| Powdery mildew （PM）, caused by the fungus Uncinula necator （Schw.） Burr, is one of thefungal diseases of grapevine which harm Vitis vinifera L. and can cause severe yield loss.Researches have identified that Chinese wild Vitis pseudoreticulata is highly resistant topowdery mildew, cloning and expression analysis of powdery mildew resistance related genesin Chinese wild V. pseudoreticulata is important to reveal the molecular mechanism of graperesistance to PM. Based on some EST sequences related to defense in cDNA library we haveobtained from young leaves of Chinese wild V. pseudoreticulata clone Baihe35-1inoculatedwith U. necator, then Cloned the full-length by RACE, and further analyze these two genes bybioinformatic. Main conclusions of this paper are as follows:（1） Using RACE technology obtained2583bp VpRLK1full-length,2079bp ORF.VpRLK1has692amino acid, molecular weight77.7kDa, isoelectric point5.47, Total numberof negatively charged residues （Asp+Glu）:87, Total number of positively charged residues（Arg+Lys）:70. Bioinformation analysis showed that VpRLK1gene located in the sixthchromes of grapevine. The N-terminals has the Bulb-type lectin domain,46amino acid in total;The C-terminals has the protein kinase domain,217amino acid in total; The VpRLK1gene inChinese wild Vitis pseudoreticulata in genetic distance most near from European grapevine, andnear from woody plant Popμlus trichocarpa, and far from leguminous plants Glycine max andMedicago truncatμla, and most far from monocotyledon and Arabidopsis. Comparisons of theamino acid sequences between the VpRLK1gene and the putative, unknown, or hypotheticalproteins are showed44.9%homology with Arabidopsis hypothetical protein （Accession no.NM125483）,48.4%homology with Oryza sativa protein （Accession no. EEC77471） and59.4%homology with Popμlus trichocarpa hypothetical protein （Accession no.XP002299254）.（2） Using RACE technology obtained1322bp VpSTART1gene full-length,1206bpORF, code401amino acid, molecular weight45.3kDa, isoelectric point9.55, Total number ofnegatively charged residues （Asp+Glu）:38, Total number of positively charged residues （Arg+Lys）:57. Bioinformation analysis showed that VpSTAT1gene located in the ninth chromes ofgrapevine. VpSTAT1has the StAR-related lipid-transfer （START） domain. The phylogenetic tree analysis showed that START1gene can be classified into four subfamilies, the Arabidopsis andSolanum lycopersicum individually categorize in one subfamily, Oryza sativa, Sorghum bicolorand Zea mays categorize in one subfamily, Vitis pseudoreticulata, Popμlus trichocarpa, andVitis vinifera categorize in one subfamily. Comparisons of the amino acid sequences betweenthe VpSTAT1gene and the putative, unknown, or hypothetical proteins are showed99%homology with Vitis vinifera hypothetical protein,59%homology with Arabidopsis,33%homology with Oryza sativa protein. |
