Journal of Henan Agricultural Sciences

Research Progress on Cold Resistance of Wheat

FANG Yuhui, HAN Liupeng, HUA Xia, ZHAO Mingzhong, GUO Rui, QI Xueli, HU Lin

2022, 51(4): 1-10. DOI: 10.15933/j.cnki.1004-3268.2022.04.001

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The freeze injury is a frequent natural disaster in wheat production，which is one of the important environmental factors restricting wheat yield and quality. In this paper，the type of wheat freeze injury，the physiological mechanism（cell membrane system，osmotic adjustment substances，antioxidant system，photosynthetic characteristics and endogenous hormones）and molecular mechanisms（cold resistant genes，cold resistant omics） of wheat cold resistance were summarized. The problems and solutions in the study of wheat cold resistance were also discussed，which provided an important reference for the research of cold resistance mechanism and breeding of wheat.

Research Advances in Biological Control of Pepper Fusarium Wilt

ZHAO Zhixiang, YAN Wanrong, WANG Bao, HE Shu, ZENG Xiangping, XIAO Tongbin

2022, 51(4): 11-21. DOI: 10.15933/j.cnki.1004-3268.2022.04.002

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Fusarium wilt of pepper caused by Fusarium oxysporum is one of the soil‑borne diseases restricting the production of pepper.At present，effective disease‑resistant materials are still lacking，and chemical agents could not control it completely，at the same time causing problems such as environmental pollution，pesticide residues，and bacterial resistance and so on.The use of microorganisms and their metabolites，plant extracts and other biological pesticides for prevention and control can not only control the occurrence of diseases，but also not damage the ecological environment and endanger the health of humans and animals.Biological control meets the requirements of agricultural sustainable development，and has become a hot research topic.In this paper，the research progress of microbial species，plant sources and their biocontrol mechanisms used for the biological control of pepper Fusarium wilt in recent twenty years was reviewed. The existing problems of biocontrol products were analysed，and the future development direction of biocontrol of pepper Fusarium wilt was prospected.

Effects of Shallow‑buried Drip Irrigation without Film on Maize Yield and Economic Benefit in Northern Xinjiang

CHEN Jianglu, DING Bianhong, ZHANG Xiaowei, ZHANG Tingjun, SUN Zhiyuan, CHEN Yongsheng, JIANG Xingxing

2022, 51(4): 22-29. DOI: 10.15933/j.cnki.1004-3268.2022.04.003

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The early maturing（KWS2030，P1），medium maturing（KWS9384，P2） and late maturing（KWS3564，P3）maize varieties were used as materials，and the effects of different drip irrigation methods（film covered drip irrigation，F1；shallow‑buried drip irrigation without film，F2）on maize growth，yield and economic benefit were studied，so as to provide theoretical basis and technical guidance for high yield，water saving and high efficiency of maize cultivation and sustainable development of irrigated agriculture in arid areas of northern Xinjiang. The results showed that the growth period of maize of F2 treatment was 4—5 days longer than that of F1 treatment. The plant height，ear height，height of gravity center，ear coefficient，gravity center coefficient，basal internode stem diameter，basal internode length and ratio of basal internode length to basal internode diameter of maize of F2 treatment were generally higher than those of F1 treatment. The dry weight of maize was not significantly different between F2 treatment and F1 treatment except for silking stage（F2 treatment was lower than F1 treatment）.Compared with F1 treatment，grain number per panicle of F2 treatment increased by 3.1%—7.3%；the panicle number of F2 treatment was slightly lower，but the difference was not significant. For P1，the thousand grains weight of F2 treatment was higher than that of F1 treatment，while P2 and P3 were the opposite. For P1，the yield of F2 treatment was lower than that of F1 treatment；for P2 and P3，the yields of F2 treatment increased by 0. 9% and 6.0% compared with F1 treatment，respectively.For P1，the output value of F2 treatment was lower than that of F1 treatment；for P2 and P3，the output values of F2 treatment increased by 0.6% and 5.7% compared with F1 treatment，respectively. Compared with F1 treatment，the economic benefit of F2 treatment increased by 2.8%—13.1%，the ratio of output to input of F2 treatment increased by 12.0%—23.1%. In conclusion，compared with film covered drip irrigation，shallow‑buried drip irrigation without film can generally improve grain number per panicle，yield，economic benefit and ratio of output to input of maize based on the premise that maize can mature normally，and middle‑late maturing varieties perform better.

Overexpression of miR528 Enhances K⁺ Homeostasis in Rice Seedling under Salt Stress

LEI Zhenshan, WANG Tongtong, JI Xin, ZHANG Tianhai, WANG Fujuan, DONG Liping, ZHAO Yafan, LIU Juan

2022, 51(4): 30-38. DOI: 10.15933/j.cnki.1004-3268.2022.04.004

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In order to investigate whether miR528 is involved in maintaining K+ homeostasis in rice seedlings under salt stress and its possible regulatory mechanism，transgenic lines overexpressing miR528（miR528‑OE1 and miR528‑OE2），T‑DNA insertion mutant mir528 and their wild types（Zhonghua 11and Dongjin）were used as experimental materials.The expression pattern of miR528 was explored under salt stress，and the growth，K⁺ content in shoot and root，K⁺ flux in root and the expression levels of K⁺channel related genes in different rice materials were studied under salt stress.The results indicated that the expression of miR528 in rice seedlings was induced by salt stress.Compared with the control without salt stress，the fresh and dry weights of shoots and roots were less inhibited by salt stress in miR528‑OE1 and miR528‑OE2 than those in wild type Zhonghua 11，but the fresh and dry weights of roots were more inhibited by salt stress in mir528 than those in Dongjin. Under salt stress，the K⁺ content in shoot and root of Zhonghua 11 decreased to 43.41% and 23.67% of the control，respectively；the K⁺ contents in shoot and root of miR528‑OE1 and miR528‑OE2 were significantly higher than those of Zhonghua 11，while the K⁺content in root of mir528 was significantly lower than that of Dongjin.Instantaneous addition of salt rapidly induced K⁺ efflux from roots.The peak value appeared at 3—4 min after salt treatment.The peak value of K+ efflux from root of miR528‑OE was obviously lower than that of Zhonghua 11，while the peak value of K⁺efflux from root of mir528 was obviously higher that of Dongjin. Under the normal condition，the expression levels of GORK（guard cell outward rectifying K⁺ channel） in roots of miR528‑OE1 and miR528‑OE2 seedlings significantly increased compared with Zhonghua 11，but the expression level of GORK in root of mir528 seedling significantly decreased compared with Dongjin.At 1 h after salt treatment，the expression levels of GORK in roots of miR528‑OE1 and miR528‑OE2 seedlings significantly decreased，but the expression level of GORK in root of mir528 seedling significantly increased compared with Dongjin.In conclusion，overexpression of miR528 can improve salt tolerance of rice and enhance K⁺ retention ability under salt stress，which may be achieved by down‑regulating GORK expression under salt stress.

Effects of Acid Environment on Rice Seedling Growth under Cadmium Stress

REN Xinyu, LUO Sheng, WEI Hongyu, CHENG Xin,

2022, 51(4): 39-47. DOI: 10.15933/j.cnki.1004-3268.2022.04.005

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In order to clarify the interaction between acid environment and cadmium（Cd）stress and provide theoretical basis for the safe production of crops，the effects of acid environment（pH values were 5.0，6.0，6.8，respectively）on the growth of rice seedling under Cd（0，0.3，0.6 mg/L）stress were studied by hydroponic experiment. The results showed that Cd stress had inhibitory effect on the growth of rice seedling shoot，but weak acid environment favored the growth of rice seedling shoot under Cd stress，when the pH value was 6.0，the growth of rice seedling shoot，and the contents of chlorophyll and carotenoid in leaves were the highest. However，when the pH value decreased to 5.0，the growth of rice seedling shoot was strongly inhibited，and the contents of chlorophyll and carotenoid in leaves were the lowest，indicating that the acid environment intensified the inhibition of Cd stress on growth of rice seedling shoot. When the Cd concentration was below 0.3 mg/L，the root growth of rice seedling（total length，surface area，diameter，total volume and root tip number）was the best at pH value of 6.0，the worst at pH value of 5.0，and malondialdehyde（MDA）content and peroxidase（POD）activity generally increased with the decrease of pH value.Also，the results of evans blue staining test and reactive oxygen species（ROS）detection test showed that the damage of rice seedling root was strengthened with the decrease of pH value under 0.6 mg/L Cd stress. In conclusion，the acidic environment can aggravate the toxic effect of Cd on rice seedling when the pH value is less than 6.0.

Genome‑wide Identification and Expression Analysis of CCCH Type Zinc‑finger Gene Family in Nicotiana tabacum

TANG Chungui, DENG Zhaolong, LIU Qiong, JIN Weihuan, XIANG Shipeng, XIE Pengfei, GUO Hongxiang, LIU Weiqun

2022, 51(4): 48-58. DOI: 10.15933/j.cnki.1004-3268.2022.04.006

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In order to understand the characteristics of tobacco CCCH type zinc‑finger gene family，bioinformatics methods were performed in the tobacco genome database using Arabidopsis CCCH typeprotein motif and 86 CCCH type zinc‑finger family members were identified. Protein physicochemical properties，systemic evolution，chromosome distribution，motif，promoter cis‑acting elements，gene expression pattern and the response to drought and phytohormone were analyzed. These results showed that 37 of 86 tobacco CCCH type zinc‑finger genes were distributed on 17 chromosomes，all members of tobacco CCCH type zinc‑finger proteins contained C‑X_7/8‑C‑X₅‑C₃‑H motif and some novel CCCH motifs were also found，such as C‑X₆‑C‑X₄‑C‑X₃‑H.Systemic evolution analysis indicated that 51 of 86 CCCH type zinc‑finger genes were divided into nine groups.Multiple cis‑regulatory elements including stress response elements and phytohormone response elements were found in the promoter region of NtC3H-39 gene. Results of gene expression patterns and stress response analysis revealed that NtC3H-39 gene showed obvious tissue specific expression and responses to drought stress and phytohormone stress.

Effects of Shading on Growth and Physiological Characteristics of Rehmannia glutinosa

LIU Chunyan, HUANG Yong, GUO Meng, ZHANG Hongrui, ZHOU Yan, LI Hemin

2022, 51(4): 59-67. DOI: 10.15933/j.cnki.1004-3268.2022.04.007

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In order to explore the effect of shading on the growth and physiological characteristics of Rehmannia glutinosa，with the main variety Jinjiu in the authentic area of Rehmannia glutinosa as the experimental material and normal light as the control（CK），three shading treatments of 20% shading，50% shading and 80% shading were set up to study the differences of agronomic characters，physiological indexes and effective component content among the treatments. The results showed that compared with the control，20% shading had no significant effect on root length，root diameter，number of roots and number of tubers of Rehmannia glutinosa.50% shading and 80% shading inhibited the growth of root length，root diameter，number of roots and number of tubers of Rehmannia glutinosa，and the inhibitory effect of 80% shading was greater than that of 50%. The fresh root weight per plant and dry root weight per plant of Rehmannia glutinosa under three shading treatments were significantly lower than those ofthe control.The higher the shading degree，the greater the decline. On October 20，the fresh root weight per plant of Rehmannia glutinosa under 80% shading，50% shading and 20% shading decreased by 95.10%，45.81% and 24.84% respectively.The contents of soluble protein and free proline in the root tubers of shaded Rehmannia glutinosa were significantly higher than those of the control（except for July 11）；50% shading and 80% shading decreased the content of soluble sugar in root tubers.Shading inhibited the root activity of Rehmannia glutinosa，and the higher the shading degree，the stronger the inhibition effect；shading treatment decreased the activity of superoxide dismutase in the early and late growth stage，and increased its activity in the middle growth stage. Under 80% shading，50% shading and 20% shading，the content of acteoside in Rehmannia glutinosa root tubers roots increased by 50.44%，37.47% and 26.11% respectively，but the contents of catalpol and rehmannioside D decreased，and the content of rehmannioside D decreased by 38.70%，26.36% and 7.69% respectively.In conclusion，the physiological metabolism of Rehmannia glutinosa root is disordered，the growth is inhibited，the material accumulation is reduced and the quality becomes worse under shading conditions.In production，it is suggested to cultivate Rehmannia glutinosa in the environment with sufficient light，or reduce the degree of shading as far as possible.

Response of Soil Microecology to Quinclorac Contamination and Remediation by Enterobacter ludwigii EM1

ZHANG Zhichao, GAO Ming, ZHAO Hao, WANG Xiaoya, CAO Zijing, WU Kun, XU Shuxia

2022, 51(4): 68-76. DOI: 10.15933/j.cnki.1004-3268.2022.04.008

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In order to clarify the remediation effect of the degrading bacterial strain Enterobacter ludwigii EM1 on quinclorac contamination in soil，the effects of EM1 on culturable microorganisms，soil enzyme activity and bacterial community diversity were studied by adding EM1 to artificially simulated quinclorac‑contaminated soil quantitatively and setting up three treatments：non‑contamination control（CK）， quinclorac‑contaminated treatment （QNC） and quinclorac‑contaminated treatment+EM1remediation treatment（QNC+EM1）. The results showed that compared with CK and QNC，the number ofsoil bacteria in QNC+EM1 treatment increased by 36. 5% and 15.6% respectively at the end of culture.Compared with CK and QNC，the number of actinomycetes increased by 6. 7% and 29.4%，respectively.The number of fungi increased by 7.6% compared with CK，and decreased by 14. 1% compared with QNC.The urease activity in QNC+EM1 had no significant change at the end of culture compared with CK，but decreased by 6.8% compared with QNC；at the end of culture，the activities of phosphatase and sucrase increased by 135. 8% and 224. 4% respectively compared with CK，and 149.7% and 199.9% respectively compared with QNC. At the end of culture，the concentration of quinclorac in soil treated with QNC+EM1 decreased by 76. 7% compared with that treated with QNC. The analysis on soil bacterial community diversity showed that the abundance of Proteobacteria increased by 15.1 percentage points，the abundance of Gemmatimonadetes and Acidobacteria decreased by 10.7 and 5.5 percentage points respectively，and the dominant bacteria，such as Verrucomicrobia，Nitrospirae and Planctomycetes，disappeared after adding EM1.The results of principal component analysis showed that the bacterial community structure changed significantly after adding EM1，and the correlation number among bacteria decreased.In general，addition of the degrading bacterial strain EM1 can effectively remediate quinclorac‑contaminated soil，improve soil flora and increase soil enzyme activity，so as to provide certain reference significance for the remediation of quinclorac‑contaminated soil.

Degradation Ability and Influencing Factors of Cellulose‑degradation Microbial Community PN‑8 on Wheat Straw

WU Fengxia, SUN Yue, XIAO Qiang, ZHANG Shubin, LI Yufei, LIU Jianbin

2022, 51(4): 77-86. DOI: 10.15933/j.cnki.1004-3268.2022.04.009

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In order to clarify the characteristics and key influencing factors of thermophilic lignocellulose‑degradation microbial community PN‑8，we analyzed the effects of different chemical pretreatments（0.5%，1.5%，10.0% sodium hydroxide，3.0%，12.0%，18.0% hydrochloric acid，2.0%，20.0% acetic acid），initial inoculation amount，glucose content and pH on the degradation of wheat straw by PN‑8.The main products of PN‑8 in the process of degrading wheat straw were analyzed by GC‑MSQP2010，and the degradation ability of PN‑8 on wheat straw was detected by scanning electron microscope.The results showed that chemical pretreatment of wheat straw affected the degradation ability of PN‑8.Alkali treatment（sodium hydroxide）could reduce lignin content and improve the degradation rate of PN‑8，while acid treatment（hydrochloric acid，acetic acid）could increase lignin content and reduce the degradation rate. Among them，after 1.5%，10.0% sodium hydroxide treatment，the degradation rate of wheat straw reached 71.61%，87.33% respectively，which were significantly higher than that without chemical pretreatment（44.33%）.When the initial inoculation amount of PN‑8 was less than 5.0%，the degradation rate was positively correlated with the inoculation amount，and there was no significantly difference when the inoculation amount was higher than 5.0%.When the initial glucose concentration was higher than 0.1%，or the initial pH value exceeded the range of 5—9，PN‑8 lost the degradation ability of wheat straw.Glycerol was the dominant fermentation product，which could reach the maximum content of 0.28 g/L on the first day of fermentation，followed by acetic acid and ethanol.Both carboxymethyl cellulase activity and filter paper enzyme activity showed a change regular of first increasing and then decreasing during the fermentation process.The results of scanning electron microscope showed that PN‑8 could destroy the structure of wheat straw and degrade lignocellulose.

Screening of Fungicides for Controlling Fusarium Root Rot of Tobacco

YAO Chenxiao, LIU Chang, LI Xiaojie, BAI Jingke, XU Min, QIU Rui, CHEN Yuguo, KANG Yebin, LI Shujun

2022, 51(4): 87-94. DOI: 10.15933/j.cnki.1004-3268.2022.04.010

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In order to screen out the high‑efficacy and safe mixtures or seed coating agents for the control of Fusarium root rot of tobacco，the indoor activity of 10 chemical fungicides and 10 biopesticides（a total of 20 species）against F.oxysporum，the dominant causal agent of tobacco root rot，was determined by mycelial growth rate method，and their control effect was verified by potting experiment.The results showed that the 20 fungicides all presented different levels of inhibitory effects on F.oxysporum，and the overall inhibitory effect of chemical fungicides was higher than that of biopesticides significantly.Among the chemical fungicides，the EC50 values of 500 g/L fluazinam SC，25% tebuconazole WP，40% difenoconazole SC，and 20% imazalil EW were 0.057 1 μg/mL，0.118 7 μg/mL，0.328 0 μg/mL，and 0.445 9 μg/mL，respectively.Among the biopesticides，the EC50 values of 1.8% abamectin EC，5% carvacrol AS，and 0.01% brassinolide EC were 10.387 0 μg/mL，18.873 2 μg/mL，and 32.343 7 μg/mL，respectively. The results of the pot experiment showed that the control efficacies of fluazinam 8 000 timesdilution，tebuconazole 2 000 times dilution，difenoconazole 1 200 times dilution and brassinolide stock solution were higher，which were 51.51%，75.75%，62.12% and 62.12%，respectively.Therefore，fluazinam，tebuconazole，difenoconazole and brassinolide could be selected as candidates for the further research and development of mixtures and seed coating agents and the rotation use of conventional chemical fungicides and integrated management for tobacco Fusarium root rot in fields.

Screening of Botanical Pesticides for Controlling Ditylenchus destructor

PENG Jingwen, ZHAO Yuanzheng, WANG Zhen, WANG Yufeng, ZHANG Niu, WANG Dong, ZHOU Hongyou

2022, 51(4): 95-102. DOI: 10.15933/j.cnki.1004-3268.2022.04.011

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To explore the control effect of botanical pesticides on Ditylenchus destructor，the indoor toxicity of six botanical pesticides was tested by the direct contact method，and their effects on morphology，behavior change and body fluid penetration of D.destructor were observed.The pesticide with the highest toxicity to D.destructor was selected for pot and field study.Indoor virulence assay showed that carvacrol 5% AS，with lethal concentration of 50%（LC50）of 39. 542 mg/L，had the strongest inhibitory activity to D.destructor in 24 h.After the treatment of nematodes with carvacrol 5% AS and eucalyptol 5% EC，the nematode body stiffened and the dead nematodes showed bubble structures or cavities.In addition，six botanical pesticides increased the fluid leakage of D.destructor under the condition of LC50，and the suspension conductivity of the carvacrol 5% AS treatment was the highest in 24 h，reaching 70.40 μS/cm.The control effect tests showed that carvacrol 5% AS had control effects of 65.73% indoors and 53.55% in fields，respectively. In conclusion，carvacrol 5% AS with higher inhibitory activity and control effect，can be used as a recommended agent for the control of D.destructor.

Effects of Deep Tillage and Border Dividing Cultivation on Soil Air Permeability and Ginger Seedling Growth

JIA Qie, ZHANG Tianxiao, LI Gang, ZHU Yongxing, CAI Xiaodong, LIU Yiqing

2022, 51(4): 103-110. DOI: 10.15933/j.cnki.1004-3268.2022.04.012

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For the reason that soil is easy to accumulate water resulting in ginger rotten seeds and retting roots in rainy weather during spring and summer in the middle and upper reaches of the Yangtze River，this research proposed a healthy cultivation model of deep tillage and border division and evaluated its effect on soil physical properties，ginger germination，growth and photosynthetic efficiency compared with general cultivation（CK）.The results showed that the air permeability of 0—30 cm soil by deep tillage and border division increased by 316.3% to 585.0% compaired with that by CK，the rotten seed rate of ginger was 35.7 percentage points lower than that of CK，and the seedling emergence rate was 94.4%.The total length，surface area and tip number of ginger seedling roots were significantly higher than those of CK，which increased by 88.5%，96.1% and 62.7% compaired with those by CK respectively.Theheight，stem diameter，leaf number，fresh weight and dry weight of ginger seedlings were significantly higher than those of CK，and increased by 90.9%，80.0%，70.4%，54.5% and 52.4% compaired with those by CK respectively.The net photosynthetic rate，transpiration rate，stomatal conductance and intercellular CO₂ concentration of ginger seedling leaves were significantly higher than those by CK，and increased by 158. 8%，210.9%，558.2% and 73.1% compaired with those by CK respectively.Therefore，cultivation by deep tillage and border division improved soil air permeability and water conductivity，avoided soil ponding and seed rot caused by rainy weather，and promoted the robust growth of ginger seedling and its roots.

Multivariate Analysis of the Relationship between Soil Nutrients and Fruit Quality and Optimization of Soil Nutrients in Soft‑seed Pomegranate Orchards

ZHANG Huan, GAO Xiaofeng, LEI Mengyao, TAO Aili, LI Yuying, YAO Gang

2022, 51(4): 111-119. DOI: 10.15933/j.cnki.1004-3268.2022.04.013

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To optimize soil nutrients for the best fruit quality，the multiple relationships between soil nutrients and fruit quality were studied in pomegranate orchards in Xichuan County，Henan Province，China in 2020.Partial least squares regression（PLSR）and linear programming methods were used to establish regression equations and optimize soil nutrients for the best fruit quality. The results showed that the weight of single fruit was mainly affected by the contents of organic matter，nitrate nitrogen and alkali‑hydrolyzable nitrogen，and 100‑grain weight was mainly affected by soil pH value，total nitrogen and available potassium contents.Seed hardness was mainly affected by soil pH value，nitrate nitrogen，ammonium nitrogen and alkali‑hydrolyzable nitrogen contents，juice yield was mainly affected by soil pH value，organic matter and available phosphorus contents，and the content of vitamin C was affected by all indexes except for soil pH value and available potassium content. The content of soluble solids was mainly affected by soil pH value，ammonium nitrogen and available potassium contents，and the content of titratable acid was mainly affected by soil pH value，ammonium nitrogen，alkali‑hydrolyzable nitrogen and available potassium contents. The content of soluble sugar was mainly affected by soil pH value，organic matter，total nitrogen，ammonium nitrogen，alkali‑hydrolyzable nitrogen and available potassium contents. When the parameters ranged as soil pH of 6.70—7.50，soil organic matter content of 27.49—65.20 g/kg，total nitrogen content of 8.73—12.91 g/kg，nitrate nitrogen content of 7.51—28.93 mg/kg，ammonia nitrogen content of 42.43—50.03 mg/kg，alkali-hydrolyzable nitrogen content of 65.50 mg/kg，available phosphorus content of 19.33—73.53 mg/kg，available potassium content of 160.31 mg/kg，thefruit quality parameters of soft‑seed pomegranate were the best：fresh weight per fruit of 776.46 g，100‑seed weight of 59.56 g，seed hardness of 0.44 kg/cm²，juice yield of 82.58%，vitamin C content of15.90×10^‑2mg/g，soluble solids content of 17.90%，titratable acid content of 0.97 mg/mL，and solublesugar content of 23.34%.

Effects of Different LED Reflecting and Lighting Methods on the Growth of Tissue Culture Seedlings of Gerbera jamesonii

WANG Zheng, ZHANG Chunling, HE Songlin, SHANG Wenqian, HE Dan, SHEN Yuxiao, LIU Yannan, SONG Yinglong, SUN Yuke

2022, 51(4): 120-129. DOI: 10.15933/j.cnki.1004-3268.2022.04.14

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Gerbera jamesonii was used as the experimental material to study the effects of different LED reflecting and lighting methods on the morphological and physiological characteristics of Gerbera jamesonii tissue culture seedlings，in order to provide technical support and theoretical support for industrialized seedling cultivation of Gerbera jamesonii.The results showed that among the eight reflective and lighting methods treatments （Different light source inclination angles+different shape reflective films+nano light guide plates：76.39° +M‑shape+nano light guide plate，79.89° +M‑shape+nano light guide plate，82.59°+convex+nano light guide plate，86.87°+arch+nano light guide plate，69.77°+plane+nano light guide plate，90°+plane+nano light guide plate，180°+nano light guide plate，180°），the plant height，leaf number，leaf width，whole plant fresh weight，aboveground fresh weight，root fresh weight，whole plant dry weight，aboveground dry weight，underground dry weight and soluble protein content of treatment 1—4 showed a trend of increasing first and then decreasing with the increase of the inclination angle of the light source，while leaf length，longest root length，aboveground dry matter rate，underground dry matter rate，total dry matter rate，stomatal size under leaf epidermis，stomatal area and root activity showed a trend of decreasing first and then increasing or decreasing. Among them，the photosynthetic related indexes such as stomatal density，stomatal size，stomatal area and chlorophyll content of treatment 4（86.87°+arch+nano light guide plate） were better than those of other treatments. The longest root length，root activity，underground dry matter rate and total dry matter rate of treatment 1（76.39°+M‑shape+nano light guide plate）were better than those of other reflective treatments. In summary，treatment 4 was conducive to the photosynthesis of tissue culture seedlings of Gerbera jamesonii and could promoted the growth of stems and leaves.Treatment 1 was beneficial to root growth and material accumulation.

Haplotype Diversity and Phylogenetic Relationships of Theileria luwenshuni from Western Hunan

LI Zhongbo, HOU Qianghong, LI Hui, SHU Ming

2022, 51(4): 130-137. DOI: 10.15933/j.cnki.1004-3268.2022.04.015

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The aim of the study was to explore the haplotype diversity and phylogenetic relationships of Theileria luwenshuni（T.luwenshun）from goats in western Hunan.A total of 16 strains of T.luwenshun from goats in western Hunan were used as study samples，and the polymerase chain reaction was applied to amplify the ribosomal 18S rRNA and cytochrome c oxidase subunit 1（cox1）genes of T.luwenshun to study their haplotype diversity and phylogenetic relationships. Results of our study showed that all the sequences of 18S rRNA and cox1 genes of T.luwenshun were 384 bp and 1 092 bp，respectively.The 18S rRNA and cox1 gene sequences contained 9 and 2 variable sites，respectively，and included 7（A1—A7，Hd=0.775）and 4（B1—B4，Hd=0.625）haplotypes，respectively；there were 11 variable sites and 10（C1—C10，Hd=0.925） haplotypes in the combined sequences of 18S rRNA + cox1.Out of these haplotypes，the A1，A4，B1，B3，B4，C1，C8 and C9 were ancient haplotypes.In addition，all the 16 strains of T.luwenshun from goats were located at a clade of phylogenetic tree constructed by using 18S rRNA+cox1 gene sequences.The results of our study show that although there is a certain degree of gene variation among 16 strains of T.luwenshuni from western Hunan，they share several ancient haplotypes and have a close genetic relationships，and there is no obvious genetic differentiation.

Cloning，Sequence Analysis and Prokaryotic Expression of Three Spliceosomes of Abd‑A from Silk Gland of Bombyx mori

LU Qingjun, LUO Zhuxing, LIU Huawei, WEI Meng, WANG Yuan, ZHANG Zhaofeng, LI Youshan

2022, 51(4): 138-150. DOI: 10.15933/j.cnki.1004-3268.2022.04.016

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In order to explore the splicing form and sequence characteristics of transcription factor Abdominal‑A （Abd‑A） from silk gland of Bombyx mori，clarify the relationship with the Abd‑A sequences of other insects，the cDNA of silk gland tissue was used as the template to amplify Abd‑A by PCR.The sequence characteristics of Abd‑A and the evolutionary relationship of Abd‑A among different insects were analyzed by bioinformatics methods.The Abd‑A was recombinantly expressed in vitro by prokaryotic expression system，and its protein expression level was detected by SDS‑PAGE and Western blot.The results showed that three spliceosomes of Abd‑A were cloned from the silk gland of B.mori，with
coding regions of 1 059，1 044，1 032 bp，named BmAbd-A L（A59T），BmAbd-A S，BmAbd-A 3，respectively.Multiple sequence alignment of Abd‑A in different insects indicated that their amino acid sequences differed greatly，but all had the DNA‑binding domain（homeobox domain） and the transcriptional activation domain（Abd‑A domain）.The homeobox domain was completely conserved，while the carboxyl terminal sequence of Abd‑A domain was quite different. The three‑dimensional structure prediction indicated that the proteins encoded by three spliceosomes of Abd‑A had four α‑helix and random coil structures，among which the random coil structures varied greatly.Phylogenetic analysis showed that the amino acid sequences encoded by Abd‑A of 35 different insects were mainly divided into four orders，which were Diptera，Lepidoptera，Coleoptera and Hymenoptera，respectively.The Abd‑Asequences of B.mori were closer to Galleria mellonella（Lepidoptera：Pyralidae） in evolutionary relationship，followed by Ostrinia furnacalis.The results of SDS‑PAGE and Western blot showed that the fusion proteins of BmAbd‑A L（A59T），BmAbd‑A S，BmAbd‑A 3 were mainly expressed in soluble form in Escherichia coli，and their theoretical molecular weights were 52.99，52.35 and 51.91 ku，respectively，which were consistent with the expectation.In conclusion，three spliceosomes of Abd‑A were successfully cloned from the silk gland of B.mori.Their sequences and three‑dimensional structures were different，and their molecular evolution process was consistent with the phylogenetic evolution of species.Soluble Abd‑A recombinant proteins were obtained by prokaryotic expression system.

Physicochemical Properties Change and Nutrient Deposition Process in Purple Soil Pond Mud

LI Tiancai, YU Mi, JIANG Rui, ZHAO Qingyun, FU Mei, SU Shengqi

2022, 51(4): 151-159. DOI: 10.15933/j.cnki.1004-3268.2022.04.017

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This paper aimed to monitor the physicochemical properties changes and nutrient deposition process in calcareous purple soil pond mud among the fishing process. Purple soils collected in Tongliang District，Chongqing（N29. 5130°，E106.4010°），were poured on the bottom of culture tank as foundation of the simulative pond.In the half year of fishing process，the mud temperature，pH，redox potential（O.R.P），organic matter（SOM）content，total nitrogen（TN） content，alkali‑N content，total phosphorus（TP）content，olsen‑P content，total potassium（TK）content，avail‑K content，Mg content，S content and the carbon nitrogen ratio（C∶N）were tested termly.The results showed that calcareous purple soil had a pH of 8.5 and was highly alkaline.In general，it was not fertile. After being flooded，the pH and O.R.P of pond mud reduced quickly，the pH approached to 7.3 and the O.R.P fluctuated around 180.0 mV.But feeding made pH and O.R.P reduce to 7.1 and -210.0 mV.In the process of feeding，the contents of SOM，TN，TP，alkali‑K and olsen‑P increased with time and were significantly higher than the flooded group. By the end of the experiment，deposit amounts of SOM，TN，TP，alkali‑K and olsen‑P were 5.10 g/kg，0.13 g/kg，0.45 g/kg，78.88 mg/kg and 88.50 mg/kg.The other nutrients contents of the feeding group pond mud increased in varying amounts，except that only the content of Mg reduced. Despite the continuous accumulation of SOM and anaerobic reaction in the feeding group pond mud，the pH did not change much in the middle and late period and was still suitable for fishing，showing that the calcareous purple soil had strong buffering ability as the foundation of ponds. According to SOM deposition ratio［5.1 g（/kg·a）］and C∶N increase rate，calcareous purple soil pond’s fish productivity would change to be low in three years. Thus，the pond mud should be managed carefully，and the sectional silt should be removed no more than every three years at the off‑fishing season to keep its fish productivity.

Identification of Farmed Aquatic Animals Types in Freshwater Aquaculture Ponds Based on Sentinel‑2 Imagery

2022, 51(4): 160-170. DOI: 10.15933/j.cnki.1004-3268.2022.04.018

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In order to realize the rapid extraction of the spatial distribution of inland freshwater aquaculture based on remote sensing images，this study chose Yixing City，Jiangsu Province as the study area and proposed a method to identify inland freshwater aquaculture ponds by combining the convolutional neural network and random forest algorithm based on the Sentinel‑2 satellite images.Firstly，the study built a convolutional neural network model for semantic segmentation of aquaculture ponds based on deep learning.Then，normalized difference vegetation index（NDVI） and normalized difference water index（NDWI）of the patches in the aquaculture areas were analyzed.Finally，the random forest algorithm was used to distinguish the types of aquaculture ponds.The results showed that there were 121.25 km² of freshwater aquaculture ponds in Yixing in 2021，and the areas of crab ponds and fish ponds were 74.48 km²and 46.77 km²，respectively.The overall accuracy of the method was 88.33%，and the kappa coefficient was 0.824 3.

The Relationship between Color Change and Energy Metabolism of Tobacco Leaves after Harvest in Different Leaf Positions

NIU Hao, LI Jinghao, LI Pengcheng, SUN Guangwei, BAI Tao, CHEN Zhenguo, SONG Zhaopeng, WANG Tao

2022, 51(4): 171-180. DOI: 10.15933/j.cnki.1004-3268.2022.04.019

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This research aimed to study the relationship between color change and energy metabolism of tobacco leaves after harvest in different leaf positions. By choosing tobacco leaves with different leaf positions as the research object，the yellowing and browning process of postharvest tobacco leaves was analyzed by dark test.The yellowing degree，browning degree，malondialdehyde content，sugar（total sugar，reducing sugar，starch）content，energy metabolism‑related substances（ATP，ADP，AMP）content，energy charge and cytochrome oxidase（CCO），succinate dehydrogenase（SDH） activities were determined during experiment.The research results showed that：（1）As the leaf position was elevated，the yellowing and browning of tobacco leaves were slower with the lower yellowing index and browning index，and the MDA content and the conductivity of the extract increased more slower.（2） In comparison，the starch content of the middle and upper leaves was higher and the degradation peak was later，and the total sugar and reducing sugar content maintained at high level in the middle and late stages.（3）Compared with the lower leaves，the ATP content and energy charge level of the middle and upper leaves were higher and the peaks appeared later，and CCO and SDH activities were relatively higher.（4）The degree of yellowing of tobacco leaves showed a significant positive correlation with total sugar，reducing sugar content and energy charge（P<0.05），respectively；the degree of browning showed a significant negative correlation with reducing sugar and ADP content（P<0.05），respectively.In summary，the yellowing and browning of the middle and upper leaves develop more slowly than the lower leaves.The difference in color changes of tobacco leaves in different leaf positions may be related to their energy metabolism activities.

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