0306878 - ÚEB 2008 RIV NL eng J - Journal Article
Kertbundit, Sunee - Pongtanom, N. - Ruanjan, P. - Chantasingh, D. - Tanwanchai, A. - Panyim, S. - Juříček, Miloslav
Resistance of transgenic papaya plants to Papaya Ringspot Virus, Thai isolate.
[Resistence transgení papáje k thajskému izolátu viru kroužkovitosti papáje.]
Biologia Plantarum. Roč. 51, č. 2 (2007), s. 333-339. ISSN 0006-3134. E-ISSN 1573-8264
Grant - others:BIOTEC, NASDA(TH) BT-B-06-PG-14-4503
Institutional research plan: CEZ:AV0Z50380511
Source of funding: V - Other public resources
Keywords : biolistic transformation * coat protein * plant regeneration
Subject RIV: EF - Botanics
Impact factor: 1.259, year: 2007

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.

Byla generována papája rezistentní k viru kroužkovitosti papáje (PRSV) a to biolistickou transformací somatických embryí genem pro obalový protein (CP) tohoto viru pod kontrolou promotoru 35S. Z celkem 1980 embryogeních kalusů bylo pouze 8 vyselektováno na půdě s kanamycinem. U všech těchto linii byl detegován CP pomocí PCR a Southern blot hybridizace. Avšak pouze linie G2 byla 100 % rezistentní k viru PRSV.
Permanent Link: http://hdl.handle.net/11104/0159782