Table of Content

15 November 2023, Volume 25 Issue 11

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AGRICULTURAL INNOVATION FORUM

Development Strategy on Agricultural Microbial Industry in China

Jingli GUO, Dlxat Nilufar, Daqing WANG

2023, 25(11):  1-7.  DOI: 10.13304/j.nykjdb.2023.0682

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The international market share of the agricultural microbial industry is increasing， making the development of germplasm resources， key technology research， safety monitoring and protection gradually become a battlefield in the field of agricultural microbial industry. It was found that the development of China's agricultural microbial industry still had limitations such as weak ability to tap and utilize resources， little breakthrough in key core technologies， weak international competitiveness of industrial development， and imperfect safety prevention and control system. In order to ensure national food security， improve the market share of biological agriculture economy， achieve green and sustainable development of agriculture， and consolidate the foundation of agricultural power. The government should accelerate the construction of the national agricultural microbial germplasm bank， strengthen the breakthrough of the key core technologies of agricultural microbial industry， optimize the development layout of agricultural microbial industry， increase support for agricultural microbial enterprises， improve the safety protection and monitoring system of agricultural microbial industry， and comprehensively enhance the international market share and international competitiveness of China's agricultural microbial industry.

Analysis of Global Transgenic Oilseed Rape Patent Information and Technology Outlook

Mingdi CHEN, Xingru CHENG, Bo XU, Haiwen ZHANG, Wangtian WANG, Youhua WANG

2023, 25(11):  8-19.  DOI: 10.13304/j.nykjdb.2023.0508

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In order to promote the application of biological breeding technology in agriculture and understand the development status， cultivation， import and export， and industrialization of global genetically modified （GM） oilseed rape crop， the analysis of global GM oilseed rape patent information through the Intelligent Bud Database （PatSnap） showed the global GM oilseed rape development trend， key technology areas， key acceptance countries， recearch and development （R&D） capability of each country， technology of R&D institutions in each country， competitiveness of major applicant institutions， legal status of patents， etc. The outlook and suggestions were proposed on GM oilseed rape R&D， layout of agricultural fields， global competition and promotion of oilseed rape industrialization.

Current Situation and High Quality Development Countermeasures of Watermelon and Muskmelon Industry in Beijing

Lin ZHANG, Yantao YANG, Lili SONG, Shiping MAO

2023, 25(11):  20-27.  DOI: 10.13304/j.nykjdb.2023.0313

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As one of the characteristic crop industries in Beijing， the high-quality development of watermelon is important for promoting the revitalization of rural industries， achieving wealth and income increase. In order to conduct a comprehensive and in-depth study of the current development status of the entire chain of Beijing’s watermelon industry， the systematic analysis has been conducted from the aspects of production scale， variety structure， facility cultivation， green development， brand construction， circulation mode， market price， etc. In addition， problems such as increasing external risk factors， urgent breakthroughs in core technologies， imbalance in labor supply and demand， difficulty in implementing supporting policies， and incomplete development of industrial chains was analyzed. The following countermeasures and suggestions was proposed： strengthening technology investment support， comprehensively attracting rural talents， promoting land transfer and moderate scale operation， promoting the integration and improvement of the “three chains”， and improving precise support policies for the entire chain， in order to promote the upgrading and high-quality development of the Beijing watermelon industry throughout the entire chain.

BIOTECHNOLOGY & LIFE SCIENCE

Evolution of Wheat （Triticum aestivum L.） Germplasm Traits and Evaluation of Breeding Value in Southern Huang-Huai Winter Wheat Region in Recent 20 Years

Huifang ZHANG, Jianhong ZHANG, Haijiao LIU, Yan SUN, Hongzhi QI, Nan WANG, Junzhi DUAN, Yan GUO, Haiyan YIN

2023, 25(11):  28-41.  DOI: 10.13304/j.nykjdb.2022.0817

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To study the evolution of wheat traits and genetic diversity in southern Huang-Huai winter wheat region， the wheat varieties in the recent 20 years in southern Huang-Huai winter wheat region were as materials. The evolution of wheat traits and genetic diversity were explored from a new perspective by using Shannon-Wiener diversity index （H\mathrm{\text{'}}） and coefficient of variation analysis， fitting trend line after dimensionless traits， principal component analysis， comprehensive score （D′ value） evaluation， regression model construction and other methods. The indexes used for the evaluation of wheat varieties in southern Huang-Huai winter wheat region and the germplasm with excellent performance were screened. The results showed that a total of 443 wheat varieties were bred in southern Huang-Huai winter wheat region from 2003 to 2022， and it could be divided into 3 stages based on the number of bred varieties： basic stable stage （from 2003 to 2017）， temporary high stage after obvious uplift （from 2018 to 2019）， and blowout eruption stage （from 2020 to present ）. In recent 20 years， plant height of wheat in southern Huang-Huai winter wheat region decreased slowly with time-going； water absorption remained unchanged； spike number， grain number per spike， 1 000-grain weight， yield， basic seedlings， test weight， protein content， wet gluten content and stability time showed upward trend， among which 1 000-grain weight， yield， basic seedlings， test weight and wet gluten content had relatively strong upward trend. The increase of yield was mainly due to the increase of 1 000-grain weight. The proportion of high-quality special wheat （strong gluten wheat， medium strong gluten wheat， weak gluten wheat） increased in recent years， mainly due to the increase in the proportion of medium strong gluten wheat. The proportion of strong gluten wheat has not increased significantly， which was mainly limited by water absorption and stability time. The dispersion degree and evolution trend of agronomic traits and quality traits were quite different， but their genetic diversity was constantly improving with the year，and genetic stability should be further strengthened. The results of regression model construction showed that 5 traits could be used as the evaluation indexes of the comprehensive performance of wheat in southern Huanghuai winter wheat region， including spike number， yield， test weight， protein content and water absorption. Among 443 varieties， ‘Jimai 336’‘Anke 1801’‘Jimai 44’‘Gushenmai 19’ and ‘Huawei 305’， which had higher D\prime， performed well and could be used as excellent germplasm resources in the breeding of strong gluten wheat. In the future， the breeders should strengthen breeding new varieties with high-quality special wheat， especially weak gluten wheat， and enhance the protection and utilization of germplasm resources.

Effects of Plant Growth Retarders on Lodging Resistance and Yield of Quinoa

Qiujing CHEN, Zhaodi YANG, Shiyu WANG, Fenggen GUO, Xiaoxue ZHAO, Fan CHEN, Yang FENG

2023, 25(11):  42-48.  DOI: 10.13304/j.nykjdb.2022.0594

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In order to improve the lodging resistance of quinoa in agricultural production and select the best concentration of plant growth retarder to reduce the lodging rate of quinoa， the randomized block experiment was conducted to investigate the effects of paclobutrazole （PP333）， enisobutrazole （S3307） and binomide （MEP） on the growth and yield of quinoa at the concentrations of 50， 100， 200 and 350 mg·L^-1， with spraying water as the control. The results showed that the 3 plant growth retarders could effectively reduce the plant height of quinoa， improve the breaking resistance， and reduce the lodging index and lodging rate in a certain concentration range. Among 3 plant growth retarders， the treatment of S3307 was optimum with concentration 100 mg·L^-1， whose plant height decreased by 6.8% and 4.2% and the flexural resistance increased 17.4% and 25.2%， respectively， compared to PP333 and MEP with same concentration. When the concentration of PP333 was 100 mg·L^-1， the anti-lodging ability was better than other concentrations， and the lodging rate significantly reduced by 79.9% compared with CK. The MEP concentration in the range from 100 to 200 mg·L^-1 was more effective than other concentrations. The above results provided theoretical reference for chemical control measures of high-quality planting of quinoa.

Effects of FeNPs on Cotton Roots Growth and Its Response to Drought Stress at Seedling Stage

Da CHEN, Jisheng JU, Qi MA, Shouzhen XU, Juanjuan LIU, Wenmin YUAN, Jilian LI, Caixiang WANG, Junji SU

2023, 25(11):  49-57.  DOI: 10.13304/j.nykjdb.2023.0625

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Nano iron （FeNPs） is a new type of nano metal fertilizer， but its regulation effects on cotton growth and stress resistance are rarely studied. 4 FeNPs treatment levels （0， 1.0， 2.5， 5.0 mg·L^-1） were set to study its effect on root morphology and physiological indexes of hydroponic cotton seedlings under PEG stress. The results showed that total root length， total root surface area， projected area， number of root tips and fresh and dry weight of roots， stems and leaves cultured with 5.0 mg·L^-1 FeNPs were significantly higher than those cultured with 1.0 and 2.5 mg·L^-1 FeNPs. Cotton seedlings were cultured in Hogland nutrient solution with 0 and 5.0 mg·L^-1 FeNPs， and then 10% PEG-6000 was applied at the four-leaf stage. The results showed that the cotton seedlings without FeNPs were more wilting than those treated with 5.0 mg·L^-1 FeNPs， and the fresh weight of root， stem and leaf was significantly lower than that with FeNPs. In addition， FeNPs significantly increased SOD， POD and CAT activities and significantly decreased MDA content in cotton leaves within 6 h of stress. After 9 h， SOD and CAT activities decreased and MDA content increased significantly. In summary， appropriate level of FeNPs would be beneficial to root growth， dry matter accumulation and antioxidant enzyme activity of cotton seedlings， thus enhancing the drought resistance of cotton seedlings.

Identification of Soybean Quality Traits and Screening of Elite Germplasms in Huang-Huai-Hai Ecoregion

Zhenqi SHAO, Wenlong LI, Youbin KONG, Hui DU, Zhanjun LI, Xihuan LI, Caiying ZHANG

2023, 25(11):  58-69.  DOI: 10.13304/j.nykjdb.2022.1057

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Soybean is an important economic crop in the world， which provides plant protein， oil and a variety of active substances beneficial to human health. In order to comprehensively identify the quality characters of soybean resources abundant in China， 16 kinds of seed quality traits of 520 soybean germplasms in Huang-Huai-Hai ecoregion were evaluated in 6 environments across 2 years with reference to the national standard method. The results showed that the genetic variations of the tested seed quality traits were abundant in the soybean germplasms， and the variation coefficients of isoflavone and tocopherol contents were relative high， while the variation coefficients of protein， oil and fatty acid components were relatively low. Meanwhile， there were significant correlations among the 16 seed quality traits. Principal component analysis showed that the 16 quality traits could be attributed to 5 principal components. Based on the results of principal component analysis， 520 germplasms could be divided into 3 categories by cluster analysis with significant differences. Based on the above results and high stability coefficient analysis， 35 germplasms with multiple or single elite seed quality traits were screened out， which provided important resources for genetic improvement of seed quality traits in soybean.

Physiological and Biochemical Responses of Peach Self-rooted Rootstock to NaCl and Analysis of Salt Tolerance Threshold

Fan ZHANG, Hong WANG, Xuebing ZHANG, Jianjun CHEN

2023, 25(11):  70-79.  DOI: 10.13304/j.nykjdb.2023.0271

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In order to explore the growth and physiological and biochemical responses of the peach self-rooted rootstock under NaCl stress and determine the salinity tolerance threshold， the cutting seedlings of one-year-old 5 rootstocks were treated with different NaCl contents （0.2%， 0.4% and 0.6% NaCl）. The results showed that， with the increase of NaCl content， the activity of superoxide dismutase （SOD） and content of malondialdehyde （MDA） increased. With the increase of NaCl content， the activity of peroxidase （POD） increased first and then decreased in Li 1 and Li 3， while decreased first and then increased in Li 5 and GF677， and showed increasing trend in RA. With the increase of NaCl content， the activity of catalase （CAT） showed increasing trend in Li 1， Li 3 and RA， showed decreasing trend in Li 5， and increased first and then decreased in GF677. The content of Pro increased in Li and RA， decreased first and then increased in Li 3 and Li 5， and increased first and then decreased in GF677. The content of soluble sugar increased in Li 3 and Li 5， increased first and then decreased in Li 1 and RA， and decreased first and then increased in GF677. The correlation analysis showed that Na⁺ content was positively related to NaCl content in media， while K⁺ content was negative correlation with NaCl content. The Na⁺/K⁺ was the highest in Li 1， while was the lowest in GF677. Under NaCl stress， the chlorophyll synthesis of 5 peach rootstocks was significantly inhibited. Fluorescence kinetic parameters and light energy absorption， capture and transfer parameters were reduced， and the curve of OJIP changed with the decrease of J， I， P phases and ΔK<0. Considering net photosynthetic rate （P_n） as a tolerance index， the salt tolerance thresholds of Li 1， Li 3， Li 5， RA and GF677 were 0.37%， 0.43%， 0.38%， 0.46% and 0.49% NaCl， respectively. According to comprehensive membership function， salt tolerance coefficient index and salt tolerance thresholds resistance， the salt tolerance showed GF677>RA>Li 3> Li 5>Li 1.In summary， self rooting stocks of peach adapted to salt stress mainly through active oxygen scavenging and osmotic adjustment mechanisms， rootstocks with high salt tolerance could better stabilize the photosynthetic reaction center and alleviate the damage of salt stress to their photosynthetic system， which provided theoretical support for revealing the mechanism of salt tolerance and screening the varieties of peach rootstock with high salt tolerant.

Authentication of Suzhou Yangcheng Lake Hairy Crab Based on Protein Fingerprint

Yahan ZHANG, Youyong ZHANG, Dan YUAN, Min XU, Yezhong CAO

2023, 25(11):  80-89.  DOI: 10.13304/j.nykjdb.2023.0115

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Crab chelate muscle tissue protein is rich in species and appears in the mass spectrum with different intensity peaks. Crab chelate muscle tissue protein is extracted， and the characteristic index of crab chelate muscle tissue protein spectrum was analyzed by matrix-assisted laser desorption/ionization time-of-flight mass spectrometry （MALDI-TOF MS） technology， and the characteristic index is found to achieve the qualitative and quantitative detection of crab chelate muscle tissue protein. In this study， protein molecular fingerprint databases of different categories of crabs were established by analyzing the histoprotein MALDI-TOF MS technology， and the database was deeply analyzed with machine deep learning algorithm to establish the homology analysis method of Yangcheng Lake hairy crabs. The results showed that the protein profiles of hairy crabs in different growing environments showed different expressions.There was an obvious difference when m/z（mass-charge ratio，m/z） was less than 10 000，the scanning results showed that the low m/z region （<10 000） had higher peak intensity and higher sub-marking rate，this region could be used as the mass spectrum resolution region to distinguish Yangcheng Lake hairy crabs from other lake crabs. Above results showed MALDI-TOF MS technique could be used to analyze the different protein expressions of hairy crabs from different places， which could play a role in identifying hairy crab from different places.

INTELLIGENT AGRICULTURE & AGRICULTURAL MACHINERY

Study on Tillage Fertilizer Mixing Effect Under Vertical Smashing Rotary Tillage and Rotary Tillage Based on EDEM

Fake SHAN, Shuo KANG, Jianxi ZHU, Yongwei WANG, Jun WANG

2023, 25(11):  90-102.  DOI: 10.13304/j.nykjdb.2022.0237

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The distribution of nutrient components of chemical fertilizer in soil directly affects crop yield， which is different under different tillage methods. In order to explore the mechanism and reveal the distribution rule of chemical fertilizer particles in soil under different tillage methods， a discrete element simulation model was established based on EDEM. A test platform using force sensor was built to calibrate the material attribute parameters and contact parameters required in the establishment process. The drag force was measured through signal acquisition and analysis system， and it was compared with that obtained by simulation during same working period to verify the accuracy of the simulation model. The results showed the error was below 10%，indicating it could be inferred that the established model had certain prediction accuracy. Through orthogonal experiments the optimal working parameters of 2 tillage methods were obtained and the results were verified through field experiments. Through the comparison of the final results， it was found that when different tillage methods were both under the optimal mixing effect working state， the maximum error between the predicted value of the fertilizer distribution proportion in each layer of the simulation model and the actual value is 8.65%， and the minimum error is 0.59%. In conclusion， the prediction model established in this study could be used to explore the particle distribution rule of the mixing effect of surface fertilizer under different tillage methods， and had a certain prediction accuracy. The results provided references for selecting the optimal tillage method， improving agricultural fertilization efficiency and reasonably controlling fertilization amount.

Calibration of Discrete Element Parameter of Rice Stubble Straw Based on EDEM

Zitao MA, Zhihao ZHAO, Wei QUAN, Fanggang SHI, Chen GAO, Mingliang WU

2023, 25(11):  103-113.  DOI: 10.13304/j.nykjdb.2022.1047

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In order to improve the accuracy of the parameters during the process of rice straw shredding and returning to the field in discrete element simulation， the stubble straw in the field after rice harvest were taken as the research object， and the contact parameters between straw and steel， straw and straw were measured through the slope test.Then based on simulation model and through the stacking angle and shear test， the bonding radius between straw model particles， tangential critical stress， normal critical stress， coefficient of restitution， static friction coefficient and dynamic friction coefficient were taken as the test factors， taking the shear resistance and radial stacking angle of straw as evaluation indicators， a multi-objective optimization simulation calibration experiment were conducted using the Box-Behnken experimental scheme.The results showed that the bonding radius between straw particles was 1.06 mm， and the normal critical stress of straw was 4.77×10¹⁰ Pa， tangential critical stress was 4.67×10⁶ Pa， the collision restitution coefficient between straw and straw was 0.21， the static Friction coefficient was 0.19， and the rolling friction coefficient was 0.09. The results of the simulation test results under the optimal parameter combination conditions and the physical test showed that the relative errors of the shear resistance and radial stacking angle of straw in the simulation test and physical test were 1.7% and 2.8%， respectively. This study could provide reference for the discrete element simulation analysis between straw and straw， as well as between straw and agricultural machinery.

ANIMAL AND PLANT HEALTH

Isolation and Identification of Pathogen of Rubber Tree Phytophthora Leaf Fall Disease and Screening of Control Fungicides

Yuping SHI, Yixian LIU, Guowei LI, Yi TANG, Liming DAI, Lanlan LI, Zhiying CAI

2023, 25(11):  114-122.  DOI: 10.13304/j.nykjdb.2023.0101

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In the last 2 years， rubber tree abnormal leaf fall occurred over large area in Xishuangbanna rubber plantation， and the symptoms were similar to those of rubber tree Phytophthora leaf fall disease. In order to clarify the pathogenic species and screen fungicides for effective control of this disease， the pathogen was isolated and purified by tissue isolation and mycelia cusp purification method， and the pathogenicity was determined by Koch’s postulates test， morphological and physiological characteristics， rDNA-ITS and COX Ⅱ gene sequence analysis. The results showed that the pathogen caused rubber tree abnormal leaf fall was identified as Phytophthora botryosa， which was a new record pathogen of rubber tree Phytophthora leaf fall disease in China. The antibiotic activities of 15 fungicides was tested in vitro with mycelial growth rate method. 48% dimethomorph-cyazofamid SC （suspension concentrate）， 80% dimethomorph WG （water dispersible granule）， 30% metalaxyl-dimethomorph WG， 35% dimethomorph-cymoxanil SC， 30% floridomide-metalaxyl WG and 0.3% tetramycin AS （aqueoces solution） could effectively inhibit the mycelial growth of the pathogen， with EC₅₀ of 0.137 1， 0.175 9， 0.201 7， 0.257 7， 0.265 1， 0.351 0 mg·L^-1， respectively， which could be used as candidates for control of the disease in the field.

Drug Resistance and Genotype Analysis of Escherichia coli in Healthy Chickens from Zhaosu， Yili， Xinjiang

Hongbo LI, Yueyue CHEN, Yujie YANG, Qiqi XU, Lei QIN, Xin CAI, Lining XIA

2023, 25(11):  123-131.  DOI: 10.13304/j.nykjdb.2022.0268

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In order to understand the drug resistance phenotype and genotype carried of Escherichia coli in healthy chickens from three villages in Zhaosu area， Yili， Xinjiang， and guide drug pertinently， 300 chicken anal swab samples were collected from Kemarkgar village， Sentas village and Uklegar village in Zhaosu， Yili and Escherichia coli was isolated and identified. The agar dilution method was used in order to check the drug resistance （MIC） of 9 antibiotics in 6 categories， and PCR was applied to test the corresponding drug resistance genes. The results showed that 289 strains were isolated from the 3 villages， which the separation rates of Sentas village， Kemagar village and Uklegar village were 100.0%， 97.0% and 93.8%. 289 strains of Escherichia coli had high resistance rates to ampicillin， florfenicol and tetracycline， which were 31.5%， 26.0% and 20.1%， respectively. The severity of drug resistance from high to low was Sentas village， Kemakgar village， Uklegar village. The drug resistance rates of Escherichia coli from chicken in Sentas village to ampicillin， cefazolin， florfenicol， enrofloxacin and colistin sulfate were 40.0%， 35.0%， 45.0%， 40.0% and 30.0%， respectively. The drug resistance rate of Escherichia coli from chicken in Uklegar village was the lowest， no more than 20.0%. The richness of drug resistance spectrum of Escherichia coli in different villages was different. Among them， the drug resistance spectrum of Escherichia coli from chicken in Kemakgar village was the most abundant， up to 32 species. Also， the drug resistance spectrums of other 2 villages were more than 10. Gene test results showed that 9 drug resistance genes were detected， of which the detection rates of floR， tet（A） and bla_TEM genes were 25.6%， 18.0% and 18.0%， respectively. 9 kinds of drug resistance genes were detected in Escherichia coli from chicken in Kemakgar village， including bla_TEM， bla_CTX-M， floR， oqxA， oqxB， tet（A）， tet（M）， ant（3''）-Ⅰa， mcr-1； 6 kinds of drug resistance genes were detected in Escherichia coli from chicken in Uklegar Village， including bla_TEM， bla_CTX-M， floR， oqxA， oqxB， tet（A）； however， only tet（A）， floR， bla_CTX-M and bla_TEM were detected in Escherichia coli from chicken in Sentas village. To sum up， Escherichia coli from healthy chicken in Zhaosu， Yili， Xinjiang had the characteristics of high isolation rate， high drug resistance rate， rich drug resistance spectrum and low carrying rate of drug resistance genes. At the same time of strengthening the supervision of antibiotics， farms could adopt different drug regimens according to drug sensitivity test results.

FOOD QUALITY & PROCESSING AND STORAGE

Amino Acid Content Analysis of Different Fresh Corn Varieties at Suitable Harvest Time

Baishan LU, Hui DONG, Jiuran ZHAO, Li XU, Yanli FAN, Yaxing SHI, Ronghuan WANG

2023, 25(11):  132-142.  DOI: 10.13304/j.nykjdb.2023.0680

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The amino acid content of 26 different fresh corn varieties was analyzed by high performance liquid chromatography （HPLC）， and the similarity between different types of fresh corn and WHO/FAO standard amino acid model value was compared， and the amino acid content was analyzed by amino acid ratio coefficient method. The results showed that，the total amino acid content of the tested fresh corn varieties ranged from 6.85 to 16.66 mg·g^-1， and Jingketian 533 （16.66 mg·g^-1）， Jingkenuo 623 （12.01 mg·g^-1） and Nongkenuo 336 （14.44 mg·g^-1） was higher than other varieties of sweet， waxy， sweet+waxy type， respectively. Essential amino acids accounted for 10.38%~22.43%， and Jingketian 816 （17.39%）， Jingkenuo 656 （22.43%） and Nongkeyu 368 （18.16%） was higher than other varieties of sweet， waxy， sweet+waxy type， respectively. Analysis of flavor amino acids showed the content of fresh and sweet amino acids of Jingketian 533 and Nongkenuo 336 were higher than that of other varieties. The content of methionine+cysteine was higher than that of WHO/FAO standard mode， while the content of other amino acids were lower than the standard amino acid model value， belonging to the relative lack of amino acids. There were 6 varieties with amino acid ratio coefficient score （SRC） more than 50， including sweet corn varieties Jingketian 183 （58.17）， Jingketian 608 （57.79）， Jingketian 533 （53.32） and Jingketian 816 （53.03）， sweet+waxy corn varieties Nongkeyu 368（51.05） and Nongkenuo 336 （50.63）， indicating that their protein nutritional value was balanced and easy to be absorbed and utilized by the human body.

Evaluation of Nutritional Composition and Quality of Phallus impudicus L. from Different Producing Areas in Guizhou

Yupeng LIU, Fang CHEN, Shuhong GU, Fang WANG

2023, 25(11):  143-153.  DOI: 10.13304/j.nykjdb.2022.0461

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In order to analyze the nutritional compositions and quality differences of Phallus impudicus L. in Guizhou，the contents of amino acids， mineral elements， ash， fat， crude fiber and total sugar in dry Phallus impudicus L. from 4 producing areas were determined， the protein quality was evaluated by amino acid ratio coefficient method and the comprehensive quality was evaluated by principal component analysis. The results showed that， the contents of N， P， K and Ca， Cu， Fe， Mg， Zn were 18.70～22.45， 3.55～4.83， 29.63～39.64 g·kg^-1 and 508.64～686.89，12.37～17.54，92.30～497.54，1.55～1.98，24.84～51.61 mg·kg^-1， respectively. The contents of ash， fat， crude fiber and total sugar were 5.83%～6.27%， 0.83%～1.49%，0.25%～0.47%，28.92%～34.90%， respectively. 17 amino acids were detected in Phallus impudicus L. from different producing areas， content of Glu was the highest， the contents of essential amino acids， total amino acids were 4.24%～5.55%， 10.70%～12.93%， respectively. Dejiang had the highest index of all the above indicators. Amino acid evaluation results showed that Lys was the first limiting amino acid of Phallus impudicus L.， and the essential amino acid index （EAAI） values of Phallus impudicus L. from 4 producing area were all more than 1， which could be used as high quality protein source. The amino acid ratio coefficient score （SRCAA） value showed that Dejiang had the highest nutritional value of protein. The result of principal component analysis showed that Dejiang had the best quality whose comprehensive score was the highest. This study provided scientific basis for quality control， development and utilization of Phallus impudicus L. in Guizhou province.

Effect of Different CO₂ Contents on Temperature Distribution in Potato Storage Room

Qi ZHEN, Guangze YAN, Na TA, Zhiyong ZHAO, Huimin YU

2023, 25(11):  154-165.  DOI: 10.13304/j.nykjdb.2023.0349

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Within the context of storing potatoes in traditional semi-underground storage room， the greenhouse effect is caused by the increase of CO₂ content. In order to clarify the distribution law of the temperature in the storage room due to the change of CO₂ content caused by potato respiration， taking the typical semi-underground potato storage room in western China as the research carrier， with the help of computational fluid dynamics method， the porous media model， k-ε turbulence model， and internal heat source model were used to construct a three dimensional numerical solution model of the heat and mass exchange with gas flow between the potato pile and the environment in the storage room. To ensure the accuracy of numerical simulation results， the numerical method and grid independence were verified. The results showed that through a comparative experiment monitoring the temperature at specific locations of potato stacks， it was found that the average relative error between experimental values and simulated values was 8.26%， with a maximum relative error of 9.77%， further confirming the reliability of the numerical heat transfer model for the storage room and the equivalent thermal model for potato stacks. Based on the environmental factor data inside and outside the storage room on a certain day in January 2022， when the CO₂ content was 0.15%， the indoor temperature distribution decreased layer by layer along the potato pile， and the CO₂ vent made a high temperature zone appear behind the pile. When the CO₂ content increased from 0.00% to 0.30%， the average indoor temperature increased from 1.34 to 1.36 ℃， and the average temperature of the potato pile increased from 1.93 to 1.94 ℃. Comparing the indoor temperature distribution under the carbon dioxide content of 0.00%， 0.15% and 0.30% components， the overall temperature in the storage room was effectively increased with the increase of the CO₂ content in the storage room. The results of this paper had important reference value for the establishment of mathematical models of semi-underground storage rooms and the selection of suitable carbon dioxide concentration control devices.

BIO-MANUFACTURING & RESOURCE AND ECOLOGY

Trichoderma Affects Crop Growth and Soil Ecological Environment

Xin ZHAO, Yifei WANG, Jiajia WANG, Peiyao WANG, Guiduan WANG, Lixia ZHU, Lili LI

2023, 25(11):  166-172.  DOI: 10.13304/j.nykjdb.2022.0456

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Trichoderma is a genus of common filamentous fungi and is widely used in agriculture as biofungicides， which can be used to control a variety of plant diseases. As is known， Trichoderma can not only well control crop diseases， but also improve soil physical-chemical properties and enhance soil productivity. However， its control effects on crop growth and soil ecological environment are not clear. Therefore， this work summarized the effects of Trichoderma on disease resistance， yield and growth of crops， the effects of Trichoderma on soil ecological environment were also reviewed from the perspective of soil physical， chemical and microbial properties. Additionally， this paper pointed out the shortcomings of the existing research on the impact of Trichoderma on crops and soil properties， and stressed corresponding prospects to promote crop growth and improve soil fertility in the future. This review could provide a theoretical basis for the in-depth promotion and application of Trichoderma in agricultural production.

Soil Initial Available Phosphorus and Exchangeable Magnesium Shaping the Response of Wheat Growth to pH

Lu ZHANG, Lei ZHENG, Siru LIU, Zejiang CAI, Nan SUN, Qiang ZHANG, Minggang XU

2023, 25(11):  173-181.  DOI: 10.13304/j.nykjdb.2022.0496

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The critical pH was the target pH for soil acidity improvement， which has important guiding significance for agricultural production and sustainable development. A pot experiment was conducted to investigate the effect of soil initial properties affecting the response of wheat growth to pH. 5 soils derived from plate shale， quaternary red earth， river alluvium， tertiary red sandstone， and granite were used in this study. The results showed that the maximum shoot biomasses of wheat were 2.71， 1.17， 2.06， 1.57 and 1.70 g·pot^-1 for soils derived from plate shale， quaternary red earth， river alluvium， granite， and tertiary red sandstone， respectively. The critical values of pH_95%， pH_50%， or pH_5% were defined as shoot biomass reaching to 95%， 50%， or 5% of maximum values. The pH_95%， pH_50%， or pH_5% were 5.31， 4.42， and 3.53 for soil derived from plate shale， 5.79， 4.79， and 3.80 for quaternary red earth， 5.25， 4.71， and 4.16 for river alluvium， 4.94， 4.61， and 4.29 for tertiary red sandstone granite， and 5.72， 4.66， and 3.59 for granite， respectively. The maximum values of shoot biomass had significant positive correlations with soil initial pH， available P， exchangeable Mg and exchangeable Ca. The pH_50% of wheat shoot biomass was negatively correlated with soil initial available P； and the pH _50% of wheat height was negatively correlated with soil initial available P and exchangeable Mg. The data indicated that soil initial available P and exchangeable Mg shaped the response of wheat growth to pH and the maximum values of crop biomass were determined by both soil nutrient status and acidity. Therefore， we recommend that co-amelioration of soil fertility and acidity was a practical method to reach a best condition for plant growth.

Effect of Phosphorus and Selenium Combined Application on Selenium Uptake， Translocation and Organic Selenium in Grain of Winter Wheat

Caixia HU, Hongen LIU, Chang LI, Shiyu QIN, Yuhuan ZHAO, Yupeng ZHANG, Haiyang LIU, Jiayang XU, Peng ZHAO, Zhaojun NIE, Qiuhong WANG

2023, 25(11):  182-191.  DOI: 10.13304/j.nykjdb.2022.0656

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In order to explore the effects of combined application on selenium（Se） and phosphorus（P） fertilizer on selenium uptake， translocation and organic selenium species in grains of winter wheat， a wheat cultivar‘Zhengmai 379’was chosen as the test material. A pot experiment with 3 P levels of 0（P₀）， 80（P₈₀） ，160 mg·kg^-1（ P₁₆₀） and 2 Se levels of 0（Se₀）， 1 mg·kg^-1（Se₁） were set. A total of 6 treatments （P₀Se₀， P₈₀Se₀， P₁₆₀Se₀， P₀Se₁， P₈₀Se₁， P₁₆₀Se₁， respectively） were used to analyze the yield， P and Se concentration， accumulation in each part of winter wheat， transport coefficient and Se species in grain. The results showed that the yield of winter wheat was significantly increased by the combination application of P and Se， and the highest yield of winter wheat was 46.25 g·pot^-1 under P₈₀Se₁ treatment. Se application could significantly reduce the P content and accumulation of each part of the winter wheat， but increase the P migration coefficient of glume to grains under P₈₀ and P₁₆₀ treatments. Each part of the winter wheat had high Se concentration under P₀Se₁ treatment. P application reduced the Se concentration of each part of the winter wheat and the Se migration coefficient of root to stem leaf， but increased Se accumulation in grains as well as in whole plant. Besides grain， each tissue as well as in whole plant had high Se accumulation under P₈₀Se₁ treatment. Se application increased the Se migration coefficient of glume to grain and the proportion of total Se accumulated in the grains and roots of winter wheat under P₈₀ and P₁₆₀ treatment， but decreased the ratio of total Se accumulated in glumes. The analysis on selenium species showed that Se application increased the organic Se concentration in the grains， Se in winter wheat grains was mainly in the organic form including selenocystine and selenomethionine. Selenomethionine and inorganic Se concentration in grains decreased with the increasing P application. Therefore， appropriate P combined with Se application enhanced grain yield of winter wheat， promoting the transfer of selenium in the winter wheat to the grain， increased Se accumulation and organic Se concentration of grain under P₈₀Se₁ treatment. The results provided guidance for rational application of phosphorus fertilizer and selenium fertilizer in selenium-enriched wheat production.

Effect of Graphene on Soil Nutrient Transformation and Root Growth of Maize Seedlings

Shiya WANG, Xinyi WANG, Ying LIU, Huiying HU, Haiyan SUN, Wei GUO

2023, 25(11):  192-206.  DOI: 10.13304/j.nykjdb.2022.0500

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Graphene is now widely used in agricultural production because of its unique structure and excellent properties. To investigate the effects of graphene on soil nutrients， enzyme activities and maize root development， the pot experiment using ‘Zhengdan 958’ as the test material was carried out to examine the effects of single-layer graphene （TS） and few-layer graphene （TF） at 0 （CK）， 25， 50， 100 and 150 g·kg^-1 on the transformation of soil nutrients and the root development of maize seedlings， respectively. The results showed that， compared with the control （CK）， the soil available nutrient content increased significantly with the increase of graphene concentration and reached the maximum at the concentration of 150 g·kg^-1， in which， the available nitrogen， available phosphorus and available potassium of single-layer graphene treatment significantly increased by 21.85%， 76.56% and 30.69%， and those of few-layer graphene treatment significantly increased by 23.05%， 82.16% and 47.20%， respectively. Meanwhile， the addition of graphene also contributed to the increase of soil enzyme activity， but the soil enzyme activity showed a trend of increasing firstly and then decreasing with increasing concentration， and the sucrase and urease activities of few-layer of graphene at a concentration of 50 g·kg^-1 significantly increased by 6.67% and 21.26% compared to CK. Low levels （25， 50， and 100 g·kg^-1） promoted inhibition at high level （150 g·kg^-1）. Low levels of graphene inhibited the rate of superoxide anion （{\mathrm{O}}_{\mathrm{2}}^{·-}） production and the accumulation of malondialdehyde （MDA）， and increased the contents of soluble protein and proline， which helped to timely scavenge reactive oxygen species， inhibit membrane lipid synthesis， lessen the harm done to the root system. In conclusion， the applied properly graphene could enhance soil nutrients， boost soil enzyme activity， and encourage the development of maize plants. And the effect of few-layer graphene was better than that of the single-layer graphene.

Study on Branching Architecture of Clipping Hedysarum scoparium in Desert-oasis Ecotone of Jilantai

Yajuan WEI, Jing GUO, Yunhu XIE, Xiangfei WANG, Shan JIN

2023, 25(11):  207-217.  DOI: 10.13304/j.nykjdb.2022.0417

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In order to investigate the response and adaptation mechanism of Hedysarum scoparium to different site types, typical sampling method was used to analyze the characteristics of shoot system and biomass allocation of Hedysarum scoparium in different site types (hilltop, gentle sandy land and interdune lowland) in the desert-oasis transition zone of Jilantai. The results showed that， the plant height and basal diameter of clipping H. scoparium in interdune lowland were significantly larger than that in hilltop (P<0.05)，and the above ground biomass of clipping H. scoparium showed as follows: interdune lowland>gentle land>hilltop. Overall bifurcation ratio increased gradually from interdune lowland to hilltop, indicating branching ability of H. scoparium in interdune lowland was weaker than that in hilltop and gentle land. Grade 2 and grade 3 bifurcation ratio （SBR_2∶3） in hilltop was greater than that in interdune lowland (P<0.05), indicating clipping H. scoparium grew in a spread-out branch architecture characteristics in hilltop. The branching length gradually decreased from interdune lowland to hilltop, branch diameter ratio of grade 2 and 3 (RBD_3:2) in hilltop was significantly higher than that in the gentle land and interdune lowland, indicating that the branching capacity of clipping H. scoparium in the hilltop was better than that in the gentle land and interdune lowland. Correlation analysis revealed that soil water content, soil temperature and atmospheric temperature had significant effects on plant height, crown width, basal diameter and aboveground biomass of clipping H. scoparium. The results showed that clipping H. scoparium could adapt to the change of external environment by increasing branch diameter ratio, reducing branch number and aboveground biomass in habitats with poor water conditions. Moreover, in the interdune lowland, chipping measure was more conducive to the rejuvenation and renewal of degenerated H. scoparium. The results were useful for the regeneration of H. scoparium plantations and ecological restoration in the region.

INNOVATIVE METHODS AND TECHNOLOGIES

Optimization on Extraction of Polysaccharide from Honeysuckle （Lonicera japonica） Leaves and Its Antibacterial Activity

Ruiwu SHI, Sufang ZHANG, Lina NIU, Tian SU, Xiaoqian ZHAO, Dongchen NA, Wei CHEN

2023, 25(11):  218-226.  DOI: 10.13304/j.nykjdb.2023.0312

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In order to clarify the antibacterial and moisturizing effects of polysaccharide from honeysuckle leaves in vitro， the polysaccharide was extracted by the optimized water extraction and alcohol precipitation method in this experiment. The antibacterial experiments against Escherichia coli， Bacillus subtilis and Staphylococcusaureus were carried out by the pouring plate with pre-additive bacterial solution and punching method. The polysaccharide from Lonicera japonica leaves was used to prepare the polysaccharide moisturizing cream so as to explore its moisturizing effect. The results showed that the content of polysaccharide was （17.29%±0.41%） using the optimized extraction method. The polysaccharide of Lonicera japonica leaves had inhibitory effects on the 3 bacteria， among them， the inhibitory effect on Escherichia coli was the best， with an average diameter of （15.80±0.58） mm， which was highly sensitive. The skin moisture test showed that the polysaccharide from Lonicera japonica leaves had good skin rehydration and moisturizing properties， which had potential development value. The extraction process of polysaccharides from Lonicera japonica leaves， laying a foundation for the deep development of Lonicera japonica and continuously improving its added value.

Construction of Chicken Embryo Fibroblast Cell Line with TET2 Gene Knockout Based on CRISPR-Cas9 Technology

Qiangzhou WANG, Shiyu PAN, Mengya FANG, Wei LI, Jiaxing WANG, Yinyin LIU, Shihao CHEN

2023, 25(11):  227-233.  DOI: 10.13304/j.nykjdb.2022.0497

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In order to studying the role of tet methylcytosine dioxygenase 2 （TET2） and its mediated hydroxymethylation in regulating innate immune responses， CRISPR-Cas9 mediated gene editing systems were used to construct a TET2 knockout chicken embryonic fibroblast cell line. 4 pairs of single guide RNAs （sgRNAs） were designed for targeting the conserved CDS sequences of the two RNA spliced isoforms of chicken TET2 gene. The plasmids expressing sgRNA was constructed， and were co-transfected with Cas9-T2A-GFP into DF-1 cells. DF-1 cells were transfected with a higher cleavage activity sgRNA. Then， the cells expressing GFP fluorescent were sorted by flow cytometry， and cultured and screened by limiting dilution method. The positive monoclonal cells were identified by DNA sequencing. When large numbers of the monoclonal knockout cells were genearated， the TET2 protein expression was detected by western blot and the level of hydroxymethylation （5hmC） was detected by dot blot. The results of DNA sequence showed that 2 and 28 bp deletion were detected in TET2 gene at position of the target site of TET2-gRNA-2. Both 2 deletions caused a reading frame shift and a premature stop codon. The clonal knockout cells were named DT-6. DT-6 cells were passaged for more than 15 times， the cell morphology was stable and the TET2 protein was not expressed. Compared with the DF-1 cells， the hydroxymethylation level of the cells was significantly reduced in DT-6 cells. In this study，the TET2 gene knockout chicken embryo fibroblast cell line was successfully constructed by applying CRISPR-Cas9 technology， which would provide a foundation for the further study of TET2 and its hydroxymethylation-mediated pathways and molecular mechanism in regulating innate immune response.