Science Letters:Optimization of soybean (Glycine max (L.) Merrill) in planta ovary transformation using a linear minimal gus gene cassette  

作　　者：Ming LIU Jun YANG Yun-qing CHENG Li-jia AN 

机构地区：[1]Department of Bioscience and Biotechnology, Dalian University of Technology, Dalian 116024, China [2]Institute of Eco-environmental Sciences, Jilin Normal University, Siping 136000, China

出　　处：《Journal of Zhejiang University-Science B(Biomedicine & Biotechnology)》2009年第12期870-876,共7页浙江大学学报（英文版）B辑（生物医学与生物技术）

基　　金：Project (No. JY03-B-18-02) supported by the National R & D Project of Transgenic Crops of Ministry of Science and Technology of China

摘　　要：Soybean transformation by ovary-drip was improved by optimizing the length of the transformation pathway by cutting the styles. These modifications facilitated soybean transformation manipulation and improved transformation reproducibility and efficiency. Using a linear minimal gus gene cassette as the foreign DNA, a maximum transformation frequency of 11% was obtained in flowers of the soybean cultivar ‘Liaodou 14’ with their styles mostly removed, whereas removal of only the stigma, partial style cutting and partial ovary cutting gave transformation frequencies of 0%, 1%, and 2%, respectively. An average transformation frequency of 8.2% was obtained when 619 flowers from three soybean cultivars (‘Liaodou 14’, ‘Liaodou 13’, and ‘Tiefeng 29’) were transformed by this optimized method. Southern blotting analysis showed that the gus reporter gene (encoding β-glucuronidase) was stably inherited with a simple pattern. Reverse transcription-polymerase chain reaction (RT-PCR) and GUS staining confirmed the expression of the gus gene in transgenic plants.Soybean transformation by ovary-drip was improved by optimizing the length of the transformation pathway by cutting the styles. These modifications facilitated soybean transformation manipulation and improved transformation reproducibility and efficiency. Using a linear minimal gus gene cassette as the foreign DNA, a maximum transformation frequency of 11% was obtained in flowers of the soybean cultivar ＇Liaodou 14＇ with their styles mostly removed, whereas removal of only the stigma, partial style cutting and partial ovary cutting gave transformation frequencies of 0%, 1%, and 2%, respectively. An average transformation frequency of 8.2% was obtained when 619 flowers from three soybean cultivars （＇Liaodou 14＇, ＇Liaodou 13＇, and ＇Tiefeng 29＇） were transformed by this optimized method. Southern blotting analysis showed that the gus reporter gene （encoding β-glucuronidase） was stably inherited with a simple pattern. Reverse transcription-polymerase chain reaction （RT-PCR） and GUS staining confirmed the expression of the gus gene in transgenic plants.

关 键 词：SOYBEAN Glycine max （L.） Merrill In planta ovary-drip transformation Optimization Style cutting Minimal gus renorter gene cassette 

分 类 号：S792.950.4[农业科学—林木遗传育种] S565.1[农业科学—林学]