Journal of Henan Agricultural Sciences

Review of Aminopeptidase N as the Receptor of Porcine Coronavirus Infection

WEI Xinxin, LI Zehui, JIN Xiaohui, WEI Zhanyong

2021, 50(8): 1-8. DOI: 10.15933/j.cnki.1004-3268.2021.08.001

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Porcine aminopeptidase N （pAPN） is a type Ⅱ zinic‑dependent membrane‑bound transmembrane glycoprotein，which is widely found on the surface of porcine tissue cells. Porcine aminopeptidase is a functional receptor，and its interaction with porcine coronavirus has become one of the hot‑spots of coronavirus research. As receptors of some coronaviruses，pAPN mediates coronavirus infection through recognizing and binding the receptor binding domain（RBD）of coronavirus S protein. In this review，we summarized the interaction mechanism of APN with viral proteins of different porcine coronavirus，analyzed the similarities and differences of their interactions，and discussed the influence of pAPN inhibitors on coronavirus infection. This review can offer a useful reference for studying the mechanism of porcine coronavirus infection，and provide a foundation for exploiting drugs which can block coronavirus infection.

Effect of Rice‑Fish‑Chicken Symbiosis on Paddy Soil Nutrient and Rice Yield under Ridge Cultivation

MA Weiwei, CHEN Can, HUANG Huang, REN Bo, ZHOU Jing, LI Guixiang, WANG Ren

2021, 50(8): 9-17. DOI: 10.15933/j.cnki.1004-3268.2021.08.002

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Four treatments were set up，including rice‑fish‑chicken pattern under ridge cultivation（RFC），rice‑fish pattern under ridge cultivation（RF），rice‑chicken pattern under ridge cultivation（RC）and rice ridge cultivation（CK），and the effects of rice‑fish‑chicken symbiosis on soil nutrient and rice yield were studied under ridge cultivation，so as to provide scientific basis for the development and popularization of rice‑fish‑chicken pattern. The results showed that compared with CK，RC and RFC treatments increased the contents of soil total nitrogen and total phosphorus at mature stage in two‑year experiments，and the total phosphorus contents of RC and RFC treatments were significantly higher than those of CK；the contents of alkali‑hydrolyzable nitrogen and available phosphorus of RC and RFC treatments were significantly higher than those of CK at mature stage in two‑year experiments，the contents of available potassium of RFC treatment were significantly higher than those of CK at full heading stage in two‑year experiments；the soil organic matter contents of RFC，RC and RF treatments were significantly higher than those of CK at mature stage in two‑year experiments. In 2018，the rice yield of RF treatment increased by 5. 75% compared with CK，and there was no significant difference among the treatments. In 2019，the rice yields of RFC，RC and RF treatments significantly increased by16. 29%，14. 03% and 9. 35% compared with CK respectively. In conclusion，RC and RFC treatments can improve soil nutrient content and rice yield，and could be used as the optimal patterns for comprehensive cultivation in paddy field，especially RFC treatment.

Effects of Different Nitrogen Topdressing Models on Root Physiological Characters and Yield of Direct Seeding Rice

FU Jing, WANG Ya, YANG Wenbo, YIN Haiqing, WANG Yuetao, BAI Tao, WANG Fuhua, WANG Shengxuan

2021, 50(8): 18-25. DOI: 10.15933/j.cnki.1004-3268.2021.08.003

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A field experiment was conducted to reveal the effects of different nitrogen topdressing models in the middle and later periods on root physiological characteristics and grain yield of direct seeding rice with Zhenghan No. 9 and Zhenghan No. 10 as materials，so as to provide a scientific basis for high‑yield cultivation of direct seeding rice and high‑efficiency utilization of nitrogen. Under the condition of total nitrogen application rate of 270 kg/ha，three nitrogen fertilizer topdressing models were set after jointing.The ratios of base and tillering fertilizer，flower‑promoting fertilizer，flower‑preserving fertilizer were 6∶4∶0（T1），6∶2∶2（T2），6∶0∶4（T3），and the treatment with the same amount of base and tillering fertilizer but without nitrogen topdressing（T0）in the middle and later periods was used as control.The results showed that dry root weight，root oxidation activity，auxin content，zeatin and zeatin nucleoside content，total root absorption area and active absorption area of Zhenghan No. 9 and Zhenghan No. 10 all showedT2>T1>T3>T0 at the heading stage and 10，20，30 d after heading stage. There was no significant difference in the number of panicles of Zhenghan No. 9 and Zhenghan No. 10 among different nitrogen topdressing models，the spikelet number per panicle showed T2>T1>T3>T0，the seed setting rate showedT2>T3>T0>T1，1 000‑grain weight showed T2>T1>T0>T3，and yield showed T2>T1>T3>T0.The spikelet number per panicle，seed setting rate，1 000‑grain weight and yield of T2 treatment were higher than those of the other treatments，and the yields of Zhenghan No. 9 and Zhenghan No. 10 increased by41. 4% and 40. 7% respectively compared with T0 treatment. In summary，nitrogen topdressing at appropriate time（flower ‑ promoting fertilizer and flower‑preserving fertilizer）after jointing of direct seeding rice can promote root growth in the middle and later periods，maintain high physiological activity of roots，delay root degeneration and senescence，and improve the yield of direct seeding rice.

Alleviating Effect of Spraying Carbon‑Adsorbed Polyglutamic Acid on Growth of Maize Seedling under Drought Stress

DANG Yongfu, CAO Liru, LU Xiaomin, DANG Zun, QIU Tian, DUAN Enzhong, QIU Junzheng, TIAN Yunfeng

2021, 50(8): 26-35. DOI: 10.15933/j.cnki.1004-3268.2021.08.004

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Zhengdan 958，Zhengdan 6122 and Zhengdan 819 were used as test materials，after drought stress，water and carbon‑adsorbed polyglutamic acid were sprayed on maize seedlings respectively，and the effects of carbon‑adsorbed polyglutamic acid on the morphology，physiology and biochemistry of maize seedlings under drought stress were studied. The results showed that the root length，root surface area，root volume，plant height，leaf area，dry aboveground weight，dry underground weight and root‑shoot ratio of maize seedlings under drought stress overall significantly increased after sprayed with carbon‑adsorbed polyglutamic acid；the relative water content（RWC），chlorophyll content，Fv/Fm，soluble protein content，activities of superoxide dismutase（SOD），peroxidase（POD） and catalase（CAT） of maize leaves significantly increased after sprayed with carbon‑adsorbed polyglutamic acid. Principal component analysis showed that RWC，chlorophyll content and soluble protein content of maize leaves were the key indexes in alleviating the damage caused by drought stress to maize after spraying carbon‑adsorbed polyglutamic acid. Grey correlation analysis showed that the chlorophyll content，root‑shoot ratio，Fv/Fm，POD and SOD activities all had higher correlation coefficients with the total dry weight，which were 0. 566 4，0. 454 9，0. 449 1，0. 429 9 and 0. 418 2，respectively. The results showed that carbon‑adsorbed polyglutamic acid enhanced the tolerance of maize to drought stress at seedling stage，and could be used as drought stress reliever.

Effect of Different Film‑uncovering Period on the Growth and Yield of Cotton in Northern Xinjiang

ZHAN Dongxia, KELAIMU Alimujiang, ZHAO Qiang, Yierdengbilike

2021, 50(8): 36-43. DOI: 10.15933/j.cnki.1004-3268.2021.08.005

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In order to determine the most appropriate uncovering time for cotton field in northern Xinjiang，the experiment set four plastic film treatments：uncovering plastic film at 10 days before first irrigation（T10），uncovering plastic film at 1 day before first irrigation（T1），uncovering plastic film at 1 day before second irrigation（E1），and plastic film mulched in the whole growth period（CK）in 2018—2019. The effects of uncovering film on the growth and yield of cotton（Xinluzao 42）were studied by measuring agronomic trait，dry matter，yield and residual plastic film weight in cotton field. The results showed that，the water content in 0—40 cm soil layer was reduced by film uncovering treatments，and the soil moisture content was lower in earlier treatments than later treatments. At boll‑opening stage,compared with CK，the plant heights of T10 and T1 treatments were decreased by 4.4%—18.8% and 4.2%—19.7%，the numbers of main stem leaves of T10 and T1 treatments were decreased by 4.8%—16.3% and 2.7%—22.8%，the numbers of fruit branches of T10 and T1 treatments were decreased by2.8%—26.0% and 4.7%—18.0%，respectively. Compared with CK，the dry matter of boll per plant of T10 and T1 treatments decreased by 28. 0% and 11. 7% respectively（boil‑opening stage in 2018），33. 9% and 25. 8% respectively（early boll‑opening stage in 2019）. The dry matter showed little difference between E1 and CK treatments. The lint yield of E1 was significantly higher than CK with increase of 15. 1% in 2018. There was no significant difference in lint yield between E1 and CK treatments in 2019. Compared with CK，the lint yield decreased by 4.4%—23.2% in T10 and by 11.8%—17.2% in T1. The residual film amount at 0—10 cm soil layer of T10，T1 and E1 treatments decreased by 54. 6%，54. 8% and 58.6%，respectively. The uncovering treatments had no significant influence on the residual film amount at 10—20 cm soil layer and 20—30 cm soil layer.The suitable period for film mulching in cotton field was from sowing to the second irrigation in northern Xinjiang.

Effect of Grafting on Curing Characteristics of Flue‑cured Tobacco

DAI Zhuoyi, ZHANG Xi, XU Zhiqiang, YAO Yifan, XUE Gang

2021, 50(8): 44-50. DOI: 10.15933/j.cnki.1004-3268.2021.08.006

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In order to explore the effect of grafting on the curing characteristics of flue‑cured tobacco and improve the curing characteristics of flue‑cured tobacco NC89，Zhongyan 100（easy to cure）and NC89（not easy to cure）were used as controls，two grafting combinations of Zhongyan 100（scion）/NC89（rootstock）and NC89/Zhongyan 100 were set up，and the water content，chlorophyll content，polyphenol oxidase（PPO）activity of the middle leaves of each treatment during the key period of curing，and the appearance quality，sensory quality and chemistry composition of the cured tobacco leaves were measured and analyzed. The results showed that the proportionality of water loss rate，chlorophyll degradation proportion，yellowing period（curing 0—72 h） average chlorophyll degradation rate，and mean PPO activity of the control Zhongyan 100 were the best，the proportionality of water loss rate was 1.26，the chlorophyll degradation proportion was 95. 25% after 72 h，the average degradation rate was 1. 32%/h during 0—72 h，and the mean of PPO activity was 0. 251U during 0—72 h. The tobacco leaves of all treatments were ranked in the curing ability and curing tolerance as Zhongyan 100> Zhongyan 100/NC89 >NC89/Zhongyan 100> NC89. The potassium content of NC89/ Zhongyan 100 was 1.67%，which was higher than that of the parent. The curing characteristics of the middle leaves of the grafted tobacco plant were between the two parents，in which the scion played a decisive role. When varieties with good curing characteristics were used as rootstocks，the curing characteristics of the scion could be improved to a certain extent.Using varieties with low nicotine content and high potassium accumulation efficiency as rootstocks could increase the potassium content and reduce the nicotine content of the scion.Therefore，reasonable selection of rootstock and scion grafting combination can improve the curing characteristics of flue‑cured tobacco and improve the quality of flue‑cured tobacco leaves.

Effect of Nitrogen，Phosphorus and Potassium Formula Fertilizer on Matter Accumulation and Quality of Angelica sinensis under Direct Seeding

GONG Chengwen, MI Yongwei, SHAO Wuping, XIE Zhijun

2021, 50(8): 51-56. DOI: 10.15933/j.cnki.1004-3268.2021.08.007

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In order to determine the reasonable fertilization schedule of Angelica sinensis seeds sowing，the characteristics of root morphology，yield and quality（ferulic acid，volatile oil，alcohol‑soluble extract and ash） of direct seeding Angelica sinensis were studied by three‑factor and three‑level combination experiment under three different fertilizer rates［N（75，150，225 kg/ha），P2O5（37.5，75.0，112.5 kg/ha）,K2O（37.5，75.0，112.5 kg/ha）］.The results showed that different N，P and K treatments had significant effects on root branch number，plant biomass and contents of extracts，ferulic acid and volatile oil of Angelica sinensis. Among them，N1P3K2 treatment had the largest root fresh weight（65.25 g）and the largest number of branches（11.9），followed by N2P3K1，of which the root fresh weight and number of branches were 63.28 g and 10.2 respectively，and the contents of alcohol‑soluble extract and ferulic acid respectively were 66.31% and 0.108%. The N3P2K1 treatment had the highest volatile oil content（1.137%）. By considering the yield and quality indexes of Angelica sinensis，the optimal NPK treatment is N₂P₃K₁，that is，the application rates of N，P₂O₅ and K₂O respectively are 150，112.5 and 37. 5 kg/ha.

Ecological Stoichiometry Characteristics of Soil and Microbial Biomass C，N and P in Vineyards with Different Planting Years

LIU Yifeng, TIAN Yaowu

2021, 50(8): 57-65. DOI: 10.15933/j.cnki.1004-3268.2021.08.008

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The soil C，N，P and microbial biomass C，N，P（MBC，MBN，MBP）contents in 0—100 cm soil layer were measured in vineyards planted for 5，8，13 and 20 years in Yanshi City grape reserve，and their ecological stoichiometric characteristics were analyzed，so as to provide scientific basis for the sustainable management of the vineyards. The results showed that the contents of C，N and P in soil increased obviously with the increase of planting years. The contents of C，N and P in soil with grape planted for 20 years increased by 20. 38%，64. 91% and 105. 26% respectively compared with the soil with grape planted for 5 years. The contents of MBC，MBN and MBP in soil also increased with the increase of planting years. The contents of MBC，MBN and MBP in soil with grape planted for 20 years increased by 205. 61%，191. 86% and 81. 56% respectively compared with the soil with grape planted for 5 years. Soil C∶N increased with the increase of soil depth，the increase trend weakened with the increase of planting years，when the grape was planted for 20 years，C∶N tended to be stable. Overall，soil C∶P（except the soil with grape planted for 13 years）and N∶P（except the soil with grape planted for 8 years）increased firstly and then decreased with the increase of soil depth. There were significant or extremely significant correlations between planting years and soil C，N，P，MBC，MBN，MBP contents and their ecological stoichiometry ratios.

Effects of Returned Straw Types on N2O Emission and nosZ‑denitrifying Bacterial Community Structure in Paddy Soil

LÜ Zefang, WANG Rong, GUO Xianjin, GAO Zhenzhen, LIU Zhangyong, JIN Tao

2021, 50(8): 66-75. DOI: 10.15933/j.cnki.1004-3268.2021.08.009

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To investigate the effects of straw returning types on N₂O emission and nosZ‑denitrifying bacterial communities in paddy soils，N₂O fluxes and community structure of nosZ‑denitrifying bacteria were observed through the static closed chamber‑gas chromatography method and high‑throughput sequencing approach under the treatments of conventional fertilization（NS），conventional fertilization plus rape seed straw（RS）and conventional fertilization plus broad bean straw（BS），respectively. Results showed that under the NS，RS and BS treatments，the N2O cumulative emissions from paddy soils were4.28，5.07 and 1.51 kg/ha，respectively. And the N₂O cumulative emission of BS was significantly lower than that of RS and NS（P<0. 05）. But there was no significant difference between NS and RS in the N₂O cumulative emission. Veen analysis showed that BS increased the groups of nosZ‑ denitrifying bacteria at the order，genera and species levels compared with NS and RS. The relative abundance of Rhizobiales and norank_p__environmental_samples at the order level in NS，RS and BS treatments were 4.30%，3.33%， 7.59% and 0.57%， 0.22%， 1.89%， respectively. And both Rhizobiales an norank_p__environmental_samples of BS were significantly higher than that of RS at the order level（P＜0.05）. At the family level，the relative abundance of norank_p__environmental_samples of BS was significantly higher than that of RS（P<0.05），and the values of NS，RS and BS were 0.22%，0.11%，and 1.67%，respectively.The correlation analysis indicated that the N2O cumulative emission during rice growing season was significantly negatively correlated with the relative abundance of Rhizobiales，norank_p__environmental_samples at the order level（P＜0. 01），and the relative abundance of norank_p__environmental_samples at the family level（P＜0.05），respectively.In conclusion，compared with NS and RS，BS significantly increased the nosZ‑denitrifying bacterial groups at the order，genus and species levels，and the relative abundances of Rhizobiales norank_p__environmental_samples at order level，and norank_p__environmental_samples at family level，thereby reducing the N2O cumulative emission.

Effects of Straw Returning on Soil Nutrients and Salinity in Luoyang Tobacco‑Growing Areas

HAN Jin, HAN Qiujing, MA Jing, LU Jian, YAO Pengwei, WU Qinyang, YE Xiefeng, WU Furu

2021, 50(8): 76-84. DOI: 10.15933/j.cnki.1004-3268.2021.08.010

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In order to explore the effects of rotten straw returning on nutrients and salinity of Luoyang tobacco‑growing soil，a field experiment with cultivated tobacco plants was organized in a completely randomized block design，Zhongyan 100 was used as the test variety. On the basis of conventional fertilization，no straw returning（CK），and the rotten straw returning treatments at rates of 6 000 kg/ha（T1），9 000 kg/ha（T2）and 12 000 kg/ha（T3），respectively were set.The contents of soil nutrients and salinity during tobacco growing stages for all the treatments were measured. The results showed that the addition of straw could significantly improve the contents of soil available phosphorus and potassium. The increase of soil available phosphorus and potassium was the largest when the amount of straw returning was 12 000 kg/ha. Compared with CK，addition of straw could decrease the soil pH value，and the soil pH value further decreased with the rates of straw returning increased. The contents of soil Ca²⁺ and SO₄^2‑decreased under the treatments with straw added at rates of 6 000 kg/ha and 12 000 kg/ha，respectively.However，straw returning at a rate of 12 000 kg/ha significantly increased the contents of K⁺，Mg²⁺ and Na⁺.Therefore，application of straw could increase soil nutrients. The straw returning at a proper rate could reduce the contents of Ca²⁺ and SO₄^2‑ in soil，whereas it also increased the contents of K⁺，Mg²⁺ and Na⁺.

Isolation and Evaluation of the Antagonistic Bacterium HJ‑5 against Orchid Stem Rot Disease

LÜ Ang, WANG Cheng, TANG Yafang, WANG Lihua, JIANG Yiquan

2021, 50(8): 85-93. DOI: 10.15933/j.cnki.1004-3268.2021.08.011

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In order to screen antagonistic bacterial strains for the biocontrol of orchid stem rot caused by Fusarium oxysporum,antagonistic bacteria were isolated and screened from the soil around the roots of healthy orchid seedlings in Xiaogan City，Hubei Province，and their biological control potential was evaluated. The results showed that 8 strains of antagonistic bacteria were isolated，which had good inhibitory effect on the growth of F.oxysporum.16S rDNA sequencing and phylogenetic tree construction showed that all the 8 antagonistic strains were Bacillus spp. B. megaterium HJ‑5 showed good and stable antagonism against F. oxysporum in plate confrontation. Its sterile fermentation broth was added into the virulent plate prepared by PDA medium at 5%. The sterile fermentation broth showed strong inhibitory effect on mycelial growth of Botrytis cinerea，Colletotrichum siamense，F.moniliforme，F.oxysporum，Monilia fructigena，Mucor hiemails，Rhizopus stolonifer，Sclerotinia sclerotiorum and Sclerotium rolfsii，and the inhibition rates were higher than 50%.The sterile fermentation broth of HJ‑5 could maintain the antifungal activity under high temperature，strong acid and alkali,and long‑term UV irradiation.In addition，the key genes sfp and fenB for the biosynthesis of antimicrobial substances surfactin and fengycin were amplified in the genome of HJ‑5.The above results indicate that B. megaterium strain HJ‑5has excellent biocontrol potential against orchid stem rot.

Screening and Control Effect of Endophytic Fungi for Brown Spot Disease of Schisandra chinensis

ZHANG Yuqi, REN Yueying, WEN Xiangsui, WANG Yumeng, FANG Ping, LI Baoquan, LENG Shihe

2021, 50(8): 94-102. DOI: 10.15933/j.cnki.1004-3268.2021.08.012

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In order to screen the excellent endophytic fungi with better biocontrol effects on the pathogen of Schisandra chinensis brown spot disease，Phoma glomerate（corad）Wollena ex Hoehapf，the conventional culture method in plant pathology was used to culture Phoma glomerate（corad）Wollena ex Hoehapf and observe its colony characteristics and micromorphology，at the same time，the plate antagonistic method was applied to screen the endophytic biocontrol fungi. By measuring the change of cell membrane permeability，soluble protein content，ergosterol content，and chitin content of Phoma glomerate（corad）Wollena ex Hoehapf，the bactericidal mechanism of WG9 fermentation liquid was preliminarily explored. Morphological observation，fungal ITS gene sequencing，and the construction of phylogenetic tree were combined to identify the screened excellent endophytic biocontrol fungi.The results showed that one strain of biocontrol fungi WG9 screened from the endophytic fungi of Schisandra chinensis had better biocontrol effect. The plate antagonistic experiment indicated that its antagonistic rate reached 64. 73% and the antagonistic rate of fermentation liquid was 48. 21%. With the increase of the concentration of the WG9 fermentation broth，the cell membrane permeability and soluble protein content of the pathogenic fungi hyphae increased，while the ergosterol content and chitin content decreased. Morphological observation and molecular biology identified the excellent endophytic fungus as Purpureocillium lilacinum. The present study showed that the endophytic biocontrol fungus WG9 screened from the rhizome of Schisandra chinensis had a good bactericidal effect on Phoma glomerate（corad）Wollena ex Hoehapf，and the WG9 fermentation broth had a certain damage effect on the cell wall of pathogenic fungi.

Relationship between Cold Tolerance and Leaf Structure of Different Pear Varieties (Strains)

WEI Jie, ZHANG Qi, LIN Caixia, JIANG Yuan

2021, 50(8): 103-112. DOI: 10.15933/j.cnki.1004-3268.2021.08.013

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In order to provide some references for the selection of parents，early identification of hybrid progeny during the breeding of cold‑resistant pear varieties and the regionalization of pear introduction and cultivation，relationship between leaf anatomic structure and cold tolerance of pear was discussed and the cold tolerance evaluation indexes were screened. In this study，8 anatomical structure indexes were measured of 13 pear varieties（strains）by using paraffin section methods.Typical representative indexes of leaf anatomical structure were screened via coefficient of variation，correlation analysis and hierarchical cluster analysis，and comprehensive evaluation on cold resistance of each varieties（strains）was conducted by the subordinate function method combined with the investigation of natural freezing injury in the field. The results showed that pear leaf transection of 13 varieties（strains）was composed of upper and lower epidermis，palisade tissue and spongy tissue and the leaves belonged to typical bifacial leaf.Cell layer number of palisade tissue was most 2—3 layers. The leaf anatomic structure of different pear varieties（strains）was similar on the whole，but there were some quantitative differences. Combined with cluster analysis and correlation index，three typical indexes of the cold‑resistant anatomical structure of leaves were selected，which were tightness of palisade tissue，sponge tissue thickness and leaf thickness. The cold resistance of the 13 varieties（strains）showed Pingguoli＞Xinli No.6＞Yali＞Xinli No.9＞84‑1‑1＞Xinli No.7＞Zaosuli＞Xinli No.8＞Dangshansuli＞Xinli No.11＞Xinli No.10＞Korla fragrant pear. Main indexes of leaf anatomical structure that affect the cold resistance of pear varieties（strains）are tightness of palisade tissue，sponge tissue thickness and leaf thickness. According to the anatomical characteristics of leaf tissue structure，the method of subordinate function combined with the performance of field freezing injury can better reflect the cold resistance of different pear varieties（strains）.

Comparison of Canopy Structure，Photosynthetic Characteristics，Yield and Quality of Korla Fragrant Pear with Different Tree Shapes

CHEN Jiuhong, MA Jianjiang, LI Yongfeng, WEI Jie, WANG Yan, HUANG Guohui

2021, 50(8): 113-123. DOI: 10.15933/j.cnki.1004-3268.2021.08.014

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Comparing the differences of canopy structure,photosynthetic characteristics,yield and quality between delayed‑open central leader system and cylindrical shape of Korla fragrant pear will provide a basis for the training and pruning of Korla fragrant pear orchard in labor‑saving and high‑efficient dense planting cultivation. Using the 6‑year‑old fragrant pear tree as the test material，and the experiment wascarried out for two consecutive years. The CI‑340 handheld photosynthesis measurement system，the CI‑110 canopy analyzer and the tree survey method were used to compare the effects of the tree structure,daily processes of photosynthetic parameters of each canopy part and canopy structure on fruit yield and quality between the two tree shapes. The number of branch groups directly attached to the cylindrical shape trunk of the 7‑year‑old tree was 37.33，equivalent to 509 975 branches per hectare. The number of main branches of the 7‑year‑old was 5.33，equivalent to 317 487.5 branches per hectare.The daily changes of the net photosynthetic rate of the two tree shapes showed asymmetric bimodal curve in the upper periphery，upper inner，middle periphery，middle inner，and lower periphery，and a single peak curve in the lower inner. The net photosynthetic rates of outer periphery of the two shapes were both higher than those of inner，and different tree shapes and different canopy azimuths had different peaks and valleys at different time. Except for the lower inner，the other canopies of the two tree shapes had midday depression of photosynthesis. Comparing the daily average value of the net photosynthetic rate of each canopy in the two shapes found that the value of the delayed‑open central leader system was higher of 6.972 6 μmol/（m²•s）. The leaf area index of the delayed‑open central leader system was significantly lower than that of the cylindrical shape，and the transmission coefficient for radiation penetration was significantly higher than cylindrical shape. The single fruit quality，fruit hardness，soluble sugar content and yield per plant of the delayed‑open central leader system fragrant pear were significantly higher than that of the cylindrical shape. The soluble solids in the upper periphery，upper inner，middle periphery，middle inner and lower periphery were significantly higher than the cylindrical shape. The results of correlation analysis showed that the net photosynthetic rate was extremely significantly positively correlated with transpiration rate and stomatal conductance（P<0.01），and was extremely significantly negatively correlated with intercellular CO₂ concentration（P<0.01），leaf area index and transmission coefficient for radiation penetration showed a very significant negative correlation（P<0.01）. 7‑year‑old Korla fragrant pear of delayed‑open central leader system was better than the cylindrical shape in terms of photosynthetic utilization efficiency,its transmission coefficient for radiation penetration was higher than that of the cylindrical shape,and it was also better than the cylindrical shape in terms of quality and yield per plant，but the yield per hectare was lower than that of the cylindrical shape.

Study on the Methods for Overcoming Interspecific Hybridization Barriers in Tulipa

ZHANG Yanqiu, XING Guimei, CUI Yuehan, YANG Di, QU Lianwei

2021, 50(8): 124-132. DOI: 10.15933/j.cnki.1004-3268.2021.08.015

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In order to overcome the interspecific hybridization barriers in Tulipa,and improve the seed setting rate，the interspecific hybridization was performed between the Tulipa wild species Tulipa altaica and three Tulipa cultivars Eskimo Chief,World’s Favorite and Strong Gold.The effects of different pollination treatments and different plant growth regulators on cross‑compatibility were studied.The results showed that the seed setting rate of interspecific hybridization in Tulipa was greatly influenced by different cross combinations and treatments. Under normal pollination condition,the ovary of the two
crossing combinations with World’s Favorite and Strong Gold as female parents could not swell，while the ovary swelling rate was 20.00% with Eskimo Chief as female parent,but no seeds were obtained.Bud pollination was not obvious to overcome the barrier of interspecific hybridization，and the ovary of three hybrid combinations could not swell and no seed was obtained. Delaying pollination，scratching stigma pollination，plant growth regulator treatments etc. had positive effects on overcoming the interspecific hybridization barriers. When diploid cultivar Eskimo Chief was used as female parent,26,19 hybrid seeds were obtained by delaying pollination and scratching stigma pollination treatments respectively，with the seed setting rate of 13.33%,6.67%,and the embryo rate of 42.31%,21.05% respectively.When the triploid cultivars World’s Favorite and Strong Gold were used as female parents,no seed was obtained despite a certain proportion of enlargement in all the treated ovaries，indicating that the interspecific hybridization between different ploidy had more serious hybridization barrier.

Physiological and Biochemical Mechanisms of Flower Color Change in Two Plants of Solanaceae Family

JIANG Duoduo, LI Lin

2021, 50(8): 133-145. DOI: 10.15933/j.cnki.1004-3268.2021.08.016

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By way of measuring the flower color phenotype，the pH value，the content of soluble sugar and soluble protein，the enzymatic activity of PAL（phenylalanine ammonia‑lyase） and CHI（halcone isomerase），the content of anthocyanin，flavonoid and carotenoid in six flowering phases of Brunfelsia acuminata and Solanum wrightii，this study explored their color changing mechanism，to provide a theoretical basis for flower color breeding. Results showed that Brunfelsia acuminata and Solanum wrightii displayed similar change trend of increasing first and then decreasing in the content of soluble sugar and soluble protein，the enzymatic activity of CHI，the content of anthocyanin during flowering，
while displayed different change trends in the pH value，the enzymatic activity of PAL，the content of flavonoid and carotenoid. The contents of flavonoid（except initial flowering phase）and anthocyanin（except late full‑bloom phase and end of the flowing phase）of Brunfelsia acuminata were higher than those of Solanum wrightii，and all reached the peak value in the late flower bud phase. Six chromatographic components were detected in the anthocyanin extract of both plants. The a2 and a3 components of Solanum wrightii were identified as cyanidin chloride and petunia chloride，and the b4
component of Brunfelsia acuminate was identified as petunia chloride，and the rest components needed to be further determined. The correlation analysis showed that the chromaticity value indexes（L*，a*，b*，C*and h） were significantly correlated with the content of anthocyanin during flowering process in Brunfelsia acuminata，but not with the content of flavonoid. The chromaticity indexes L*，a*，b*，and C*were significantly correlated with the content of anthocyanin and flavonoid during flowering process in Solanum wrightii. The results indicated that the flower color of Brunfelsia acuminata was connected with the content of anthocyanin，and the flower color change was caused by the decline of anthocyanin content.Some anthocyanin which was synthesized and accumulated during the bud stage degraded under the intense sunlight，resulting in the color fading. However，the flower color of Solanum wrightii was controlled by the content of anthocyanin and flavonoid，the flower color change was not only related to the decline of anthocyanin content，but also to the proportion changes of anthocyanin components.

Genome Sequencing of Two Pseudorabies Virus Variants and Amino Acid Mutation Analysis of Major Protective Antigen

GUO Zhenhua, XING Guangxu, WENG Maoyang, JIN Qianyue, QIAO Songlin, ZHANG Gaiping

2021, 50(8): 146-153. DOI: 10.15933/j.cnki.1004-3268.2021.08.017

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To investigate the genetic variations of pseudorabies virus（PRV）prevalent strains in China，we sequenced the full‑length genomes of two PRV（HeNLH/2017 and HeNZM/2017）isolated strains.Then，we conducted bioinformatics analyses，including phylogenetic analysis and amino acid mutation assay.The results showed that the length of genome was about 143 kb with a very high GC content of 73. 8%.The whole genomic sequences displayed 95. 28%—95. 37%，98. 70%—98. 80% and 98. 90%—99. 42%nucleotide homology with those of Bartha，classical PRV strains（Ea and Fa）and PRV variants（HeN1 and HN1201），respectively. Phylogenetic analysis indicated that the two PRV strains belonged to Genotype 2. 2 and were very close to the PRV variants. Furthermore，compared with the Bartha strain，extensive amino acid mutations were indentified in the major protective antigens of gB，gC and gD among the two PRV isolates and other PRV prevalent strains in China. They contained deletion of ⁷⁵SPG⁷⁷ for gB，insertion of ⁶³AAASTPA⁶⁹ for gC and ²⁷⁸S/RPRP²⁸¹ for gD. However，the gB，gC and gD were still very conserved between PRV variants（including the two PRV isolates）and early prevalent strains（Ea and
Fa），except that only a few amino acid substitutions were found.

Prokaryotic Expression of African Swine Fever Virus K145R Protein and Preparation of Monoclonal Antibody

GENG Xiaolin, SUN Jie, WANG Yanwei, HUANG Tian, LIU Peng, CAO Hongmei, PANG Wenqiang, HAO Liying, DENG Junhua, HUANG Yuxin, TIAN Kegong

2021, 50(8): 154-159. DOI: 10.15933/j.cnki.1004-3268.2021.08.018

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In order to develop an immunodiagnostic reagent for African swine fever virus（ASFV），the recombinant ASFV K145R protein was expressed by E. coli system，and the monoclonal antibody against K145R protein was prepared and identified. The results showed that the pET28a‑K145R expression vector was successfully constructed，and the soluble recombinant protein of K145R with the size of about 17 ku was obtained. Western blot analysis confirmed that recombinant K145R protein had a good
reactivity with ASFV positive serum. A monoclonal antibody 1D4 was obtained by cell fusion and screening. The titer was 1∶5 120 000 by ELISA. The heavy chain was identified as IgG2a and the light chain as κ. Western blot analysis showed that it could specifically recognize recombinant K145R protein.IFA test showed that it could react with ASFV. In summary，the soluble expression of K145R protein in E.coli and the preparation of monoclonal antibody were achieved.

Influence of Different Feeding Styles on Slaughter and Meat Quality Traits of Huainan Pigs

XING Baosong, WANG Jing, BAI Xianxiao, CHEN Junfeng, ZHANG Jiaqing, REN Qiaoling, GUO Hongxia, ZHANG Hua, CAO Hai

2021, 50(8): 160-165. DOI: 10.15933/j.cnki.1004-3268.2021.08.019

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To explore the influence of free‑range and captivity feeding styles on slaughter and meat quality of Huainan pigs，twenty well‑developed Huainan pigs with similar birth weight were randomly selected，and divided into two groups，free‑range group and captivity group. Five pigs were randomly selected in each group for slaughtering test when their body weight achieved around 100 kg. Slaughter traits，meat quality traits，the content of amino acid，fat，vitamin and mineral elements in longissimus dorsi（LD）muscle were compared between these two groups. The results indicated that compared with the
captivity group，the backfat thickness and intramuscular fat content were reduced by 33. 74%（P<0. 05）and 53. 25%（P<0. 05）in free‑range group，respectively. But single kidney weight was increased by 18. 85% in free‑range group compared to captivity group（P<0. 05）. Compared with captivity group，the eye muscle area and ham percentage in the free‑range group were increased by 2. 45% and 23. 37%，respectively. Drip loss in free‑range group was 45. 83% lower than that of captivity group. The content of umami amino acid was higher in free‑range group，but the content of essential amino acid was higher in
captivity group. In free‑range group，the content of vitamin B1 was 24. 24%（P<0. 05）higher than that in captivity group. The calcium and potassium contents in free‑range group were 44. 94%（P<0. 05）and 5. 54%（P>0. 05）higher than those in captivity group，respectively. In summary，the free‑range feeding style significantly reduced fat deposition，increased the eye muscle area and hind leg ratio，promoted the deposition of vitamin B1，calcium，potassium，and umami amino acid，and improved umami taste and tenderness. The free‑range breeding style could improve the meat quality traits of Huainan pigs.

Screening，Identification and Fermentation Properties of Aroma Producing Yeast for Kiwi Fruit

LU Yunfeng, LIU Zhixing, ZHANG Sipu, NIU Jiajia, HU Ruiying, TIAN Long, YANG Yongfeng

2021, 50(8): 166-173. DOI: 10.15933/j.cnki.1004-3268.2021.08.020

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In order to develop kiwi wine fermentation strains，aroma producing yeast was isolated from the leaves，branches，peel and soil under the wild kiwifruit trees in this study.A strain of aroma producing yeast was isolated by CO2 weight loss comparison，aroma smelling and kiwi fruit juice fermentation. It was identified as Hanseniaspora vineae，named X‑5，by morphological observation，physiological and biochemical tests and 26S rDNA gene sequence homology analysis. Then the tolerance tests of SO₂，alcohol and sugar and fermentation test were carried out. The results showed that strain X‑5 had strong aroma producing ability，could tolerate 12% alcohol，300 g/L sugar and 0. 20 g/L SO2. After fermentation，the liquor was clear and transparent，the color was yellowish and green，the aroma was strong，the alcohol content was 9. 6%，the vitamin C content was 0. 327 7 g/L，and the sensory quality was high. It can be used as a special strain for kiwi fruit wine fermentation or the starting strain for breeding and domestication.

Design of Automatic Control System for Greenhouse Environment

SHAN Huiyong, ZHANG Chenghao, LI Chenyang, ZHAO Hui, WEI Yong, GUO Xucun

2021, 50(8): 174-180. DOI: 10.15933/j.cnki.1004-3268.2021.08.021

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In order to realize the adaptive adjustment of greenhouse environmental parameters，based on the greenhouse light intensity and carbon dioxide optimization model，an automatic control system for greenhouse environment was designed. Real time acquisition of temperature，light intensity，CO₂ and other parameters in greenhouse was done by PLC，remote communication was realized through OPC（OLE for process control）communication protocol，combined with data read in real time，according to the optimization model，the optimal CO₂ concentration and light intensity of crop growth in different temperature environment were analyzed and predicted，gas fertilizer generator and fill light lamp were controlled. At the same time，for the convenience of operators monitoring greenhouse real‑time environmental change and making corresponding decision control，a human‑computer interaction GUI interface based on MATLAB was designed.The preliminary test results showed that the control system could be used for crop growth adaptive light and air replenishment.The increment of photosynthetic rate during the regulation period was 1 176. 918 μmol/（m²·s）.Data transmission was steady and real‑time feedback was good.The control system can automatically regulate the greenhouse environment，beneficial to the intelligent development of facility greenhouse.

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