D1 and OTX2 transcription factors related to muscle growth and cell differentiation.The relative mRNA expression trends of PVALB gene in the leg and longest dorsal muscles of Tibetan and Yorke pigs were basically the same，at 30 and 90 days of age，PVALB gene expression in the longest dorsal muscle of York pigs was highly significantly higher than that of Tibetan pigs（P<0.01），significantly higher than Tibetan pigs at 180 days of age（P<0.05）.In conclusion，PVALB gene expression may be positively correlated with muscle growth and development，and PVALB gene may be a new candidate gene for early skeletal muscle development and meat quality trait studies.

Effect of Traditional Chinese Medicine Residues On Growth Performance and Digestive Enzyme Activity of Earthworms

LI Yang, KANG Jungang, MA Zhanfei, WANG Yabo, ZHAO Bowei, JIANG Guojun

2023, 52(5): 156-161. DOI: 10.15933/j.cnki.1004-3268.2023.05.018

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In order to solve the problem of environmental pollution caused by livestock and poultry feces and drug residues，and explore a new way of resource utilization of traditional Chinese medicine residues and cow dung，healthy earthworms with average body weight of 239.40 mg and average body length of 5.51 cm were selected and randomly divided into groups A，B，C and D，with three replicates in each group.On the basis of fermented cow dung，0，10%，20% and 30% pharmaceutical residues of Wuwei Jianpi granules were added，respectively，and earthworms were fed outdoors for 28 days.On the 0th and 28th days of feeding，9 earthworms were randomly selected from each group，and their body weight and body length were measured. At 0，7，14，21 and 28 days of feeding，9 earthworms were randomly selected from each group to extract crude enzyme solution，and enzyme linked immunosorbent assay was used to detect the activities of protease，cellulase and phosphatase.On the 28th day，compared with group A，the body weight and body length of earthworms in group C were significantly increased，the weight gain rate was increased by 55.50 percentage points，and the body length was increased by 1.83 cm.The activities of protease，cellulase and phosphatase in group C reached the highest（P<0.05）on the 28th day.The above results show that the drug residues of Wuwei Jianpi granules promote the growth and development of earthworms and enhance the digestive enzyme activities，and the best effect is obtained by adding 20% of the drug residues.

Research and Application of Lightweight Yolov7‐TSA Network in Tea Disease Detection and Identification

LI Weihao, ZHAN Wei, ZHOU Wan, HAN Tao, WANG Peiwen, LIU Hu, XIONG Mengyuan, SUN Yong

2023, 52(5): 162-169. DOI: 10.15933/j.cnki.1004-3268.2023.05.019

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Aiming at the problems of low accuracy，slow model running speed and no detection function of the existing tea disease identification methods，a novel Yolov7‐TSA（Yolov7‐Tiny‐SiLU‐Attention）lightweight network architecture was proposed to detect and classify tea diseases.Specifically，the LeakReLU activation function in the Yolov7‐T network was replaced with the SiLU activation function to improve detection accuracy and prevent overfitting.Meanwhile，the feature perception of target contour and spatial location was further improved by fusing the coordinate attention mechanism. Experiments on the dataset containing eight tea diseases（including healthy tea leaves） showed that the recognition accuracy of Yolov7‐TSA network reached 94.2%，which was 3.2 and 1.2 percentage points higher compared to Yolov7‐T and Yolov7 networks，respectively.Furthermore，it showed significant results in terms of parameters，floating point operations，model size and inference time per image，which were reduced by 83%，87%，83% and 34%，respectively，compared to the Yolov7 network. The network model achieves the detection and classification of tea diseases when balancing recognition accuracy and real‐time performance.

Physicochemical Properties and Microstructure Changes of Tomato Straw Hydrothermal Biochar under Different Reaction Conditions

HUANG Jiaqing, LIU Cenwei, YE Jing, WANG Yixiang, REN Lihua

2023, 52(5): 170-180. DOI: 10.15933/j.cnki.1004-3268.2023.05.020

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To investigate the effects of carbonization temperature and pyrolysis time on the microstructure，physicochemical properties，principal components，and carbonization efficiency of tomato straw hydrothermal biochar（TSHB）and obtain efficient carbonization conditions，tomato straw was decomposed into TSHB by hydrothermal carbonization method with low energy consumption.This experiment set up three carbonization temperature（180，220，and 260℃）and four pyrolysis time（2，4，6，and 8 h），with a total of twelve experimental treatments，to study the effects of different combinations of carbonization
temperature and pyrolysis time on the morphology，structure，functional groups，pH value，conductivity（EC）and biochar composition of TSHB，and to analyze the principal components changes of TSHB.The results showed that the porous and hollow vascular bundle structure of tomato straw was conducive to the rapid transfer of water and heat within the tomato straw.Increasing the hydrothermal carbonization temperature could accelerate the destruction of the vascular bundle structure of tomato straw and carbonize the strip‐shaped tomato straw into granular hydrothermal carbon. The functional groups of TSHB were mainly fat ether C-O，alkane C-H，saturated fatty acid ester C-O，amine C-N，and the groups of C-O，C=O，C-N，and C-O-C in the cell wall. Increasing the carbonization temperature could increase the functional groups of TSHB，but those functional groups could be destroyed and reduced significantly if the carbonization temperature exceeded 260℃.TSHB prepared by hydrothermal pyrolysis was acidic biochar（pH value 5.13—5.33）.Under the same pyrolysis time，compared to the carbonization temperature of 180℃ or 260℃，the TSHB prepared at 220℃ had the highest EC value（6 213—7 899 μs/cm），but the EC values of TSHB significantly decreased by 18.19%—26.92% as the pyrolysis time was extended from 2 h to 8 h.Increasing the carbonization temperature and prolonging the pyrolysis time could significantly increase the fixed carbon content of TSHB（2.23—27.99 percentage points），but significantly reduce the volatile content of TSHB（2.46—30.97 percentage points）.Increasing the carbonization temperature could increase the carbonization yield，total nitrogen content，and ash content of TSHB by 10.63—29.91，0.58—0.87，and 1.82—3.09 percentage points，respectively.In summary，compared with prolonging the pyrolysis time，increasing the hydrothermal carbonization temperature could more effectively improve the carbonization efficiency of tomato straw and increase the content of nitrogen‐oxygen functional groups，fixed carbon，and total nitrogen of TSHB.The hydrothermal carbonization temperature was the critical factor determining the microstructure，functional groups，pH value，and principal components of TSHB.The carbonization temperature of 220℃ and the pyrolysis time of 4—6 h were suitable carbonization conditions for preparing tomato straw biochar by hydrothermal carbonization method.

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