caa a22 4500 463160800 CHVBK 20180406164747.0 cr unu---uuuuu 170326e20070601xx s 000 0 eng 10.1007/s10535-007-0065-1 doi (NATIONALLICENCE)springer-10.1007/s10535-007-0065-1 Resistance of transgenic papaya plants to Papaya ringspot virus [Elektronische Daten] [S. Kertbundit, N. Pongtanom, P. Ruanjan, D. Chantasingh, A. Tanwanchai, S. Panyim, M. Juříček] 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. Institute of Experimental Botany, ASCR, 2007 biolistic transformation nationallicence coat protein nationallicence plant regeneration nationallicence AMV : alfalfa mosaic virus nationallicence CP : coat protein nationallicence CPMR : coat protein mediated resistance nationallicence GUS : glucuronidase nationallicence NTR : non translated region nationallicence PCR : polymerase chain reaction nationallicence PRSV : Papaya ringspot virus nationallicence PTGS : post-transcriptional gene silencing nationallicence RT-PCR : reverse transcription PCR nationallicence Kertbundit S. Department of Plant Science, Faculty of Science, Mahidol University, Rama VI Road, 10400, Payathai, Bangkok, Thailand aut Pongtanom N. Institute of Molecular Biology and Genetics, Mahidol University, Salaya Campus, 73170, Nakhon Pathom, Thailand aut Ruanjan P. Institute of Molecular Biology and Genetics, Mahidol University, Salaya Campus, 73170, Nakhon Pathom, Thailand aut Chantasingh D. Institute of Molecular Biology and Genetics, Mahidol University, Salaya Campus, 73170, Nakhon Pathom, Thailand aut Tanwanchai A. Institute of Molecular Biology and Genetics, Mahidol University, Salaya Campus, 73170, Nakhon Pathom, Thailand aut Panyim S. Department of Plant Science, Faculty of Science, Mahidol University, Rama VI Road, 10400, Payathai, Bangkok, Thailand aut Juříček M. Institute of Experimental Botany, Academy of Sciences of the Czech Republic, Na Pernikářce 15, CZ-16000, Prague, Czech Republic aut Biologia Plantarum Kluwer Academic Publishers-Consultants Bureau 51/2(2007-06-01), 333-339 0006-3134 51:2<333 2007 51 10535 https://doi.org/10.1007/s10535-007-0065-1 text/html Onlinezugriff via DOI 1 research-article jats NATIONALLICENCE 856 40 https://doi.org/10.1007/s10535-007-0065-1 text/html Onlinezugriff via DOI NATIONALLICENCE 700 1- Kertbundit S. Department of Plant Science, Faculty of Science, Mahidol University, Rama VI Road, 10400, Payathai, Bangkok, Thailand aut NATIONALLICENCE 700 1- Pongtanom N. Institute of Molecular Biology and Genetics, Mahidol University, Salaya Campus, 73170, Nakhon Pathom, Thailand aut NATIONALLICENCE 700 1- Ruanjan P. Institute of Molecular Biology and Genetics, Mahidol University, Salaya Campus, 73170, Nakhon Pathom, Thailand aut NATIONALLICENCE 700 1- Chantasingh D. Institute of Molecular Biology and Genetics, Mahidol University, Salaya Campus, 73170, Nakhon Pathom, Thailand aut NATIONALLICENCE 700 1- Tanwanchai A. Institute of Molecular Biology and Genetics, Mahidol University, Salaya Campus, 73170, Nakhon Pathom, Thailand aut NATIONALLICENCE 700 1- Panyim S. Department of Plant Science, Faculty of Science, Mahidol University, Rama VI Road, 10400, Payathai, Bangkok, Thailand aut NATIONALLICENCE 700 1- Juříček M. Institute of Experimental Botany, Academy of Sciences of the Czech Republic, Na Pernikářce 15, CZ-16000, Prague, Czech Republic aut NATIONALLICENCE 773 0- Biologia Plantarum Kluwer Academic Publishers-Consultants Bureau 51/2(2007-06-01), 333-339 0006-3134 51:2<333 2007 51 10535 Metadata rights reserved Springer special CC-BY-NC licence nationallicence BK010053 XK010053 XK010000 NATIONALLICENCE NATIONALLICENCE NL-springer