Table of Content

Volume 51 Issue 3
15 March 2022
Contents
Reviews
Crop Cultivation & Genetic Breeding
Agricultural Resources and Environment
Plant Protection
Horticulture
Animal Science Veterinary Medicine
Agricultural Information and Engineering Agricultural Product...

Article List
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Reviews

Research Progress on Effects of Combined Application of Organic and Inorganic Fertilizers on Soil Fertility and Soil Environment

LIANG Lu, ZHANG Weijie, XU Bohan, ZHUANG Qiuli, JIANG Xiufang, HUANG Yubo

2022, 51(3):  1-11.  DOI: 10.15933/j.cnki.1004-3268.2022.03.001

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The technology of replacing part of chemical fertilizer with organic fertilizer is one of the important measures to develop green agriculture and plays a more and more important role.The effects and mechanism of combined application of organic and inorganic fertilizers on soil physical and chemical properties，soil microorganisms，soil nitrogen and phosphorus loss and heavy metals in farmland were reviewed，in order to provide theoretical reference for the rational combined application of organic and inorganic fertilizers.


Review of Crop Row Recognition Algorithms Based on Image Processing

DIAO Zhihua, YAN Jiaonan, ZHAO Suna, HE Zhendong

2022, 51(3):  12-19.  DOI: 10.15933/j.cnki.1004-3268.2022.03.002

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Reliable crop row recognition is the cornerstone of reliable navigation by intelligent agricultural robot，which can effectively reduce the damage of agricultural robot to crops.Traditional crop row recognition technology based on image processing includes image preprocessing，feature extraction and crop row fitting.The traditional crop row recognition methods such as Hough transform，least square method and vertical projection method are summarized，and other traditional crop row recognition methods are also summarized.With the development of smart agriculture，deep learning has attracted more and more attention in the agricultural field. Better collection of various features of images and effective combination with agricultural machinery are the differences between deep learning crop row recognition and traditional methods.From the above two aspects，the research on crop row recognition algorithm at home and abroad is systematically analyzed，and the existing problems of crop row recognition algorithm based on image processing are pointed out. Finally，the future research direction is prospected according to the present research situation.


Crop Cultivation & Genetic Breeding

Effect of Side Deep and Reduced Application of Slow⁃Release Urea on Yield and Quality of Rice in Middle and Lower Reaches of the#br# Yangtze River

JIANG Hengxin, HUANG Heng, WANG Yuan, ZHAO Can, WANG Weiling, HUO Zhongyang

2022, 51(3):  20-29.  DOI: 10.15933/j.cnki.1004-3268.2022.03.003

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The high⁃quality and delicious japonica rice Nanjing 9108（late maturing variety）and Nanjing 5718（medium maturing variety）were used as materials，and the conventional nitrogen application was used as control with nitrogen rate of 300 kg/ha，and the ratio of base and tiller fertilizer to panicle fertilizer of 7∶3.The side deep basal application amounts of slow⁃release urea were 0（A），10%（B），15%（C），20%（D），25%（E）and 30%（F）lower than base and tillering nitrogen rate of control，tiller fertilizer was not applied，and panicle fertilizer rate was the same to control.The effect of side deep and reduced application of slow⁃release urea on rice yield and quality in the middle and lower reaches of the Yangtze River was studied，so as to provide a basis for the better popularization and application of side deep application technology of slow⁃release fertilizer in agriculture.The results showed that under the same nitrogen application rate，the side deep application of slow⁃release urea could promote rice growth and significantly improve rice yield，improve rice processing quality，but reduce rice taste value. The yields of two rice varieties increased by 11.63% and 10.61% respectively compared with control.For the five nitrogen reduction treatments，with the decrease of nitrogen application rate，the growth of two rice varieties slowed down，the yield and its components decreased，the processing quality and nutritional quality decreased，and the appearance quality and eating quality improved. Compared with the control，B，C and D treatments promoted the growth of two rice varieties，significantly increased the yield，improved the processing and appearance quality；B and C treatments reduced the eating quality，and improved the nutritional quality；D treatment improved the eating quality，and reduced the nutritional quality；E and F treatments delayed the growth of two rice varieties，significantly decreased the yield，reduced the processing quality and nutritional quality，and improved the appearance quality and taste quality.Compared with control，the yields of two rice varieties of D treatment increased by 3.54% and 2.16% respectively，and the processing，appearance and eating quality improved. D treatment is a fertilization method with high quality and stable yield.



Effect of Nitrogen Application Rate on Yield，Nitrogen UtilizationEfficiency and Quality of Different Indica Rice Varieties（Lines）with High Quality

NIE Xinxing, ZHANG Minmin, DUAN Xiaoli, WU Maoqian, FENG Jingyun, LIU Bo, ZHOU Lei, YANG Li

2022, 51(3):  30-37.  DOI: 10.15933/j.cnki.1004-3268.2022.03.004

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In order to investigate the variation of yield，nitrogen utilization efficiency and quality of three indica rice varieties（lines）which were newly bred or mainly cultivated in Hubei Province under different nitrogen application rates and provide a theoretical basis for selecting nitrogen efficient varieties and rational application of nitrogen fertilizer in high⁃quality rice production，a split plot experiment was set.In the experiment，the nitrogen application rates（0，100，140，180 kg/ha，named as N0，N100，N140，N180 respectively）were used as the main plot，and three high⁃quality indica rice varieties（lines）including Fengliangyou 4，Ju2you60 and Eliangyou 263 were used as split block. The results showed that the application of nitrogen increased yields of Fengliangyou 4，Ju2you60 and Eliangyou 263 by 12.3%—18.8%，33.5%—50.3% and 24.1%—39.4% respectively.The yield order was Ju2you60≈ Eliangyou 263>Fengliangyou 4 under N140 and N180 treatments，and the main reason for the increase of yield was the increase of effective panicles number. With the increase of nitrogen application rate，only the partial factor productivity of nitrogen of the three indica rice varieties（lines）decreased significantly.On the whole，the agronomic efficiency and physiological use efficiency of nitrogen of Ju2You60 were higher than those of Fengliangyou 4 and Eliangyou 263，and the nitrogen use efficiency of Eliangyou 263 was higher than that of Fengliangyou 4 and Ju2you60. The partial factor productivity of nitrogen of Ju2you60 and Eliangyou 263 were almost the same，which were higher than that of Fengliangyou 4.The application of nitrogen significantly increased the protein content of three indica rice varieties（lines），and the protein content of Eliangyou 263 was the highest under N180 treatment，reaching 9.7%.In addition，nitrogen application significantly reduced the gel consistency of Fengliangyou 4；significantly increased the brown rice rate and polished rice rate，and decreased the chalkiness degree of Ju2you60；significantly increased the head rice rate and chalkiness degree of Eliangyou 263 under N140 treatment；significantly increased the polished rice rate of Fengliangyou 4，head rice rate of Ju2you60，and decreased the gel consistency of Ju2you60 under N180 treatment.However，the length⁃width ratio and amylose content of the three indica rice varieties（lines）did not change significantly after application of nitrogen.As a whole，Eliangyou 263 had the highest chalky grain rate，chalkiness degree，gel consistency and protein content，followed by Fengliangyou 4；Fengliangyou 4 and Ju2you60 had similar amylose content，which was higher than that of Eliangyou 263；the three indica rice varieties（lines）were similar in brown rice rate，polished rice rate，head rice rate and length⁃width ratio. In summary，Ju2you60 and Eliangyou 263 combined high yield and high nitrogen efficiency compared with Fengliangyou 4，but the response of the rice quality indexes to nitrogen application rate among the three rice varieties（lines）was different.


Effect of Sowing Date on Main Agronomic Characters and Grain Mineral Element Content of Quinoa

LIU Junna, KONG Zhiyou, ZHANG Ping, WANG Qianchao, HUAN Xiuju, LIU Yongjiang, LI Li, QIN Peng

2022, 51(3):  38-46.  DOI: 10.15933/j.cnki.1004-3268.2022.03.005

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The two lines of Dianli 1（red quinoa）and Dianli 2（white quinoa）were used to study the effect of different sowing dates（April 1，April 15，May 1，May 15，June 1，June 15，July 1 and July 15）on main agronomic characters and grain mineral element content of quinoa，so as to clarify the best sowing date.The results showed that with the delay of sowing date，the growth periods of red quinoa and white quinoagenerally first decreased and then increased，and reached the shortest under the sowing date of June 15；The plant height of red quinoa and white quinoa first decreased and then increased，finally tended to be stable，reached the lowest under the sowing date of May 15，the highest under the sowing date of April 1，and the plant height of red quinoa was higher than that of white quinoa under all sowing dates；The yield per plant of white quinoa generally first increased and then decreased，while that of red quinoa showed an increase⁃decrease⁃increase⁃decrease trend，both of which were the highest under the sowing date of May1，and the yield per plant of red quinoa was generally higher than that of white quinoa；The 1 000⁃grain weight of white quinoa and red quinoa generally first increased and then decreased，the 1 000⁃grain weight of red quinoa was the highest under the sowing date of April 15，while that of white quinoa was the highest under the sowing date of May 15，and the 1 000⁃grain weight of white quinoa was generally higher than that of red quinoa；The calcium content of grain generally first increased and then decreased，the calcium content of white quinoa grain was the highest under the sowing date of May 1，and that of red quinoa grain was the highest under the sowing date of May 15；The magnesium content of red quinoa grain generally first increased and then decreased，reached the highest under the sowing date of June 15，the magnesium content of white quinoa grain was relatively stable，reached the highest under the sowing date of May 15，and the magnesium content of red quinoa grain was higher than that of white quinoa grain；The ferrum content of white quinoa grain was the highest under the sowing date of April 1，that of red quinoa grain was the highest under the sowing date of April 15；The zinc content of red quinoa grain was the highest under the sowing date of June 15，for white quinoa grain，except the sowing date of May 15，the zinc content of early⁃sowing and early⁃ripening quinoa grain was higher than that of late⁃sowing and late⁃ripening quinoa grain，and the highest was under the sowing date of May 1.To sum up，the yield per plant and the contents of calcium，magnesium and zinc in grain of red quinoa are higher than those of white quinoa；The suitable sowing date can effectively improve the yield and grain mineral element content of quinoa，and the suitable sowing date is from May to June.



Effect of Different Priming Treatments on Seed Germination and Seedling Growth of Waxy Maize under Low Temperature Stress

CUI Zhengguo, CAO Qingjun, HU Bo, WANG jingyu, YANG Hao, LI Gang

2022, 51(3):  47-54.  DOI: 10.15933/j.cnki.1004-3268.2022.03.006

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In order to explore the effects of different priming treatments on low temperature resistance of waxy maize，Wannuo 2000 was used as material，early sowing was adopted in field，and the effects of seed priming with 0.5 mmol/L melatonin（MT），0.5 mmol/L spermidine（Spd），and 0.5 mmol/L MT + 0.5 mmol/L Spd（MT+Spd）on seed germination and seedling growth of waxy maize were studied under low temperature stress. The results showed that compared with the control without priming agent，the emergence rates of priming treatments（MT，Spd and MT + Spd）significantly increased by 26.05，27.09 and 31.25 percent points respectively，the emergence time significantly reduced by 3.75、4.50、5.25 days，the shoot height and uniformity of shoot height also significantly improved under low temperature stress，and MT+Spd treatment had the best effect. Meanwhile，priming treatment significantly increased the root surface area，root length，root diameter，dry weight of shoot，dry weight of root and root⁃shoot ratio of waxy maize seedling under low temperature stress，and MT+Spd treatment had the best effect.Correlation analysis showed that there were significantly positive correlations between dry weight of shoot and dry weight of root，root surface area，as well as root length.Thus，seed priming is a feasible way to enhance low temperature resistance under field condition，and MT+Spd treatment has the best effect.




Genetic Diversity Analysis and Elite Allele Mining in Main Sesame Cultivars Released in China

LI Chun, DUAN Yinghui, JU Ming, MIAO Hongmei, DU Hua, ZHANG Haiyang

2022, 51(3):  55-64.  DOI: 10.15933/j.cnki.1004-3268.2022.03.007

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Researching the diversity of cultivars and mining the elite alleles underlying important traits are critical for sesame（Sesamum indicum L.）molecular breeding. In order to analyze the diversity of sesame cultivars in China，and to accelerate the progress of molecular breeding in sesame，the genome sequencing data of 705 sesame germplasms（including 95 modern sesame cultivars in China，405 landraces and 205 foreign germplasms）were analyzed in this study.As a result，12 704 SNP/InDel variants，which commonly existed in sesame lines，were identified，and by comparing the nucleotide diversity of 95 modern cultivars and 405 landraces，the cultivars were found to have a lower diversity level（0.158）than the landraces（0.246）.Further，by using the permutation test method，2 483 and 115 loci were found to possess significantly lower and higher nucleotide diversity respectively in the cultivars，and 1 290 loci were found to be significantly divergent between the cultivars and landraces using the genetic differentiation index（Fst）as the proxy. By selecting the intersection of the loci that were significantly divergent between the cultivars and landraces and the loci that possessed significantly higher nucleotide diversity in the 95 modern cultivars，80 loci were harvested. Of the 80 loci，14 were annotated to have high impact effect on gene functions，including the two loci Chr2：8176782 and Chr10：3441593，which were located in the disease resistance related genes（C_2_2.176 and C37.81）. Further analysis showed that the allele frequency of the two loci increased by 12.93 and 3.44 folds respectively in the cultivars compared with the landraces，and thus might contribute to the breeding for disease resistance. To sum up，the findings above suggest that some loci are significantly divergent between the cultivars and landraces，and the loci may relate to the improvement of sesame cultivars.



Effects of Overwintering Mulching on Freezing Injury of Tea Plant and Spatio⁃Temporal Variation of Soil Temperature in Tea Plantation

YU Paolan, SUN Yongming, WU Yan, XIAO Xiaojun, LIN Xiaobing, ZHOU Ying, WANG Haihong

2022, 51(3):  65-72.  DOI: 10.15933/j.cnki.1004-3268.2022.03.008

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The objective of this study was to explore the effect of different surface mulching on low temperature freezing injury of tea plant，in order to guide tea farmers to effectively prevent freezing injury.The three materials including rice straw，white film and grass⁃proof cloth were used to mulch the surface of tea plantation，and no mulch was used as the control（CK），the spatio⁃temporal variation of soil temperature and the occurrence of tea plant freezing injury were compared，and then，the relationships between minimum surface temperature，daily range，duration time of low temperature and freezing rate of tea plant were anaylzed. The results showed that，compared with CK，the soil surface minimum temperatures mulched with white film，rice straw and grass⁃proof cloth were increased by 2.9，2.6，0.9℃；the freezing rates of tea plant mulched with white film and rice straw were significantly lower than CK and mulched with grass⁃proof cloth. White film mulching could increase the minimum and maximum soil temperature at different depths averagely by 1.0—2.9℃，1.0—4.7℃ compared with CK due to warming effect，and the warming effect was greater than other mulch materials. The rice straw mulching had a bidirectional regulation effect of decreasing high temperature and increasing low temperature，which increased the stability of soil temperature at the same depth. Among all the treatments，the daily range was the smallest and the cooling effect was the best in rice straw mulching.The cooling effect of grass⁃proof cloth mulching was lower than rice straw mulching，and its heating effect was lower than the white film mulching.The spatio⁃temporal variation of soil temperature was mainly reflected in the surface soil，and the temperature variation of underground soil was more gentle than that of surface soil.The peak time of soil temperature was delayed along with the increasing of soil depth.The freezing rate of tea plant was significantly correlated with the lowest mean surface temperature and the longest duration time of the surface temperature less than 0.Furthermore，the freezing rate of tea plant was extremely significantly correlated with the longest duration time of the surface temperature less than 4 ℃ .The surface temperature less than 0℃ was not the critical temperature of freezing for tea plant. When the surface temperature of tea plantation was from 0 to 4℃，the duration time was more than 7 d，and the daily duration time was more than 13.5 h，the tea plant could suffer from freezing.Therefore，white film and rice straw are recommended as mulching materials for preventing low temperature freezing injury in tea plantation. Moreover，rice straw is recommended for maintaining the stability of soil temperature.



Agricultural Resources and Environment

Microflora and Major Metabolic Pathways in Rhizosphere Soil of Rice⁃Crab Symbiosis System

SONG Yu

2022, 51(3):  73-83.  DOI: 10.15933/j.cnki.1004-3268.2022.03.009

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In order to clarify the microbial community composition and major metabolic pathways of ricerhizosphere soil in rice⁃crab symbiosis system，with rice monoculture applied with bio⁃organic fertilizer as the control，the rhizosphere soil of two kinds of rice fields at mature stage was sequenced by Illumina NovaSeq sequencing platform to analyze the difference of species and abundance of bacteria，fungi and archaea，and the difference of main metabolic pathways of microorganisms. The results showed that the bacterial community accounted for 98.09%，archaeal community accounted for 1.76%，and fungal community and others accounted for 0.15%. At the level of bacterial phylum，the dominant phyla of the two paddy fields were Proteobacteria，Actinobacteria，Chloroflexi and Acidobacteria，and the sum of the relative abundance of the four dominant phyla in two paddy fields was 90.86%；At the level of bacterial genus，the relative abundances of the top four bacterial genera in rice⁃crab symbiosis system were higher than those in the control group. At the level of archaeal phylum，there were four dominant phyla.The relative abundance of Thaumarchaeota was 4.41% in rice⁃crab symbiosis system，which was 9.19 times of the control group（0.48%）；At the level of archaeal genus，Methanospirillum and Methanothrix accounted for 53.63%，which were the main genera in the samples. At the level of fungal phylum，Ascomycota was the dominant phylum.The relative abundance of Ascomycota was 72.58% in rice⁃crab symbiosis system，which was 1.12 times of the control group（64.81%）；Among the top 10 fungal genera，the relative abundance of Chaetomium in rice⁃crab symbiosis system was 23.62%，which was 1.99 times of the control group（11.88%）；Trichoderma was 2.56%，which was 2.78 times of the control group（0.92%）.The results of metagenomic sequencing analysis showed that the number of species in rice⁃crab symbiosis system was higher than that in control group，regardless of bacteria，archaea or fungi.A total of 1 594 699 non⁃redundant genes were annotated into the KEGG metabolic pathway，belonging to 421 metabolic pathways.The relative abundance of the first 20 metabolic pathways in rice⁃crab symbiosis system was higher than that in the control group，indicating that the introduction of crab into the bio⁃organic fertilizer paddy field enriched the species composition of microorganisms and enhanced the metabolic capacity of species.




Effects of Organic Materials with Different Carbon Sources on Soil Carbon and Nitrogen and Bacterial Communities in Tobacco⁃Planting Soil

ZHANG Rui, WANG Huanhuan, ZHAO Yuanyuan, CHENG Yuyuan, WU Jiang, SHI Hongzhi

2022, 51(3):  84-94.  DOI: 10.15933/j.cnki.1004-3268.2022.03.010

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In order to explore the effects of organic materials with different carbon sources on carbon and nitrogen，bacterial communities and tobacco growth and development in tobacco⁃planting soil under the same carbon⁃nitrogen ratio，a pot experiment was conducted，adding biochar，decomposed wheat straw，decomposed cow dung and biogas residue，respectively.Four kinds of organic materials were mixed with inorganic nitrogen（nitrogen fertilizer）and adjusted to the carbon⁃nitrogen ratio of 30，and their effects on carbon and nitrogen and bacterial communities in tobacco⁃growing soil were studied.The results showed that the application of organic materials reduced soil bulk density and nitrate nitrogen content，increased the content of alkaline hydrolyzed nitrogen and ammonium nitrogen，and increased soil total carbon content and total nitrogen content.The application of decomposed wheat straw and decomposed cow dung increased soil microbial biomass carbon and nitrogen. It provided good environmental conditions for the growth and development of soil microorganisms，which in turn affected soil bacterial diversity and community structure. In terms of the overall effect of adding organic materials with different carbon sources，the combined application of decomposed wheat straw and inorganic nitrogen had better effects on improving the physicochemical properties and microbial diversity of tobacco⁃growing soil. Compared with the control（single application of nitrogen fertilizer），the soil bulk density decreased by 9.22% at 60 days after tobacco transplanting，and the total carbon，total nitrogen，microbial biomass carbon，microbial biomass nitrogen and bacterial OTUs（Operational taxonomic units） increased by 44.39%，21.06%，49.22%，53.37%，6.70% respectively；The dominant flora of soil bacteria were Actinobacteriota，Proteobacteria，Chloroflexi，Acidobacteriota，Gemmatimonadota，Firmicutes，Bacteroidota and Myxococcota.



Plant Protection

Effect of Fusarium verticillioides Dihydroorotate Oxidase on the Production of Fumonisins

LI Shaokuan, LIU Jing, ZHAO Nan, PANG Minhao, TANG Bowen, ZHAO Bin, LIU Yingchao

2022, 51(3):  95-103.  DOI: 10.15933/j.cnki.1004-3268.2022.03.011

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Fusarium verticillioides is one of the main pathogenic fungi that cause maize ear rot.Fumonisins（FBs）are a group of carcinogenic mycotoxins，produced as secondary metabolites by F.verticillioides，which severely reduce corn quality and then pose a threat to food security.DHOD gene encodes dihydroorotate oxidase，which is involved in the pyrimidine synthesis pathway and affects DNA and RNA synthesis in bio⁃orgamisms，thereby regulating the metabolism of secondary substances.Nevertheless，the mechanism underlying the regulation of DHOD gene on secondary metabolites remains unclear.This research aims to investigate the role of DHOD gene in the biosynthesis of fumonisins in F.verticillioides.Firstly，we analyzed the phylogenetic relationship of dihydroorotate oxidase across species，and then created DHOD-knockout mutants of F.verticillioides by homologous recombination. Secondly，we used HPLC（high performance liquid chromatography）to detect the difference in fumonisin production between the wild type and the DHOD-knockout mutants of F. verticillioides.Thirdly，by using RT⁃qPCR，we analyzed the expression levels of DHOD gene and associated genes of fumonisins synthesis in the wild type and knockout mutants.The sequence analysis results showed that the DHOD gene was conserved in Fusarium spp.，and we obtained the DHOD⁃knockout mutants of F. verticillioides using homologous recombination.Compared to the wild type，a significant reduction（88.32%—93.56%）in the fumonisin production was found in the DHOD⁃knockout mutants. Compared with the wild type，the expression levels of fum1，fum6，fum8 and fum13 were significantly reduced in the DHOD knockout mutants. In summary，the expression level of DHOD gene has a positive correlation with the fumonisins production.



Isolation and Identification of the Pathogen of Catalpa bungei Stem Rot and Differences in Disease Resistance of Different Catalpa Species

YANG Xiao, WANG Shuguang, LIANG Shen, LOU Changcheng, WANG Limin, SHI Man, ZHANG Hechen

2022, 51(3):  104-111.  DOI: 10.15933/j.cnki.1004-3268.2022.03.012

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In order to clarify the pathogenic cause of Catalpa bungei stem rot，and to understand the differences in resistance of different Catalpa species to the pathogen，fungi strains were isolated and purified from the diseased Catalpa tree plants by tissue separation method to verify their pathogenicity，and then morphological and molecular identification was carried out. The pathogen was inoculated into different Catalpa species by in vitro leaf puncture method and root dipping method，and activity changes of β⁃1，3⁃glucanase，superoxide dismutase and peroxidase were measured.The results showed that the pathogen of Catalpa bungei stem rot was Myrothecium roridum.In the in vitro leaf puncture test，the disease index of Catalpa bungei Luoqiu 1（CL1）was the lowest，which was 39.17.In the root dipping test，the disease index of Catalpa bungei var. Jinsi（CJS）was the lowest，which was 26.67.Among thethree Catalpa species，the β⁃1，3⁃glucanase activity of CJS changed the fastest，and its peak activity was 14.89% higher than that of CL1 and 7.12% higher than a hybrid of Catalpa bungei Yuqiu 2 and Catalpa fargesii Bur.，which was called CZ2.The SOD activity of CL1 increased the fastest. The POD activities of CL1 and CJS reached their peaks on the 3rd and 6th days respectively，and the peak value of CJS was 11.11% higher than that of CL1 and 25.00% higher than that of CZ2.By comprehensive consideration of disease index and enzyme activity changes，CJS was more resistant to stem rot.



Horticulture

Cloning and Expression Analysis of EIN3/EIL Family Genes in Paeonia ostii cv‘.Phoenix White’

HAN Bo, XU Lingxia, CAO Xiaoying, ZHANG Guangqin

2022, 51(3):  112-117.  DOI: 10.15933/j.cnki.1004-3268.2022.03.013

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Ethylene regulates plant nonbiological stress response and tolerance，and the EIN3/EIL family genes are key genes in the ethylene pathway. In this study we synthesized primers of three EIL genes referring to Paeonia suffruticosa and obtained three EIN3/EIL genes of Paeonia ostii cv.‘Phoenix White’，respectively named PoEIL1，PoEIL2，PoEIL3.Sequence analysis showed that they had high similarity to the Arabidopsis EIN3/EIL family genes，PoEIL1 clustered together with Arabidopsis AtEIL3 and PoEIL2 and PoEIL3 together with AtEIN3 and AtEIL1.Conservative motif analysis showed that each sequence had at least five conservative motifs at the same location.qRT⁃PCR analysis indicated that the three genes were expressed in each tissue，but the transcriptional levels varied.The expression level of the three genes in the petals of different flowering periods was also inconsistent.The expression level of PoEIL1 showed up⁃regulation at the opening stage，so it is speculated that PoEIL1 is related to flower development and flowering phase can be regulated by regulating PoEIL1 expression.


Study on Cell Immobilization Technology and Storage Conditions of Liquid Strains of Pleurotus eryngii

ZHOU Tingting, ZHOU Rongxiang, ZHANG Yao, CHEN Wencai, LI Jihong, MA Guizhen

2022, 51(3):  118-126.  DOI: 10.15933/j.cnki.1004-3268.2022.03.014

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In order to solve the shortcomings of the liquid strains of Pleurotus eryngii that are not easy to be stored and transported，the immobilization technology of Pleurotus eryngii liquid strain cells was studied by screening suitable immobilization carrier types and their mass concentrations，determining the effects of the volume ratio of liquid strain to carrier，the mass concentration of CaCl₂，and the crosslinking time on the formation of immobilized beads，and optimizing the immobilization conditions.The biological activity and storage stability of immobilized strains were determined by measuring the survival rate，mycelium growth rate，respiratory intensity and cellulase activity of immobilized strains stored at different temperatures for different time. The results showed that the immobilization conditions of Pleurotus eryngii liquid spawn were as follows：the mass ratio of sodium alginate to polyvinyl alcohol was 3∶7 as carrier，the mass concentration of carrier was 80 g/L，the volume ratio of liquid spawn to immobilization carrier solution was 5∶5，the mass concentration of CaCl₂ was 20 g/L，and the crosslinking time was 6 h.The immobilized strain was produced with regular spherical shape and fast mass transferability of 15 min，the mechanical strength was 60.1 g，and the mycelial growth rate was 200.1%.The survival rate and mycelial growth rate of the immobilized strains did not change significantly after 30 days of storage at 20℃ or 4℃，but the respiratory intensity decreased and the cellulase activity increased，which indicated that the immobilized strains still had high biological activity and it was conducive to the stability of the biological activity of the immobilized strains. The survival rate and growth rate decreased by 50 percentage points and 72.73%，respectively，after 10 days of storage at -20℃.Under this condition，the respiration intensity and cellulase activity were significantly reduced，and the biological activity of the strain was reduced，which was not conducive to the preservation of the strain.




Temperature Suitability Evaluation and Run Analysis of Tomato Cultivation in Wide⁃span Plastic Greenhouse with External Thermal Insulation

DONG Xiaoxing, YU Luming, NIU Xuxu, ZHANG Zengguang, LI Shengli

2022, 51(3):  127-138.  DOI: 10.15933/j.cnki.1004-3268.2022.03.015

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In order to make scientifically use of the wide⁃span plastic greenhouse with external thermal insulation（hereinafter referred to as WSTP）for production of tomato and carry out targeted environmental regulation，this study evaluated the tomato cultivation temperature suitability of three stubbles in the greenhouse by using the temperature suitability model based on the observed temperature data.The run analysis was carried out for the unsuitable period.The environmental limiting factors and suitable cultivation plan were screened，and the corresponding environmental control measures were put forward.The results showed that among the three stubbles in WSTP greenhouse in Henan Province，the suitability of winter⁃spring stubble was the highest，and the cumulative suitability（day air temperature+night air temperature+day ground temperature+night ground temperature）was 3.50.Over⁃winter stubble followed it，autumn⁃winter stubble had the lowest suitability，but there was little difference in cumulative suitability between them. The unsuitable periods in the shed were from August 31 to September 9 and from December 5 to February 23 of the following year. The most unfavorable environmental factor in the shed was night air temperature，which had the lowest suitability，the highest run intensity and the longest run time，followed by day air temperature，day ground temperature and night ground temperature.For tomato planting，the production plan in WSTP greenhouse should be arranged twice a year，which was more conducive.



Effect of Reduced Application of Chemical Fertilizer Combined with Application of Bio⁃organic Fertilizer and Irrigation Amount on Yield and Quality of Garlic

ZHANG Dan

2022, 51(3):  139-145.  DOI: 10.15933/j.cnki.1004-3268.2022.03.016

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The effect of reduced application of chemical fertilizer［675 kg/ha（F3），607 kg/ha（F2）and 540 kg/ha（F1）］combined with application of bio⁃organic fertilizer［3 000 kg/ha（O1），5 000 kg/ha（O2）and 8 000 kg/ha（O3）］on the yield and quality of garlic sprout and garlic was studied with Mousuan No.1 as material，so as to determine the optimal fertilizer combination，and on this basis，the effect of irrigation amount［800 m³/ha（I1）and 1 600 m³/ha（I2）］on the yield and quality of garlic was studied，so as to determine the best water and fertilizer management measures.The results showed that with the increase of the application rate of bio⁃organic fertilizer，the yields of garlic sprout and garlic increased first and then decreased，and the O2 treatment was the highest. Under O1 condition，with the increase of chemical fertilizer application rate，the yields of garlic sprout and garlic significantly increased；under O2 and O3 conditions，with the increase of chemical fertilizer application rate，the yields of garlic sprout and garlic decreased. O2F1 treatment had the highest yields of garlic sprout and garlic，which significantly increased by 2.79%—28.28% and 1.84%—21.44% compared with other treatments，respectively.With the increase of the application rate of bio⁃organic fertilizer，the contents of soil organic matter，alkali⁃hydrolyzed nitrogen，available phosphorus and available potassium all significantly increased.With the increase of bio⁃organic fertilizer application rate and the decrease of chemical fertilizer application rate，the allicin，soluble sugar，soluble protein and vitamin C（Vc）contents of garlic and garlic sprout gradually increased，and the nitrate content gradually decreased.The quality of garlic and garlic sprout of O3F1 treatment was the best，O3F2 and O2F1 treatments were better，and there was no significant difference among the three treatments in general. By considering the yield and quality of garlic and garlic sprout，O2F1 treatment was the best. Irrigation test was carried out on the basis of O2F1 treatment，and the results showed that compared with I1O2F1 treatment，the garlic yield of I2O2F1 treatment significantly increased by 4.4%，the contents of allicin，Vc and nitrate significantly decreased by 10.40%，4.19% and 9.38%，and the contents of soluble sugar and soluble protein significantly increased by 5.52% and 3.83%.By considering the yield and quality of garlic，I2O2F1 treatment was the best.



Animal Science Veterinary Medicine

Molecular Epidemiological Investigation and Genetic Evolution Analysis of ORF5 Gene of Porcine Reproductive and Respiratory Syndrome Virus in Henan Province in 2020

GUO Zhanda, DU Jimei, GUO Ziyi, WANG Xingang, ZHANG Yanhua

2022, 51(3):  146-153.  DOI: 10.15933/j.cnki.1004-3268.2022.03.017

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In order to monitor the prevalence and variation of porcine reproductive and respiratory syndrome virus（PRRSV）in Henan Province，a total of 401 clinical samples of porcine reproductive and respiratory syndrome were collected in Henan Province in 2020 and analyzed by TaqMan real⁃time quantitative PCR（FQ⁃PCR）.The results showed that a total of 100 positive samples were detected，with a positive rate of 24.9%. The ORF5 genes of 18 PRRSV epidemic strains were sequenced for homologous alignment and genetic evolution analysis. Nucleotide homology analysis showed that the ORF5 gene nucleotide homology among 18 strains was above 83.4%，and the nucleotide homology with the reference strain was 82.9%—99.7%.The results of genetic evolution analysis showed that all the 18 PRRSV strains belonged to American genotype，including lineage 1，lineage 5 and lineage 8.Two PRRSV strains belonged to lineage 8 represented by JXAl，one PRRSV strain belonged to lineage 5 represented by VR2332 and 15 PRRSV strains belonged to lineage 1 represented by NADC30. The results of amino acid sequence analysis showed that the 13th amino acid virulence related site of HN⁃TH1⁃2020，HY⁃PY1⁃2020，HN⁃XY1⁃2020 and the 151st amino acid virulence related site of HN⁃XC1⁃2020 had different degrees of mutation.

Establishment of Two⁃step Fluorescence Quantitative PCR Method for Detection of Deformed Wing Virus in Honeybee

WANG Hongkun, WANG Yihua, ZHOU Danyin, DONG Kun, ZHANG Xuan

2022, 51(3):  154-161.  DOI: 10.15933/j.cnki.1004-3268.2022.03.018

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In order to establish a more sensitive two⁃step fluorescence quantitative PCR detection method for deformed wing virus in honeybee，primers were designed according to the conservative gene fragment of deformed wing virus，and a two⁃step real⁃time fluorescence quantitative PCR detection method based on SYBR dye was established by conducting reverse transcription process and fluorescence quantitative PCR step by step，and its specificity，sensitivity，stability and feasibility were verified.The results indicated that the primer specificity of the two⁃step real⁃time fluorescence quantitative PCR method established in this study was good，and there was a good linear relationship in 6.58×10¹—6.58×10⁸copies/μL of plasmid standard.R² was 0.999 7 and the amplification efficiency was 100.8%.The sensitivity of the method was 6.58 copies/μL，and there were no cross⁃reactions between deformed wing virus and black queen cell virus，Israeli acute paralysis virus.The coefficient of variation was 0.20%—0.66% within the group and 0.08%—0.98% between the groups.The deformed wing virus infection rate of honeybee samples detected by this method was higher than that by ordinary PCR.The method showed good specificity，repeatability and practicability.In conclusion，a two⁃step real⁃time quantitative PCR method with higher sensitivity and better applicability was successfully established，which provides new technical support for epidemiological investigation，epidemic surveillance and early warning mechanism construction of bee deformed wing virus disease in China.



Optimization of Preparation Technology of Livestock and Poultry Qingfei Granules by Response Surface Methodology#

ZHANG Yan, HAO Jingyou, XUE Zhanmei, LIU Xuesong, SU Jing, TANG Wei, LIU Yankai, SUN Hongjie, SHI Tongrui

2022, 51(3):  162-172.  DOI: 10.15933/j.cnki.1004-3268.2022.03.019

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In order to solve the technical problems of low extraction efficiency of traditional Chinese herb，single factor experiment combined with response surface methodology was carried out to improve thepreparation process of livestock and poultry Qingfei granules，including fermentation extraction，extract extraction，alcohol precipitation，granule formulation，granulation，drying and straightening granulation，etc.，and the content of active ingredients of the modified livestock and poultry Qingfei granules was determined.The results showed that the optimal preparation conditions of livestock and poultry Qingfei granules were as follows：inoculation ratio was 3%，culture temperature was 37℃ and culture time was 72 h；the fermented traditional Chinese medicine was extracted twice with 8 times of water，1.5 h each time；the extractum was soaked in 70% ethanol under the condition of relative density of 1.35 for 24 h；the extractum was mixed with auxiliary materials at a ratio of 1∶7.6，and the auxiliary materials were lactose and dextrin with the ratio of 1∶2；the wet mixing time was 30 min with 80% ethanol，and then they were passed through a 1.40 mm aperture sieve，and mixed for 30 min；the wet particles were dried at 70℃ for 4 h and coarse granules could be prepared with a 1.40 mm aperture sieve of a rocking granulator. Compared with traditional livestock and poultry Qingfei granules，the contents of epigoitrin and quercetin 3⁃O⁃β⁃D⁃glucose⁃7⁃O⁃β⁃D⁃gentiobioside were increased by 22.67% and 25.39% respectively，and the contents of liquiritin and ammonium glycyrrhizinate were doubled in modified livestock and poultry Qingfei granules.



Agricultural Information and Engineering Agricultural Product...

Influence of Fan Frequency on Leaf Temperature，Yellowing and Dehydration during Tobacco Bulk Curing

LI Yulin, DING Yingfu, WANG Shenglei, DUAN Weidong, LI Jingchao, WANG Tingxian, SONG Zhaopeng, SONG Jiangyu

2022, 51(3):  173-180.  DOI: 10.15933/j.cnki.1004-3268.2022.03.020

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In order to provide a theoretical basis for the screening of fan frequency，influence of different fan frequency on leaf temperature，yellowing and dehydration during tobacco bulk curing was studied.The fan frequencies of 30 Hz，40 Hz and 50 Hz in the whole process were set，and the non⁃uniformity of leaf temperature，color parameters and water loss index of tobacco leaves at different temperatures were measured，and the relationship between non⁃uniformity of leaf temperature and yellowing dehydration coordination was analyzed.The results showed that the frequencies of the minimum leaf temperature unevenness at each temperature were 30 Hz（36℃），40 Hz（38℃），50 Hz（42℃），50 Hz（48℃），40 Hz（54℃），30 Hz（68℃）.In terms of yellowing of tobacco leaves，the yellowing degree and yellow⁃blue value of 30 Hz were the highest when the sampling temperature was 36—42℃，and the yellowing degree，brightness value and yellow⁃blue value of 50 Hz were the highest when the sampling temperature was48—68℃. In terms of water loss of tobacco leaves，the water loss degree of 50 Hz was the highest at the sampling temperature of 38—48℃ ；the thickness shrinkage rate of 30 Hz was the highest at the sampling temperature of 38—68℃ ；the horizontal curling degree and vertical curling degree first decreased and then increased，and overall reached the maximum at 50 Hz. The best frequencies of yellowing and dehydration coordination at each temperature were 30 Hz（36℃），40 Hz（38℃），and 50 Hz（42—68℃）.The uniformity of leaf temperature is a relatively coordinated feature of tobacco leaves turning yellow and losing water. The degree of non⁃uniformity of leaf temperature during the curing process can be used as an important indicator for screening the frequency of fans at different temperatures.