Journal of Henan Agricultural Sciences

Research Progress on Pretreatment Technology of Straw Biomass

WANG Yuanyuan, HAN Xiuli, YAN Zhenli, DU Chaojun, CHEN Yujie, CHANG Chun

2022, 51(11): 1-10. DOI: 10.15933/j.cnki.1004-3268.2022.11.001

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Straw biomass is a kind of cheap，sustainable and abundant renewable raw material，and its high‑value utilization is a research hotspot in the world.Pretreatment process is a key step in the conversion of biomass to sugar，biofuel and other value‑added products.The researches on straw biomass pretreatment technology were reviewed，and the physical，chemical and biological pretreatment methods were introduced to effectively separate hemicellulose，cellulose and lignin.The advantages and disadvantages of various pretreatment methods were summarized，and the future research direction of pretreatment was prospected.

Combining Ability Analysis of Quality Traits of Maize Inbred Lines from Different Sources

ZHANG Fengqi, DING Yong, ZHANG Jun, LU Yuanfang, MA Zhiyan, MU Xinyuan, QI Jianshuang, XIA Laikun, TANG Baojun

2022, 51(11): 11-20. DOI: 10.15933/j.cnki.1004-3268.2022.11.002

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Fourteen maize inbred lines from China and America were used to cross 182 combinations according to complete diallel cross design.The combining ability of protein，starch，fat and lysine contents in maize grains and the relationship between the above four traits and combining ability were analyzed for three consecutive years，so as to provide materials and theoretical basis for the improvement of quality traits and breeding of high‑quality maize varieties.The results showed that the contents of protein，starch and fat in grains of 14 maize inbred lines and their 182 hybrid combinations were significantly positively correlated among three years.The variances of general combining ability（GCA）of protein，starch，fat and lysine contents were obviously higher than those of special combining ability （SCA）.The genetic effects of protein，starch and lysine contents were mainly additive；starch content had the highest heritability（71.82%），followed by lysine content（69.74%），and fat content（42.83%）was the lowest.There were significant differences in GCA of four quality traits among different inbred lines，the effect values of GCA of protein，starch and fat contents were larger，and that of lysine content was lower.The SCA effect values of quality traits of the hybrid combinations with LH202，ZhengA88，PHR55 and ZhengH71 as parents were higher，and the quality was better. The starch content in maize grain was significantly negatively correlated with protein，fat and lysine contents，respectively，while there were significantly positive correlations among protein，fat and lysine contents；the four quality traits of hybrid combinations were extremly significantly and significantly positively correlated with the sum of parental GCA effect values and SCA effect value，respectively.

Effects of Nitrogen，Phosphorus and Potassium Application Methods and Rates on Agronomic Traits and Yield of Sweet Potato

LIU Yangyang, SONG Jiale, JI Zhixin, ZHANG Kai, QIAO Yake, JIN Hengxing, LIU Yunfeng, SI Zengzhi

2022, 51(11): 21-34. DOI: 10.15933/j.cnki.1004-3268.2022.11.003

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The primary sweet potato varieties Jishu 25，Tengfei，Xuzishu No. 8 and Pushu 32 in eastern Hebei were utilized as materials，together with the sweet potato lines Jikezi 18（JK274），Jikezi 19（JK142），and Jikeshu 20（JK7）.The effects of application methods of nitrogen（N），phosphorus（P）and potassium（K）fertilizers［one‑time base application（1），1/2 base application and 1/2 topdressing（1/2+1/2），1/3 base application and 2/3 topdressing（1/3+2/3）］and application rates（N：0，75.27，150.55，225.82 kg/ha；P₂O₅：0，39.27，78.55，117.82 kg/ha；K₂O：0，85.09，170.18，255.27，340.36 kg/ha）on the number of branches per plant，the longest vine length per plant，aboveground and underground fresh weight，number of potato per plant and yield of seven sweet potato varieties（lines）were studied，and the appropriate N，P and K fertilizers application methods and rates were selected，so as to provide reference for the cultivation of sweet potato with high yield and efficiency.The results showed that suitable application methods and appropriate application rates of N，P，K fertilizers could promote the growth of sweet potato，and increase yield.However，the best application methods and rates of N，P，K fertilizers varied among different sweet potato varieties（lines）.By considering comprehensively，the N application method was N₁>N_1/2+1/2>N_1/3+2/3，the P application method was P_1/3+2/3>P_1/2+1/2>P₁，and the K application method was K_1/2+1/2>K₁>K_1/3+2/3；the appropriate N application rate was 75.27—157.37 kg/ha，P application rate was 39.27—117.82 kg/ha，and K application rate was 170.18—294.12 kg/ha.The requirement order of sweet potato for N，P and K was K>N>P.The yield order of seven sweet potato varieties（lines）was Pushu 32>Tengfei>Jishu 25>Jikezi 19>Jikezi 18>Jikeshu 20>Xuzishu No.8 under the optimum application rates of N，P and K.To get high yield，N，P and K should be supplied precisely in the above acceptable application rate range according to the sweet potato varieties（lines）.

Analysis of Function and Expression Characteristics of GmFULa Gene

WU Gengxue, LU Mengdan, LI Zhen, CHEN Yu, YUE Yanlei

2022, 51(11): 35-41. DOI: 10.15933/j.cnki.1004-3268.2022.11.004

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To clarify the function of GmFULa［Glycine max（L.）Merr.FRUITFULLa］gene in plant vegetative and reproductive growth，genetic transformation of Arabidopsis thaliana was performed by Agrobacterium mediated floral dip method，and phenotype（flowering time，number of rosette leaves at the beginning of flowering，number of branches and pods at maturity）of Arabidopsis thaliana overexpressing GmFULa gene was analyzed.Meanwhile，the soybean varieties Zigongdongdou（photoperiod sensitive variety）and Heihe 27（photoperiod insensitive variety）were used as materials to analyze the expression of GmFULa gene during the whole growth period under long day and short day conditions.Finally，the tissue expression specificity and circadian rhythm of GmFULa gene were analyzed in Zigongdongdou under short day condition.The results showed that overexpression of GmFULa gene increased the rosette leaves，branches and pods number，and might make the flowering of Arabidopsis thaliana advance.Gene expression analysis showed that the expression level of GmFULa gene at reproductive growth stage was higher than that at vegetative growth stage.The expression level of GmFULa gene in pods was the highest，far higher than that in other tissues.GmFULa gene showed a diurnal rhythm of high expression level at day and low expression level at night，and was induced by optical signal.In conclusion，GmFULa gene affects plant vegetative and reproductive growth，and plays an important role in pod development.

Effects of Exogenous Hormones on Growth，Yield and Physiological Characteristics of Tigernut（Cyperus esculentus L.）under Drought Stress

LIU Jiayao, CHENG Yan, WEI Zunmiao, LONG Wei, WANG Liang, MU Zhongsheng

2022, 51(11): 42-49. DOI: 10.15933/j.cnki.1004-3268.2022.11.005

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In order to determine the optimal spraying concentration of exogenous hormones on tigernut under drought stress，Jisha No.2 was used as material，drought stress was simulated by 15% polyethylene glycol（PEG 6000）to study the effects of different concentrations of gibberellin（GA3，0.1，0.2，0.3，0.4，0.5 μg/L），melatonin（MT，20，40，60，80，100 μmol/L）and salicylic acid（SA，0.25，0.50，0.75，1.00，1.25 mmol/L） on the growth，yield，relative conductivity，chlorophyll content，proline content，malondialdehyde（MDA）content，superoxide dismutase（SOD）and peroxidase（POD）activities of tigernut seedlings.The results showed that under drought stress，spraying the three kinds of exogenous hormones could promote the growth and yield of tigernut，G3（0.3 μg/L），M3（60 μmol/L），S2（0.50 mmol/L）treatments had the best effect，the yields of which significantly increased by 38. 32%，31.73% and35.90% compared with the drought stress treatment（CK2）respectively. Under drought stress，spraying the three kinds of exogenous hormones could reduce the relative conductivity and MDA content in leaves，and increase the chlorophyll content，proline content，SOD and POD activities，alleviating the damage caused by drought stress to the seedlings of tigernut，the best treatments were G3，M3 and S2，respectively.Compared with CK2，for G3，M3 and S2 treatments，the relative conductivities significantly decreased by 54.92%，34.44% and 27.86%，chlorophyll contents obviously increased by 33.83%，19.03% and 15.84%，proline contents significantly increased by 478.88%，627.62% and 412.87%，MDA contents significantly decreased by 53.51%，53.43% and 54.75%，SOD activities increased by 0.95%，13.51%and 16.25%，POD activities increased by 1.80%，10.36% and 12.84%，respectively.In conclusion，under drought stress，spraying appropriate concentrations of GA3，MT and SA could promote the growth of tigernut，alleviate the damage caused by drought stress，and then increase the yield，overall the best concentrations of GA3，MT and SA were 0.3 μg/L，60 μmol/L and 0.50 mmol/L，respectively.

Effect of Colchicine Mutagenesis on Emergence Rate and Quality of Tigernut（Cyperus esculentus L.）

WEI Zunmiao, LIU Jiayao, CHENG Yan, WANG Zhanhai, XU Fangda, CHEN Xin, MU Zhongsheng

2022, 51(11): 50-55. DOI: 10.15933/j.cnki.1004-3268.2022.11.006

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In order to determine the half‑lethal dose for colchicine mutagenesis of tigernut（Cyperus esculentus L.）tuber and obtain high quality variant germplasm resources，the tubers before and after germination were mutagenized with different concentrations（0，1，2，3，4 g/L）of colchicine，the emergence rate was calculated one month later，and the quality was tested by the near‑infrared（NIR）grain analyzer after maturity.The results showed that whether mutagenesis was done before or after germination，the emergence rate of tigernut decreased with the increase of colchicine concentration and mutagenesis time，colchicine concentration had extremely significant effect on the emergence rate，mutagenesis time had significant effect on the emergence rate.Before germination，tigernut tuber was more sensitive to colchicine，the half‑lethal dose of colchicine was lower than that of mutagenesis after germination，and mutagenesis before germination could cause bud deformity，but buds of mutagenesis treatment after germination did not obviously change.On the principle of low mutagenesis concentration and short mutagenesis time to half‑lethal dose，the optimal half‑lethal dose of colchicine was colchicine concentration of 1 g/L，mutagenesis for 24 hours before germination of tigernut tuber.By testing，one germplasm resource with starch content higher than 32% and six germplasm resources with total sugar content higher than 36% in tuber were obtained，which layed the foundation for high‑quality breeding.

Differential Analysis of Chlorion Absorption，Accumulation and Distribution in Different Genotypes of Tobacco

RAO Wei, LI Chunping, WEN Xiaoyang, QIN Dehua, GUO Fangyang, WANG Hui, CHEN Mingcan

2022, 51(11): 56-64. DOI: 10.15933/j.cnki.1004-3268.2022.11.007

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To clarify the differences of chlorion absorption，accumulation and distribution in tobacco with different genotypes，and to screen tobacco genotypes with high and low chlorion absorption，in this study，nine flue‑cured tobacco varieties were selected as test materials，and pot experiments were conducted with three chlorine application levels（ 0，50，100 mg/kg ）to determine the characteristics of chlorion absorption，accumulation，transportation and distribution in different parts of tobacco at maturity stage.The results showed that the content of chlorion in different parts of tobacco increased with the increase of chlorine application rate，and the content of chlorion in different parts of tobacco was foot leaf> lower leaf>middle leaf>upper leaf>stem>root.Among them，the total chloride content and accumulation in K326 leaves were higher，which were 206.4 mg/kg and 16.58 mg，respectively. The content of chloride in the middle and upper leaves of Yujinxiang No.1 was lower，which were 150.09 mg/kg and 95.00 mg/kg，respectively.The chlorion accumulation in the foot leaves（1.35 mg ）and lower leaves（4.63 mg）was higher.Under 50 and 100 mg/kg chlorine treatments，the content，accumulation and translocation coefficient of chlorion in leaves of Liaoyan 17 increased by 59.62%，95.67%；37.04%，52.49% and 39.88%，89.43% compared with CK，respectively.The chlorine transport coefficient of Yujinxiang No.1 only increased by 7.88% and 12.90%.The accumulation and distribution rate of chlorion in Qinyan 96 increased only by 1.73%，2.04% and 0.05%，0.07%，with no significant difference compared with CK.In general，among the nine tobacco varieties tested，Qinyan 96 and Yujinxiang No.1 have less chlorion absorption and accumulation，less chlorion transport and distribution in the middle and upper leaves，and can be planted in tobacco areas with higher soil chloride content. K326 and Liaoyan 17 have larger absorption of chlorion，and the chlorine content in each part of tobacco plant is higher，so corresponding chlorine control measures should be taken in production.

Spatial Heterogeneity of Available Kalium in Cultivated Layer of Typical Albic Soil

HOU Yonghua, WANG Xia, ZHAO Yinghui, LUO Chong, LIU Huanjun, MA Shinai, WANG Shuo

2022, 51(11): 65-72. DOI: 10.15933/j.cnki.1004-3268.2022.11.008

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Understanding the spatial heterogeneity of soil nutrients is an important basis for achieving precise fertilization and sustainable management of agriculture. In order to accurately obtain the spatial distribution information of soil nutrients in cultivated land in typical albic soil area，the content of available kalium（AK）in 0—20 cm topsoil of typical albic soil area was taken as the research object，and the spatial heterogeneity of AK in typical albic soil area was analyzed by comparing the interpolation accuracy of different auxiliary variables with field sampling points and cokriging interpolation method.The results showed that the AK content in the study area was high in the south and low in the north，ranging from 92.00 to 364.00 mg/kg，with an average of 171.78 mg/kg，showing a high level.Soil factors such as available phosphorus（AP），topographic factors such as elevation（E），and remote sensing indexes such as normalized difference vegetation index（NDVI）were all significantly correlated with AK content，and the accuracy of the kriging interpolation method with the above auxiliary variables（cokriging，COK） was higher，that is，COK>OK（ordinary kriging）. Among them，the accuracy was the highest when AP was used as an auxiliary variable，and the cross‑validation RMSE was 36.225 mg/kg.In summary，selecting the best auxiliary variables is more helpful to analyze the spatial heterogeneity of AK in the study area.

Optimization of the Conditions of an Efficient Phosphate‑Solubilizing Bacterium and Study of Its Growth‑Promoting Effect

LI Sisi, FU Yunhui, LUO Yu, ZHOU Jia, ZHAO Shuai, QU Jianhang

2022, 51(11): 73-81. DOI: 10.15933/j.cnki.1004-3268.2022.11.009

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An efficient phosphate‑solubilizing bacterial strain 1416X3 was isolated from lake sediments，To provide a basis for improving the utilization of available phosphorus in soil and the rational development of microbial fertilizers，the classification status of the strain was identified by colony morphology，physiological and biochemical characteristics and 16S rRNA gene phylogenetic analysis，and its phosphorus solubilizing conditions were optimized.The phosphate‑solubilizing process in soil and its ability to promote plant growth were studied by soil phosphate‑solubilizing test and pot experiment.The results showed that strain 1416X3 was Pseudomonas sp..The optimal carbon source for phosphorus dissolution was glucose，the optimal nitrogen source was ammonium sulfate，the optimal initial pH value was 7.0，the optimal inoculation amount was 1%，the optimal fermentation temperature was 28℃，and the maximum phosphorus dissolution amount could reach 827.12 mg/L. Strain 1416X3 could colonize effectively in vegetable and garden soils，and significantly increase the content of available phosphorus in soil.Compared to the control group that was not inoculated with 1416X3，the plant height，root length，stem diameter，fresh weight，dry weight and total phosphorus of maize inoculated with strain 1416X3 in pot were increased by 16.90%，114.02%，43.03%，80.28%，80.85% and 88.99%，respectively，and the content of available phosphorus was 111.80% higher than that of the control.In summary，phosphate‑solubilizing strain 1416X3 can effectively improve soil available phosphorus content，promote the growth of maize seedlings，and has high application potential.

Effects of Chinese Milk Vetch Combined with Chemical Fertilizer on Rice Growth and Phosphorus Utilization

ZHANG Jishi, LIU Chunzeng, LÜ Yuhu, WANG Shougang, CAO Weidong, ZHANG Yuting

2022, 51(11): 82-90. DOI: 10.15933/j.cnki.1004-3268.2022.11.010

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Chinese milk vetch（MV）returned to the field to replace some chemical fertilizers is a main technical measure for saving fertilizer and increasing efficiency in paddy fields in southern Henan rice‑growing area.A field experiment was conducted in southern Henan rice‑growing area，to explore the effect of different fertilizer reduction ratio on soil phosphorus supply and rice phosphorus uptake under the condition of equal amount of MV returned to the field. The location experiment of MV combined with chemical fertilizer（started in 2008）included six treatments：No fertilization（CK），single application of chemical fertilizer（F100），equal amount of MV returned to the field（22.50 t/ha）combined with different amount of chemical fertilizer（100% chemical fertilizer，MVF100；80% chemical fertilizer，MVF80；60% chemical fertilizer，MVF60；40% chemical fertilizer，MVF40）.The effects of MV returned to the field combined with chemical fertilizer on soil available phosphorus content，shoot phosphorus content and accumulation，leaf area index and dry matter accumulation at maturity were preliminarily studied in the year of 2021.The results showed that MVF100 and MVF80 treatments significantly increased the total dry matter accumulation by 8.52% and 7.14% respectively，compared with the F100 treatment at maturity.The dry matter accumulation of straw under MVF100 at maturity was 12.84% higher than that in the F100 treatment.Compared with F100，MVF80 treatment increased the dry matter accumulation of panicle by 11.05% at the mature stage，but no significant difference was observed.The leaf area index at the booting stage under MV returned to the field combined with chemical fertilizer was lower than that in the F100 treatment，while the leaf area index at maturity in MVF100 and MVF60 treatments was higher than that in the F100 treatment.The soil available phosphorus，shoot phosphorus content and accumulation of all fertilization treatments were significantly higher than CK，but there was no significant difference among all fertilization treatments on the whole. The highest soil available phosphorus content，shoot phosphorus content and accumulation were showed in the MVF100 treatment at the booting stage and in MVF60 treatment at maturity.By comprehensive consideration，it is appropriate that MV is returned to the field（22.50 t/ha）to replace 20%—40% chemical fertilizer，and the effect is not significantly different between MV returned to the field combined with chemical fertilizer treatments and the F100 treatment in southern Henan rice‑growing area. Based on the significantly correlated quadratic curve relationship，appropriate amount of soil available phosphorus content and leaf area index could improve the dry matter accumulation of panicles at maturity.To achieve the maximum dry matter accumulation of panicles，the optimal soil available phosphorus content at maturity was 7.56 mg/kg，and the leaf area index was 7.02.The MVF60 treatment was close to the optimal value in this study. In conclusion，it is suggested that MV returned to the field（22.50 t/ha）combined with 60% chemical fertilizer is more suitable in this area.

Effect of 6‑Phosphate‑Trehalose on Grain Yield and Fusarium wilt Resistance in Common Beans

XUE Renfeng, HUANG Yuning, JIANG Shan, ZHAO Yang, CHEN Jian, GE Weide

2022, 51(11): 91-97. DOI: 10.15933/j.cnki.1004-3268.2022.11.011

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6‑phosphate‑trehalose is an important signaling molecule that regulates the plant growth and development，and responds to the stress. To study its function in regulating growth，development and resistance‑related reaction，the spraying of exogenous 6‑phosphate‑trehalose solution was used to reveal the effect of 6‑phosphate‑trehalose on the grain yield and Fusarium wilt resistance.The results of this study showed that exogenous T6P praying could significantly improve the relevant grain yield traits. The 100‑grain weights of Longfandou No.1 and Red kidney bean reached 42.7 g and 43.2 g，increased by 18.6% and 20.7%，respectively，and the yields were 2 250.5 kg/ha and 2 549.6 kg/ha，increased by 14.5% and 19.2%. After treatment by 6‑phosphate‑trehalose，the content of soluble carbohydrates in leaves of common beans significantly increased，Red kidney bean had the highest levels of trehalose，glucose，sucrose and starch，respectively reaching 37.4 nmol/g，4.3 μmol/g，5.5 μmol/g，3.6 μmol/g.Longfandou No.1 had the highest fructose content，reaching 8.4 μmol/g；Longfandou No.1 had the highest net photosynthetic rate，intercellular CO₂ concentration and transpiration rate，reaching 18.6，249.6，and 6.5 μmol/（m²·s），respectively.The stomatal conductance of Red kidney bean was maximum，reaching 0.57 μmol/（m²·s）.Spraying of exogenous 6‑phosphate‑trehalose solution promoted plant growth，development and carbohydrate synthesis，improved the photosynthetic efficiency，and also promoted the transport of nutrients from leaves to grains. In addition，the activity or content of the defense factors in the bean roots treated with 6‑phosphate‑trehalose reached the highest level at 48 h after inoculation. T6P induced the significant increase in POD，SOD and PAL activities，reaching 78.3，32.9 and 0.054 nk at/mg，H₂O₂ content was 0.33 ng/mg and the expression levels of defense genes PR1，PR2，and PR5 in the roots of common beans reached peak，which were 8.3，13.2 and 9.4 times of that at 0 h post inoculation，respectively.The disease score was only 4.2 after 28 days post inoculation.In conclusion，6‑phosphate‑trehalose probably increases the yield by regulating soluble sugar content and leaf photosynthetic efficiency，and activates host disease resistance through the expression of plant defense factors.

Differences of Bacterial and Fungal Communities in the Root Tissues of Healthy and Root Rot‑Diseased Tree Peony

JI Junhua, YANG Ruixian, LIU Ping, WANG Aoyi, LI Jingyi, SUN Gaofeng, HE Yanhong, DU Fan

2022, 51(11): 98-108. DOI: 10.15933/j.cnki.1004-3268.2022.11.012

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In order to effectively prevent and control tree peony root rot，Illumina MiSeq sequencing technology was used to conduct high‑throughput sequencing analysis of the V5—V7 variable region of bacterial 16S rRNA gene and fungal ITS gene in healthy and diseased tree peony root samples，so as to clarify the structural differences of bacterial and fungal flora in the root tissues of diseased plants and healthy plants of tree peony，and analyze the occurrence mechanism of tree peony root rot.The results showed that the abundance of bacterial and fungal communities in diseased plants was significantly lower than that in healthy plants. The diversity of fungal population was significantly decreased，but the diversity of bacterial population was not significantly different. The relative abundance of dominant bacteria changed at the levels of phyla and genus.Compared with healthy plants，the relative abundance of Proteobacteria in diseased plants significantly increased by 34.03 percentage points，while that of Firmicutes significantly decreased by 65.00 percentage points. At the genus level，the relative abundance of Streptomyces and Steroidobacter increased by 3.56 and 0.38 percentage points respectively，while that of Pseudonocardia decreased by 13.42 percentage points. The relative abundance of dominant fungi also changed significantly at the phyla and genus levels. Compared with healthy plants，the relative abundance of Basidiomycota in diseased plants increased significantly，with an increase of 9.21 percentage points，while the relative abundance of Ascomycota decreased significantly，with a decrease of 1.61 percentage points.The relative abundance of Hymenula and Dactylectria was significantly increased by 30.71 and 11.69 percentage points，respectively.The relative abundance of Unclassified‑p‑Ascomycota and Exophiala fungi was significantly decreased by 18.28 and 4.41 percentage points，respectively.At the same time，it was speculated that the unique fungus genera in the diseased samples，such as Kurtzmanomyces，Unclassified_f__Nectriaceae and Unclassified_o__Hypocreales might be closely related to the occurrence of tree peony root rot.The prediction results of fungal functional groups showed that the groups related to plant pathogens and saprotrophs in diseased plants were significantly higher than those in healthy plants.To sum up，tree peony root rot changes the bacterial and fungal flora structure in the root tissues of tree peony plants，especially the abundance of fungi related to plant pathogens in the diseased root tissues is significantly higher than that in healthy tissues.

Differences of Enrichment Characteristics of Arsenic and Mineral Elements in Swamp Cabbage Varieties under Different Typical Cultivation Modes

WU Zhichao, YANG Xiuli, LI Furong, XU Shoujun, SHI Hanzhi, DU Ruiying, WANG Xu

2022, 51(11): 109-118. DOI: 10.15933/j.cnki.1004-3268.2022.11.013

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In order to compare and evaluate the differences in the enrichment characteristics of arsenic（As）and mineral elements in swamp cabbages under different typical cultivation modes，this study investigated the enrichment regularity of As，Ca，Mg，Fe，Mn，Cu and Zn in edible parts of 28 swamp cabbage varieties grown in As‑contaminated soils（68.9 mg/kg）under aquatic and dryland cultivation modes.The results showed that significant differences were found in the enrichment capacities of As and mineral elements in swamp cabbage varieties under different cultivation modes.The contents of As，Fe，Mn，Cu and Zn in edible parts of swamp cabbage varieties were higher under aquatic cultivation mode than those of dryland cultivation mode，while the contents of Ca and Mg showed the opposite trends.The average enrichment coefficients of As and Fe，Mn，Cu，Zn in aquatic cultivation mode were 0.043，0.002，0.067，0.401，0.052，and 0.004，0.001，0.042，0.093，0.040 in dryland cultivation mode respectively. These indicated that the enrichment efficiency of As，Fe，Mn，Cu and Zn in aquatic cultivation mode was significantly higher than that at dryland cultivation mode，but the enrichment efficiency of Ca and Mg showed an opposite trend.The enrichment of As，Ca，Mg，Fe，Mn，Cu，and Zn showed significant differences among different swamp cabbage varieties，with 26.0，1.8，1.3，4.1，1.4，53.7，0.9 times higher between the maximum and minimum values under aquatic cultivation mode，and 11.0，1.4，1.0，8.4，2.9，7.8，1.1 times higher for dryland cultivation mode.According to the cluster analysis，under the two cultivation modes，the As enrichment trends in the same group were not consistent due to different cultivation modes，which indicated that the influencing factor of cultivation mode should be fully considered in the selection of swamp cabbage cultivars with low As accumulation.In comparison with the limit standard of As in vegetables（0.5 mg/kg）of GB 2762—2017，the over‑limit rate of As was 89.3% under aquatic cultivation mode，and 7.1% under dryland cultivation mode.Overall，in As‑contaminated soil，the aquatic mode was beneficial to the enrichment of As and microelements in swamp cabbage，while the dryland mode was beneficial to the enrichment of medium elements.

Preparation and Identification of Monoclonal Antibody Broadly Recognizing HA Protein of H7 Subtype Avian Influenza Virus

WANG Xun, LI Ge, LI Qingmei, LÜ Rongzhou, MENG Zekun, CHAI Shujun, YANG Jifei, GUO Junqing, ZHANG Gaiping

2022, 51(11): 119-126. DOI: 10.15933/j.cnki.1004-3268.2022.11.014

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The rapid evolution and mutation of H7N9 subtype avian influenza virus（AIV）made it escape the recognition of existing antibodies. Monoclonal antibodies （mAbs） were prepared that broadly recognized the HA protein of H7 subtype AIV，which would provide effective antibodies for the diagnosis of H7 subtype AIV. BALB/c mice were immunized with H7N9 subtype AIV（A/Chicken/Guangdong/SW154/2015）purified by differential centrifugation，and hybridoma cells were prepared by hybridoma technology.MAbs were screened by hemagglutination inhibition assay （HI），immunoperoxidase monolayer assay（IPMA）and indirect ELISA. Five strains of hybridoma cells that stably secreted mAbs anti‑HA protein were obtained，and the ELISA titers of ascites were all 1∶1 000 000.Among them，the heavy chain of four mAbs was IgG1，the heavy chain of one mAb was IgG2b，and the light chains were all κ chain.The specificity assay showed that the five mAbs only reacted with H7 subtype AIV and did not react with other subtype of influenza viruses.The results of broad‑spectrum assay showed that all five mAbs could react with H7 subtype AIV isolated in different years. The HI results showed that the HI titers of ascites against the H7‑Re3 antigen strain were between 7 log2 and 12 log2.The results of the micro‑neutralization test of MDCK cells showed that all five mAbs had neutralizing activity.Dot blot results showed that all five mAbs could recognize H7‑Re2 antigenic strain and H7‑Re3 variant strain.Western blot showed that a specific protein band appearing in the culture supernatant of five hybridoma cells was about 63 ku，indicating that these five mAbs recognized linear epitopes.In summary，five mAbs that broadly recognize the HA protein of H7 subtype AIV have been successfully prepared，which not only lays the foundation for the detection of influenza virus and the identification of neutralizing antibody epitopes，but also provides ideas for the design and immunological evaluation of new influenza vaccines.

Immunogenicity Evaluation of Porcine Epidemic Diarrhea Virus S1 Protein

CHEN Weicong, LIU Yunchao, ZHOU Chuanjie, YANG Suzhen, WEI Qiang, CHAI Shujun, ZHANG Gaiping

2022, 51(11): 127-134. DOI: 10.15933/j.cnki.1004-3268.2022.11.015

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In order to evaluate the immunogenicity of porcine epidemic diarrhea virus（PEDV）S1 protein，Drosophila melanogaster embryo S2 cells were used to express recombinant S1 protein.The 4‑week old Kunming mice were immunized with purified S1 protein，and the serum and spleen lymphocytes were collected.The immunogenicity of the expressed recombinant S1 protein was analyzed by ELISA，virus neutralization test and flow cytometry. The results showed that，compared with the control group（PBS），the level of specific IgG antibody in serum of mice immunized with recombinant PEDV S1 protein was significantly increased. At 14 days after the second immunization，the neutralizing antibody titers of mice immunized with recombinant PEDV S1 protein was 1∶320.The splenic lymphocytes of immunized mice had stronger proliferative ability.The percentage of peripheral blood CD3⁺T cells increased significantly，and the ratio of CD4⁺/CD8⁺T cells was higher than that of the control group. The content of IL‑4 and IFN‑γ in mice serum increased significantly. In summary，the recombinant PEDV S1 protein is successfully expressed，and it can induce mice to produce high titer neutralizing antibody and promote cellular immune response.

Prokaryotic Expression，Purification and Immunogenicity Study of Outer Membrane Protein AHA1182 of Aeromonas hydrophila

JIAN Sijie, CHAO Jia, SUN Wei, CHEN Rui, DING Rui, CHEN Chen, LIU Xiang

2022, 51(11): 135-144. DOI: 10.15933/j.cnki.1004-3268.2022.11.016

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To evaluate the immunogenicity of outer membrane protein AHA1182 of Aeromonas hydrophila，bioinformatics method was firstly performed to analyze the relationship among A.hydrophila，A.salmonicida，A.schubertii，A.diversa，A.jandaei，Providenci alcalifaciens and Edwardsiella ictaluri.Recombinant expression strain of AHA1182 was constructed by molecular cloning and the optimal expression conditions were identified. AHA1182 was purified by inclusion body washing and SDS‑PAGE，and was used to immunize Carassius auratus.Western blot was used to assess the specificity and titer of the C.auratus serum，and enzyme‑linked immunosorbent assay（ELISA） was used to simulate the recognition effect between anti‑AHA1182 serum and A.hydrophila. Nonspecific immunity was assessed by acid phosphatase（ACP），alkaline phosphatase（AKP）and leukocyte phagocytosis.Furthermore，the toxicity of AHA1182 immunization to C.auratus was investigated by histopathological sections of visceral organ.The results showed that AHA1182 was genetically related among different bacteria，especially in Aeromonas，which meant that AHA1182 serum might have cross‑immune protective ability.AHA1182 was cloned，expressed and purified successfully，and the optimal expression conditions for AHA1182 included strain OD₆₀₀ value of 1.0，isopropyl‑β‑D‑thiogalactoside（IPTG）concentration of 0.5 mmol/L，inducing temperature of 28 ℃ and inducing time of 8 hours.Western blot showed that AHA1182 serum had a high specificity and could recognize A.hydrophila in vitro with the titer of 1∶3 200.The results of AKP，ACP，and leukocyte phagocytosis indicated that AHA1182 stimulated the non‑specific immune of C.auratus. AHA1182 immunization had no toxicity to the visceral structure of C.auratus.The results suggest that A.hydrophila AHA1182 has good immunogenicity.

Research on Tobacco Shred Image Recognition Method Based on Efficient Channel Attention Mechanism and Multi‑Scale Feature Fusion

LIU Jiangpeng, NIU Qunfeng, JIN Yi, CHEN Xia, WANG Li, YUAN Qiang

2022, 51(11): 145-154. DOI: 10.15933/j.cnki.1004-3268.2022.11.017

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To address the problems of poor generalization ability and low accuracy of existing methods in identifying tobacco shred，a method for identifying tobacco shred types based on efficient channel attention mechanism and multi‑scale feature fusion is proposed.Noise reduction is performed on the four types of collected tobacco images，cut stem，expanded tobacco silk，tobacco silk and reconstituted tobacco shred，and K‑means clustering is performed on the processed images to obtain the foreground and hindground of images and complete segmentation to improve the environmental interference resistance and feature extraction capability of the input images. An efficient channel attention mechanism is introduced into the Inception‑ResNet‑V2 network to strengthen the model’s ability to extract features；at the same time，the improved module outputs feature maps for multi‑scale fusion to increase feature representation and reduce the risk of overfitting. Finally，the ReLU activation function and Adam optimizer are replaced by PReLU and AdaBound when comparing convergence and accuracy.The experimental results showed that the proposed algorithm had good generalization ability and enabled efficient identification of four types of tobacco shred，with final recognition accuracy of 97.23% and detection time of 0.107 s for a single image.

Detection Method of Sesame Capsules Based on Improved YOLOX Model

WANG Chuan, ZHAO Hengbin, LI Guoqiang, ZHANG Jiantao, GAO Tongmei, ZHAO Qiaoli, ZHENG Guoqing

2022, 51(11): 155-162. DOI: 10.15933/j.cnki.1004-3268.2022.11.018

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In order to achieve accurate detection of sesame capsules under dense conditions，this study proposes a sesame capsule detection and localization method based on the YOLOX model（CE‑YOLOX model）.In this model，CSPDarknet‑53 is used as the backbone feature extraction network，and a 104×104 large‑scale feature layer is added to the path aggregation network PANet to strengthen the acquisition of the target fine‑grained feature information.By introducing the convolutional block attention module，the important contour features and spatial location information of the object are obtained.The classical NMS is replaced by the Soft‑NMS algorithm，which is more conducive to overlapping target detection，to decrease the missed detection.The results showed that the F₁ average of CE‑YOLOX tested on all datasets at IoU threshold of 0.5 was 0.99，0.05 higher than that of YOLOX.The recall rate and average accuracy of CE‑YOLOX were 98.65% and 99.71%，6.27 and 3.28 percentage points higher than that of YOLOX.The counting accuracy of CE‑YOLOX was 96.84%，5.28 percentage points higher than YOLOX.Consequently，the improved model can recognize sesame capsules under dense conditions.

Study on Automatic Extraction Algorithm of Farmland Flood Disaster Information Based on Remote Sensing

WANG Laigang, XU Shaobo, LI Shimin, GUO Yan, CHENG Yongzheng, HE Jia

2022, 51(11): 163-170. DOI: 10.15933/j.cnki.1004-3268.2022.11.019

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In order to improve the efficiency of information extraction of farmland flood disaster，an effective method of automatic extraction of water body from remote sensing images was explored.The flood disaster in Xunxian County，Henan Province in late July 2021 was taken as the research object，and the NDWI（normalized difference water index），MNDWI（modified normalized difference water index），MBWI（multi‑band water index） and B12 band of Sentinel‑2 remote sensing data were taken as the multi‑dimensional characteristics before，during and after the disaster，and the multi‑dimensional unsupervised water body automatic extraction method was used to extract the water body area.At the same time，Canny‑Edge‑Otsu automatic water body extraction method was used to segment MBWI，MNDWI，NDWI and the HV polarization band of GF‑3 data respectively to extract farmland flood disaster information，and the accuracies of different data sources and methods to extract farmland flood disaster information were compared and analyzed.The results showed that the multi‑dimensional unsupervised water body automatic extraction method integrated various water body indexes and bands as multi‑dimensional features，and the extraction errors of pre‑disaster and disaster water bodies were 6.99% and 7.45% respectively，which were lower than those of Canny‑Edge‑Otsu automatic water body extraction method. By comparing NDWI，MBWI and MNDWI，MBWI had the smallest extraction error and NDWI had the largest extraction error，but buildings and cloud shadow areas were easily mistaken for water bodies.The error of flood extraction based on GF‑3 after the disaster was 15.57%，which was larger than Sentinel‑2 image.However，GF‑3 remote sensing image was not affected by cloud and rain weather，so it provided a strong data support in emergency monitoring of flood disaster.

Microbial Changes and Difference Analysis of Domestic Cigar Tobacco during Stacking Fermentation

PAN Yong, WU Qiaoyin, LI Linlin, TAN Zaiyu, LI Junyao, ZHANG Juan, SHI Youzhi

2022, 51(11): 171-180. DOI: 10.15933/j.cnki.1004-3268.2022.11.020

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To explore the changes and differences of microorganisms among different tobacco leaf parts and origins during the stacking fermentation of domestic cigars，high‑throughput sequencing technology was used to study the microbial community diversity and species composition of tobacco leaves in the middle and lower parts of Shiyan（Hubei）and Deyang（Sichuan）origins.The results showed that the early stage（0—7 days）and the middle stage（7—14 days）were the key nodes of bacterial community changes.During the fermentation process，the microbial community diversity of the middle leaves in Shiyan and Deyang origins basically showed an upward trend，while the microbial community diversity of the lower leaves in different origins basically showed a downward trend.Staphylococcus was the dominant bacterium in the early stage of fermentation.As the fermentation progressed，the dominant bacteria in the middle and lower leaves in Shiyan origin were transformed into Aerococcus and Corynebacterium_1；Staphylococcus was always the dominant bacterium in the lower leaves of Deyang origin，and the dominant bacterium in the middle leaves was changed to Halomonas.Asperguillus was the dominant fungus in the early stage and the end of fermentation.The relative abundances of Alternaria and Microascus increased in the middle and late stages of fermentation，and Talaromyces was the dominant fungus in the middle leaves of Shiyan origin at the end of fermentation.To sum up，during the stacking fermentation process，the community diversity of tobacco leaves in the middle gradually increased，while the community diversity of lower tobacco leaves gradually decreased.The composition and succession of the dominant microbes such as Staphylococcus，Corynebacterium_1，Aerococcus，Asperguillus，Alternaria and Microascus were quite different in different tobacco leaf parts and origins during the stacking fermentation，these dominant microbes might affect the aroma and quality of tobacco leaves through the degradation of harmful substances or the transformation of flavor compounds.

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