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Correlation Analysis of New Soybean[Glycine max(L.)Merr]Gene Gm15G117700 with Oleic Acid

Phyton-International Journal of Experimental Botany 2021年第4期90卷 1177-1192页

作者： Shuo Qu Yaolei Jiao lamboro abraham Piwu Wang Plant Biotechnology Center Jilin Agricultural UniversityChangchun130018China College of Agronomy Jilin Agricultural UniversityChangchun130018China

The fatty acid dehydrogenase gene plays an important role in regulating the oleic acid content in ***-wide association study screened out soybean oleic acid related gene Gm15G117700.A fragment size of 693bp was obtained by PCR amplification of the gene and,it was connected by seamless cloning technology to the pMD18T cloning *** on the gene sequence cloned,bioinformatic analysis of gene protein was *** overexpression vector of Gm15G117700 and the CRISPR/Cas9 gene editing vector were *** positive plants were obtained by Agrobacterium-mediated transformation of soybean cotyledon nodes and T2 plants were identified by conventional PCR,QT-PCR and Southern blot hybridization.10 copies of high and low oleic acid seeds were selected for QT-PCR to identify the expression content of Gm15G117700 gene in different soybeans,and finally near-infrared spectroscopy analyzer was used to identify the oleic acid quality of soybeans.T2 RT-PCR identification showed that overexpression was reduced by 3.94%,and gene editing was increased by 3.49%.It is determined that the Gm15G117700 gene may belong to a regulatory gene,a minor gene that can promote the conversion to linoleic acid content in soybean oleic acid *** gene cloning and its functional verification was not reported *** is the first report by PCR amplification of soybean Gm15G117700 genes and gene expression *** the content of oleic acid in soybean lay a foundation for ***;this study clearly identified the function of soybean Gm15G117700 gene and its role played in oleic acid synthesis and metabolism.

关键词： Soybean Gm15G117700 genes cloning expression vector

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Bioinformatics Analysis of Disease Resistance Gene PR1 and Its Genetic Transformation in Soybeans and Cultivation of Multi-resistant Materials

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Phyton-International Journal of Experimental Botany 2022年第7期91卷 1445-1464页

作者： Huimin Cui Shuo Qu abraham lamboro Yaolei Jiao Piwu Wang College of Agronomy Jilin Agricultural UniversityChangchun1300118China

In agricultural production,a single insect-resistant and disease-resistant variety can no longer meet the *** this study,the expression vector pCAMBIA-3301-PR1 containing the disease-resistant gene PR1 was constructed by means of genetic engineering,and the PR1 gene was genetically transformed to contain the PR1 gene through the pollen tube *** CryAb-8Like transgenic high-generation T7 receptor soybean,a new material that is resistant to insects and diseases is *** T2 transformed plants,routine PCR detection,Southern Blot hybridization,fluorescence quantitative PCR detection,indoor and outdoor pest resistance identification and indoor disease resistance identification were *** results showed that there were 9 positive plants in the routine PCR test of T2 *** Southern Blot hybridization,both PR1 and CryAb-8Like genes are integrated in soybeans in the form of single *** quantitative PCR showed that the expression levels of PR1 and CryAb-8Like genes are different in different *** average expression levels of PR1 gene in plant roots,stems,and leaves are 2.88,1.54,and 5.26,***-8Like genes are found in roots,stems,and *** average expression levels were 1.36,1.39,and 4.25,*** insectivorous rate of the CryAb-8Like gene in outdoor plants with positive insect resistance identification was 3.78%.The disc partition method was used indoors for pest resistance identification,and the bud length of transformed plants increased *** average mortality rate of untransformed plants in indoor disease resistance identification was as high as 56.66%,and the average mortality rate of plants transformed with PR1 gene was 10.00%,and disease resistance was significantly ***,a new material with resistance to diseases and insects is obtained.

关键词： Soybean PR1 gene CryAb-8Like gene insect resistance disease resistance

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Evaluation of Beta-Lactam Antibiotics on the Regeneration of Peanut Plants and Their Inhibitory Effect on Agrobacterium Growth

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Phyton-International Journal of Experimental Botany 2023年第9期92卷 2489-2501页

作者： abraham lamboro Songnan Yang Xueying Li Dan Yao Jun Zhang College of Agronomy Jilin Agricultural UniversityChangchun130118China College of Life Science Jilin Agricultural UniversityChangchun130118China Department of Biology College of Natural and Computational ScienceWachemo UniversityHossana667Ethiopia

The effect of beta-lactam antibiotics on shoot induction and plantlet regeneration from cotyledonary nodes was tested using two peanut *** culture media contained 4 mg/L 6-benzylaminopurine(BAP)as the main growth *** concentrations(100–600 mg/L)of cefotaxime,carbenicillin,and timentin were applied in the culture *** all the tested media,there were no significant differences in the shoot induction as compared to the ***,little phytotoxic effect was observed at higher concentrations of these antibiotics in the shoot elongation *** shoot elongation medium,shoots turned brownish and partly died at higher concentrations where shooting rates were not affected by the *** cefotaxime,timentin,and carbenicillin-containing media,levels of antibiotics greater than 400,300,and 200 mg/L,respectively resulted in the brown coloration of ***,the mean shoot number and shoot weight significantly decreased as their dosage *** results indicate that maximum levels of antibiotics have an adverse effect on the growth and development of ***,cefotaxime(100–300 mg/L)and timentin(100–300 mg/L)will be sufficient in controlling Agrobacterium growth in the culture media with the least phytotoxic effect on the peanut plants.

关键词： Peanut antibiotics shoot cotyledonary node regeneration

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Bioinformatics and Functional Analysis of High Oleic Acid-Related Gene GmSAM22 in Soybean [Glycine max (L.) Merr.]

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Phyton-International Journal of Experimental Botany 2023年第2期92卷 501-519页

作者： Shuo Qu Qi Cai Huimin Cui lamboro abraham Yaolei Jiao Guilong Ma Piwu Wang College of Agronomy Jilin Agricultural UniversityChangchun1300118China College of Plant Protection Agricultural UniversityChangchun1300118China

High yield,high quality,stable yield,adaptability to growth period,and modern mechanization are the basic requirements for crops in the 21st *** oleic acid is a natural unsaturated fatty acid with strong antioxidant properties and *** as a safe fatty acid,it has the ability to successfully prevent cardiovascular and cerebrovascular *** the fatty acid composition of soybean seeds,can not only speed up the breeding process of high-quality high-oil and high-oleic soybeans,but also have important significance in human health,and provide the possibility for the development of soybean oil as a new energy ***,the aim of this study was to analyze the high oleic acid elated gene GmSAM22 in *** this research the soybean oleic acid-related gene GmSAM22 was screened out by Genome-wide association analysis,a 662 bp fragment was acquired by specific PCR amplification,and the pMD18T cloning vector was linked by the use of a seamless cloning *** analysis of the signal peptide prediction,subcellular localization,protein hydrophobicity,transmembrane region analysis,a phosphorylation site,protein secondary and tertiary structure and protein interaction analysis of the protein encoded by the SAM22 gene was carried *** plasmid of the gene editing vector is *** overexpression vector was transformed from pCAMBIA3301 as the base vector,and overexpression vector were *** plants were obtained by genetic transformation by the pollen tube channel *** quantitative PCR was performed on the T2 generation plants to detect the relative expression levels in different *** Blot was used to detect the presence of hybridization *** genes BAR,35S,and NOS in plants were identified by conventional PCR.10 seeds with high and low oleic acid content were chosen for quantitative PCR identification,and finally,the concentration and morphology of soybean fatty acids were

关键词： Soybean SAM22 gene oleic acid cloning CRISPR/Cas9 expression vector

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Combination of 6-Benzylaminopurine and Thidiazuron Promotes Highly Efficient Shoot Regeneration from Cotyledonary Node of Mature Peanut (Arachis hypogaea L.) Cultivars

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Phyton-International Journal of Experimental Botany 2022年第12期91卷 2619-2631页

作者： abraham lamboro Xiao Han Songnan Yang Xueying Li Dan Yao Baixing Song Jun Zhang Department of Crop Genetics and Breeding College of AgronomyJilin Agricultural UniversityChangchun130118China Department of Biochemistry and Molecular Biology College of Life ScienceJilin Agricultural UniversityChangchun130118China

Efficient in vitro plantlet regeneration is an important step to successfully transform genes for the improvement of agronomic traits.A combination of 6-benzylaminopurine(BAP)and thidiazuron(TDZ)plant growth regulators was applied to evaluate shoot regeneration capacity whereasα-naphthalene acetic acid(NAA)combination with 6-benzylaminopurine(BAP),and 2,4-dichlorophenoxyacetic acid(2,4-D)with 6-benzylaminopurine were tested to optimize root induction for two peanut *** result showed combination(BAP with TDZ)was found to be effective in promoting *** highest shoot regeneration frequency(93%)was obtained on a medium supplemented with 4 mg/L BAP and 0.5 mg/L TDZ while an average regeneration frequency(87%)was achieved in a medium containing combinations of 2 mg/L BAP with 1 mg/L *** shooting rate increased for both cultivars as the concentrations of BAP increased and TDZ *** highest rooting rate(93%)was obtained on a medium supplemented with 3.5 mg/L NAA with 2.5 mg/L BAP for both *** rooting rate increased as the concentration of auxin to cytokinin ratio *** maximum rooting rate(83%)was obtained on MS medium supplemented with 0.3 mg/L 2,4-D with 0.2 mg/L BAP for the cultivar *** result indicated that BAP with NAA was much better than BAP with 2,4-D in rooting ***,the protocol developed was genotype independent and effective for peanut tissue culture.

关键词： Auxin cotyledonary node cytokinins in vitro peanut

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Construction and Functional Analysis of CRISPR/Cas9 Vector of FAD2 Gene Family in Soybean

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Phyton-International Journal of Experimental Botany 2022年第2期91卷 349-361页

作者： Zhifeng Xiao Yukun Jin Qi Zhang abraham lamboro Baozhu Dong Zhuoyu Yang Piwu Wang College of Agronomy Jilin Agricultural UniversityChangchun130118China

Soybean oleic acid content is one of the important indexes to evaluate the quality of soybean *** the synthesis pathway of soybean fatty acids,the FAD2 gene family is the key gene that regulates the production of linoleic acid from soybean oleic *** this study,CRISPR/Cas9 gene editing technology was used to regulate FAD2 gene ***,the CRISPR/Cas9 single knockout vectors GmFAD2-1B and GmFAD2-2C and double knockout vectors GmFAD2-2A-3 were ***,the three vectors were transferred into the recipient soybean variety Jinong 38 by Agrobacterium-mediated cotyledon node transformation,and the mutant plants were *** analysis and comparison of the mutant plants of the T2 and T3 generations were carried *** results showed that there was no significant difference in agronomic traits between the CRISPR/Cas9 single and double knockout vectors and the untransformed CRISPR/Cas9 receptor *** oleic acid content of the plants that knocked out the CRISPR/Cas9 double gene vector was significantly higher than that of the single gene vector.

关键词： Soybean FAD2 gene family CRISPR/Cas9 oleic acid content

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