NDLI: Agrobacterium–mediated genetic transformation of Elymus breviaristatus with Pseudomonas pseudoalcaligenes insecticidal protein gene

Content Provider	Springer Nature Link
Author	Li, Daxu Sun, Qun Huang, Min Zhang, Jie Bai, Shiqie Zheng, Linyong Zhao, Jian Qiu, Dunlian Li, Li Yang, Zhirong You, Minghong Liu, Gang Zhang, Yu Zhang, Changbing Li, Shaocai
Copyright Year	2007
Abstract	An efficient and repeatable protocol for generating transgenic Elymus breviaristatus plants was first established by Agrobacterium-mediated transformation: calli of Elymus breviaristatus were induced from mature seeds, and proliferated on callus induction and maintenance medium containing kinetin 0.05 mg/l and 2, 4-D 8.0 mg/l under dim-light condition (10–20 μmol/m$^{2}$s,16 h light) at 26°C. The calli produced were used for transformation mediated by Agrobacterium tumefaciens, EHA105, which carries the binary vector pCAMBIA 1304-ppip, coding for the Pseudomonas pseudoalcaligenes insecticidal protein gene (ppip) and hygromycin phosphotransferase II gene (hptII). The calli transformed were selected and grown in the presence of 60 mg/l hygromycin. Approximately 90 transgenic plants were produced, and transformation frequency of the calli reached 10.52%. The presence of ppip gene in the genomic DNA of regenerated plants was detected by means of PCR and Southern-blot analysis, and the expression of the transgenes was verified by reverse transcription-PCR. Approximately 90% of the tested plants appeared to have only one or two copies of the T-DNA inserts. Verification of anti-grasshoppers activity shows the mortality of grasshoppers reached 16.73%. These results indicated that Agrobacterium-mediated transformation in our condition was effective for transferring foreign genes into Elymus breviaristatus.
Starting Page	159
Ending Page	168
Page Count	10
File Format	PDF
ISSN	01676857
Journal	Plant Cell, Tissue and Organ Culture (PCTOC)
Volume Number	89
Issue Number	2-3
e-ISSN	15735044
Language	English
Publisher	Kluwer Academic Publishers
Publisher Date	2007-06-14
Publisher Place	Dordrecht
Access Restriction	One Nation One Subscription (ONOS)
Subject Keyword	Agrobacterium–mediated transformation Callus regeneration Mature embryo ppip gene Transgenic plant Plant Physiology Plant Sciences
Content Type	Text
Resource Type	Article
Subject	Horticulture

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