Genetic transformation of the actinorhizal tree Allocasuarina verticillata by Agrobacterium tumefaciens

被引：53

作者：

Franche, C ^{[1
]}

Diouf, D ^{[1
]}

Le, QV ^{[1
]}

Bogusz, D ^{[1
]}

NDiaye, A ^{[1
]}

Gherbi, H ^{[1
]}

Gobe, C ^{[1
]}

Duhoux, E ^{[1
]}

机构：

[1] UNIV PARIS 07, F-75005 PARIS, FRANCE

来源：

PLANT JOURNAL | 1997年 / 11卷 / 04期

关键词：

D O I：

10.1046/j.1365-313X.1997.11040897.x

中图分类号：

Q94 [植物学];

学科分类号：

071001 ;

摘要：

We have developed an efficient transformation system for the tropical actinorhizal tree Allocasuarina verticillata using Agrobacterium tumefaciens mediated gene transfer. Mature zygotic embryos were inoculated with the disarmed strain C58C1 carrying, in the binary vector BIN19, the nptll gene, providing kanamycin resistance as a selectable marker, and the reporter gene beta-glucuronidase containing an intron. The transformed embryos were cultivated on nutrient medium supplemented with 0.5 mu M NAA, 2.5 mu M BA, 100 mg l(-1) kanamycin and 250 mg l(-1) cefotaxime. After 2 months, a 21% transformation frequency was obtained. Within 6-9 months, transgenic plants were recovered from 70% of the transformed calli. The presence of the transgenes was demonstrated by PCR analysis and by the expression of the beta-glucuronidase; integration of the T-DNA was confirmed by Southern hybridization. More than 100 transgenic plants from a total of 23 independent transformation events have been successfully established in soil. The possibility to obtain nitrogen-fixing nodules after inoculation of transgenic A. verticillata plants by the actinomycetal strain of Frankia Allo2 was established.

引用

页码：897 / 904

页数：8

共 32 条

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