Cloning And Analysis Of Tomato & Pepper Eukaryotic Initiation Factor 4E Related Genes

The plant virus causes decline of production and the quality of grain,oil,vegetables, fruit and flowers,tomato and pepper are our country important vegetable crops,have been suffered by virus disease,cause severe losses in the production.The growing knowledge on eukaryotic initiation factor 4E has provided the novel strategy for genetic engineering for virus resistance.The protein translation mechanism in host is required for virus infection and multiplication.Recent research demonstrates that virus resistance mediated by eIF4E and its isoform eIF(iso)4E occurs in several plant-virus interactions, thus indicating the potential new role of eIF4E/eIF(iso)4E in resistance strategy against plant viruses.Interfering the process of virus utilizing of the protein translation via genetic engineering not only improves the virus resistance of host plants,but also helps to deepen insights into molecular interaction of plant-virus.So another part of this thesis was to clone the eukaryotic initiation factor 4E from tomato and pepper,and regulate the eIF4E and eIF(iso)4E expression in tomato and pepper via sense and antisense strategy.This would not only improve virus resistance in plants but also help to elucidate the function of eIF4E and eIF(iso)4E in plant virus interaction.Main experiments were carried out as follows:1.cDNA of Eukaryotic initiation factor 4E(eIF4E) from tomato ZS5,this cDNA contains a ORF length in 687 bp,encoding 228aa.and show 98%and 100% nucleotide and amino acid sequence identities to the reported gene AY723735.2.Sense construct for tomato eIF4E and antisense construct for tomato eIF4E was respectively constructed.3.Agrobacterium-mediated transformation was carried out on tomato(ZS5) with tomato eIF4E sense construct and antisense construct.Thirteen and four kanamycin resistant plants were obtained respectively.4.cDNA of Eukaryotic initiation factor 4E(eIF4E) from pepper SS5,this cDNA contains a ORF length in 687 bp,encoding 228aa.and show 100%and 100% nucleotide and amino acid sequence identities to the reported gene AF521963.5.Sense construct for pepper eIF4E and antisense construct for pepper eIF4E was respectively constructed. 6.cDNA of Eukaryotic initiation factor(iso)4E(eIF(iso)4E) from tomato ZS5,this cDNA contains a ORF length in 603 bp,encoding 200aa.and show 99%and 100%nucleotide and amino acid sequence identities to the reported gene BT014561.7.Sense construct for tomato eIF?(iso)4E and antisense construct for tomato eIF(iso)4E was respectively constructed.8.Agrobacterium-mediated transformation was carried out on tomato(ZS5) with tomato eIF(iso)4E sense construct and antisense construct.Five and three kanamycin resistant plants were obtained respectively.9.cDNA of Eukaryotic initiation factor(iso)4E(eIF(iso)4E) from pepper SS5,this cDNA contains a ORF length in 609 bp,encoding 202aa.and show 99%and 100%nucleotide and amino acid sequence identities to the reported gene TC5256.