Journal of Henan Agricultural Sciences

Research Progress on Chemical Structure，Toxic Effects and Detection Methods of Alternaria Toxins

WANG Ran, LI Yuke, YI Yin, TANG Ming

2023, 52(3): 1-11. DOI: 10.15933/j.cnki.1004-3268.2023.03.001

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Alternaria sp.is ubiquitous in nature and a causal agent of various crops，vegetables，and fruits diseases.Alternaria toxins are a general term for different secondary metabolites produced during the Alternaria infection process.The disease mechanism caused by Alternaria toxins is a hot issue in the current research.Alternaria toxins negatively impact subcellular structures，including chloroplast，mitochondria，plasma membrane，nucleus，etc.Based on interaction characteristics of Alternaria and host，Alternaria toxins are classified into host‑specific toxins（HST）and non‑host‑specific toxins（nHST）.This review focuses on the chemical structure，pathogenicity，and detection techniques of important Alternaria toxins，including the similarities and differences of chemical structure characteristics，differences of targets of pathogenicity，and advantages and disadvantages of different detection methods，which could provide reference for the control of Alternaria disease and the development of toxin detection methods.

Effect of Different N Application Rates on Absorption，Accumulation and Distribution of N and Zn in Different Organs in Wheat at Different Growth Stages

GUO Fangfang, HAN Yaqian, ZHOU Sumei, DU Xihe, SONG Miao, ZHANG Jiemei, HUANG Yuan, HU Naiyue, ZANG Hecang, YANG Xiwen

2023, 52(3): 12-25. DOI: 10.15933/j.cnki.1004-3268.2023.03.002

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Four wheat cultivars were used，including Shangmai 156（Henan provincial authorized cultivar 2015011），Bainong 418（Henan provincial authorized cultivar 2015014），Bainong 207（National authorized cultivar 2013010）and Kaimai 21（Henan provincial authorized cultivar 2011017），the effect of nitrogen（N）rates［0 kg/ha（N1），135 kg/ha（N2），270 kg/ha（N3）］on the absorption，accumulation and transfer of N and zinc（Zn） of wheat in different organs at different stages in 2019—2021 was investigated，so as to reveal the change pattern of N and Zn accumulation in different organs of wheat under different N rates at different growth stages.The results showed that N accumulation was the highest in leaf before anthesis stage in aboveground part，and Zn accumulation in aboveground part was the highest in leaf before jointing stage，and in stem at anthesis.Aboveground N and Zn accumulation was the highest in spike at filling and mature stages.N accumulation in roots was concentrated in seminal roots before the regreening stage，and gradually shifted to nodal roots after the regreening stage.Zn accumulation in roots was concentrated in seminal roots before pre‑winter，and gradually shifted to nodal roots after pre‑winter. N and Zn absorption had synergistic effect.N and Zn accumulation in different organs at different growth stages increased significantly with the increase of N rate.The sums of the contribution rates of different nutrient organs to grain N under N1，N2 and N3 treatments were 84.32%，77.36% and 91.32%，respectively，and the Zn accumulation obtained through redistribution in the upper，middle and lower grains of spike accounted for 35.68%，32.87% and 34.50% of the total Zn absorption in the upper，middle and lower grains of spike，respectively.Under the condition of this experiment，when N rate<270 kg/ha，the accumulation of N and Zn was mainly concentrated in organs such as roots，leaves，stems and glumes in the early and middle growth stages，and transferred to the grain through redistribution in the later growth stages；increase of N rate significantly increased N and Zn accumulation in different organs of wheat at different growth stages；the grain obtained 84.02% and 33.94% of N and Zn，respectively，through redistribution.Grain N mainly came from the redistribution of nutrient organs，and the mobility of N was stronger；the mobility of Zn was weaker，and increase of N rate increased the redistribution of Zn from different nutrient organs to the upper，middle and lower grains of spike，and the redistribution shifted less Zn to the middle grain of spike than the upper and lower grains of spike.

Cloning of Maize ZmHY5 and ZmHY5L Genes and Their Transcriptional Abundance in Responses to Different Light Treatments

YAO Shuaitao, WU Wenjing, WANG Shaoci, ZHAN Weimin, LIU Tong, JIANG Liangliang, YANG Jianping

2023, 52(3): 26-35. DOI: 10.15933/j.cnki.1004-3268.2023.03.003

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In order to explore the function of maize（Zea mays）HY5（elongated hypocotyl 5）gene，two HY5 genes were cloned from maize inbred line B73，named ZmHY5 and ZmHY5L，respectively.Real‑time quantitative PCR（qRT‑PCR） was used to analyze the expression levels of ZmHY5 and ZmHY5L genes in different tissues of maize and their transcriptional abundance in responses to different light，conversion light and long and short sunshine treatments.The results showed that HY5 of Zea mays had similar domains and high amino acid sequence identity with those of Sorghum bicolor，Miscanthus lutarioriparius，Setaria italica and Arabidopsis thaliana，suggesting that they may have similar functions.The tissue expression patterns of ZmHY5 and ZmHY5L genes were similar，with the highest expression level in leaves，but the expression level of ZmHY5 gene was significantly higher than that of ZmHY5L gene.The response patterns of ZmHY5 and ZmHY5L genes to different continuous light were basically similar.ZmHY5 gene had the highest expression level under blue light condition，and more obvious response to darkness to white light.The response ability of ZmHY5L gene under long‑day condition was stronger than that of ZmHY5 gene，while ZmHY5 gene had stronger response ability in darkness phase under short‑day condition.Overall，ZmHY5 and ZmHY5L genes may play a role in maize photomorphogenesis and flowering regulation.

Effects of Different Cultivation Modes on Yield and Resource Utilization Efficiency of Summer Maize

CHEN Yan, WU Jian, MA Haibin, YANG Li, LIU Yingxue, LI Chengyu, LIU Tianxue, LI Chaohai, ZHAO Yali

2023, 52(3): 36-47. DOI: 10.15933/j.cnki.1004-3268.2023.03.004

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The split‑plot test design was adopted，the main plot was sowing date，with three sowing dates of June 10（T1），June 17（T2）and June 24（T3）；the sub‑plot was planting density，the planting densities were 60 000 plants/ha（D1），75 000 plants/ha（D2）and 90 000 plants/ha（D3）；the nitrogen amount was used as sub‑sub‑plot，which were 0 kg/ha（N0），180 kg/ha（N1），270 kg/ha（N2）and 360 kg/ha（N3）.The effects of sowing date，planting density and nitrogen amount on the leaf area index，dry matter accumulation，yield，resource utilization efficiency and the contribution rate of different factors were studied，and thus the suitable cultivation mode of summer maize in the south of Huanghuai region was defined.The results showed that sowing date，planting density and nitrogen amount had extremely significant effects on leaf area index，dry matter accumulation，yield and resource utilization efficiency of summer maize. Summer maize leaf area index，dry matter accumulation and transportation，yield and resource utilization efficiency（partial productivity of nitrogen fertilizer，radiation utilization efficiency，heat utilization efficiency）generally decreased with the delay of sowing date，and increased with the increase of planting density，reached the highest value under the nitrogen amount of 270 kg/ha.Compared with the treatment with planting density of 60 000 plants/ha，nitrogen amount of 180 kg/ha and on June 24，the treatment with planting density of 90 000 plants/ha，nitrogen amount of 270 kg/ha sowing on June 10 increased the yield，partial productivity of nitrogen fertilizer，radiation utilization efficiency and heat utilization efficiency by 101.5%，34.2%，116.6% and 97.4% respectively. However，Howing on June 17，the yield of the treatment with planting density of 90 000 plants/ha and nitrogen amount of 270 kg/ha increased by 90.6%，the partial productivity of nitrogen fertilizer，radiation utilization efficiency and heat utilization efficiency increased by 26.6%，100.0% and 87.2%，respectively.The contribution rates of sowing date，planting density and nitrogen amount to the yield and resource utilization efficiency of summer maize were different，nitrogen amount had the largest contribution rate，and sowing date had the smallest contribution rate. Therefore，it was suitable for summer maize in the south of Huanghuai region when sowed during the first twenty days of June combined with planting density of 90 000 plants/ha and nitrogen amount of 270 kg/ha.

Effect of NtCPS2 Gene on Development and Secretion Synthesis of Tobacco Gland Hair

HAN Wenlong, ZHENG Cong, CAO Kexin, ZHANG Luyang, XU Xiaowen, XIE Ke, ZU Qiongyao, XU Shixiao

2023, 52(3): 48-55. DOI: 10.15933/j.cnki.1004-3268.2023.03.005

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To investigate the relationship between tobacco（Nicotiana tomentosiformis L.）cis‑abisol synthesis gene NtCPS2（ent‑copalyl diphosphate synthase 2）and tobacco gland hair development and secretion synthesis，the 8306 homozygous editing line（knock‑down，KD）with the key synthetic gene NtCPS2 of cis‑abisol edited and 8306 wild‑type（WT）were used as materials，the glandular hair density，contents of cis‑abisol，cembranoids and gibberellic acid（GA₃）and related gene expression levels of KD and WT were compared under the normal growth conditions（CK），application of GA₃ and tetraphyllin.The results showed that for CK，compared with WT，the endogenous GA₃ content of KD tobacco plant significantly increased by 25%，the glandular hair density of leaves significantly increased by 28%，the content of cembranoids secretion in leaves significantly increased by 25%，and the expression levels of key synthetic genes of cembranoids CYC（cembratrien‑ol synthetase）and CYP71D16（cytochrome P450 hydroxylase）significantly increased. After external application of GA₃，the content of GA₃ in KD and WT increased by an average of 50%，the density of glandular hairs in leaves increased by 50% on average，and the content of cembranoids in leaves increased by 50% on average，and the overall increase in KD was greater than that of WT.After the external application of GA₃ inhibitor tetraphyllin，the content of endogenous GA₃ of KD and WT decreased by 25% on average，the glandular hair density of leaves significantly reduced by 56% on average，and the content of cembranoids in leaves significantly reduced by 46% on average，and the overall decrease in KD was greater than that of WT.After editing the NtCPS2 gene，the GA₃content of KD tobacco plant increased，the density of leaf glandular hair increased，and the content of cembranoids secretion increased，which was consistent with the result of external application of GA₃，and opposite to the result of external application of tetraphyllin，indicating that NtCPS2 gene affects the development of tobacco glandular hair and the synthesis of cembranoids by affecting GA₃ biosynthesis.

Traits Observation and SSR Identification of New Panax ginseng Variety Fuxingniu

YANG Zhongliang, XU Huaiyou, HOU Yubing, ZHANG Rui, LEI Xiujuan, WANG Yingping

2023, 52(3): 56-63. DOI: 10.15933/j.cnki.1004-3268.2023.03.006

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In order to comprehensively understand the phenotypic traits of the new Panax ginseng variety Fuxingniu and screen molecular markers that can be used for accurate identification，the characteristics of stem，leaf，fruit，seed，root，saponin content and root resistance of Fuxingniu and two high‑yield control varieties，Fuxing 2 and Damaya were investigated and compared；Precise identification primers were screened and SSR molecular identification was carried out.The results showed that the traits of stem，leave，fruit，seed and root yield of Fuxingniu were significantly higher than those of Fuxing 2 and Damaya Panax ginseng types（P<0. 05）；The saponin Rg₁+Re total content in the root of Fuxingniu was 1.76%，and the content of Rb₁ was 0.56%，which were the highest among the three，while the incidence indexes of rust disease and red skin disease were the lowest among the three；The primer Pg83678 could be used to accurately identify Fuxingniu.To sum up，the new Panax ginseng variety Fuxingniu has good characters，and has good promotion foundation and application prospects.

Effect of IAA on Seed Germination and Physiological Characteristics of Polygonatum sibiricum Red.

YUE Hewei, LI Lianzhen, WANG Yuqing, WANG Rongrong, LIU Yang, WEI Junjie, CHANG Jianfeng

2023, 52(3): 64-72. DOI: 10.15933/j.cnki.1004-3268.2023.03.007

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In order to explore the effect of indoleacetic acid（IAA）on the germination and physiological characteristics of Polygonatum sibiricum Red.seeds，50 mg/L（T1），100 mg/L（T2），200 mg/L（T3）IAA and distilled water（CK） were used to treat Polygonatum sibiricum Red.seeds，the germination and growth of the seeds were investigated，the activities of peroxidase（POD），superoxide dismutase（SOD），catalase（CAT），lipase and the content changes of malondialdehyde（MDA），fat，free amino acid，protein，starch and soluble sugar were determined in Polygonatum sibiricum Red.seeds.The results showed that the germination rates of Polygonatum sibiricum Red. seeds in T1，T2，T3 treatments were 52.82%，174.98% and 68.05% higher than CK on the 33rd day.Bulb diameter，main root length and dry mass were significantly higher in T1，T2，T3 than in CK，and the number of lateral roots，main root length and dry mass of T2 treatment were significantly higher than those of other treatments.The activities of POD，SOD and CAT reached the highest on the 22nd day after treatment.The activities of POD in T1，T2 andT3 were 16.36%，30.12% and 14.07% higher than CK，the activities of SOD were 16.36%，32.33%and 9.79% higher than CK，and the activities of CAT were 22.62%，63.43% and 32.33% higher than CK，respectively.MDA content during the entire seed germination process was CK>T1>T3>T2.The contents of starch，soluble sugar，protein and free amino acid were the lowest on the 22nd day，and T2 was significantly lower than other treatments. Fat content in each stage was CK>T1>T3>T2.The lipase activity reached the highest level on the 36th day，and T1，T2 and T3 were 149.15%，175.42% and 137.20% higher than CK respectively. Principal component analysis showed that starch content，soluble sugar content，free amino acid content，protein content，POD activity，SOD activity，CAT activity and lipase activity could be used as characteristic indicators to reflect the growth of Polygonatum sibiricum seeds.To sum up，IAA can improve the germination rate，substance metabolism level and antioxidant enzyme activity of Polygonatum sibiricum Red. seeds，and 100 mg/L is the best concentration to promote the germination of Polygonatum sibiricum Red.seeds.

The Cd Absorption Characteristics and Edible Safety Evaluation of Different Varieties of Potato Tubers

CHAI Guanqun, ZHOU Lixing, WANG Li, LIU Guihua, QIN Song, CAO Yan, FAN Chengwu

2023, 52(3): 73-81. DOI: 10.15933/j.cnki.1004-3268.2023.03.008

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Field experiments were conducted in Cd‑contaminated farmland in T and Y locations of northwest Guizhou. Twenty one common potato varieties in northwest Guizhou were used as research objects to analyze the effect of varieties and environment，and to explore the Cd absorption characteristics and edible safety of different varieties of potato tubers，in order to provide practical reference for the safe planting of potatoes in northwest Guizhou. The results showed that the yield of different potato varieties in Y location ranged from 3 564.3 kg/ha to 15 271.8 kg/ha，with an average of 9 200.1 kg/ha.The yield of different potato varieties in T location ranged from 1 825.9 kg/ha to 16 305.6 kg/ha，with an average of 6 754.0 kg/ha. Zhuanxinwu in T and Y locations had the highest potato tuber yield.Variety，environment and their interaction had significant effects on potato tuber yield.The Cd content in tubers of different potato varieties in Y location ranged from 0.061 to 0.153 mg/kg，with an average of 0.094 mg/kg.The Cd content in tubers of different potato varieties in T location ranged from 0.055 to 0.111 mg/kg，with an average of 0.080 mg/kg.Variety，environment and their interaction had significant effects on Cd content in potato tubers.The standard exceeding rates of Cd in 21 potato varieties tested in Y and T locations were 38.09% and 14.29%，respectively.The Cd content in tubers of Qingshu 9 was the lowest in both Y and T.The edible safety evaluation showed that eating Cd‑excessive potato varieties had less risk to human health，but considering the Cd content in tubers，it is suggested that the varieties of potato with excessive Cd content should be used as industrial raw materials more safely.

Effects of Phosphorus Supply Level on Dry Matter Weight and Phosphorus Uptake and Utilization Characteristics of Switchgrass

QU Xiaoyu, WU Na, ZHAO Cong, CHEN Juan, LIU Jili

2023, 52(3): 82-90. DOI: 10.15933/j.cnki.1004-3268.2023.03.009

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In order to explore the effects of different phosphorus supply levels on the dry matter weight and phosphorus uptake and utilization characteristics of switchgrass，a two‑factor randomized block design was adopted to conduct a pot experiment.Three varieties of switchgrass（Alamo syndiploid，Alamo and Pathfinder）were set up，and three phosphorus supply levels P0（0 mg/kg phosphorus），P10（10 mg/kg for low phosphorus）and P100（100 mg/kg for high phosphorus）were used to analyze the growth indexes and phosphorus utilization characteristics of switchgrass，and comprehensively evaluate the phosphorus uptake and utilization characteristics of switchgrass under different phosphorus supply levels.The results showed that the plant height，tiller number，leaf area and dry matter weight of Alamo syndiploid，Alamo and Pathfinder increased with the increase of phosphorus supply level，and reached the maximum under P100 treatment.The root‑shoot ratio of switchgrass decreased with the increase of phosphorus supply level，compared with P0 treatment，the root‑shoot ratio of switchgrass in P100 treatment was significantly decreased by 16.67%，17.24%，20.69%，respectively.With the increase of phosphorus supply level，the leaf phosphorus content，root phosphorus content and phosphorus uptake of three switchgrass varieties showed an increasing trend，and reached the maximum under P100 treatment.Compared with P0 treatment，phosphorus utilization efficiency in P100 treatment decreased by 11.32%，18.31%，18.38%，respectively. The phosphorus transfer efficiency of both Alamo syndiploid and Alamo reached the lowest value in P100 treatment，while that of Pathfinder was the lowest in P10 treatment.The results of membership function analysis showed that the means of membership function values related to phosphorus uptake and utilization characteristics of three switchgrass varieties under P100 treatment were the largest. In conclusion，100 mg/kg（P100）phosphorus supply condition can not only promote the plant growth and dry matter weight of three switchgrass varieties，but also improve the phosphorus uptake and utilization characteristics to a certain extent.

Effects of Different Herbicide Reduction and Adjuvant Combination on Weed Control，Growth，Yield and Quality of Sweet Potato

LIU Yajun, WANG Wenjing, LI Min, HU Qiguo, WANG Honggang, CHU Fengli

2023, 52(3): 91-100. DOI: 10.15933/j.cnki.1004-3268.2023.03.010

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Through the field experiment，eight test treatments were set up，including no herbicide and no artificial weeding（CK1），no herbicide and artificial weeding（CK2），100% metolachlor emulsifiable concentrate 4.50 L/ha（T1），70% metolachlor emulsifiable concentrate 3.15 L/ha + Jijian 150 mL/ha （T2），70% metolachlor emulsifiable concentrate 3.15 L/ha+ Maisi 100 mL/ha（T3），100% pendimethalin emulsifiable concentrate 5.00 L/ha（T4），50% pendimethalin emulsifiable concentrate 2.50 L/ha+ Jijian 150 mL/ha（T5） and 50% pendimethalin emulsifiable concentrate 2.50 L/ha + Maisi 100 mL/ha（T6），the effects of different herbicide reduction and adjuvant combination on weed control，growth，yield and quality of sweet potato were studied，and the appropriate herbicide reduction and additive combination scheme was determined，so as to provide scientific basis for the safe selection of sweet potato herbicides in eastern Henan.The results showed that，at 5 days and 10 days after applying herbicide，compared with the other treatments，the plant control effect of T2 at treatment significantly increased by 16.94%—40.22% and 5.71%—21.79%，and the fresh weight control effect increased by 8.54%—36.35% and1.50%—14.99%，followed by T5 treatment.At 15 days after applying herbicide，compared with the other treatments，the plant control effect of T5 treatment increased by 3.39%—11.71%，and the fresh weight control effect increased by 3.60%—11.10%，which were significantly higher than T1，T3 and T4 treatments. Compared with CK2，the herbicide treatments had certain adverse effects on plant height，aboveground fresh weight，leaves SPAD value，net photosynthetic rate，activities of antioxidant enzymes［catalase（CAT），peroxidase（POD），superoxide dismutase（SOD）］，content of malondialdehyde（MDA），yield and quality of sweet potato.Among them，T5 treatment had the least effect，and there was no significant difference compared with CK2 treatment；T4 treatment caused obvious damage to the growth of sweet potato. Compared with CK1，the herbicide treatments had certain promoting effect on the growth and development，yield and quality of sweet potato.The plant height and plant fresh weight significantly increased by 5.60%—37.30%，17.05%—57.13%，the SPAD value significantly increased by 7.60%—23.78%，the net photosynthetic rate increased by 1.33%—15.98%，the activities of SOD，POD and CAT increased significantly by 11.80%—33.38%，5.59%—20.20%，8.80%—41.40% respectively，the content of MDA decreased by 0.83%—17.02%，the sweet potato yield and rate of commercial sweet potato significantly increased by 21.95%—46.72% and 7.63%—17.28% respectively，the contents of starch，reducing sugar，soluble sugar，carotene and crude protein increased by 6.72%—17.00%，19.44%—33.10%，2.45%—7.83%，9.16%—20.64% and 4.81%—16.55% respectively.Overall，on the premise of ensuring weed control effect，the treatment of 50% pendimethalin emulsifiable concentrate 2.50 L/ha + Jijian 150 mL/ha had the least impact on the growth of sweet potato，and was a suitable combination scheme of sweet potato herbicide and adjuvant agent.

Isolation and Identification of Dominant Mold and Its Moldy Volatile Metabolites in Tobacco Strips during Storage

WANG Lin, WANG Yudong, ZHU Bao, LI Hu, ZHOU Hongshen, WANG Kaiyue, YAN Tiejun, ZHOU Ping, XIA Hai, ZHOU Min

2023, 52(3): 101-108. DOI: 10.15933/j.cnki.1004-3268.2023.03.011

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To establish a rapid identification method for tobacco mildew during storage，pure culture and 18S rDNA sequencing were used to isolate，purify and identify the molds in warehouse moldy pieces of tobacco. The dominant mold was tested for mildew.The differences of volatile components in tobacco strips during different infection periods of the dominant mold were analyzed by gas chromatography‑ion mobility spectrometry（GC‑IMS）.It was found that Rhizopus was the most abundant in tobacco leaf samples，followed by Aspergillus，Mucor and Penicillium.Further identification showed that Rhizopus oryzae was the dominant mold in tobacco leaves.The mildew experiment showed that Rhizopus oryzae had certain mildew ability.Tobacco leaves were inoculated with Rhizopus oryzae for artificial contamination.After 18 days of culture at 35℃ and 70% relative humidity，the amount of Rhizopus oryzae reached 5.0×104 cfu/mL，and mild mildew occurred in the tobacco leaves，and 42 characteristic volatile compounds were identified. Among them，the concentrations of 1‑octene‑3‑one，3‑octanone，2‑furan methanethiol，alpha‑terpineol，1‑octanol，3‑ethyl‑phenylpropanoate，pentanoic acid，and alpha‑pinene decreased significantly during the infection stage of Rhizopus oryzae，the concentrations of（Z）‑3‑hexen‑1‑ol，methyl butanoate，2‑pentanone，and hexyl propionate increased significantly in the mildew stage of tobacco，which could be used as potential labeling molecules for mold in tobacco strips.

Effects of Alcoholic Waste Mash Liquid Organic Carbon Fertilizer on Root Growth，Yield and Quality of Small Tomato

SHAO Zeqiang, BU Xintong, GOU Qiandong, GUO Xiaoxiao, ZHOU Linyuan, WANG Jingbo, LI Yinghua

2023, 52(3): 109-117. DOI: 10.15933/j.cnki.1004-3268.2023.03.012

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In order to verify the effect of alcoholic waste mash liquid organic carbon fertilizer on small tomato，a randomized group trial was conducted in pots with five treatments：CK（blank control），T1 （conventional fertilizer），T2（alcoholic waste mash liquid organic carbon fertilizer），T3（commercial liquid fertilizer A）and T4（commercial liquid fertilizer B），T2—T4 applied at a rate of 100 mL/pot. Plant height，stem diameter，SPAD value，root morphology，tomato yield，vitamin C content，nitrate content，soluble sugar content and organic acid content were determined，and the effects of alcohol waste mash liquid organic carbon fertilizer were compared with chemical fertilizers and two commercially available liquid fertilizers on agronomic traits，root morphology，yield and quality of small tomato. The results showed that compared with the T1 treatment，the T2 to T4 treatments could significantly promote the growth of small tomato，and yield per plant increased ranging from 8.3% to 16.7%，with the T2 treatment being the highest；root length，root surface area，and average root diameter decreased by 2.8% and 4.3%，0.8% and 4.5%，and 2.0% and 0.1% in the T3 and T4 treatments，respectively，while only the T2 treatment increased by 25.7%，23.3%，and 18.8%，respectively；root volume increased by 29.4% and 2.9% in the T2 and T3 treatments，respectively，and decreased by 8.8% in T4 treatment；vitamin C and soluble sugar content increased by 21.5% and 3.0%，15.1% and 4.8% in T2 and T3 treatments，respectively，both of which were the highest in the T2 treatment；nitrate content decreased by 15.4% to 34.1%，with the lowest in the T3 treatment；the reduction in organic acid content ranged from 29. 7% to 43.8%，with the lowest in the T2 treatment.Correlation analysis indicated that root length，root surface area，root volume and root mean diameter of small tomato showed significantly positive correlations with yield，vitamin C content and soluble sugar content，while root length and root surface area showed significantly negative correlations with organic acid content.Therefore，alcoholic waste mash liquid organic carbon fertilizer improves fruit yield and quality by promoting the growth of the root system of small tomato.

Effects of Chloride Varieties and Concentration on Watermelon Growth and Root Vitality

LIU Xue, WANG Xina, LI Xuefang, HAO Wenyue, HUO Qingzhu, TAN Junli

2023, 52(3): 118-126. DOI: 10.15933/j.cnki.1004-3268.2023.03.013

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To explore the toxic mechanism of chlorine on watermelon，and to provide theoretical basis for the safe utilization of chlorine‑containing fertilizer and chlorine‑containing brackish water，the effects of different chloride varieties and concentration on root biomass，malondialdehyde content，root activity，Clcontent and NO₃^-content of watermelon Jincheng No.5 were studied through hydroponic experiment.The results showed that the growth of watermelon root system was affected by both different chlorine varieties and concentrations.Compared with the control without chlorination，the NO₃^-content in roots treated with NaCl，NH₄Cl，CaCl₂ and KCl decreased by 44.16%—47.95% and 35.96%—49.62%，respectively，at medium and high concentrations（80 mmol/L and 160 mmol/L）.The content of Cl− increased by 0.71—1.34 times and 1.24—1.86 times，respectively，and the aboveground and underground fresh weight decreased by 41.39%—57.78%，31.31%—44.73% and 57.28%—72.13%，49.35%—67.38%.NaCl treatment did not significantly affect the content of MDA.Other chlorides significantly reduced MDA content by 31.87%—45.05% and 25.27%—28.57%.For different chlorides，NaCl treatment aggravated oxidation loss compared with other chlorides，and NH₄Cl treatment promoted Cl^-absorption to a certain extent.Watermelon root activity significantly increased by 41.18%—104.17% under medium‑concentration CaCl₂treatment compared with other chlorides.The underground dry weight increased by 36.00%—112.50% compared with other chlorides.Correlation analysis showed that fresh weight and dry weight were significantly correlated with Cl⁻and NO₃^-content.We can conclude that exogenous chlorine can reduce NO₃^-content by increasing Cl⁻content in roots，thus reducing root vitality and inhibiting root growth.With different ions，the inhibition degree and mechanism of chlorine on watermelon are different. The toxic effect of CaCl₂ and KCl on watermelon root growth was less than that of NaCl and NH₄Cl.

Eukaryotic Expression of Porcine Pseudorabies Virus gE Protein and Identification of Its Activity

WANG Qinglong, WANG Ruining, WANG Xun, LI Qingmei, LI Ge, ZHANG Zhenzhen, YANG Suzhen, GUO Junqing, ZHANG Gaiping,

2023, 52(3): 127-134. DOI: 10.15933/j.cnki.1004-3268.2023.03.014

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In order to obtain the recombinant gE protein of porcine pseudorabies virus（PRV）and evaluate its antigenic activity，the optimized extracellular region of gE gene was inserted into the eukaryotic expression vector pCMV and the Hexa‑His tag was added to its C‑terminal to construct the eukaryotic expression plasmid pCMV‑gE.HEK293F cells were transfected with the recombinant plasmid，and secretory expression of the gE recombinant protein in the transfected cells was identified by Dot blot.The expressed gE protein was then purified from supernatant of the transfected cells by nickel affinity chromatography and gel chromatography，which was identified by SDS‑PAGE and Western blot.Furthermore，the antigenic activity of the gE recombinant protein was analyzed by ELISA using the PRV positive serum，and its glycosylation modification as well as its effect on the antigenic activity was analyzed by PNGase F treatment.Dot blot results showed that the gE recombinant protein was secreted and expressed in supernatant of the transfected cells through its own signal peptide，which reached its peak at 96 h post‑transfection.The results of SDS‑PAGE and Western blot showed that the purity of the gE recombinant protein purified from the cell supernatant was about 90%.Molecular weight of the gE recombinant protein after treated with disaccharidase PNGase F was significantly lower than that of the untreated protein.The indirect ELISA results showed that the gE recombinant protein detected PRV positive serum at 1∶12 800，while the detection limit of PRV positive serum for the expressed gE protein was determined as 31.25 ng/mL，whose antigen activity was four times that of the prokaryotically expressed protein.The detection of clinical serums showed that the coincidence rate between the gE‑based ELISA and IDEXX antibody detection kit was 96.5%.In conclusion，the recombinant gE protein with high expression level，high purity，high activity and glycosylation modification was secreted and expressed in eukaryotic cells，showing a good application potential in the differential detection of PRV infection.

Development and Application of TB Green Ⅱ Real‑time PCR Method for Detection of Porcine Circovirus Type 3

LI Peng, SUN Yanju, WANG Yinbiao, JIN Qianyue, LIANG Xiaoxiao, YIN Mei, WANG Xuannian, LIU Xingyou, WANG Liping

2023, 52(3): 135-142. DOI: 10.15933/j.cnki.1004-3268.2023.03.015

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In order to establish a rapid，sensitive and specific detection method for porcine circovirus 3（PCV3），specific primers were designed based on the conservative region of the whole genome of PCV3，and the standard plasmid was constructed.By optimizing the reaction system and procedure，a real‑timequantitative PCR（qPCR）method based on TB Green Ⅱ was established，and its specificity，sensitivity，repeatability and feasibility were verified.The results showed that the primer specificity of the established TB Green Ⅱ qPCR detection method was good，and there was a good linear relationship（R²=0.999）when the plasmid standard samples were from 4.74×102 to 4.74×107 copies/μL.The detection limit of the method was 10 copies/μL，and there was no cross reaction with classical swine fever virus（CSFV），porcine reproductive and respiratory syndrome virus（PRRSV），porcine parvovirus（PPV）and porcine circovirus type 2（PCV2）.The intra‑group variation coefficient was 0.33%—0.62%，and the inter‑group variation coefficient was 0.40%—0.73%.A total of 132 clinical samples were tested，the detection rate of TB Green Ⅱ qPCR was 9.10%（12/132），which was higher than that of conventional PCR（4.55%，6/132）.The established TB Green Ⅱ qPCR method has good sensitivity，specificity and repeatability，and can be used for the diagnosis，epidemiological monitoring and laboratory research of PCV3 infection in clinical samples.

Analysis of Biofilm Culturing，Microbial Community Composition of Three Biological Fillers in Recirculating Aquaculture System and#br# Their Effects on Growth of Largemouth Bass（Micropterus salmoides）

LI Qian, SUN Lihui, GUO Jianlin, ZHENG Gang, JIANG Jianhu, CHEN Jianming, GAO Lingmei

2023, 52(3): 143-152. DOI: 10.15933/j.cnki.1004-3268.2023.03.016

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In order to explore practical water‑treating biological fillers for largemouth bass（Micropterus salmoides）in recirculating aquaculture system，with square sponge，Mutag Biochip 30 and fluid sphere as biological fillers，largemouth bass with initial body weight of（20.46±0.46）g as experimental objects，the biofilm maturing time and microbial community composition were analyzed，and the growth of largemouth bass was evaluated.The results showed that the maturing time of the three biological fillers was 17—21 d under natural conditions.16S rDNA high‑throughput sequencing results showed that the dominant bacteria of biological fillers at the phylum level were the same as those in the corresponding cultured water samples，namely Proteobacteria and Actinobacteriota.At the genus level，the bacterial microbial community composition of biological fillers was different from that of corresponding cultured water samples.Leifsonia and Incerta‑sedis were the dominant bacteria in the water samples.Thermomonas was the dominant genus in the square sponge biological filler，while Mutag Biochip 30 and fluid sphere had the same dominant genus as Nitrospira.The results of breeding experiments indicated that after 44 days，the final body weight and weight gain rate of largemouth bass in the square sponge group were significantly higher than those in the other experimental groups，the feed conversion ratio was significantly lower than that of Mutag Biochip 30 group，and there was no significant difference in the survival rate among different biological fillers.In conclusion，in the short term，square sponge is a practical biological filler for recirculating aquaculture water treatment.

Research on Improved U‑Net Method for Rice Leaf Cell Segmentation

WEI Gaixing, YI Wenlong, LIU Yucheng, ZHAO Yingding, CHEN Tingzhuo

2023, 52(3): 153-160. DOI: 10.15933/j.cnki.1004-3268.2023.03.017

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Aiming at the issues of blurred boundaries，low signal‑to‑noise ratio，mutual adhesion and stacking of mesophyll cells in the image of rice leaf cells，which lead to low segmentation accuracy，we propose an improved U‑Net method for rice leaf cell segmentation. Firstly，the bridge attention（BA）module is introduced into the ResNeXt network to form the BAResNeXt module as the network encoder to improve the network’s attention to mesophyll cells when extracting deep semantic features；secondly，a channel cross‑attention mechanism is added between the encoder and the decoder to ease the semantic ambiguity between the decoder and the encoder to enhance the information fusion of the segmented image features；finally，the SE attention mechanism is used in the upsampling phase of decoder to filter the interference information of image background. In order to verify the effectiveness of the method，it was compared with deep learning networks such as U‑Net，Res‑UNet，U‑Net++ and Deeplabv3+.The results showed that our method had the best performance in rice leaf cell segmentation. The Precision（96.03%），Recall（97.67%），Acc（97.47%），IoU（93.96%）and Dice（96.78%）of our method were all higher than other networks.

Study on Canopy Chlorophyll Estimation Model of Buckwheat with Different Selenium Levels Based on UAV Multispectrum

MA Wei, WU Zhiming, YU Kesong

2023, 52(3): 161-172. DOI: 10.15933/j.cnki.1004-3268.2023.03.018

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In order to verify the possibility of estimating the chlorophyll content of buckwheat canopy by using low‑altitude unmanned aerial vehicle（UAV）with multi‑spectral camera，and to explore the effect of selenium（Se）on the chlorophyll and spectral characteristics of buckwheat canopy，Jinsage No.6 sweet buckwheat and Jinsage No.9 bitter buckwheat were selected as the research objects，and a experimental study was conducted in Taigu District，Jinzhong City，Shanxi Province.Buckwheat multispectral images were collected under different selenium levels at different stages by using UAV with multi‑spectral camera，and the relative chlorophyll content（SPAD value）was synchronously measured in the field.Firstly，the SPAD values of buckwheat canopy under different selenium application levels were analyzed.Secondly，the reflectance of buckwheat canopy under five bands was obtained by extracting the spectral information of multi‑spectral images.On this basis，the spectral characteristics of buckwheat at full bloom and filling stage were analysed，11 vegetation indices were constructed using the reflectance in five bands，and the absolute magnitude of the correlation coefficients between the 16 spectral variables and the measured SPAD values were obtained by Pearson correlation analysis.Partial least square regression（PLSR），principal component regression（PCR），support vector machine regression（SVR）and back propagation neural network（BPNN）were used to construct a buckwheat canopy SPAD monitoring model，and the optimal estimation model was determined by accuracy test.The results showed that moderate application of selenium fertilizer could increase SPAD value of buckwheat，while excessive application could inhibit SPAD value.The blue，red，red‑edge and NIR bands showed strong correlation among the five bands，and the NIR band showed high and stable correlation. In terms of vegetation index，the correlation coefficients（|r|）of standardized precipitation index（SPI），green chlorophyll index（GCI），green normalized difference vegetation index（GNDVI），normalized green light index（NGI），transformed optimized soil‑regulated vegetation index（TOSAVI），transformed chlorophyll absorption ratio index（TCARI），and triangular vegetation index（TVI）ranged from 0. 50 to 0. 91，which indicated a good correlation.At the full‑bloom stage，BPNN had the best prediction effect，the correlation coefficient of prediction set（R²_P）was 0.97，and the root mean square error（RMSE）of prediction set was 0.95.In the filling stage，the prediction effect of SVR was better than other models，R²_Pwas 0.96，and RMSE was 0.45.At the full‑bloom stage and filling stage，PLSR showed the best performance，R²_P was 0.98，RMSE was 0.28.By comparing all the models，SVR showed higher stability and accuracy（R²_P and RMSE ranges were 0.94—0.96 and 0.45—0.82，respectively，and RPD values were greater than 3.00）.These results show that UAV with a multispectral camera can achieve rapid monitoring of buckwheat canopy chlorophyll content in the field，providing a reference for optimization of the model algorithm for low altitude prediction of chlorophyll content by UAV.

Difference Analysis of Main Quality Indicators of Upper Leaves of Flue‑cured Tobacco with Different Colors

HAO Xianwei, LI Qi, BI Yiming, TIAN Yunong, LI Yongsheng, WANG Hui, ZHONG Yongjian, WANG Fang, XU Xiuhong, FU Qiujuan, SUN Tingting, HAN Xiao

2023, 52(3): 173-180. DOI: 10.15933/j.cnki.1004-3268.2023.03.019

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To clarify the differences of main quality indicators of upper leaves of flue‑cured tobacco with different colors，analysis of variance（ANOVA）was applied to compare the 41 main chemical components and 9 sensory evaluation indexes of upper leaves with light orange，orange and dark orange colors from 27 production areas of 12 provinces（cities，districts）.The results showed that，with the deepening of tobacco color，the contents of potassium，ether extracts，scopletin，2，3‑bipyridine，megastigmatrienone，damascenone，ionone，neophytadiene，geranyl geraniol，oxalic acid and succinic acid increased，while the contents of total sugar，calcium，chlorogenic acid，caffeic acid，fumaric acid，palmitic acid，stearic acid and pH decreased.The color of upper tobacco leaves mainly affected the delicacy，clarity，miscellaneous gas，moistening feeling，irritation，aftertaste，concentration and strength of sensory quality indicators.Light orange tobacco leaves had good delicacy，high definition，low irritation and miscellaneous gas，comfortable aftertaste，and lower smoke concentration and strength than dark orange tobacco leaves.

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