RESUMEN
Gerbera (Gerbera jamesonii L.) has its immense importance to the floriculture industry worldwide. The gerbera flower production has been hampered by various viruses, among them cucumber mosaic virus (CMV) has shown considerable damage.As natural resistance to CMV is absent in gerbera, here, we have made an attempt to develop transgenic gerbera plants expressing coat protein (CP) gene of CMV via Agrobacterium mediated transformation of base petiole explants for genetic resistance to CMV infection. Among the 44 putative transgenic gerbera plant acclimatized, 39 were found positive for integration of CP gene by polymerase chain reaction and southern hybridization assay using their specific primer and probe respectively. Northern hybridization assay using CP gene specific probe confirmed the transcription of transgene in all 39 transgenic plants. These plants showed translation of CP during DAS-ELISA when tested with antiserum specific to CP of CMV. These 39 plants when challenged by mechanical inoculations with CMV gerbera isolate showed virus resistance in 53% (21 out of 39) plants, virus tolerance (delayed mild symptom) in 33% (13/39) plants, while rest 12.8% (5/39) plants showed severe disease symptoms. The CP mediated resistance of CMV in transgenic gerbera is being reported for the first time from India.
RESUMEN
Stable transgenic rice line (named KRSV-1) with strong resistance against rice stripe virus was generated using the genesequence of disease-specific protein by RNA interference. Comprehensive safety assessment of transgenic plants has turnedinto a significant field of genetic modification food safety. In this study, a safety assessment of KRSV-1 was carried out in astepwise approach. The molecular analysis exhibited that KRSV-1 harbored one copy number of transgene, which wasintegrated into the intergenic non-coding region of chromosome 2 associated with inter-chromosomal translocations of 1.6-kb segments of chromosome 8. Then, transcriptomics and proteomics analyses were carried out to detect the unintendedeffects as a result of the integration of the transgene. Although 650 dramatically differentially expressed genes (DDEGs)and 357 differentially expressed proteins were detected between KRSV-1 and wild-type (WT) by transcriptomics andproteomics analyses, no harmful members in the form of toxic proteins and allergens were observed. Encouragingly, thenutritional compositions of seeds from KRSV-1 were comparable with WT seeds. The results of this entire study ofmolecular analysis, transcriptome and proteome profile of KRSV-1 revealed that no detrimental changes in the form of toxicproteins and allergens were detected in the transgenic rice line due to the integration of the transgene.
RESUMEN
MP-Rep fusion gene which size is about 1000bp was constructed by recombinant PCR technique.In order to construct fusion gene,Tobacco mosaic virus(TMV) partial movement protein gene(MP) and Cucumber mosaic virus(CMV) partial replicase gene(rep) were ligated.Two copies of MP-Rep fusion gene were ligated with soybean intron in inverted repeat manner,the recombinant fragments were then inserted into binary vector pBIN438 under the control of 35S promoter.Recombinant clone pBIN438-MP-Rep(i/r) which contained two different virus derived genes was constructed.Recombinant clone pBIN438-MP-Rep(i/r) in accord with expected design was certified by restriction endonuclease enzymes digestion and PCR analysis.This approach provides a basis for Broad-spectrum plant virus resistance mediated by RNA Silencing.