Resistance of transgenic papaya plants to Papaya ringspot virus

被引:11
|
作者
Kertbundit, S.
Pongtanom, N.
Ruanjan, P.
Chantasingh, D.
Tanwanchai, A.
Panyim, S.
Juricek, M.
机构
[1] Mahidol Univ, Fac Sci, Dept Plant Sci, Bangkok 10400, Thailand
[2] Mahidol Univ, Inst Mol Biol & Genet, Nakhon Pathom 73170, Thailand
[3] Acad Sci Czech Republ, Inst Expt Biol, CZ-16000 Prague, Czech Republic
关键词
biolistic transformation; coat protein; plant regeneration;
D O I
10.1007/s10535-007-0065-1
中图分类号
Q94 [植物学];
学科分类号
071001 ;
摘要
The coat protein gene isolated from Papaya ringspot virus, Thai isolate, was used to generate transgenic papayas. A binary vector containing the coat protein gene under the control of a 35S promoter, was constructed and transformed into somatic embryos of papaya cultivar Khak Dum by microprojectile bombardment. Eight transgenic lines were identified from 1980 bombarded calli of papaya somatic embryos under kanamycin selection. Integration of the transferred genes into kanamycin resistant papaya calli was verified by PCR amplification of the coat protein gene, GUS assays and Southern blot hybridization. Although the coat protein gene was detected in all transgenic lines, only line G2 was found to be highly resistant to virus. This resistant line showed high degree of rearrangement of the inserted coat protein expression cassette while the coat protein gene itself had a deletion of 166 bp on the 3' end of its sequence. Although the transcription of the coat protein gene was detected in all transgenic lines by RT-PCR, only two transgenic papayas expressed the intact coat protein. Moreover, in the resistant line G2 the amount of the truncated coat protein mRNA was significantly decreased. These results point to an RNA mediated mechanism of coat protein mediated resistance in papaya, probably based on post-transcriptional gene silencing.
引用
收藏
页码:333 / 339
页数:7
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