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Nguyen Xuan Dung. 2017. Orchid Breeding To Improve The Dendrobium Resistance To Cymbidium Mosaic Virus VIA Rnai – Assessment of Technical Efficiency . PhD Thesis. Institute of Agricultural Sciences for Southern Vietnam.
Thứ tư, 10-01-2018 | 11:42:44

PhD Thesis submitted in September 21 2017, at the IAS, Ho Chi Minh City, Vietnam.

ABSTRACT

Dendrobium is a major orchid species growing in Vietnam farms. Two viruses CyMV (Cymbidium mosaic virus) and ORSV (Ondotoglossum rinspot virus) have become severe damages to orchids. However, the infection to Dendrobium by CyMV in Ho Chi Minh City offered 67.5% more severely than ORSV. Variety Sonia’ s PLDs were successfully transformed via Agrobacterium tumefaciens – mediated with strain C58 (EHẠ and LBA4404). MS medium plus 100 µM AS was used for three and five days with strain C58, EHẠ and LBA404, respectively. The CP (coat protein) sequences by CyMV, which were identified from various places in Vietnam, expressed their similarity of 95-99%. Vectors transferred RNAi were designed from single CP sequences (RNAi-CP) or designed from the association by ORF2-4 (RNAi – ORF2-4 + CP) by CyMV with Gateway protocol. Both of them successfully offered good transfomants. PLBs carried RNAi-CP/ORF2-4+CP were then selected in the medium plus kanamycin (500 mg/L). PCR analysis was used to examine the target gene. After six months; orchid plants in 1/2MS culture medium could be harvested with three distinctive phenotypes as compared to ordinary phenotype. The plant resistance to CyMV could be evaluated in vitro through virus inoculation. The transformants exhibited better resistance to CyMV than wild type.

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