|Table of Contents|

Rapid And Efficient Selection For TransgenicSoybean Plants With The ImprovedGlufosinate Selection System*(PDF)

《大豆科学》[ISSN:1000-9841/CN:23-1227/S]

Issue:
2006年04期
Page:
373-378
Research Field:
Publishing date:

Info

Title:
Rapid And Efficient Selection For TransgenicSoybean Plants With The ImprovedGlufosinate Selection System*
Author(s):
Xue Rengao 　Xie Hongfeng
(Laiyang Agricultural College , Qingdao 266109)
Keywords:
Agrobacterium tumef aciens ; Cotyledonary node; Glufosinate; Soybean; Transfor-mation
PACS:
-
DOI:
10.11861/j.issn.1000-9841.2006.04.0373
Abstract:
　A rapid and efficient glufosinate selection system for obtaining high frequency of trans-formants insoybean [ Glycine max (L. ) Merrill] was developed. The cotyledonary node cellswere wounded and inoculated with Agrobacterium tumefaciens strain LBA4404 harboring a bina-ry vector pCAMBIA3201 that contained a selectable bar gene and a gus reporter gene. The pres-ent standard selection based on glufosinate was performed at 5 mg /L glufosinate during shoot in-duction and at 3～ 5 mg /L glufosinate during shoot elongation. Many non-transformants were es-caped with this standard selection system which resulted in low transformation efficiency of1. 6%, and a delayed shoot elongation with majority of transformed shoots elongation occurredfrom 21 to 41 weeks after Agrobacterium inoculation , and thus an improved glufosinate selectionsystem was provided in this paper. After 3 weeks on shoot induction medium without glufosi-nate , the explants were transferred to shoot elongation medium containing 4 mg /L glufosinate forthe first selection. The elongation of majority of transformed shoots occurred only from 7 to 12weeks without glufosinate selection during shoot induction. The transgenic shoots were effective-ly screened by placing the excised shoots on the root induction medium (RIM containing 3 mg /Lglufosinate to facilitate direct uptake of the selective agent that resulted in the high transforma-tion efficiency of 6. 7%. Shoots on the RIM rapidly responded to the selective agent applied , allthe glufosinate-sensitive shoots were completely necrotic within 10 days after selection. The ma-jority of transgenic plantlets were obtained only 8 ～ 16 weeks under the improved selection sys-tem. Genomic Southern blot analysis confirmed stable integration of the transgenes in the genomeof soybean. Stable expression was confirmed by GUS expression and herbicide application.

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Last Update: 2016-12-16