Journal of Agricultural Science and Technology

Research on Change Trend and Risk of China’s Food Production

Lili SONG, Ruitao ZHANG

2023, 25(2): 1-9. DOI: 10.13304/j.nykjdb.2022.0855

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Food production is crucial to the national transportation and people’s livelihood， and food security is the top priority in governing the country. From grain production capacity and grain business entities， China’s food rations are self-sufficient and the planting structure has been further optimized. However， there are still realistic difficulties in grain production， such as tighter constraints on ecological resources， low enthusiasm of grain business entities to grow grain， and increase risks of uncertainty in international trade， which have brought considerable impact on China’s grain production. So this paper proposed to further implement the strategy of “storing grain on the ground， storing grain in technology”， cultivate new food production and operation entities to adapt to the new era， and improve the ability of risk prevention and control and subsidy guarantee to ensure the long-term sustainable food security in China.

Overall Framework and Strategic Value of Agricultural and Rural Scientific Research Collaborative Innovation Infrastructure

Lihua JIANG, Song FANG, Dandan ZHANG, Ruixue ZHAO, Xiaoyan CHANG

2023, 25(2): 10-18. DOI: 10.13304/j.nykjdb.2022.0702

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The CPC Central Committee and the State Council attach great importance to agricultural scientific and technological innovation. The “bottleneck” problem in agriculture has become a national strategy. Collaborative innovation is an effective way to solve the “bottleneck” problem. It faces the main battlefield of modern agricultural construction， the resolution of major problems facing the development of agricultural industry and the breakthrough of core technology. The construction and development of agriculture and rural areas put higher demands on scientific and technological innovation. As an important organizational form of agricultural science and technology innovation， collaborative innovation will play a pivotal role in the construction of innovative countries in China， and it is also of great significance for the construction of modern agricultural research institutes. Starting from the policy background of building agricultural and rural collaborative innovation infrastructure and the research status at home and abroad， this paper deeply analyzed the bottleneck problems faced by China’s agricultural scientific and technological innovation， such as agricultural scientific research big data， high-performance computing， original algorithm model， data application innovation and data security protection， and built an emerging agricultural collaborative innovation infrastructure of “data” + “algorithm” + “computing power” based on advanced information technologies such as artificial intelligence （AI）， blockchain， cloud computing and big data. Furthermore， a collaborative innovation platform for agricultural and rural scientific research was constructed to serve the interdisciplinary integration. The platform simplified and speed up the iteration and flow of AI models and data， and implemented large-scale heterogeneous computing resource scheduling technology， multi-model data fusion method and friendly development support environment. The platform was dedicated to assist researchers in different disciplines to deeply integrate data， models， algorithms and computing power resource management. It created an out-of-the-box interactive cloud development environment， which was applied to the teaching， scientific research， scientific data sets and algorithm research and other scenarios， and formed an ecological closed-loop of artificial intelligence integration with multiple disciplines. It was of great strategic significance to enhance the original innovation ability in China’s agricultural and rural fields， realize the leap in key fields， ensure the long-term development of science and technology， and transform from a large agricultural country to an agricultural power.

Research Progress on Application of Lactic Acid Bacteria in Pig Production

Xin XU, Kun MENG, Hongying CAI, Peilong YANG, Xianren JIANG

2023, 25(2): 19-26. DOI: 10.13304/j.nykjdb.2022.0041

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Lactic acid bacteria （LAB） is a general name of bacteria that can produce a large amount of lactic acid by using fermentable carbohydrates. It is a kind of microorganism beneficial to human health and its way to obtain energy is to ferment carbohydrates （mainly glucose） to produce lactic acid. The biological characteristics of LAB and its application in pig production were summarized， which should provide theoretical basis and reference for the better application of LAB in pig production.

Identification and Expression Analysis Under Abiotic Stress of IQM Gene Family in Wheat （Triticum aestivum L.）

Ting LIANG, Jinghong ZUO, Qing LU, Dong YANG, Yimiao TANG, Chunman GUO, Dezhou WANG, Weiwei WANG

2023, 25(2): 27-37. DOI: 10.13304/j.nykjdb.2022.0612

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IQM genes are important branch of the calmodulin-binding protein family that plays an important role in plant growth，development and stress response. In this study， 23 IQM gene family members were identified in the whole wheat genome using bioinformatics methods， and their chromosomal location， physicochemical properties， phylogenetic relationships， gene structure， protein conserved structural domains， promoter cis-acting elements and gene expression characteristics were systematically analysed. The results showed that the TaIQM gene family members were randomly distributed on the 18 chromosomes of wheat， and their subcellular localization showed that all genes were located in the nucleus. Phylogenetic analysis classified them into three subclasses with high similarity in gene structure and protein conserved structural domains among， and presumably similar functions； cis-acting element analysis showed that the promoters of TaIQM gene family members contained a variety of cis-acting elements related to stress， growth and development. Transcriptome analysis showed that the TaIQM gene family members were significantly differentially expressed in roots， stems， leaves， spikes and seeds at different times. qRT-PCR analysis showed that TaIQM genes were up- or down-regulated in response to various stresses such as drought， low and high temperature， NaCl and ABA in the above- and below-ground parts of wheat seedlings. It was tentatively concluded that these genes might be involved in abiotic stress regulatory through the Ca²⁺ signalling pathway. The results of this study provided clues for a comprehensive analysis of the structure and biological functions of TaIQM genes， the molecular mechanisms of abiotic stress response.

Comprehensive Evaluation of Low Temperature Tolerance of Waxy Maize Varieties at Germination Stage

Shengwei GUO, Siwen BIAN, Jianwen DING, Xiaochen ZHANG, Xing YANG, Jin DU, Chunyang XIANG

2023, 25(2): 38-47. DOI: 10.13304/j.nykjdb.2022.0550

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In order to construct evaluation system of different waxy maize varieties during germination for low temperature tolerance， 48 waxy maize varieties were used as test materials to measure 10 indicators on germination and growth under low temperature stress. And the low temperature tolerances of waxy maize varieties were comprehensively evaluated by principal component analysis， membership function method， cluster analysis and regression analysis. The results showed that 10 measurement indexes were transformed into 2 independent comprehensive indexes by principal component analysis， including root growth factor and seed germination factor. The comprehensive score model of low temperature tolerance of waxy maize was established， Y=0.647 7Y₁+0.144 2Y₂， where Y₁ and Y₂ were scores of the two principal components， respectively. Y₁ included relative root dry weight， relative root surface area， relative root volume， and relative root length， which could reflect the state of root growth of waxy maize under low temperature stress； and Y₂ included relative seedling dry weight， relative germination rate， relative vitality index， and relative germination potential， which could reflect the germination of waxy maize under low temperature stress. The 48 waxy maize varieties were ranked by affiliation function method for low temperature tolerance， and the five varieties with the strong low temperature resistance were screened， including ‘Runcainuo 686’‘ Jingpo 82’‘Runnuo 175’‘Runnuo 605’ and ‘Tianguinuo 932’. The cluster analysis classified 48 waxy maize varieties into 4 groups. The regression analysis showed that the relative root dry weight， relative seedling length， relative germination potential， relative germination rate， relative seedling dry weight and relative root surface area could be used as important indicators for the evaluation of low-temperature tolerance of waxy maize. The comprehensive evaluation of multiple methods could effectively screen and classify waxy maize varieties with different low-temperature tolerances， which provided reference for the evaluation of waxy maize low-temperature tolerance and the screening of low-temperature tolerance germplasm resources.

Genome-Wide Identification and Analysis of BGLU Genes Family in Gossypium hirsutum

Man ZHANG, Zhicheng WANG, Zhengwen LIU, Guoning WANG, Xingfen WANG, Yan ZHANG

2023, 25(2): 48-59. DOI: 10.13304/j.nykjdb.2021.0699

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BGLU genes play crucial role in the defense of biotic and abiotic stresses， this study identified the members of the BGLU family in cotton via bioinformatics methods. The sequence features， conserved domain structure， chromosome location， promoter elements and other family features were analyzed， and the transcriptome data and qRT-PCR were used to analyze the pathogenic stress response pattern of BGLU family genes. The results showed a total of 53 BGLU genes were identified and divided into 6 sub-families according to the phylogenetic relationship in upland cotton， and some genes were specifically expressed under pathogen stress. qPCR analysis and RNAseq data showed that GhBGLU5， GhBGLU14， GhBGLU18， GhBGLU26 and GhBGLU34 maintained higher expression level in resistant and susceptible cottons， implying that these genes positively regulated cotton Verticillium wilt resistance. This study provided theoretical basis for further analyzing the functions and molecular mechanisms of the BGLU genes in cotton.

Physiological and Biochemical Characteristics of Coffea arabica Seedling Under Low Temperature Stress

Lifang HUANG, Yuzhou LONG, Jinqin LI, Yunping DONG, Xiaoyang WANG, Peng CHEN, Xianwen WANG, Lin YAN

2023, 25(2): 60-67. DOI: 10.13304/j.nykjdb.2021.0640

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To investigate the mechanism of cold-resistance in Coffea arabica， a few physiological indexes including osmotic adjustment substance content， antioxidant enzyme activities， and cell membrane permeability for leaves of one-year-old seedlings under different low temperatures were measured， and which were treated with correlations and principal component analysis. The results indicated that the contents of proline， soluble sugar， soluble protein， and the activities of superoxide dismutase and catalase were increased steadily at 8 ℃， while they were showed a trend of increasing first and then decreasing at 4， 0 and -2 ℃. For peroxidase activities， they also had the same trends with decreasing of temperatures and extension of processing time. The malondialdehyde content and relative conductivity had similar trends， manifested as increasing gradually with temperature decreasing， and maximum values were both reached at 0 ℃ for 48 h and -2 ℃ for 48 h， which were 132.00 and 124.58 nmol·g^-1， 91.29% and 98.35%， respectively. Correlation analysis showed that there was a highly significant positive correlation between malondialdehyde content and relative conductivity. In summary， Coffea arabica seedlings could alleviate damage from low temperature stress and showed strong cold-resistance ability by regulating its own antioxidant enzyme activities and osmotic adjustment substance contents. This study would provide theoretical support for resource protection， introduction， domestication and variety breeding of coffee.

Research Progress on Flowering Regulation and Flower Morphogenesis of Rose

Zhiying XIONG, Rui YU, Ziyi HU, Yuman ZHANG, Qinglin LIU

2023, 25(2): 68-75. DOI: 10.13304/j.nykjdb.2022.0098

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Rose has great ornamental value and economic value. Flowers is one of the important economic ornamental organs， flowering time and flower shape on the production and application have very important significance. As one of the few woody flowers continuous blooming， the research of rose flowering regulation and flower formation is of great significance. This article summarized the research progress from the flowering regulation pathway， flower bud differentiation， morphogenesis， and flowering regulation approach in recent years， hoping for ornamental flower crop development， flowering regulation and providing reference for design modification.

Catalytic Properties of the Active Pocket Key Tryptophan on Chitinase Chi304

Xiaoqian TIAN, Haiqiang LU, Ningfeng WU, Jian TIAN, Feifei GUAN

2023, 25(2): 76-82. DOI: 10.13304/j.nykjdb.2021.0877

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There are binding sites and hydrolytic active sites in chitinase， and the binding of key amino acids to substrate chitin can appropriately regulate the hydrolytic activity of the enzyme. The thermophilic chitinase Chi304 was as experimental material. Swiss-Model and analysis of its advanced structure revealed the presence of two tryptophans （W140 and W272） near its substrate binding pocket. And these two tryptophans were site-directed mutated to alanine. High performance liquid chromatography was used to detect the hydrolysis products of the enzyme. The ratio of product triacetyl chitosaccharide to diacetyl chitosaccharide （DP₃/DP₂） was used to evaluate the effect of the mutants. The mutants （W140A， W272A and W140/272A） hydrolyzed colloidal chitin， and the proportions of product （DP₃/DP₂） were increased by 23.3%， 45.7% and 80.0% compared with that of wild type， respectively. The results showed that W140 and W272 were the key amino acids affecting the binding of enzyme to substrate， and the mutation of alanine to Chi304 increased the endogenous activity and decreased the exogenous activity.

Research on Characteristics and Coverage Extraction of Rice Multi-phase Vegetation Index

Linjiang YIN, Wei LI, Weiquan ZHAO, Zulun ZHAO, Sisi LYU, Xiaoqiong SUN

2023, 25(2): 83-98. DOI: 10.13304/j.nykjdb.2022.0256

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In order to accurately and quickly obtain the vegetation index characteristics and vegetation coverage information of rice， the UAV was used to collect the multispectral image data of rice tillering stage， heading stage and seed setting stage， select different types of commonly used vegetation indexes， and extract and explore the vegetation index characteristics of three growth stages of rice at the scale of plot and pixel by using the ideas of sample statistics method and vegetation index intersection method， Then the threshold segmentation method was used to extract and calculate the rice vegetation information and coverage information. The results showed that in the three growth periods of rice， there were obvious phenological characteristics at the pixel and plot scales， which had obvious phenological characteristics， and there were obvious differences between weeds and trees； the extraction accuracy of vegetation coverage of multispectral vegetation index was higher than that of visible vegetation index；（normalized difference vegetation index， NDVI） had the highest accuracy in extracting vegetation coverage in three stages of rice， with extraction errors of 0.40%， 0.43% and 0.81%， R²^{of 0.77， 0.92 and 0.98， and （root mean square error， RMSE） of 9.09%， 2.97% and 0.38%. The extraction accuracy of （visible-band difference vegetation index，VDVI） was higher than that of （excess green-red-blue difference index，EGRBDI） and （excess green-excess red index，ExG-EXR）， the extraction errors were 4.30%， 1.36% and 1.60%， respectively， R² were 0.53， 0.77 and 0.80， respectively， and RMSE were 14.62%， 3.70% and 5.50%，respectively. It could be seen that the research results can provide technical support for crop growth monitoring and vegetation coverage extraction.}

Vegetable Recognition Based on Unmanned Aerial Vehicle （UAV） Multispectral Imagery and Random Forest Algorithm

Qian GUO, Jiahao WEI, Jian ZHANG, Zhangxi YE, Houxi ZHANG, Zhengqing LAI, Hui DENG

2023, 25(2): 99-110. DOI: 10.13304/j.nykjdb.2022.0027

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Real-time and accurate information about vegetable distribution is critical for improving the management of water/fertilizer and accurate estimation of the yield. Based on spectrum features of the segmented UAV multispectral images objects， other features including index， textural and geometric features were added to build eight classification schemes （S1～S8）， and then the random forest algorithm was used to classify and analyze the classification effect. The results showed that the scheme S5 （SPEC+GLCM+INDE） performed better than the others， with overall accuracy and Kappa of 92.75% and 0.92， respectively. We discovered that adding geometric features reduced the classification accuracy， while adding index and texture features had the opposite effect. It was difficult to distinguish the Chinese cabbage and cabbage effectively only by spectrum， index and texture features， so it was necessary to introduce features such as height to improve the accuracy in the follow-up research. The feature importance ranking results showed that spectrum features， followed by the index features， had the greatest impact on classification accuracy. To summarize， the method proposed in this study， which combined object-oriented and RF algorithm， could achieve higher classification accuracy based on multi-spectral images from UAV， and the important features that affect vegetable classification could be effectively identified， providing a reference for the accurate identification of other crops.

Morphological and Polygenetic Identification of Pathogen of Wheat Leaf Blight

Tongtong ZHENG, Wendi YANG, Ning WANG, Junjie MA, Long LIU, Qingyuan GUO

2023, 25(2): 111-118. DOI: 10.13304/j.nykjdb.2021.0912

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In order to clarify the causes and pathogens of wheat leaf blight and spot phenomenon in Tacheng wheat producing area of Xinjiang， more random sampling dry leaf or with irregular disease spot disease of plants were collected from Tacheng prefecture Emin county， and pathogenic bacteria were isolated and purified from diseased leaves by routine tissue separation. The pathogenicity was determined by acupuncture inoculation. And the species of pathogens were identified by morphological observation and multi-gene sequence analysis. The results showed that 76 strains of Alternaria spp were isolated and identified， which confirmed as pathogens by pathogenicity determination. Through the morphological identification， 76 strains belonged to A. alternata， A.rosae and A. infectoria， among which， A. alternata as the main pathogen with 50% frequency of separation. Based on sequences of EF-1 and Alt genes and ITS， the evolutionary tree was constructed， and the three kinds of representative strains YC3， LK19， LK3 showed 100%， 100% and 99% homology with A. alternata， A. rosea and A.infectoria， respectively， which was consistent with morphological identification results. The above results showed that， in Tacheng prefecture， Xinjiang winter wheat field of area， the phenomenon of wheat leaf blight for chain and spore wheat leaf blight was caused by Alternaria infection， and A. alternata， A. rosae and A. infectoria were the main pathogens.

Fermentation Products Analysis of Producing β-glucosidase Endophytic Bacteria in Martianus dermestoides Transformated to Ginsenoside and Its Anti-tumor Activity

Qian HU, Yining WANG, Pengfei SHEN, Yaqian LI, Yang YANG, Yancheng WANG, Wenxiu JI, Weiwei DONG

2023, 25(2): 119-127. DOI: 10.13304/j.nykjdb.2021.0691

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In order to improve the bioavailability of ginsenosides， and clarify the transformated products of ginsenosides in Martianus dermestoides and the anti-tumor activity of their metabolites， the functional strains of high-yield β-glucosidase in the endophytes of Martianus dermestoides were screened by MRS （esculin， ferric citrate） medium to prepare compound fermentation agent liquid fermentation ginsenoside extract. The chemical composition and dynamic changes of ginsenosides in fermentation products were analyzed by HPLC-MS/MS. The anti-tumor activities of the fermentation product in low （0.5 mg·L^-1）， medium （2 mg·L^-1）， and high （5 mg·L^-1） dose groups on A549 cells were clarified using methylthiazoletrazolium （MTT） blue staining method. A total of 7 saponins （Re， Rg1， Rf， Rb1， Rc， Rb2， Rd） were detected in ginseng extract. Among the metabolites， deglycosylated ginsenosides Rh1， Rg3 and compoud K were formed. The MTT experiment showed that the inhibition rate of tumor cells was 85% after treated 72 h in the high does group with the does 5 mg·L^-1. These results suggested that a variety of high-activity stimulating ginsenosides were transformed by endophytic bacteria of Martianus dermestoides， which improved the medicinal value of Chinese herbal medicine ginseng and onion， and provided a new idea for fermentation and modification of Chinese herbal medicine.

Review of Absorption and Utilization of Different Nitrogen Forms and Their Effects on Plant Physiological Metabolism

Yujing LI, Yuqing FENG, Yuanyuan ZHAO, Hongzhi SHI

2023, 25(2): 128-139. DOI: 10.13304/j.nykjdb.2021.0909

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The absorption and utilization processes of different forms of nitrogen and their effects on plant growth， stress resistance， quality and safety， as well as the effects of external environmental conditions on their effects were summarized. It showed that $N O 3 -$ -N could generally promote the root development of plants and promote the absorption of cations by plants. $N H 4 +$ -N could alleviate the damage of salt stress and reactive oxygen species to plants， and also improve the resistance of plants to diseases. However， $N H 4 +$ -N poisoning was easy to be caused by single application of $N H 4 +$ ， such as increase of cell acidity， reactive oxygen species damage and lignification of cell walls. In addition， the future research was prospected， which provided theoretical basis for rational application of nitrogen fertilizer in agriculture.

Temporal and Spatial Variation of Natural Grassland Vegetation Coverage and Its Relationship with Meteorological Factors in Northern Xinjiang from 1998 to 2018

Zhixiong HOU, Changqing JING, Gongxin WANG, Wenzhang GUO, Weikang ZHAO

2023, 25(2): 140-151. DOI: 10.13304/j.nykjdb.2021.0881

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In order to analyze the temporal and spatial changes of natural grassland vegetation coverage in different months of the growing season in Northern Xinjiang and its relationship with meteorological factors， the pixel dichotomy model， change rate analysis and correlation analysis were used to explore the temporal and spatial changes of natural grassland vegetation coverage and its relationship with meteorological factors in Northern Xinjiang in June， July， August and September from 1998 to 2018. The results showed that the vegetation coverages of natural grassland in Northern Xinjiang were lower in the middle and east with less precipitation and higher temperature， higher in the north， west and south with higher altitude， more precipitation and higher relative humidity， which increased from the middle and east to the north， west and south. In June， July， August and September， the natural grassland vegetation coverage in Northern Xinjiang showed no significant decreasing trend， in which the monthly average natural grassland coverage was June>July>September>August. The normalized difference vegetation index of natural grassland in Northern Xinjiang showed an increasing trend in July and decreasing trend in June， August and September. The monthly average natural grassland vegetation coverages in Northern Xinjiang were mainly positively correlated with precipitation and relative humidity in June， July and August， and were mainly positively correlated with temperature and sunshine hours in September.

Effects of Straw Biochar and Straw on Soil Nutrients and Crop Yield in Wheat-Maize Rotation System

Yunzhu ZHENG, Shuchen SUN

2023, 25(2): 152-162. DOI: 10.13304/j.nykjdb.2021.0996

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In order to explore the effects of biochar and straw on wheat-maize rotation system in calcareous fluvo-aquic soil area， the single-season returning mode was adopted with maize straw returning in winter wheat season and wheat straw not returning in summer maize season. According to the actual average dry matter mass of corn stalks in the observation plot， the amount of maize straw returned to the field was set to 0.5 （S0.5）， 1.0 （S1.0）， 1.5 （S1.5） and 2.0 （S2.0） times， the amount of biochar was used to convert the same amount of corn straw into biochar （B0.5， B1.0， B1.5 and B2.0）， and no biochar and straw was used as control （CK）. The soil nutrition content and crop yield under different treatments were determined. The results showed that biochar and straw treatments could promote the contents of total available nutrients and available nutrients in soil during the growth seasons of winter wheat and summer maize. On the whole， the improvement of soil nutrients in winter wheat and the increase of C/N ratio were higher than those in summer maize. Compared with the control， biochar and straw treatments significantly increased grain yield of winter wheat by 9.04%~21.76% and 15.31%~22.96%， respectively. In summer maize season， B0.5， S1.0， S1.5 and S2.0 treatments significantly increased grain yield by 10.86%， 8.72%， 10.89% and 12.22%， respectively. The effect of biochar and straw on winter wheat yield was higher than that of summer maize， and the effect of straw returning was better. Therefore， on the basis of normal fertilization in the calcareous fluvo-aquic soil area of the Luxi Plain， the applications of 0.5 times of corn straw carbonized biochar and 1.0 times of corn straw in the winter wheat season should not only conducive to improving soil fertility， but also increase the grain yield of winter wheat and summer maize.

Impact of Controlled-release Urea Combined with Conventional Urea on Yield and Nitrogen Utilization Efficiency of Spring Sweet Corn Under One-off Application

Qiaoyi HUANG, Yongpei WU, Xu HUANG, Ping LI, Hongting FU, Mu ZHANG, Yuwan PANG, Zhaobing ZENG, Shuanhu TANG

2023, 25(2): 163-173. DOI: 10.13304/j.nykjdb.2021.0760

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Field experiments of sweet corn were carried out in Boluo county and Huiyang county of Guangdong province simultaneously in 2017 to determine the effects of one-off application of controlled-release urea combined with common urea on the biomass accumulation and nitrogen accumulation of spring sweet corn. Four treatments were employed in the experiments， including no nitrogen application treatment（CK）， conventional nitrogen application treatment（U100， three times application） and 2 controlled-release urea combined with common urea under one-off application treatments（CCU100 and CCU80， which denoted 0% and 20% reduction of N rate based on U100， respectively）. The blending ratio of controlled-release urea and common urea in CCU100 and CCU80 was 7∶3. The results showed that nitrogen release characteristics of controlled-release urea in soil under spring sowing sweet corn were similar to the “S” line， and more than 85% of nitrogen from controlled-release urea had been released. The cumulative release rate of nitrogen from controlled-release urea was correlated with accumulated temperature positively. Comparing with U100， the biomass of sweet corn in CCU100 were increased by 4.68% （Boluo） and 6.75% （Huiyang ） averagely， the nitrogen uptake of sweet corn in CCU100 were increased by 2.74% （Boluo ） and 6.20% （Huiyang ） averagely. No significant differences in accumulative of biomass of sweet corn between CCU80 and U100 treatments were observed. During harvest， the N uptake of sweet corn at Boluo in CCU80 treatment was 10.02% lower than that of U100 treatment （P<0.05）. Compared with U100 treatment， the yield of CCU100 and CCU80 treatment did not decrease significantly. Compared with U100 treatment， the agronomic N use efficiencies and N recovery efficiency were significantly increased in CCU80 treatment（P<0.05）， with an increase of 14.20% （Boluo ） and 10.16% （Huiyang ）， and 4.92% （Boluo ） and 22.03% （Huiyang） respectively. During harvest， N contents of straw and ear were increased significantly due to nitrogen application， but no significant differences in the nitrogen， phosphorus and potassium contents of straw and ear were observed among different N fertilization treatments. To sum up， for the production of spring sowing sweet corn in Guangdong province， one-off application of controlled-release urea combined with conventional urea could increase the nitrogen use efficiency while maintained crop yield， so it’s an effective way to save labor and increase profit.

Effects of Biological Agents on Hydrothermal Status and Enzyme Activity During Composting of Corn Straw

Riping GAO, Xiaoyue LIU, Zuntian PAN, Dongxu ZHANG, Xiangjun SHEN, Yanfang LI, Jie HUANG, Yupeng JING

2023, 25(2): 174-181. DOI: 10.13304/j.nykjdb.2021.0651

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In order to solve the problems of difficult decomposition of returning corn straw and long decomposition period in Xiliaohe Plain of Inner Mongolia， the composting experiments of field straw were carried out in 2020. Four treatments were set including straw decomposition agent+straw （SRT）， fresh cow dung+straw （SRN）， straw decomposition agent+fresh cow dung+straw （SRW） and single straw （CK）. The temperature， water content and the activities of cellulase， urease and lignin catalase in different treatments during corn straw composting were determined. The results showed that adding straw decomposition agent and fresh cow dung could improve the decomposition rate of straw and shorten the decomposition time. Compared with single straw composting， the addition of fresh cow dung and straw decomposition agent could significantly improve the temperature and water content of straw composting. Compared with CK， the cellulase activities of SRW， SRT and SRN treatments increased by 15.64%， 4.87% and 7.96%， respectively； the urease activities increased by 10.29%， 3.43% and 6.86%， respectively； and the lignin catalase activities increased by 71.43%， 48.05% and 53.25%， respectively. Therefore， the addition of straw decomposition agent and fresh cow dung could improve the hydrothermal conditions of straw composting and the activity of straw hydrolase， and accelerate the process of straw decomposition. So the coupling effect of straw decomposition agent and fresh cow dung was better， which could be applied as an efficient straw composting method in the Xiliaohe Plain of Inner Mongolia.

Effect of Bio-organic Fertilizers on Quality and Soil of Continuous Crop Chinese Flowering Cabbage

Juxian GUO, Bishan OUYANG, Guihua LI, Mei FU, Wenlong LUO, Shanwei LUO, Meilian LU

2023, 25(2): 182-191. DOI: 10.13304/j.nykjdb.2022.0553

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In order to solve the obstacles of successive crop and improve the yield and quality of successive crop， we applied bio-organic fertilizer （0， 3 000， 4 500， 6 000 and 7 500 kg·hm^-2） to Chinese flowering cabbage as the experimental material to study its effect on the growth， quality and soil of successive cabbage. The results showed that the application of bio-organic fertilizer significantly increased the plant and shoots weight of continuous crop flowering Chinese cabbage， and soil organic matter， alkali hydrolyzable N and electrical conductivity were also increased with the same treatment. In addition， bio-organic fertilizer substantially increased the activity of soil acid phosphatase， catalase and sucrase， but decreased the activity of urease. Compared with cheinicoil fertilizer， the application of 7 500 kg·hm^-2 of bio-organic fertilizer， the content of vitamin C， protein， crude fibre and soluble solids of successive vegetable shoots were increased by 6.28%， 12.39%， 5.30% and 37.38%， respectively. Moreover soil organic matter， alkali hydrolyzable N and electrical conductivity were increased by 59.67%， 97.63% and 175.76%， respectively. The activitiy of acid phosphatase， catalase and sucrase in root zone of soil were greatly increased by 23.10%， 7.40% and 116.92%， respectively， while urease was decreased by 8.39%. The numbers of bacteria， fungi and actinomycetes were noticeably increased by 80.62%， 79.26% and 46.00%， respectively， and the relative abundance of bacteria and fungi also increased. In conclusion， application of bio-organic fertilizer could improve the growth and quality of continuous cropping flowering Chinese cabbage， and increase the soil fertility and microbial diversity.

Effect of Chitosan on Alfalfa’s Cold Resistance and Content of Cold-resistant Protective Substances

Ze TENG, Yuxia ZHANG, Weidong CHEN, Baiming CONG, Yonglei TIAN, Qingxin ZHANG, Yongliang ZHANG, Dongru WANG

2023, 25(2): 192-198. DOI: 10.13304/j.nykjdb.2021.0622

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In order to explore the response of alfalfa cold resistance to chitosan application level in Horqin sandy land， taking ‘Gongnong 1’ alfalfa variety as the material， a single factor randomized block experiment design was adopted， and 0， 50，100， 150， 200 μg·g^-1 amount of chitosan were sprayed in autumn. The alfalfa overwintering materials were treated under simulated low-temperature freezing conditions （–20 ℃） and cold storage （4 ℃）， and the changes of relative conductivity of alfalfa root collar and the contents of soluble protein， proline， sucrose and fructose were detected. The results showed that under the condition of low temperature freezing， when treated with 100 μg·g^-1 chitosan， the relative conductivity of alfalfa root collar was the lowest， which was 31.49%， while the contents of proline， sucrose and fructose were the highest， which were 8.05，88.17^{and 86.32 mg·g^-1， respectively.When treated with 150 μg·g^-1 chitosan， the soluble protein content of alfalfa root collar was the highest， which was 27.19 mg·g^-1. The results of correlation analysis showed that the relationships between soluble protein， proline， sucrose， fructose and relative conductivity were negative under different low temperature stress treatments. Under the condition of low temperature freezing stress， chitosan could reduce the relative conductivity and improve the cold resistance of alfalfa by increasing the contents of soluble protein， proline， sucrose and fructose in the root collar of alfalfa. Spraying 100~150 μg·g^-1 chitosan in Horqin sandy land in autumn was conducive to the safe overwintering of alfalfa.}

Composition and Diversity of Fungal Community in Rhizosphere Soil of Camellia Oleifera

Xin LUO, Yuekai WU, Niannian ZHANG, Jie XU, Zaihua YANG

2023, 25(2): 199-210. DOI: 10.13304/j.nykjdb.2021.0675

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In order to clarify the composition and diversity of rhizosphere soil fungi of Camellia oleifera Abel.， and to explore the correlation between rhizosphere soil fungi and soil environmental factors， rhizosphere soil fungal communities from Guiyang， Tianzhu， Yuping， Wangmo， Weining， Liping and Ceheng in Guizhou were as materials. The composition and structure of C. oleifera rhizosphere soil fungal communities were analyzed by illumina high-throughput sequencing， and the physical and chemical properties of soil were determined and their correlation with rhizosphere soil fungal communities were studied. The results showed that the rhizosphere soils of C. oleifera were acidic， rich in organic matter and available potassium， but low in nitrogen and phosphorus. And there were significant differences among different regions in soil physical and chemical properties. Alpha diversity analysis showed that the rhizosphere soil fungi of C. oleifera had rich diversity， among which Wangmo area had the highest diversity， Ceheng was the second， and Guiyang was the lowest. Beta diversity analysis showed that there were significant differences in the composition of rhizosphere soil fungi among the seven regions. A total of 634 OTUs were obtained from soil samples from 7 regions， belonging to 9 phyla， 32 classes， 73 orders， 141 families and 213 genera. Basidiomycete was the dominant phyla of rhizosphere soil fungi in Yuping area， while Ascomycota was the dominant phyla of rhizosphere soil fungi in the other six areas. There were significant differences in the distribution and abundance of soil fungi at genera level， such as Saitozyma， Archaeorhizomyces， Mortierella and so on. Correlation analysis showed that the contents of total potassium， available phosphorus， available potassium and organic matter in soil were the main driving factors affecting the composition of soil fungal community in C. oleifera rhizosphere. Some soil fungi such as Penicillium， Cladophialophora and Oidiodendron showed significant positive correlation with soil environmental factors， and some soil fungi such as Ilyonectria and Fusarium with pathogenic characteristics had significant negative correlation with soil environmental factors. These results provided a scientific basis for the development of beneficial rhizosphere fungi resources from C. oleifera and the prevention of diseases by regulating soil environmental factors.

Signal Peptide Optimization Increases Glucose Oxidase Expression in Pichia pastoris

Na ZHANG, Yaru YAN, Yun WU, Yuhong ZHANG, Wei ZHANG

2023, 25(2): 211-219. DOI: 10.13304/j.nykjdb.2020.0219

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Glucose oxidase （GOD） is widely used in food processing， feed additives， biosensors and industrial production. Compared to Aspergillus， Pichia pastoris is a microorganism that has the potential to produce GOD more efficiently， with fewer by-products and simpler separation purification. However， due to factors such as exogenous genes， gene dosage， and secretion signals， the GOD yield of Pichia pastoris is still low， which limits the large-scale application of GOD in food processing and other fields. In this study， based on Pichia pastoris codon preference and mRNA secondary structure energy， the alpha factor signal peptide in Pichia pastoris recombinant expression cassette was modified to obtain 4 Pichia recombinant strains containing different signal peptides. The results showed that the GOD activity of the recombinant strain containing the highest codon frequency signal peptide α¹was 2.01 times that of the starting strain， while the GOD enzyme activity of the recombinant strain containing low codon frequency signal peptide α⁴ was only 41%. The above results also indicated that the arrangement of the signal peptide nucleic acid sequence in Pichia pastoris expression vector could affect the expression level of recombinant GOD， which provided a new research idea for increasing the expression level of Pichia pastoris recombinant protein.

Preliminary Study on Determination of β-nicotinamide Mononucleotide Content from Flammulina velutiper Extract by High Performance Capillary Electrophoresis

Xiaoying MA, Jun XIAO, Xun CHEN, Na GONG, Guoli LIU, Tao YANG, Ying ZHAO

2023, 25(2): 220-226. DOI: 10.13304/j.nykjdb.2021.0728

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Flammulina velutiper is not only a delicious food， but also a good health food which is rich in nutrition. However， there was no report on the determination of β?-nicotinamide mononucleotide （NMN） content in Flammulina velutiper. A method for the determination of β??-nicotinamide mononucleotide （NMN） in Flammulina velutiper was established by high performance capillary electrophoresis （HPCE-DAD）. The detection conditions included detection wavelength 218 nm； running buffer （30 mmol·L^-1 boron sand buffer pH 9.2； 10 mmol·L^-1 phosphate bobbin）； separation voltage of 25 kV； injection time of 5 s. The results showed that the method could detect the β-nicotinamide mononucleotide content of fruits from Flammulina velutiper with the detection limit of 0.17 μg·mL^-1， and showed a good linear relationship from 0.03 to 2.00 mg·mL^-1. The regression equation was obtained by the standard curve： y=2E+6x-345.69，R²=0.999 4， the corresponding β??-nicotinamide mononucleotide （NMN） peak area was put into the regression equation， and the calculated content was 27.57 μg·mL^-1. The relative standard deviations of migration times for β??-nicotinamide mononucleotide and migration peak areas were 0.68% and 1.44%. The relative deviations of migration time and peak area within 8 h were 1.26% and 2.82%， respectively. The recoveries of β??-nicotinamide mononucleotide in Flammulina velutiper were 89%~94%. These results indicated that the method was suitable for the determination of β??-nicotinamide mononucleotide in Flammulina velutiper and provided a new method for the determination of β-nicotinamide mononucleotide in edible fungus.

Rapid Detection of CP4-EPSPS Transgenic Products Based on RPA Technology

Xiuwen HU, Bo DENG, Jinbin WANG, Hua LIU, Xueming TANG, Yu WANG, Haijuan ZENG, Wei JIANG, Hong LI

2023, 25(2): 227-233. DOI: 10.13304/j.nykjdb.2021.0785

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Nucleic acid detection is widely used in the safety detection of agricultural products. With the commercial cultivation of genetically modified crops， the method with rapid， specific and sensitive is an urgent need for the detection of genetically modified crops. The recombinase polymerase amplification （RPA） technology was used to detect CP4-EPSPS genes in genetically modified products and their products. Five pairs of primers were designed to screen the best pair of primers， and the reaction system and temperature were optimized. The results show that CP4-EPSPS gene could be quickly detected by this method with 20 μL reaction system， 15 min reacted at 37 ℃. The sensitivity threshold for detecting transgenic DNA was 45 copies with highly sensitive and specific nucleic， which provided a new way for large-scale screening of CP4-EPSPS gene.

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