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Title:: Construction of Transformation Vector of GmAOC3 Gene and Preliminary Study on the Transformation of Soybean

Author(s):: NING Ai-ling; DU Hai-ping; YU De-yue; WANG Hui; Soybean Research Institute, Nanjing Agricultural University/National Center for Soybean Improvement, Ministry of Agriculture/Key Laboratory for Biology and Genetic Improvement of Soybean (General), Ministry of Agriculture / National Key Laboratory of Crop Genetics and Germplasm Enhancement; Nanjing 210095,China.

Keywords:: Soybean; Allene oxide cyclase; Vector construction; Genetic transformation; Molecular detection

PACS:: -

DOI:: 10.11861/j.issn.1000-9841.2015.04.0588

Abstract:: Allene oxide cyclase (AOC) plays a key role in the synthesis of jasmonic acid, which is an important defense signaling molecular in plant. In this study, we constructed a recombinant vector pBA002-GmAOC3 with bar gene using double enzymes cleavage method (SacⅠand MluⅠ) and the recombinant vector was successfully transformed into soybean via Agrobacterium transformation system. Two soybean varieties (Jack and Nannong 88.1) and explants (cotyledonary node and whole cotyledonary node) were used to study their effects on the Agrobacterium-mediated transformation of GmAOC3. The results showed that the whole cotyledonary node of Jack had the highest budding rate (79.5%) and conversion rate (2.27%) Based on the results of QuickStix PAT/bar- test paper detection, molecular detection of target DNA and Basta resistance test of leaves, we obtained three T₀generation GmAOC3 positive transgenic soybean plants and six T₁generation GmAOC3 positive transgenic soybean plants, of which five T₁generation transgenic plants from Jack showed higher expression level of GmAOC3 than Jack. We also used the fluorescence quantitative PCR to detect the copy number of target gene Besides two T₁generation transgenic plants with two copies of bar genes, the others contained one copy of bar gene. The data in this study would give some references to soybean genetic transformation and the GmAOC3 transgenic plant obtained in this study could be used in soybean insect resistance breeding in future.

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Construction of Transformation Vector of GmAOC3 Gene and Preliminary Study on the Transformation of Soybean(PDF)

《大豆科学》[ISSN:1000-9841/CN:23-1227/S]

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