Journal of Henan Agricultural Sciences

Contents

2019, 48(10): 0.

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Ｒesearch Status and Farming Development Prospects of Barbus capito

ZHU Ｒui, WU Lifang, LI Liang, YU Zhe, ZOU Ｒuixing, ZU Xiujie

2019, 48(10): 1-5. DOI: 10.15933/j.cnki.1004-3268.2019.10.001

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Barbus capito has characteristics of a wide range of adaptability,fast growth,delicious taste,high nutritional value,strong resistance to stress and saline-alkali resistance.It is a popular species of Cyprinidae.On the basis of reviewing related literature reports at home and abroad,this paper introduces the biological characteristics,research status,development prospects of Barbus capito and so on,aiming to provide reference for the healthy breeding of Barbus capito,and the adjustment of the structure of aquacul ture species in China．

Transformation of Phosphorus Efficient Gene OsPHＲ2 into Wheat and Tolerance of Transgenic Plants to Low Fertility

HUA Xia, FANG Yuhui, GAO Chong, HAN Liupeng, HU Lin, LI Yan

2019, 48(10): 6-12. DOI: 10.15933/j.cnki.1004-3268.2019.10.002

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Phosphorus is one of the major necessary elements for plant growth and development． In order to obtain transgenic wheat plants with high phosphorus utilization efficiency,OsPHＲ2(Oryza sativa L.phosphate starvation response 2) gene was transferred into Zhengmai 7698 and other five wheat varieties (lines) by particle bombardment transformation method． Genetic analysis was carried out in the positive transgenic wheat progenies.Furthermore，the yield and other traits of T1 transgenic plants under high and low fertility treatments were analyzed.PCＲ results showed that there were 180 positive transgenic wheat plants in T0 generation.Genetic analysis revealed that the OsPHＲ2 gene segregation ratio of 15 lines among 16 lines was in accordance with the law of Mendelian single gene genetic segregation.The results of high and low fertility treatments showed that OsPHＲ2 could significantly increase the wheat yield of sin gle plant under low fertility treatment，but the high fertility treatment was not.The yields of single plant of OsPHＲ2-overexpressing transgenic Zhengmai 1342 and Zhengmai 7698 significantly increased by 11.06% and 9.27% compared with wild type plants under low fertility treatment，which was mainly due to the increase of 1 000-grain weight and grain number per spike．

Association Analysis between SSＲ Markers and Salt-tolerant Traits at Seedling Stage in Japonica Ｒice Germplasm Ｒesource

XIE Feifei, TIAN Pujiang, MA Shuaiguo, BAI Tianliang, CHEN Jingru, ZHANG Qian, TIAN Ｒongrong, YANG Zhiwei, TIAN Lei, LI Peifu

2019, 48(10): 13-22. DOI: 10.15933/j.cnki.1004-3268.2019.10.003

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In order to provide excellent intermediate materials and technical support for the breeding of salt-tolerant rice varieties,thirty-eight pairs of simple sequence repeat(SSR) markers which were uni formly distributed throughout rice genome were used to detect 160 japonica rice germplasm resources,and the molecular marker sites associated with salt-tolerant traits were explored.The results showed that there was certain correlation among salt-tolerant traits.Salt tolerant score was significantly positively correlated with relative plant height,relative shoot fresh weight,relative root fresh weight and relative stem diameter.Salt tolerant score was significantly negatively correlated with leaf damage percentage and relative ratio of root to shoot.A total of 192 alleles were identified at all of 38 SSＲ loci,the effective alleles number was 109.60,and the effective alleles number per marker locus was 2.88.The average values of Shannon’s in formation index and Nei’s genetic diversity index of these SSＲ markers were 1.139 4 and 0.586 9,re spectively.Eleven sites were significantly associated with salt tolerance related traits,which were located on chromosomes 1,3,4,7,8,9,10,11 and 12,and explained the phenotypic variance from 4.10% to 18.85%.ＲM286-7 that was related with relative shoot fresh weight had the highest phenotypic contribu tion rate，and ＲM7585-1 had the highest positive phenotypic effect for salt tolerant score，whose phenotyp ic contribution rate was 6.80%,and the carrier variety was Bertone．

Effects of Organic-Inorganic Superabsorbent Polymer Composites on Soil Moisture and Growth of Maize at Seedling Stage

YU Gao, CHEN Fen, XIE Yinghe, ZHAO Xiaodong, ZHENG Haixia, TAN Jiebin

2019, 48(10): 23-29. DOI: 10.15933/j.cnki.1004-3268.2019.10.004

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To provide some scientific basis for the reasonable application of organic-inorganic superabsorbent polymer composites in corn production in the semi-arid,arid areas of northern China,the effects of organic-inorganic superabsorbent polymer composites［polyacrylamide(PAM) ,attapulgite(NA),humic acid( FA) ，straw biochar(CB) ］on soil moisture and growth of maize seedling were studied with five treatments including T1 (PAM∶ NA∶ FA∶ CB =4∶ 3∶ 2∶ 1),T2 (PAM∶ NA∶ FA∶ CB =4∶ 3∶ 1∶ 2),T3 (PAM∶ NA∶ FA∶ CB =5∶ 2∶ 2∶ 1) ，T4 (PAM∶ NA∶ FA∶ CB =5∶ 2∶ 1∶ 2) and the control (CK) without organic-inorganic superabsorbent polymer composites.The results showed that all the composite treatments significantly decreased the cumulative evaporation rate of soil water compared with CK,the order was T4＜T3＜T2＜T1＜CK,and the cumulative evaporation rate of soil water decreased with the increase of application rate of organic-inorganic superabsorbent polymer composites.Soil moisture content,plant height,leaf area,stem diameter and dry matter accumulation of maize seedling were significantly higher than CK,and the order was T4＞T3＞T1＞T2＞CK.When the application rate of organic-inorganic superabsorbent polymer composites was 0.013—0.053 g/kg,the cumulative evaporation rate of soil water decreased,the soil moisture content increased with the increase of application rate of organic-inorganic superabsorbent polymer composites.When the application rate of organic-inorganic superabsorbent polymer composites was 0.053 g/kg,the growth of maize seedling was obviously promoted，and the plant height,leaf area,stem diameter and dry matter accumulation of maize seedling were the highest.These results suggested that the T4 treatment was a best way to reduce the cumulative evaporation of soil water,increase the soil moisture content,and promote the growth of corn seedling with the superabsorbert polymer composites of 0.053 g/kg．

Genome-Wide Identification and Expression Analysis of TCP Gene Family in Sorghum bicolor L．

ZHENG Ling, BAI Xueting, LI Huiyun

2019, 48(10): 30-36. DOI: 10.15933/j.cnki.1004-3268.2019.10.005

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In order to study the function of TCP(Teosinte-branched 1/cycloidea/proliferating cell fac tors) gene family of Sorghum bicolor L.in growth and response to adversity stress,the SbTCP genes were identified by bioinformatic analysis,and their conserved domain,phylogenetic relationship,chromosome lo cation,conserved domain and expression pattern were analyzed.The results showed that a total of 27 SbTCP genes were identified,they were unevenly distributed across 9 chromosomes,and there were gene clusters on 3,6,9 chromosomes.All the SbTCP proteins could be divided into two groups,Class Ⅰ and Class Ⅱ,and Class Ⅱ could be divided into two subgroups,CIN and CYC/TB1.Class Ⅰ contained 10 SbTCP proteins,CYC/TB1 contained 3 SbTCP proteins,and CIN contained 14 SbTCP proteins.Twenty seven SbTCP proteins all had atypical bHLH(Basic-helix-loop-helix) conserved region,and two SbTCP proteins had Ｒ domain.Transcriptome analysis showed that most of SbTCP genes expressed in seed,em bryo,pistil and early inflorescence; the expression level of most of SbTCP genes increased in roots under PEG treatment,and the expression level of several SbTCP genes increased in shoots under ABA treat ment,which suggested that SbTCP genes might play important role in response to drought stress．

Effect of Intercropping Modes between Millet and Mung Bean on Their Growth and Yields

XUE Ｒenfeng, ZHAO Yang, WANG Yingjie, CHEN Jian, GE Weide

2019, 48(10): 37-40. DOI: 10.15933/j.cnki.1004-3268.2019.10.006

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In order to screen out the best intercropping mode between millet and mung bean which was suitable for Liaoning Province，this study compared the effects of different intercropping modes between millet and mung bean on their yields,agronomic traits,physiological indexes and economic benefits,which could lay important foundation for ensuring income of farmers in Northeast of China.The results showed that millets of 4 ∶ 2 intercropping mode between millet and mung bean had height of 135.8 cm,stem diameter of 0.65 cm,stem weight of 10.1 g,spike length of 21.5 cm and single ear weight of 18.3 g,which were all significantly higher than those of 2 ∶ 1 intercropping and monoculture modes.The intercropping modes could activate the antioxidant enzymes in millets.The root length and plant height of intercropped mung bean increased significantly，and the 2 ∶ 1 intercropping mode between millet and mung bean was greater than 4∶ 2 intercropping mode.The main stem branch number,pod number per plant，pod length,grain number of pod and 100-grain weight of intercropped mung bean were all lower than those of mung beans of monoculture mode.The 4∶ 2 intercropping mode between millet and mung bean had the highest profit of 22 185.93 Yuan/ha,which increased by 27.1% compared with millet monoculture mode，and was 4.95 times of mung bean monoculture mode．

Effects of Layered Bottom Application of Slow-release Fertilizer on Growth，Dry Matter Accumulation and Yield of Summer Maize

GONG Yu, DUAN Weiwei, WANG Guiyan, CHEN Zongpei, CEHN Zhaoyue, XUE Jiaxin, LI Ben

2019, 48(10): 41-46. DOI: 10.15933/j.cnki.1004-3268.2019.10.007

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This study is to find out the effect of the layered bottom application mode of slow-release fertilizer,and provide theoretical basis for the popularization and application of corn simplified fertilization technology.The corn variety Xinrui 25 was used as the experimental material，and the random block design was adopted.The slow-release fertilizer layered(0—15,15—30 cm soil layer) bottom application mode(M1),common fertilizer bottom application mode(M2) and common fertilizer bottom application and topdressing mode (M3) three treatments were set.The results showed that summer maize of M2 mode had high population growth rate and net assimilation rate at the early growth stage，while summer maize of M1 and M3 modes had higher population growth rate and net assimilation rate during the middle and later growth period.The population growth rate and net assimilation rate of M1 mode were significantly higher than those of other fertilizing modes from grain-filling stage to maturity.The dry matter accumulation of M1 mode were significantly higher than that of M2 and M3 modes after tasseling.The dry matter transfer rate and contribution rate of vegetative organs to grain of M2 mode were the highest,which of M1 mode were similar to the M3 mode.There was no significant difference in the grain number per spike and yield between M1 and M3 modes,but they were significantly higher than that of M2 mode.The yield of summer maize in M1 mode was 13.9% and 5.2% higher than that of M2 and M3 modes respectively.The results indicated that the layered fertilization mode could promote the growth and development and improve the accumulation of dry matter at the later stage of summer maize,moreover,it could increase the yield by improving the number of kernels per ear．

Comprehensive Evaluation on Drought Ｒesistance of Maize at Maturity

YUAN Chuang, ZHU Lin, XU Xing, WANG Kaiyuan

2019, 48(10): 47-53. DOI: 10.15933/j.cnki.1004-3268.2019.10.008

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In order to identify drought-resistant maize varieties,fifteen maize genotypes were adopted as experimental materials and were planted in severe drought and well-watered conditions.The plant height,stem diameter,ear length,ear weight,numbers of ear rows and grains per row,diameters of ear and axis,bald tip length,100-grain weight,and grain yield were measured under the two contrasted treatments at mature stage.The drought resistance of different maize varieties at maturity was evaluated by a series of methods,including correlation analysis,principal component analysis,membership function,cluster analysis and other methods.The results showed that severe drought stress exerted significant effect on plant height,stem diameter,ear length,kernel number,ear weight and 100-grain weight.Generally,there were significant correlations among 11 indexes in this study.Principal component analysis showed that 5 principal components could represent 87.27% of the original information of maize resistance.According to the comprehensive evaluation value of drought resistance(D),the tested maize materials could be divided into 4 drought resistant classes.ClassⅠ was highly drought-resistant type,including Baiqing 161andZhongyuandan32.ClassⅡwasdrought-resistant type,includingGuiqingzhu,A18/H237andNorth 16/5996.ClassⅢwasmoderate drought-resistant type,including C5/A18,Baiqing 131,5178/1519,J112/1528,6522/1522,H14/599-2 andH237/1506.ClassⅣwas drought susceptible type,including Baiqing178,J152/1506,andA3/A28.

Effects of Transplanting Time and Harvesting Time on Economic Characters and Quality Characteristics of Upper Six Leaves of Flue-cured Tobacco in Central Henan

GAO Zhenzhen, LI Jianhua, YANG Mingkun, DUAN Weidong, DUN Songyang, CHEN Xiaolong, HE Bing, SHI Hongzhi

2019, 48(10): 54-63. DOI: 10.15933/j.cnki.1004-3268.2019.10.009

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Field experiment was carried out using Zhongyan 100，the main cultivar as material，to investigate the effects of transplanting time and harvesting time on economic characters and quality characteristics of upper six leaves of flue-cured tobacco.The results showed that the transplanting time and harvesting time significantly affected the economic traits of upper six leaves,and there was a very significant interaction effect.The effect of transplanting time on yield and output value was greater than that of harvesting time,while the harvesting time had a greater influence on the proportion of superior tobacco，single leaf weight and average price of upper six leaves.Transplanting from April 25 to May 5 gave rise to higher yield and output value.Delaying harvest by 6 d and 12 d resulted in no difference in average price,but it was significantly higher than other treatments,with the proportion of superior tobacco increased significantly by 10.66% and 15.39% respectively compared with the normal harvest.The transplanting time was closely related to strong-aroma style of upper six flue-cured tobacco leaves,and the transplanting time no later than May 5 was beneficial to the display of the strong-aroma style of upper six leaves.With the delay of the transplanting time from April 15 to May 5,the potassium-chlorine ratio of upper six leaves increased,and the flammability was better.The content of neutral aroma components was highest when transplanting tobacco from April 25 to May 5.The harvesting time had a great influence on the quality of upper six leaves,as harvesting time delayed,the protein content,starch content,sugar-alkali ratio of upper six leaves decreased,and tended to be optimum when the harvesting time was delayed for 12 days.The sensory quality of upper six leaves showed an increasing trend as a whole as harvesting time delayed,and was the best under a delay of 12 days with significant increase of total amount of neutral aroma components.Comprehensive analysis showed that transplanting from April 25 to May 5 and harvesting with 6-12 days delayed were conducive to improvement of economic characters,quality,and the display of strong-aroma style of upper six tobacco leaves in central Henan．

The Ｒole of Hydrogen Sulphide in Improving Aluminium Tolerance of Soybean

LIU Wenwen, JI Fang, WANG Huahua

2019, 48(10): 64-69. DOI: 10.15933/j.cnki.1004-3268.2019.10.010

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Aluminium (Al) toxicity is a major constraint on agricultural production in acidic soils.To elucidate how crops cope with Al toxicity，the role of hydrogen sulphide (H₂S) ,a novel gas signaling molecule in plants,was investigated in soybean under Al stress.Ｒesults showed that Al treatment induced H₂S production in root tips.Furthermore,application of an H₂S donor significantly alleviated Al-induced inhibition of root elongation,and decreased Al accumulation,cell deathrate,malondialdehyde and reactive oxygen species ( H₂O₂and O₂) contents in root tips,whereas the treatment with an H₂S inhibitor elicited the opposite effects.Further investigation showed that H2 S donor increased the activities of antioxidant enzymes (APX,CAT,POD and SOD),while the H₂S inhibitor treatment inhibited the antioxidant enzymes activities in root tips under Al stress.These results indicate that H₂S alleviates Al induced root elongation inhibition through enhancement of antioxidant defenses in soybean,subsequently reducing reactive oxygen species accumulation.

Determination of Herbicide Atrazine Degradation Products by Strain Enterobacter sp.and Degradation Gene Cloning

LIU Dandan, SUN Wanyu, LIU Chang, MA Haoke, WANG Ｒuijiao, SUI Yufan

2019, 48(10): 70-76. DOI: 10.15933/j.cnki.1004-3268.2019.10.011

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In order to analyze the biodegradation process of herbicide atrazine，liquid chromatography mass spectrometry was used to detect the degradation products of atrazine by functional strain Enterobacter sp.The degradation gene was cloned by the PCＲ amplification technology,and the degradation gene func tion was detected by using the GO function enrichment analysis.Hydroxyatrazine,cyanuric acid and urea were successively detected in the degradation products，and the degradation genes L465-ＲS09425,L465 ＲS10465,L465-ＲS12445,L465-ＲS09100,L465-ＲS12200 and UreA were cloned.It was speculated that strain Enterobacter sp.could mineralize atrazine gradually by dechlorination,dealkylation,deamination,ring cleavage,decarboxylation and hydrolysis of allophanate under the action of degrading genes．

Identification of a Cellulase Producing Fungus from Tibet Hot Spring and Optimization of Its Enzyme Production Conditions

LU Yuxin, ZHAO Hangke, TANG Xiaofei, LIU Kaihui, DENG Baiwan, DING Xiaowei

2019, 48(10): 77-83. DOI: 10.15933/j.cnki.1004-3268.2019.10.012

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In order to deepen the development and utilization of thermophilic cellulase resources,a cellulase producing fungus THN8 from Nimu hot spring in Tibet was isolated and identified，and the cellulase producing activity of this strain was screened by Congo red culture medium.According to the morphology and ITS sequence analysis,the strain THN8 was identified as Melanocarpus albomyces.On the basis of single factor experiment,orthogonal experiment was used to optimize the cellulase producing conditions of the strain THN8． Finally，the optimal medium formula for the cellulase producing strain THN8 was as follows: lactose 92.0 g/L,(NH₄)₂SO₄ 6.0 g/L，K₂HPO₄3.0 g/L，MgSO₄·7H₂O 0.55 g/L.The optimal fermentation conditions were as follows: the fermentation temperature was 58℃，the initial pH value was 5.5,and the fermentation time was 4 days. Under these conditions,the cellulase activity of the strain THN8 reached 28.1 U/mL.The thermophilic fungus isolated in this experiment plays an important role in the degradation and utilization of cellulose agricultural waste．

Isolation，Identification and Antimicrobial Activities of Endophytic Fungi from Stephania dielsiana Y.C.Wu

YAN Zhenling, LI Guoping, LUO Haiyu, LU Baoyi, LIANG Chunxia, DENG Yecheng, DENG Zhiyong

2019, 48(10): 84-92. DOI: 10.15933/j.cnki.1004-3268.2019.10.013

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In this study，endophytic fungi were isolated and identified from the stems and leaves of Stephania dielsiana Y.C.Wu,and their antimicrobial activities were analyzed to provide a basis for the protection of Stephania dielsiana Y.C.Wu plants and the development of active natural products.The results showed that a total of 38 endophytic fungi were obtained from the stems and leaves of Stephania dielsiana Y.C.Wu,31 of them were identified and classified into 14 genera based on the morphological and molecular identification,including Alternaria,Hypoxylon,Cladosporium,Phoma,Fusarium,Stagonosporopsis,Ascomycete,Chaetomium，Pestalotiopsis，Curvularia，Aureobasidium,Annulohypoxylon,Leptosphaerulina and Colletotrichum.Among them,Alternaria and Phoma were the dominant genera.14 endophytic fungi were selected based on the antagonism results in pre-experimental test,to determine the antimicrobial activity with their extracts from the fermented culture against 9 plant pathogenic fungi and 10 animal pathogenic bacteria.The results showed that the extracts of the endophytic fungi had inhibitory activity against tested pathogens to some extent.The extracts of 7 endophytic fungi (leaf-8,Y-66，YB-12， J-12，J-34，J-36 and J-55) displayed significant antifungal activity against 5 or more phytopathogenic fungi.Among them，leaf-8 and J-12 showed the best inhibitory activity against Exserohilum turcicum and Diaporthe citri with the EC₅₀ values at 0.063 1 mg/mL and 0.006 2 mg/mL,respectively.J-34 had the best antibacterial activity against Alternaria oleracea and Ceratocystis paradoxa with the EC₅₀ values at 0.018 1 mg/mL and 0.004 2 mg/mL,respectively.In addition,the extracts of 10 endophytic fungi exhibited inhibitory activity against Staphyloccocus aureus with the MIC values ranging at 0.500—10.000 mg/mL,and 4 of them had inhibitory activity against Escherichia coli with the MIC values ranging at 0.156—4.000 mg/mL.The results indicate that there are abundant endophytic fungi distributed in the stems and leaves of Stephania dielsiana Y.C.Wu,and their secondary metabolites possess broad-spectrum antimicrobial activity．

Identification of a New Pathogen Causing Ｒice Spikelet Ｒot Disease

CHEN Lijun, WANG Chunsheng, TIAN Xueliang, ZHI Ya’nan, LIU Yankun, WEI Yunfei

2019, 48(10): 93-98. DOI: 10.15933/j.cnki.1004-3268.2019.10.014

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In China,the damage caused by rice spikelet rot disease (ＲSＲD) was increased by years.It was found that exposed grain of some rice plants with dehiscent glume had blue-green mold,which was different from ＲSＲD symptoms caused by other pathogens during the investigation of that in Xinyang City of Henan Province.The isolate from collected samples in laboratory was tested for pathogenicity and iden tified by morphological characteristics and multigene phylogenetic analysis.The results showed that the isolate Ｒ1 was identified as Penicillium oxalicum based on the same morphological characteristics and identical branch with 100% support rate in phylogenetic tree constructed with sequences of ITS,BenA and CaM.The test for pathogenicity indicated that Penicillium oxalicum was a new pathogen of ＲSＲD．

Ｒesearch on the Field Control Effect of Biocontrol Agent LZ-1 against Apple Ｒing Ｒot on Branches and Trunks

ZHANG Linyao, MENG Xianglong, WANG Wuke, ＲEN Hongmin, HU Tongle, CAO Keqiang

2019, 48(10): 99-104. DOI: 10.15933/j.cnki.1004-3268.2019.10.015

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In order to seek new fungicides and methods for controlling apple ring rot on branches and trunks,the field control effect of self-developed protective biocontrol agent LZ-1was studied.The results showed that the biocontrol agent LZ-1 could effectively inhibit the production of new warts in apple trees with high control effect on the diseased trees,and the highest prevention effect reached 89.93%,which was the same as the chemical methyl thiophanate paint.Treatments of applying biocontrol agent LZ-1 from March to May had good field control effect,which were the key months for field application of pesticides.The effective adhesion period of the biocontrol agent LZ-1 was 60 days on branches，and that was 6 months on trunks.The application of biocontrol agent LZ-1 can reduce the usage of chemical fungicides and achieve the purpose of fungicides reduction．

Effects of Different Substrates on Pepper Plug Seedling Growth

CHEN Fen, YU Gao, HOU Jianwei, TAN Jiebin, WANG Xiaojun

2019, 48(10): 105-111. DOI: 10.15933/j.cnki.1004-3268.2019.10.016

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This study used agricultural organic waste in northern Guizhou and perlite as matrix material， to screen seedling substrates suitable for pepper planting． Effects of different seedling substrates on pepper plug seedling growth were tested with nine treatments including T1［V(organic material A) ∶ V(organic material B) ∶ V(old ash) ∶ V(perlite) =1∶ 4∶ 5∶ 0) ］，T2(2∶ 3∶ 4∶ 1) ，T3(3∶ 3∶ 3∶ 1) ，T4(4∶ 3∶ 2∶ 1) ，T5 (4∶ 4∶ 1∶ 1) ，T6(3∶4∶2∶1) ，T7(3∶2∶ 4∶ 1) ，T8(4∶ 1∶ 5∶ 0) and vegetable garden soil(CK).The pepper Xiangla No.12 was used as plug seedling experimental material.The results showed that all treatments could meet the requirements of the growth of pepper except for the water-gas ratio under T1 treatment and the bulk density under CK treatment.Except T1 treatment，the emerging rate of the other seven treatments was 12.25%—36.79% higher than CK treatment，the survival rate was 12.8%—33.67% higher than CK treatment.The morphological indexes of pepper seedlings treated with different seedling substrates were better than CK treatment( except for T1 treatment).Pepper seedlings under T3 treatment had the highestplantheight,stemdiameter,freshweight per plant,root-shoot ratio,strong seedling indexandG value.TheseresultssuggestedthatT3treatmentcouldsignificantlypromotethegrowthofpepperseedlings andcouldbeanewtypeofpepper seedlingsubstrate．

Effects of Water and Fertilizer Coupling on Growth and Photosynthetic Characteristics of Young Apple Tree

ZHOU Hanmi, NIU Xiaoli, YAN Hui, ZHAO Long, ZHAO Na, XIANG Youzhen

2019, 48(10): 112-119. DOI: 10.15933/j.cnki.1004-3268.2019.10.017

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With barrel planted red Fuji apple saplings as test materials,experiments which had four levels of irrigation(W₁ ,sufficient irrigation; W₂,mild water stress; W₃,moderate water stress; W₄,severe water stress.The lower and upper limits of irrigation were 75%—90%，65%—80%，55%—70% and 45%—60% field capacity,respectively) and three levels of fertilization( F₁，higher level; F₂,medium level; F₃，lower level.The N,P₂O₅ and K₂O were 0.6，0.2，0.2 g/kg，0.4，0.2，0.2 g/kg and 0.2,0.2,0.2 g/kg,respectively) were conducted to study the effects of water and fertilizer coupling on growth,photosynthetic characteristics and water use efficiency of two-year young apple tree under the drip irrigation.The results showed that compared with F₃W₄ treatment,F₁W₁treatment increased plant growth and leaf area by 85.2% and 74.5% respectively,while F₁W₂treatment increased plant growth and leaf area by 98.7% and 87.3% respectively,which indicated that the combination of high fertilizer and mild water deficit (F₁W₂treatment) was most conducive to the plant and leaf growth of young apple tree. The maximum val ues of water use efficiency occurred in F₁W₂treatment at each growth stage(germination and flowering stage,new growth stage,fruit setting and expanding stage and maturation stage) and were 3.49,3.84,4.37 and 3.90 mmol/mol，respectively.Meanwhile，compared with F₁W₁,although the net photosynthetic rate decreased by 1.9%,7.2%,6.8% and 2.1% respectively,the water use efficiency increased by 5.8%，6.7%，4.5% and 3.2% respectively,which indicated that the moderate water deficit could im prove the water use efficiency of young apple trees.Moreover,the maximum differences of net photosyn thetic rate and transpiration rate of apple saplings under different treatments all occurred in the fruit set ting and expanding period,which indicated that the fruit setting and expanding period of young apple tree was the most critical period of water and fertilizer demand,during which reasonable regulation of water and fertilizer could promote the growth of young apple trees．

Status and Development Trends of Grape Industry in Henan Province

LV Zhongwei, ZHANG Ke, WANG Peng, LOU Yusui, FAN Hongjie, WU Wenying, ZHANG Xiaofeng

2019, 48(10): 120-126. DOI: 10.15933/j.cnki.1004-3268.2019.10.018

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To grasp the grape industry development situation in Henan,production actuality,market con ditions,problems and solutions，development trends were explored in this paper,based on analyzing the grape planting area,yield,areal distribution,cultivars sturcture,production value,production organization form,market requirement and market supply,price trends,market prediction,and so on.The results showed that the yield,area and culture technique of Henan grape have been progressively developed in re cent years,but lots of problems still existed in Henan grape industry,such as unreasonable zoning,single breed，small scale,poor standardization,low market value and low benefit,week competitiveness,labour shortage,etc.Only by adhering to the varieties marketization,production standardization,labor-saving cul tivation,economization of resources and fruit safety can the regional advantages of Henan be strengthed,which will comprehensively promote the transformation of fruit production from traditional‘quantity-orien ted benefits’to ‘quality-oriented benefits’,and promote the healthy and sustainable development of Henan grape industry．

Cloning and Expression Analysis of HSP70 Gene in Different Heat-Ｒesistant Pansy(Viola tricolor) Materials under Heat Stress

NIU Yangli, ZHU Xiaopei, YANG Yaping, GE Xiaomin, DU Xiaohua, LIU Huichao

2019, 48(10): 127-132. DOI: 10.15933/j.cnki.1004-3268.2019.10.019

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In this study,the cDNA and gDNA sequences of the VtHSP70 gene were amplified from the heat-resistant pansy(DFM-16) and heat-sensitive material(08H) by PCＲ，and the expression level of the VtHSP70 gene under heat stress was analyzed by real-time PCＲ technique.The results showed that the length of VtHSP70 cDNA sequence and the length of open reading frame were the same in the two materi als，encoding 649 amino acids and the similarity reached 98.61%.The VtHSP70 gDNA structure of the two materials was similar and contained an intron.Homology comparison and phylogenetic tree showed that VtHSP70 gene had the highest homology with cytoplasmic HSP70 gene in Arabidopsis thaliana and rice.Ｒeal-time PCＲ results showed that the expression of VtHSP70 gene increased significantly in the two materials after heat shock treatment at 42℃ for 0,1,2,6,12 hours,but the increase rate in the heat-re sistant material DFM-16 was significantly higher than that in the heat-sensitive material 08H．

Expression of Pseudorabies Virus gE Protein in Pichia pastoris and Screening of Its Monoclonal Antibodies#br#

2019, 48(10): 133-139. DOI: 10.15933/j.cnki.1004-3268.2019.10.020

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In order to obtain pseudorabies virus(PＲV) gE protein with biological activity and its specific monoclonal antibodies,in this study,the recombinant plasmid pPICZαA-gE was transformed into Pichia pastoris X-33 by electroporation,and the positive clones were screened.PＲV gE protein expression strains were screened by SDS-PAGE and Western blot.The gE secretory protein was obtained by inducing expression and purified by Ni column.The purified protein was identified by Western blot,Dot blot and ELISA.At the same time，BALB/c mice were immunized with purified protein.After cell fusion,monoclonal antibodies were screened by immunoperoxidase monolayer assay(IPMA) and the reactiveness,titer and subtype
of monoclonal antibodies were identified.The results showed that PＲV gE protein was successfully expressed in Pichia pastoris and purified by Ni column.The purity of PＲV gE protein was over 90%，and the protein had good biological activity and well reactivity with PＲV positive serum.Four strains of monoclonal antibodies of gE with high sensitivity and specificity were screened，which could recognize PＲV gE protein and virus specifically at the same time.They were named 2F10,4 C10,9E1 and 10C3 respectively.In conclusion，PＲV gE protein was successfully expressed in yeast system，and four strains of monoclonal antibodies against PＲV gE protein were screened．

Pathogenicity Analysis and Molecular Characterization of Newcastle Disease Virus Isolated from Ｒacing Pigeon

LI Peiyao, GAO Jingping, TIAN Yong, WANG Cunlian, XU Mingju, LI Kai, LI Jun, ZHANG Ｒuihua, LAN Jinping, XU Tong

2019, 48(10): 140-146. DOI: 10.15933/j.cnki.1004-3268.2019.10.021

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In order to understand the molecular genetic characteristics and variation of Newcastle disease epidemic strains in racing pigeon population,in this experiment,the disease material was collected from the suspected pigeon Newcastle disease case in Hebei Province.After routine treatment,the SPF chicken embryo was inoculated.The chicken embryo allantoic fluid was collected and identified by Hemagglutina tion and Hemagglutination inhibition test as Newcastle disease virus,and pathogenicity tests,virulence as says were conducted.The sequences of F gene were amplified by ＲT-PCＲ and genetic evolution and ami no acid sequence homology with other different species of Newcastle disease virus strains were analyzed． The results showed that a Newcastle disease virus(named HB strain) was isolated from the disease mate rial.The pathogenicity test of HB strain presented similar clinical symptoms and necropsy changes with those found in naturally diseased pigeons． The mean death time，intracerebral pathogenicity index and in travenous pathogenicity index of HB strain were 52.8 h,1.78,and 2.59，which indicated that the HB strain was a virulent strain.F gene of HB strain contained a complete open reading frame(OＲF) of 1 662 bp.The genotyping results showed that the HB strain belonged to the Ⅵ genotype.Genetic relationship of HB strain showed that it was closed to pi/CH/LGD/110208,dove/Italy/2736/00,Q-GB 506/97 based on F gene.Especially with the pigeon/Qinghai/1344/2017 strain，the closest relativities of nucleotide and amino acid sequences were 98.45% and 99.5%.However,it had far genetic relationship with traditional chicken Newcastle disease vaccine strains of B1,LaSota,V4 and Mukteswar.The F protein cleavage site amino acid sequence of HB strain was 112Ｒ-Ｒ-Q-K-Ｒ-F117,which was consistent with the characteristics of virulent strains.The results indicate that the HB strain is a virulent strain and has a large genetic differ ence from the current widely used Newcastle disease vaccine strains．

Effects of Six Traditional Chinese Medicines against Porcine Ｒeproductive and Ｒespiratory Syndrome Virus in Vitro

LIU Yun, ZHU Shanyuan, GONG Zhunan, QIN Feng, XIA Wenlong, WU Shuang, WU Xiaojie

2019, 48(10): 147-154. DOI: 10.15933/j.cnki.1004-3268.2019.10.022

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In order to study the effects of six traditional Chinese medicines，such as Nepeta cataria L.,Potentilla chinensis Ser.,Humulus lupulus Linn.,Potentilla discolor Bge.,Stemona japonica(Blume) Miq.and Ｒeynoutria japonica Houtt.,against porcine reproductive and respiratory syndrome in vitro,the Marc 145 cell model was established in this experiment.Based on the study of their maximum safe mass con centration on Marc-145 cells,combined with the observation of cell morphology and MTT method,the three actions of blocking,inhibiting and inactivating the virus were determined,respectively.The results showed that the maximum safe mass concentrations of Nepeta cataria L.,Potentilla chinensis Ser.,Humu lus lupulus Linn.,Potentilla discolor Bge.,Stemona japonica(Blume) Miq.and Ｒeynoutria japonica Houtt.on Marc-145 cells were 0.25 mg/mL，1.25 mg/mL，0.63 mg/mL，0.31 mg/mL，1.56 mg/mL， and 0.47 mg/mL.In the experiment of anti-PＲＲSV effect in vitro，it was found from the cell morphology observation that under the three modes of action on PＲＲSV，the cells treated by Potentilla chinensis Ser.and Ｒeynoutria japonica Houtt. had no obvious lesions，and the treatment of other four drugs showed dif ferent degrees of lesions.From the inhibition rate of PＲＲSV，the highest inhibition rates of blocking，inhi bition and direct inactivation of Nepeta cataria L.were 80.6%，52.2% and 56.5%,respectively.The highest inhibition rates of PＲＲSV by three modes of action of Potentilla chinensis Ser.were 115.8%,93.0% and 125.4%,respectively.The highest inhibition rates of PＲＲSV in the three modes of action of Humulus lupulus Linn.were 61.5%，32.0% and 82.4%，respectively.The highest inhibition rates of PＲＲSV in the three modes of action of Potentilla discolor Bge.were 15.6%,129.4% and 57.7%,re spectively.The highest inhibition rates of PＲＲSV in the three modes of action of Stemona japonica (Blume) Miq. were 120.2%，61.8% and 71.9%,respectively.The highest inhibition rates of PＲＲSV in the three modes of action of Ｒeynoutria japonica Houtt.were 110.4%，94.3% and 130.8%,respective ly.Combined with morphological observation and inhibition rate results，Potentilla chinensis Ser.and Ｒey noutria japonica Houtt.had the best anti-PＲＲSV effect，followed by Nepeta cataria L.and Stemona japon ica(Blume) Miq．

Effects of Aquaponics Mode on Growth，Digestive Enzyme Activity and Promotion Benefit of Cyprinus carpio haematopterus

SHI Dongjie, HU Jinyou, WANG Saisai, CHEN Xiaoxuan, ZHU Hua, ZHANG Xin

2019, 48(10): 155-161. DOI: 10.15933/j.cnki.1004-3268.2019.10.023

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In order to further promote and widely apply the aquaponics mode，thereby reducing the envi ronmental impact of traditional fish farming mode,a comparative study of aquaponics mode and traditional culture mode was carried out.The effects of aquaponics culture mode on growth and digestive enzyme ac tivity of Cyprinus carpio haematopterus were studied by measuring the weight gain rate,specific growth rate，survival rate,intestinal protease activity，intestinal lipase activity and intestinal amylase activity.The results showed that different coverage of biological floating bed of Ipomoea aquatica Forsk(5%,10%,20%,40%) had a promoting effect on the weight gain rate，specific growth rate，intestinal protease activity,intestinal lipase activity and intestinal amylase activity of C.carpio haematopterus.10% of the coverage area of the biological floating bed of Ipomoea aquatica Forsk could effectively improve the growth of C.carpio haema topterus，and significantly promote its digestive enzyme activity,which was suitable for pilot test.Judging from the indicators of economic benefits，the mode had obtained economic benefits of 10.561 4 million Yuan in the five-year economic efficiency calculation period，and the economic benefits that might be gen erated was 18.596 4 million Yuan，and the annual economic benefit reached 5.832 2 million Yuan.After the mode was applied to aquaculture production,each unit of development funds could increase the net in come by 4.17 times,and the economic benefits was significant．

Diameter and Area Measurement of Sunflower Faceplate

2019, 48(10): 162-173. DOI: 10.15933/j.cnki.1004-3268.2019.10.024

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To provide parameters and basis for subsequent studies such as flower phenotypic characteris tics，pollinator interaction，and yield estimation，the diameter and area of sunflower faceplate were meas ured.Through the special background board and shielding board，the image of sunflower faceplate was col lected non-destructively in the field，and the image was automatically transformed.The tip point of the contour of the faceplate was found，and the different radial diameters of the sunflower faceplate were meas ured by the tip wrapping method，to establish the average diameter of the sunflower faceplate.The area of the sunflower disc was established by operations such as texture segmentation，morphological opening and closing operations，and the average diameter of the sunflower disc was calculated.The recommended num ber of sampling measurements for the average diameter of the sunflower faceplate was 32.Prediction mod els for the average diameter and area of sunflower faceplate and disc were established.The results showed that,the determined coefficients of the average diameter prediction models of the sunflower faceplate and disc were 0.991 and 0.999 respectively,and the determined coefficients of the prediction models of the area surrounded by the tip contour of the faceplate and the original contour were 0.942 and 0.935.The tip wrapping method proposed measured the diameter and area of the faceplate with high accuracy and no sensitivity to scale changes．

Ｒelationship between Color Parameters and Curing Characteristics of Upper Leaves of Flue-cured Tobacco

SUN Yang, LI Qingshan, NI Chao, TAN Xiaolei, LIU Li, WANG Chuanyi, XU Xiuhong

2019, 48(10): 174-180. DOI: 10.15933/j.cnki.1004-3268.2019.10.025

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In order to clarify the relationship between color parameters of tobacco leaves and the curing characteristics，changes of color parameters and curing characteristics of upper leaves of flue-cured tobac co NC55 and NC102 with harvesting time were studied.The correlation and regression analyses of color parameters with yellowing index and browning index were carried out.Curing characteristic regression models based on color parameters of tobacco leaves were established and tested.Ｒesults showed that，with the passage of harvesting time，the color parameter L value(brightness value) ，b value(yellowness val ue) ，C value(saturation value) of flue-cured tobacco leaves of varieties NC55 and NC102 increased in a whole，H°value( color phase angle) tended to decrease totally，a value (redness value) decreased first and then increased.The yellowing index increased，and the browning index decreased first and then in creased.The curing characteristics at a certain stage were better than those in other stages.Color parame ters had extremely significant or significant correlation with yellowing index and browning index，and the curing characteristic regression models based on color parameters of tobacco frontal leaves had better pre diction effect.It was feasible to establish a regression model based on the color parameter of tobacco fron tal leaves to predict curing characteristics．

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