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Evidence for RNA-mediated resistance to PVYn in tobacco plants transformed with the viral coat protein gene(PDF)

Chinese Journal of Applied & Environmental Biology[ISSN:1006-687X/CN:51-1482/Q]

Issue:
Vol.09 No.4
Page:
372-376
Research Field:
Reviews
Publishing date:

Info

Title:
Evidence for RNA-mediated resistance to PVYn in tobacco plants transformed with the viral coat protein gene
Author(s):
GUO Xingqiet al.
( College of Life Sciences; 1 College of Plant Protection, Shandong Agricultural University, Tai an271018, China )
Keywords:
Keywords Potato virus Y coat protein gene transgenic tobacco resistance mechanism
CLC:
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PACS:
DOI:
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DocumentCode:

Abstract:
Abstract According to the published nucleotide sequence of tobacco veinal necrosis strain (PVYN), the PVYN coat protein (CP) gene was synthesized by reverse transcription-polymerase chain reaction, and constructed into the cloning vector pBSK. The PVYN CP gene was introduced into tobacco (NC89) plants via Agrobacterium tumefaciens (LBA4404)-mediated transformation and the transformed tissue was selected in the presence of Kanamycin sulfate. Regenerated plants were screened by PCR and 38 transgenic plants were obtained. Resistant test indicated that the resistant levels of the transgenic plants against PVYN varied among different transgenic plants. Four lines were highly resistant to PVYN infection. Southern blot analysis indicated that the CP gene had been integrated into the tobacco chromosomes and the resistance levels of the transgenic plants were related to copy numbers of the transgene. The highly resistant plants contained 5~6 copies of the transgene, the resistant plants contained 3~4 copies and the susceptible plants contained 1~2 copies. Northern blot analysis demonstrated that the transgene was expressed at RNA level and there was an inverse correlation between transgene transcript accumulation and virus resistance. Western blot analysis showed that no specific protein was observed in highly resistant plants, transgene CP detected in both the resistant and susceptible plants, and the CP accumulation varied among different transgenic plants. Results indicated that the resistance mechanism of the transgenic plants was similar to post-transcriptional gene silence, i.e., RNA-mediated resistance. Fig 7, Tab 1, Ref 13

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Last Update: 2001-08-26