Table of Content

15 February 2024, Volume 26 Issue 2

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AGRICULTURAL INNOVATION FORUM

Research Progress on Mechanism of Brassinosteroids Regulating Plant Salt Tolerance

Chen CHEN, Dawei CHENG, Lan LI, Hong GU, Xizhi GUO, Ming LI, Jinyong CHEN

2024, 26(2):  1-12.  DOI: 10.13304/j.nykjdb.2022.0923

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Brassinosteroid（BR）， as a novel and highly efficient plant hormone， is widely involved in various physiological processes of plants and has important roles in alleviating abiotic stresses. This article elaborated the biological functions of BR treatment in alleviating salt stress， and summarized the BR signaling pathways in Arabidopsis thaliana， rice and several horticultural plants. By introducing the salt tolerance mechanism of BR， it further analyzed the response signal transduction pathways when the plants subjected to salt stress， which provided a theoretical basis for the application of BR in improving salt tolerance in plants.

Research and Application Progress of High Voltage Electrostatic Field in Aquatic

Peiliang CHEN, Minjun XU

2024, 26(2):  13-19.  DOI: 10.13304/j.nykjdb.2022.0928

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High-voltage electrostatic field （HVEF） technology involves the integration of physics， chemistry， electronics， biology and other disciplines. It has the advantages of low energy consumption， high efficiency， cleanliness and environmental protection. It is also an important developing direction for disinfection， sterilization， low-carbon and environmental protection in the future， and becomes research hotspot in agriculture field. To promote a more comprehensive development of HVEF， this paper reviewed the main research progress of HVEF in recent 20 years and especially its application in aquaculture， which was expected to promote its application in aquatic product treatment and aquatic breeding， based on the research and application of the biological effects of HVEF in biological breeding.

BIOTECHNOLOGY & LIFE SCIENCE

Physiological and Chemical Characteristics and Transcriptome Analysis of Different Type of Wheat Seedlings Under Salt Stress

Shuang LI, Aiying WANG, Zhen JIAO, Qing CHI, Hao SUN, Tao JIAO

2024, 26(2):  20-32.  DOI: 10.13304/j.nykjdb.2022.0679

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To explore the response difference of different types of wheat to salt stress， the leaves of ‘Bainong 889’ and ‘Chinese spring’ seedlings were subjected in 200 mmol·L^-1 NaCl solution. The physiological and biochemical characteristics of wheat seedling under salt stress were analyzed， and the differences of gene expression and the occurrence of variable shearing events were investigated based on transcriptomics data. The results showed that， under salt stress， the leaves of ‘Bainong 889’ had less chlorosis than ‘Chinese spring’； the leaves of ‘Chinese spring’ stained by Evans blue were greater and the relative activity of cells was significantly reduced. In addition， ‘Chinese spring’ wheat seedlings significantly wilted under salt stress， while ‘Bainong 889’ was no significant difference compared with control in the morphology. The relative electrolyte leakage rates of both cultivars under salt stress were higher than the control， and that of ‘Chinese spring’ seedlings was the highest. Under salt stress， the concents of chlorophyll a， chlorophyll b and total chlorophyll in leaves of both ‘Bainong 889’ and ‘Chinese spring’ seedlings decreased， and the net photosynthetic rate （P_n） and stomatal conductance （G_s） of ‘Chinese spring’ seedlings were significantly inhibited， while those of ‘Bainong 889’ were similar to the control. Transcriptome sequencing results showed that 2 299 differentially expressed genes were up-regulated and 2 527 differentially expressed genes were down-regulated in ‘Bainong 889’ than ‘Chinese spring’. GO （gene ontology） enrichment analysis of differentially expressed genes suggested that functions related to chloroplast， photosynthesis and plasma membrane were significantly enriched. KEGG （Kyoto encyclopedia of genes and genomes） enrichment analysis showed that pathways related to plant hormone signal transduction and plant secondary metabolites were significantly enriched. The analysis of variable shear events in transcriptome data showed that there were more variable shear events in ‘Chinese spring’ under salt stress， which were mainly concentrated in RNA transport and protein processing in endoplasmic reticulum. Therefore， wheat could responded to salt stress by regulating the expression of genes related to chlorophyll， plasma membrane， secondary metabolites and photosynthesis. The occurrence of variable shear events also indicated that two types of varieties regulated the gene interaction network distinctively under salt stress.

Comprehensive Evaluation of 69 Wheat Germplasm Resources

Yan JIN, Quanhao SONG, Jiajing SONG, Liang CHEN, Lishang ZHAO, Jie CHEN, Dong BAI, Tongquan ZHU

2024, 26(2):  33-45.  DOI: 10.13304/j.nykjdb.2023.0583

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In order to clarify the genetic differences and characteristics of wheat germplasms at home and abroad， and analyze their utilization efficiency， 41 new wheat varieties （lines） in Huang-Huai wheat region and 28 wheat resources introduced from CIMMYT （International Maize and Wheat Improvement Center） were taking as materials. 11 agronomic and yield traits including plant height， tiller number per plant， kernels per spike， 1 000-kernel weight， yield per plant， etc.， and 6 grain quality traits including protein， gluten， etc.， were comprehensively evaluated. 69 varieties were comprehensively analyzed and evaluated. The results showed that 69 wheat germplasm resources had abundant phenotypic diversity. The variation coefficient of agronomic and yield traits ranged from 11.87% to 57.86%， with an average of 22.00%， and the diversity index ranged from 1.83 to 2.09， with an average of 2.00. The variation coefficient of quality traits ranged from 1.61% to 13.21%， with an average value of 6.10%， and the diversity index ranged from 1.83 to 2.06， with an average value of 1.98. The principal component analysis showed that the accumulating contribution rate of 6 principal components was 80.27%， which represented 80.27% of the total amount of information， and plant height， biomass， yield， quality and flag leaf could be used as the main indexes to evaluate the quality of resources. The comprehensive score （F value） showed that some excellent germplasm with excellent comprehensive performance were screened， such as CIMMYT18， CIMMYT5， CIMMYT7， Xinong 1125 and Xinmai 58 etc.；the scores of imported resources were higher than those of wheat varieties （lines） from Huang-Huai region， and the germplasms ranking 1 to 25 were imported resources， which had high utilizable value and great application prospect. The 69 wheat germplasm resources were divided into 4 groups， which the clustering results were closely related to the parental origin of the germplasm， and there were obvious differences among the various groups. Group Ⅳincluded 5 varieties in Huang-Huai region and 17 CIMMYT resources， which displayed high water absorption， protein content， gluten content， hardness and settlement value. Above results showed that 69 wheat germplasm had the genetic diversity characteristics， and the utilization value in agronomy， yield and quality， which provided a theoretical basis for the breeding of new wheat varieties and the expansion of genetic diversity.

Development and Effect Evaluation of KASP Markers for Fiber Strength in Gossypium hirsutum L.

Lihua LI, Zhengwen SUN, Huifeng KE, Qishen GU, Liqiang WU, Yan ZHANG, Guiyin ZHANG, Xingfen WANG

2024, 26(2):  46-55.  DOI: 10.13304/j.nykjdb.2022.0818

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Fiber strength （FS） is an important indicator of fiber quality traits， and developing molecular markers associated with target traits would improve the selection accuracy and accelerate the process for breeding. The selected KASP markers were genotyped among 376 varieties （lines）， and then the polymorphism and the efficacy for higher strength were analyzed. 3 SNP markers showed polymorphic in the population， and the selection rate of FS-15 for high-strength fiber was 63.2% while FS-16 and FS-29 were 60.0% and 56.1%， respectively. Furthermore， the evaluation efficacy of different haplotype combination （Hap1， Hap2， Hap3 and Hap4） suggested that the accessions harboring Hap3 （TTA） showed higher fiber strength than those harboring other haplotypes. The selection rate of Hap3 for higher strength was 72.7%. In addition， Hap3 had no adverse effect on fiber length， boll weigh， seed index and lint index. The above results indicated these KASP markers and Hap3 could be used for selection of higher strength materials by providing genotypic information and did not adversely affect the fiber length， boll weight， seed index and lint index.

Identification and Characterization of IPT Gene Family in Brassica rapa L.

Bo LIU, Wangtian WANG, Li MA, Junyan WU, Yuanyuan PU, Lijun LIU, Yan FANG, Wancang SUN, Yan ZHANG, Ruimin LIU, Xiucun ZENG

2024, 26(2):  56-66.  DOI: 10.13304/j.nykjdb.2022.0373

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Isopentenyltransferase （IPT） is the first limiting enzyme for cytokinin biosynthesis， so to analyze the function of the IPT gene family in Brassica rapa L.， a total of 13 BrIPT genes were identified from the genome of Brassica rapa L. by bioinformatics， which were heterogeneously distributed on 7 chromosomes. These BrIPT geneswere divided into 4 substrates and each BrIPT gene included 8~10 conserved motifs and 1~2 UTR regions. The promoter regions of BrIPT genes contained numerous responsive elements. The BrIPT genes were regulated by environmental factors， biohormones， defense and stress. The qRT-PCR results showed that the tRNA-IPT genes BrIPT4， BrIPT6 and BrIPT9 were expressed in various parts of Brassica rapa L.. The expressions of IPT genes in Brassica rapa L. were higher at the mature stage than those at the seedling stage. The above results provided some basis for subsequent in-depth studies on the physiological and biochemical functions of IPT gene family members.

Effects of Different Nitrogen Supply Level on Plant Growth and Development in Miniature Potted Rose

Tingting CAO, Chun LIU, Youwei FAN, Li MA, Zhiyu REN, Suxia YUAN, Junyun ZHANG, Zunyao QIAN, Guangzhao YANG

2024, 26(2):  67-79.  DOI: 10.13304/j.nykjdb.2022.0844

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In order to study the effects of different nitrogen application levels on growth and development of miniatured potted rose， ‘Juicy Terrazza’ ‘Cinderella’ ‘Rosa Terrazza Pisce’ and ‘Apollo Jewel’ 4 miniature potted roses varieties were as experimental materials， and 4 nitrogen application rates of 0 （N1）， 70 （N2）， 140 （N3） and 210 mg·L^-1 （N4） were set. 11 traits including measuring vegetative， reproductive growth， and chlorophyll fluorescence parameters were analyzed by the methods of principal component analysis and hierarchical cluster analysis. The critical nitrogen content in leaves at each growth and development stage was determined， and the critical nitrogen content dilution curve of miniatured potted rose was constructed. The results showed that the 11 traits varied in different degrees among different nitrogen application treatments. There were some correlations among traits， thus， a single trait could not objectively and comprehensively evaluate the effect of nitrogen application levels on the growth and development of miniature potted roses. Principal component analysis was used to the principal components， and the cumulative contribution rates of which in all cultivars were more than 93.500%. Hierarchical cluster analysis showed that the 4 nitrogen treatments of ‘Juicy Terrazza’ were divided into 2 groups， and the other varieties were divided into 3 groups. Based on the comprehensive evaluation score of principal component analysis and cluster analysis results， considering the principle of reducing fertilizer and increasing benefits， the optimal nitrogen content in ‘Juicy Terrazza’ ‘Cinderella’ and ‘Rosa Terrazza Pisce’ was 140 mg·L^-1 each time， but that in ‘Apollo Jewel’ was 210 mg·L^-1. According to the comprehensive analysis， the N1 and N2 treatments were divided into nitrogen-limited treatment， and the N3 and N4 treatments were divided into non-nitrogen-limited treatment， and based on leaf area index the leaf nitrogen content dilution curves were constructed. Above results provided a reference for precision nitrogen application.

Study on Recovery Characteristics of Photosynthesis in Alfalfa Leaves After Low Temperature Stress

Yu MIAO, Jie WANG, Yaoyao ZHAO, Lijia ZHANG, Meijun LIU

2024, 26(2):  80-89.  DOI: 10.13304/j.nykjdb.2022.0873

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In order to investigate the restriction sites during the recovery of photosynthesis in alfalfa after the low temperature was lifted， using‘Xinmu 4’and‘ Gannong 5’2 varieties of alfalfa as materials， the changes in photosynthetic activity of alfalfa leaves during the low temperature stress and recovery process were determined. The results showed that chlorophyll content， photosynthetic rate and photosystem Ⅱ（PSⅡ） activity of the 2 alfalfa varieties decreased significantly under low temperature stress， while non-photochemical quenching factor （NPQ） increased significantly， but had no significant effect on photosystem Ⅰ （PSⅠ） activity； the chlorophyll of‘ Gannong 5’ was more sensitive to low temperature and its content decreased more. The activity of PSⅡ and photosynthetic rate of both varieties of alfalfa increased after 72 h recovery from room temperature but did not recover to the pre-treatment level； NPQ slowly decreased and dropped to the pre-treatment level， but NPQ of ‘Gannong 5’ was higher than that of ‘Xinmu 4’ during the recovery process， which indicated that ‘Gannong 5’ still had excess light energy production to be consumed through NPQ during the recovery process. The electron transport chain of alfalfa leaves was severely restricted at low temperature， and the restriction on the PS Ⅱ acceptor side at low temperatures was largely lifted after 72 h of recovery， while the PS Ⅱ donor side was significantly restricted after 72 h of recovery. The efficiency of energy captured per unit reaction centre for electron transfer （ET_O/RC） and energy captured per unit reaction centre for reducing Q_A （TR_O/RC） of both alfalfa varieties decreased significantly at 72 h of low temperature treatment and recovered to pre-treatment after lifting the low temperature， and there was no significant difference between the 2 varieties. The energy dissipated by the unit reaction center （DI_O/RC） of both alfalfa varieties did not recover to the pre-treatment level， and the energy required for recovery of ‘Gannong 5’ at 72 h was significantly higher. In summary， low temperature stress significantly injured the photosynthetic machinery of alfalfa leaves reducing photosynthetic rate， which was not restored to pre-treatment levels and was strongly correlated with PSⅡ activity， and the site limiting photosynthetic recovery was on its donor side. Non-photochemical quenching was the main way to restore 24 h excess light energy.

Comprehensive Evaluation of 5 Cultivation Types of Medicinal Chrysanthemum morifolium Ramat. at Branching Stage

Panpan MENG, Haiyan HE, Yuxin CAO, Lixin ZHANG, Qinghao LYU, Ruilin QI, Hongrui ZHANG

2024, 26(2):  90-99.  DOI: 10.13304/j.nykjdb.2023.0519

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In order to investigate the impact of drought stress on growth of different cultivated types of medicinal Chrysanthemum morifolium Ramat. at branching stage， as well as to assess their drought resistance， 5 cultivation types of medicinal C. morifolium were used as experimental materials， and the agronomic traits， bioaccumulation， physiological characteristics and related indicators of each material were measured. The membership function method， principal component analysis， and correlation analysis were employed for a comprehensive evaluation of drought resistance. The results showed that under drought stress， plant height， stem diameter， branch number and biomass accumulation decreased in the branching stage. The contents of proline， soluble sugar and soluble protein were generally increased. The anti-stress enzyme activity was enhanced， and the degree of membrane lipid peroxidation was aggravated. The comprehensive evaluation of drought resistance showed that the drought resistance of each cultivation type of medicinal C. morifolium during the branching stage was ranked as follows： Huaiju （‘Huaibaiju’） > Chuju > Boju > Huangju > Hangju （‘Dayangju’）. The results could provide a foundation for medicinal C. morifolium germplasm selection and promoting high-yield and efficient cultivation practices.

Characteristics of Temporal Expression of Key Genes During Differentiation from Sheep Preadipocyte

Guangjun YANG, Yuan LIAO, Zhichao ZHANG, Rongzhen ZHONG, Ziyuan DUAN

2024, 26(2):  100-108.  DOI: 10.13304/j.nykjdb.2022.0930

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To analyze the expression pattern and characteristics of key genes in sheep preadipocytes during adipogenic differentiation， 3T3-L1 cell line was selected as reference， the preadipocytes were isolate from the tail adipose tissue of female lambs from Hulun Buir sheep 20-day after birth using tissue digestion method and prepared into primary cell line. The 3T3-L1 cell line and sheep preadipocyte cells were induced to differentiate into mature adipocyte by cocktail method， and the temporal expressions of key genes during differentiation were detected by real-time fluorescence quantitative PCR. The results showed that the adipogenic differentiation of sheep precursor adipocytes took longer， and some cells still had differentiation ability after 16 d of induction. The patterns of temporal expression indicate that the expressions of DLK1， SREBP1， ACC1 and FASN were basically consistence both in the sheep preadipocytes and 3T3-L1 cell lines， the expression trends of ZFP423， ADIPOQ and C/EBPα were highly similar， however， the genes of PPARγ， FABP4， LPL and HSL were different. Compared with 3T3-L1 cell line， the relative expression levels of ZFP423， ADIPOQ， C/EBPα and FABP4 in sheep preadipocytes still remained at a high level in the middle and late stages of induced differentiation， and the relative expressions of PPARγ， LPL and HSL showed a continuous increase trend， which illustrated that sheep preadipocytes had its own characteristics in adipogenic differentiation. It was speculated that the cells from the sheep adipocytes synthesized triglycerides and then deposited in mature adipocytes through ingesting fatty acids from the external environment after 16 d of induction according to the expression of related genes. It indicated that sheep adipose cells had strong ability to absorb external fatty acids， which might be one of reasons for the excessive deposition of subcutaneous fat in native sheep. So the sheep adipocyte was more suitable as a cell model in research of sheep fat metabolism， which provided reliable experimental model for research of sheep fat metabolism.

INTELLIGENT AGRICULTURE & AGRICULTURAL MACHINERY

Extraction of Winter Wheat Planting Area Based on Fusion Features of HJ-1 and GF-1 Image

Hong ZHANG, Weiguo LI, Xiaodong ZHANG, Bihui LU, Chengcheng ZHANG, Wei LI, Tinghuai MA

2024, 26(2):  109-119.  DOI: 10.13304/j.nykjdb.2022.0703

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In order to improve the accuracy of extracting large-scale winter wheat planting area from the data of domestic environment and disaster monitoring and forecasting satellite （HJ-1/CCD）. This study took Shuyang County， Suqian City， Jiangsu Province as the research area. The fusion and object-oriented classification of the 30 m×30 m HJ-1/CCD multispectral image and the 2 m×2 m GF-1 panchromatic image （GF-1/PMS） at the jointing stage of winter wheat were carried out. The GF-1/PMS panchromatic images were resampled at 8， 16 and 24 m， and panchromatic images with four spatial resolutions （including 2 m） were obtained， which were fused with HJ-1/CCD multispectral images by Gram-Schmidt （GS）， respectively. Through the quality evaluation of the fused image， the appropriate scale image suitable for the pattern of winter wheat planting fields in the study area was selected. The HJ-1/CCD multispectral image was resampled to obtain an image with the same scale as the appropriate scale fused image. In the 2 scene images， the training samples SFI （samples of fused image） and SRI （samples of resampling image） containing spectral and texture information were selected respectively， the object-oriented classification method was used to extract the planting area of winter wheat from fused image （FI） and resampling image （RI）. The results showed that the fusion effect of 16 m×16 m fused images was better than 2 m×2 m， 8 m×8 m and 24 m×24 m fused images， and the mean， standard deviation， average gradient and correlation coefficient were 161.15， 83.01， 4.55 and 0.97. After object-oriented classification， the overall accuracy of SFI for the classification of resampled image RI_16m was 92.22%， and the Kappa coefficient was 0.90. The overall accuracy of SFI for the classification of fused image FI_16m was 94.44%， and the Kappa coefficient was 0.93. The overall accuracy of SRI for the classification of resampled image RI_16m was 84.44%， and the Kappa coefficient was 0.80. The classification effect of SFI for the fused image FI_16m was the best， indicating that the object-oriented classification method combined with the fused image and the extraction samples of fused image （SFI） could accurately extract the winter wheat planting area. In addition， the object-oriented classification method combining resampling image and the extraction samples of fused image （SFI） could also better extract the winter wheat planting area. This method provided a reference for the effective extraction of large-scale winter wheat planting area information combined with domestic medium-spatial resolution HJ-1/CCD images and GF-1 satellite images.

Development and Experiment of an Electronic Forest Compass Instrument

Hening FU, Zhongke FENG, Linhao SUN, Jueying SU, Tiantian MA, Zhichao WANG

2024, 26(2):  120-126.  DOI: 10.13304/j.nykjdb.2022.0763

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Tree height and tree position are important indicators in empirical forest decision-making and are commonly used to estimate standing parameters such as forest growth， tree age， wood volume， biomass and carbon stock， and the accuracy in the measurement has a significant impact on the evaluation of standing wood quality and estimation of forest growth. This study addressed some of the drawbacks within the present forest survey， such as high device price， not easily moved， long duration of one measuring procedure， and labor-intensive. Aming at those drawbacks， a novel electronic compass device was developed for forest survey based on three-dimensional electronic compass and laser ranging radar. The device consisted of embedded hardware and software and an Android application was developed to support user to operate this device. Overall， it realized the electronic and digital measurement of tree height and tree position by one device. After then， 59 standing trees were employed to verify the accuracy and efficiency of the device. The results showed that： in terms of accuracy， the mean absolute error of azimuth was 4.5° and the accuracy of tree height was 96.95%； in terms of operational efficiency， this device was about twice as efficient as the total station. In conclusion， the developed device in this study had the features including portable， low manufacturing cost， and internal and external operations integration. Only in this way， it could accord with various technical provisions such as accuracy in the continuous forest inventory （CFI） in China， which would have broad application prospects in forestry resource inventory.

Measurement of Contact Parameters and Discrete Element Simulation Calibration of Rice Bran and Broken Rice

Lin CHEN, Nanhui YU, Lizong WANG, Jijun FAN, Gang LEI, Xiaopeng LIU, Long ZHOU, Jin ZHOU

2024, 26(2):  127-136.  DOI: 10.13304/j.nykjdb.2022.0835

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In order to determine the contact parameters required for the discrete element simulation of rice bran and broken rice separation equipment， the contact parameters of rice bran and broken rice particles were measured and calibrated through bench test and simulation test. By free-fall impact method， impact diffusion method， inclined plane sliding method and inclined plane rolling method respectively， it was calibrated that the collision recovery coefficient between broken rice and acrylic was 0.45， static friction coefficient was 0.38， rolling friction coefficient was 0.17， the collision recovery coefficient between rice bran and acrylic was 0.15， static friction coefficient was 0.76. Based on the measurement test of repose angle and MATLAB image processing technology， the repose angle of particle reactor was obtained. Through the steepest climb test and quadratic orthogonal rotation combination test， a quadratic regression model between the influencing factors and repose angle was established， and the minimum relative error of the repose angle was taken as the constraint condition to optimize the regression equation. The optimal parameter combination was obtained： static friction coefficient 0.60， rolling friction coefficient 0.40， collision recovery coefficient 0.61 for broken rice； static friction coefficient was 0.81， the rolling friction coefficient was 0.18， and the collision recovery coefficient was 0.15 for rice bran. The results provided reference for the discrete element simulation parameter setting of rice bran and broken rice separation equipment.

ANIMAL AND PLANT HEALTH

Research Progress of Cyprinid Herpesvirus Ⅱ （CyHV-Ⅱ） Vaccine

Linjiao WU, Lifang SUN, Chunhua ZHU, Hong ZHANG, Panpan DONG, Leiqing CHEN, Yunkun WU

2024, 26(2):  137-144.  DOI: 10.13304/j.nykjdb.2022.0775

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Cyprinid herpesvirus Ⅱ （CyHV-Ⅱ） is the cause of （goldfish haematopoietic necrosis， GFHN），which can infect goldfish， Crucian carp and their hybrid at any developmental stage. It is a highly contagious disease with high mortality rate， which has caused huge economic losses to goldfish and Crucian carp aquaculture in China. The research and development of fish vaccine is the most effective way to prevent and treat the disease. The research and development progress of inactivated CyHV-Ⅱ vaccine， subunit vaccine， nucleic acid vaccine， live vector vaccine and Nano-delivered vaccines were reviewed in this paper. Furthermore， the application prospect of fish vaccine nano materials was discussed， which would provide new technical methods and new ideas for the prevention and treatment of goldfish herpes disease and fish vaccine research.

Immune Effect of Nocardia seriolae Caviar Vaccine on Grouper

Haofeng GAO, Peng SHAO, Zun WU, Yating WU, Xiaoyu WANG

2024, 26(2):  145-152.  DOI: 10.13304/j.nykjdb.2022.0653

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In order to explore the immune effect of inactivated vaccine of Nocardia seriolae on grouper， the immune protection of the vaccine on grouper was analyzed by the expression of immune gene. In this study， formalin-inactivated vaccine was prepared by this bacteria， and the pearl gentian grouper was immunized by intraperitoneal injection. The expression changes of TLR2， MyD88， TNF-α and IL-1β immune gene in the kidney， liver and spleen of the fish were detected by fluorescence quantitative method. The results showed that by injecting inactivated vaccine， the expressions of TNF-α and IL-1β genes were higher in kidney tissues， MyD88 gene’s expression was higher in spleen tissues， and TLR2 gene’s expression was higher in liver tissues. Except for a trend of decreasing relative expression of TNF-α gene in the tissue spleen， and the other genes showed up-regulation in all organs compared with that in blank group. Above results provided a theoretical basis for exploring the interaction between pearl gentian grouper and Nocardia seriolae， and laid a foundation for the development and application of fishery vaccines.

BIO-MANUFACTURING & RESOURCE AND ECOLOGY

Effects of Long-term Manure Fertilizer on Saline-sodic Properties and Stoichiometric Ratio of Saline-sodic Soil in Songnen Plain

Cheyu ZHAI, Jingmei LUO, Changjie LIU, Juan ZHANG, Qingfeng MENG

2024, 26(2):  153-161.  DOI: 10.13304/j.nykjdb.2022.0549

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Saline-sodic soil has a high degree of salinization and poor nutrient availability， the application of organic fertilizer can promote soil salt leach， reduce the degree of soil salinization， and increase the nutrient content and the supply capacity of soil carbon， nitrogen and phosphorus. According to the years of applying cattle manure in the soil， 4 treatments were conducted with different application years of cattle manure for 21 （Y21）， 18（Y18）， 15（Y15） and no application of cattle manure （CK）. Soil samples in each treatment were collected from 0 to 20 and 20 to 40 cm soil layers to explore the soil saline properties and nutrient content. The results showed that soil organic carbon （SOC）， total-N （TN） and total-P （TP） contents of Y21，Y18，Y15 treatments were significantly increased than CK， but soil water-soluble salt ions， pH， electrical conductance （EC）， sodium adsorption ratio （SAR） and exchangeable sodium percentage （ESP） were significantly decreased compared to CK at both 0 to 20 and 20 to 40 cm soil layers. At 0 to 20 and 20 to 40 cm soil layers， carbon nitrogen ratios（C/N） of Y21，Y18，Y15 treatments significantly reduced compared to CK. At 0 to 20 cm soil layer， nitrogen phosphorus ratios（N/P） of Y21， Y15 treatments significantly increased from 0.95 to 1.84 and 2.11. At the 20 to 40 cm soil layers， N/P of Y21， Y15 treatments significantly increased from 1.15 to 2.25 and 2.44. The carbon phosphorus ratio（C/P） of each treatment had no similar regularization compared to CK at 0 to 20 cm soil layer and no significant difference was observed at 20 to 40 cm soil layer. The multivariable linear regression model showed that C/N was affected by SOC， TN， and ESP， while N/P and C/P were mainly regulated by TP and TN. In conclusion， the long-term application of manure fertilizer could reduce the degree of soil salinity and C/N， increase the contents of SOC， TN， TP and N/P. The nutrient supply of soil reached its maximum at 15 years of manure fertilizer application.

Effects of Different Nitrogen Sources Addition on Humification of Coconut Leaf Compost

Xiaohong HUANG, Jing JIAO, Jihua DU, Yi WU, Zunxiang LI, Xinpeng LIU

2024, 26(2):  162-170.  DOI: 10.13304/j.nykjdb.2023.0650

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In order to investigate the effect of different nitrogen sources on the humification of coconut leaf compost， adding chicken manure， pig manure， biogas residue and urea as control variables to co-compost with coconut leaves，the indexs of temperature， the ratio of carbon to nitrogen ratio（T value）， humic substances， huminification rate， humification index and degree of humus polymerization were analyzed. The results showed that the 4 treatments could maintain the high temperature period （above 55.0 ℃） for more than 15 d， reached the harmless requirements. The T value was less than 0.6， which met the requirements of maturity. Compared with pre-fermentation and post-fermentation， composting led to a decrease in total oganic carbon， but humic substances carbon of 4 treatments increased， and the degree of humification increased. Chicken manure had the best effect on promoting the humification process of compost， after the composting， huminification rate increased by 19.28%， humification index increased by 65.80%， and degree of humus polymerization was 2.38. Adding pig manure as nitrogen source was lower than adding chicken manure， higher than adding biogas residue and urea. The huminification rate of adding biogas residue compost was the highest， but degree of humus polymerization was only 1.61， and the stability of compost products was poor. The decrease of humification index and degree of humus polymerization in coconut leaf compost with urea as nitrogen source hindered its humification process. In conclusion， humification effect was best when adding chicken manure as a nitrogen to source coconut leaf compost， and the results provided a scientific basis for the resource utilization of coconut leaf waste in Hainan.

Effects of Co-incorporation of Rice Straw and Chinese Milk Vetch on Photosynthetic Characteristics， Yield and Quality of Rice

Zhongyi LI, Hongqin TANG, Wenbin DONG, Caihui WEI, Tieguang HE

2024, 26(2):  171-180.  DOI: 10.13304/j.nykjdb.2022.0627

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To evaluate the effects of co-incorporation of rice straw and Chinese milk vetch on photosynthetic characteristics， yield and quality of rice， the conducting randomized field experiments were carried out with 6 treatments of different cultivation practices， including no fertilizer （CK）， normal nitrogen fertilizer （N）， normal nitrogen fertilizer+rice straw return （RN）， normal nitrogen fertilizer+Chinese milk vetch return （MN）， normal nitrogen fertilizer+rice straw+Chinese milk vetch return （MRN）， 60% nitrogen fertilizer+rice straw+Chinese milk vetch return （MRN60%）. The chlorophyll content （SPAD value）， net photosynthetic rate （P_n）， stomatal conductance （G_s）， intercellular CO₂ concentration （C_i） and transpiration rate （T_r）， dry matter accumulation， yield and rice quality were measured under different treatments. The results showed that， compared with CK， N， RN， MN， MRN and MRN60% treatments could significantly increase the SPAD value， P_n， G_s， yield， brown rice rate， head rice rate and protein content， and decreased the amylase content. Compared with N treatment， MRN treatment significantly increased SPAD value， P_n and G_s by 2.36%， 5.95%， 7.80%， respectively. Meanwhile， MRN treatment significantly increased dry matter accumulation at tillering stage， heading stage and milk-ripe stage by 14.77%， 3.52%， 4.95%， respectively. Especially， MRN treatment significantly improved the effective spike number， grain number and rice yield by 7.38%， 3.08% and 15.96%， respectively. Additionally， MRN treatment significantly increased the head rice rate by 4.63% and significantly reduced amylase content by 6.54%. However， MN treatment only significantly increased P_n by 4.75% compared to N treatment， MRN60% treatment only significantly reduced amylase content by 7.00% compared to N treatment. The correlation analysis showed that SPAD value， P_n and G_s extremely significantly positively correlated with effective spikes， grains per panicle， yield， brown rice rate and head rice rate. However， the SPAD value， P_n and G_sextremely significantly negatively correlated with amylase content. The SPAD value and P_n extremely significantly positively correlated with protein content， and G_ssignificantly positively correlated with protein content. In conclusion， with conventional nitrogen application， the co-incorporation of rice straw and Chinese milk vetch could significantly improve SPAD value， P_n， G_s， yield and cooking quality. With 60% nitrogen application， the co-incorporation of rice straw and Chinese milk vetch could stabilize the photosynthetic characteristics and maintain the yield and quality of rice， which could provide reference in high quality rice production.

Effects of Wheat Root Exudates on Bitter Gourd Seeding Growth and Soil Environment

Jingyun ZHANG, Feng GUAN, Bo SHI, Xinjian WAN

2024, 26(2):  181-190.  DOI: 10.13304/j.nykjdb.2022.0682

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In order to solve the problem of continuous cropping in bitter gourd， the bitter gourd seedlings was treated with allelopathic wheat root exudates， and the changes of enzyme activity and microbial diversity of bitter gourd rhizosphere soil were analyzed compared with untreated varieties.The results showed that the plant height， stem diameter， root length and biomass of 3 bitter gourd materials treated by the wheat root exudates was higher than the control with a significant or extremely significant difference， and the plant height and stem diameter especially showed an increasing trend over growth days. The plant height and stem diameter of the treatment groups increased by 7.32%， 14.67%， 19.98% and 7.98%， 8.24%， 10.89% at 5， 10 and 15 d， respectively. The root length， fresh weight of aerial part and root， shoot and root dry matter weight of treatment groups increased by 16.29%， 18.08%， 33.74%， 31.70% and 27.21%， respectively. And there was a significant or extremely significant difference in the activity of urease， polyphenol oxidase， transferase and catalase in rhizosphere soil of bitter gourd seedlings between treatment group and control group with increase of 84.88%， 35.68%， 443.73% and 54.39%， respectively. The amount and richness of bacteria and actinomycetes increased， while fungi and fusarium oxysporum decreased， mainly the potentially pathogenic fusarium genus. In conclusion， wheat root exudates could improve soil enzyme activity， improve soil biological environment， promote the growth of bitter melon plants， and change the adverse ecological effects of continuous cultivation of bitter gourd.

Effect of Topdressing Different Types of Nitrogen Fertilizer on Growth and Yield of Intercropped Peanut with Wheat

Jiayu HU, Yang YANG, Hongyan ZHANG, Bingyang GAO, Linglu WANG, Junying YAN, Xiaomei SUN, Yanan ZHAO, Youliang YE

2024, 26(2):  191-197.  DOI: 10.13304/j.nykjdb.2022.0796

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To study the effects of different types of nitrogen fertilizer on the growth and yield of intercropped peanut in sandy soil， the field experiment was conducted in Xinxiang City of Henan Province. 7 treatments were set， including no fertilizer control （CK）， ammonium chloride （AN）， ammonium sulfate （AS）， calcium ammonium nitrate （CAN）， urea （UR）， humic acid urea （UHA） and controlled-release urea （SRU）. The plant height， branch number， compound leaf number， dry matter and pod yield of peanut were investigated. The results showed that， compared with CK， the plant heights of peanut in different stages were increased by all the N fertilizer types. The plant height of UR treatment was relative higher in all growth periods， and were higher in the later growth period under UHA and SRU treatments. The application of N fertilizer could promote the peanut branching， with great increase for AS， UR and SRU treatments before the needling stage. The N fertilizer application significantly affected the number of compound leaves of peanut. The number of compound leaves was less in the early growth stage but more in the late stage for AS and CAN treatments， while were more in the early stage but less in the mature stage for UHA and SRU treatments， the number of compound leaves for AN and UR were relative stable. The N fertilization significantly increased the dry matter accumulation at fruit swelling and maturity stages. Compared with CK， application of N fertilizer increased the yield by 25.0%~61.0%， with the greatest increase by CAN and UHA treatments. The yield increase for UR and SRU treatments were higher than AS and AN. The CAN， UR and SRU treatments significantly decreased bad fruit rate of peanut than other treatments. In conclusion， N fertilizer application could promote the growth and yield of intercropped peanut with wheat， with the best effect for CAN， followed by UHA and SRU.

Effects of Corncob on Capillary Water Transport and Evaporation Characteristics of Sandy Soil

Wencheng CONG, Limin YUAN, Zhongju MENG, Yu YANG

2024, 26(2):  198-207.  DOI: 10.13304/j.nykjdb.2022.0640

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In order to explore the effective utilization of corncob in arid areas and improve its utilization rate， the water absorption and evaporation experiments were carried out by mixing 1.5~6.0 mm corncob particles with aeolian sand by means of soil column simulation. The water absorption， wetting front， evaporation， evaporation rate et al. of the soil column with 10 kinds of corncob added were measured， and the soil column without corncob was used as the control. The effects of corncob and its dosage on water absorption and water retention in sandy soil were analyzed. The results showed that the rising height of capillary water in the mixed soil was 0.0%~76.3% lower than that of the control， the average rise rate of capillary level was 7.3%~78.7% lower than that of the control， and the evaporation intensity was 10.7%~64.2% lower than the control. When the addition amount of corncob≤40%， the water absorption was 8.6%~16.2% higher than the control； when the addition>40%， the water absorption was 1.1%~46.0% lower than the control. There was a power function relationship between capillary water rise height and time， a power function relationship between groundwater recharge and time， and a linear correlation between groundwater recharge and capillary water rise. After adjustment， the Green-Ampt model under water absorption could be used to simulate the capillary water rise process of mixed soils. The Rose evaporation models could express the variation characteristics of the cumulative evaporation of mixed soils with time. The results provided a theoretical basis for the corncob material in the practice of desertification land management and straw returning to the field to improve cultivated land.

Effects of Combined Application of Auxin and Molybdenum on Physiological Metabolism and Quality of Upper Leaves of Flue-cured Tobacco

Fengfeng LIU, Ming WU, Yinghui ZHOU, Yong WU, Jiashu TIAN, Jiayang XU, Zicheng XU, Jiewang HE

2024, 26(2):  208-215.  DOI: 10.13304/j.nykjdb.2023.0106

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In order to explore the effects of auxin and molybdenum on the physiological metabolism and quality of flue-cured tobacco， a field experiment was conducted to measure the photosynthesis， carbon and nitrogen metabolism， physical properties and chemical composition by using the cultivar‘Yunyan 87’as material. The results showed that compared with the control， the application of auxin and molybdenum after topping increased the contents of photosynthetic pigments （chlorophyll， carotenoids）， the values of gas exchange parameters （net photosynthetic rate， stomatal conductance， transpiration rate） and the enzymes activities of carbon and nitrogen metabolizing （amylase， invertase， nitrate reductase， glutamine synthetase）； the leaf length， leaf width， single leaf weight， filling value， tensile strength， potassium， total sugar and reducing sugar contents increased significantly， while the contents of nitrogen and nicotine decreased significantly. The total variation contribution rates of auxin and molybdenum to photosynthetic pigments of tobacco leaves were 68.85%， 13.23%， respectively； to the gas exchange parameters were 50.66% and 42.63%， respectively； to the rates of carbon and nitrogen metabolizing enzymes were 40.74%， 34.02%， respectively； to the rates of physical properties were 46.72%， 49.55%， respectively； to the rates of chemical composition were 54.94%， 36.22%， respectively. On the whole， auxin was the main effector molecule， the treatment of spraying 20 mg·kg^-1 auxin and 4 mg·plant^-1 molybdenum had the best effect on tobacco for promoting physiological metabolism and improving quality.The above results provided a theoretical basis for the rational application of hormones and trace elements in flue-cured tobacco production.

Effects of Shading Cultivation on Leaf Uniformity of Cigar-wrapper Tobacco （Nicotiana tabacum）

Hao GUO, Ronglei TAN, Jinpeng YANG, Jun YU, Wenchang HUANG, Jiuhong YANG, Baoming QIAO, Ruiwei YANG, Fangsen XU, Chunlei YANG, Guangda DING

2024, 26(2):  216-225.  DOI: 10.13304/j.nykjdb.2022.0706

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In order to explore the influence of shading cultivation on the leaf uniformity of cigar-wrapper tobacco， a cultivar‘Chuxue 26’ （CX-26） was used as material to conduct different shading treatments in the field， and the leaf thickness， leaf flatness， leaf density， main and branch vein thickness， proportion of leaf vein to leaf dry weight， and SPAD value of the upper and middle leaves at maturity were analyzed. The results showed that the leaf thickness， leaf density and SPAD value of upper leaves decreased gradually with the increase of shading degree. When the light transmittance was 70%， the leaf thickness and leaf density decreased by 35.5% and 33.3%， respectively，compared with the treatment without shading， while they increased gradually in middle leaves. Shade cultivation significantly increased the leaf flatness of upper leaves， but decreased the leaf flatness of middle leaves. When the light transmittance was 70%， the leaf flatness of the upper leaves reached the maximum， which was 0.98. In addition， shading cultivation increased the proportion of the main branches in the upper leaves to the leaf dry matter， and the influence on the proportion of the main branches in the middle leaves to the leaf dry matter showed a downward trend with the increase of shading degree， but no significant difference was observed. Moreover， shading cultivation had no effect on the thickness of the main branches of the middle leaves， but it made the main branches of the upper leaves thinner. Above results provided a theoretical basis for shading cultivation techniques of domestic cigar tobacco.

INNOVATIVE METHODS AND TECHNOLOGIES

Construction of Bifunctional Saccharomyces cerevisiae with Ability to Detoxify Mycotoxins

Yijun WANG, Huoqing HUANG, Xiaoyun SU

2024, 26(2):  226-233.  DOI: 10.13304/j.nykjdb.2023.0130

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The contamination of mycotoxins is common in feed and becomes one major hurdle preventing healthy breeding of animals. There are many types of mycotoxins， among which aflatoxins，and particularly aflatoxin B₁（AFB₁） have the highest impact and harm. Therefore， developing new methods for detoxification of mycotoxins in feeds is currently a hot research topic. In this study， Saccharomyces cerevisiae strains were first isolated and four were compared for their absorption rates of AFB₁. It was then discovered that， in comparison with other methods including heating and mannanase treatment and strains， glucanase treatment of CEN.PK had the highest absorption of AFB₁， which could reach as high as 53.3%±8.2%. Then， CEN.PK was used as the host cell to constitutively express a myo-inositol oxidase gene from Flavobacterium johnsoniae （FjMiox）. The engineered yeast could convert 1 g·L^-1 of myo-inositol into 0.273 g·L^-1 of glucuronic acid. In the liver of animals， the glucuronic acid could be conjugated with mycotoxins by enzymatic catalysis to increase their water solubility， thereby facilitating their excretion. Therefore， the engineered Saccharomyces cerevisiae able to synthesize glucuronic acid combines the functions of binding the mycotoxin in the gut and conjugating to the mycotoxin in the liver， which could provide a new solution to combat the contamination of aflatoxins in feeds.