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Received: 2009-07-14

Revision Accepted: 2009-11-03

Crosschecked: 2009-11-16

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Journal of Zhejiang University SCIENCE B 2009 Vol.10 No.12 P.870~876


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

Author(s):  Ming LIU, Jun YANG, Yun-qing CHENG, Li-jia AN

Affiliation(s):  Department of Bioscience and Biotechnology, Dalian University of Technology, Dalian 116024, China; more

Corresponding email(s):   junyang@dlut.edu.cn, ljan.bio@gmail.com

Key Words:  Soybean, Glycine max (L.) Merrill, In planta ovary-drip transformation, Optimization, Style cutting, Minimal gus reporter gene cassette

Ming LIU, Jun YANG, Yun-qing CHENG, Li-jia AN. Optimization of soybean (Glycine max (L.) Merrill) in planta ovary transformation using a linear minimal gus gene cassette[J]. Journal of Zhejiang University Science B, 2009, 10(12): 870~876.

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author="Ming LIU, Jun YANG, Yun-qing CHENG, Li-jia AN",
journal="Journal of Zhejiang University Science B",
publisher="Zhejiang University Press & Springer",

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%T Optimization of soybean (Glycine max (L.) Merrill) in planta ovary transformation using a linear minimal gus gene cassette
%A Ming LIU
%A Yun-qing CHENG
%A Li-jia AN
%J Journal of Zhejiang University SCIENCE B
%V 10
%N 12
%P 870~876
%@ 1673-1581
%D 2009
%I Zhejiang University Press & Springer
%DOI 10.1631/jzus.B0920204

T1 - Optimization of soybean (Glycine max (L.) Merrill) in planta ovary transformation using a linear minimal gus gene cassette
A1 - Ming LIU
A1 - Jun YANG
A1 - Yun-qing CHENG
A1 - Li-jia AN
J0 - Journal of Zhejiang University Science B
VL - 10
IS - 12
SP - 870
EP - 876
%@ 1673-1581
Y1 - 2009
PB - Zhejiang University Press & Springer
ER -
DOI - 10.1631/jzus.B0920204

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.

Darkslateblue:Affiliate; Royal Blue:Author; Turquoise:Article


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