Cloning Of Zinc Finger Protein Gene In Chinese Wild Vitis Pseudoreticulata And Establishing Of Grape Regenerati System

Uncinula necator is a main fungus destructive disease in grape, Chinese wild Vitis pseudoreticulata Baihe-35-1 are highly resistant to powdery mildew, so it is a great significance for cloning the resistance genes to powdery mildew and function analysis. One of these study based on T11GG/B0324-292 cDNA fragment of V. pseudoreticulata Baihe-35-1,cloning and charcterization of the Zinc finger protein gene from Baihe-35-1.Another study using the leaves and petioles of V. pseudoreticulata Baihe-35-1,Hunan-1and V. vinifera cv.Red Globe as the materails, researching the action of the hormones concentrations regulators and medium on the adventitous bud regeneration via organogenesis and embryogenesis. The results were obtained as follows:1.The specific primers designed based on the cDNA fragment from Chinese wild V. pseudoreticulata Baihe-35-1 obtained by mRNA Differential Display. The full-length cDNA of the fragment was cloned by rapid amplification of cDNA ends (RACE).The full length cDNA sequence consisted of 2663 bp (GenBank accession number HM008621), had an complete open reading frame (ORF) of 2223 bp ,encoded a zinc finger protein of 740 amino acids. the non-translation of 5′and 3′-terminal region were 395 bp and 44 bp respectively. The multiple comparison sequence analysis showed that the deduced amion acid sequences shared identity to 61%, 60% and 70% with A. thaliana, M. truncatula and O. sativa,respectively. This is the first zinc finger protein gene had been cloned from grape.2.Invitro culture and high frequency propagation of"Hunan-1": The first culture medium aseptic of"Hunan-1"was MS + IBA 0.2mg/L+NAA 0.05mg/L+AC 1.0g/L. The subculture medium was 1/2MS+IBA 0.15mg/L+NAA 0.02mg/L+AC 1.0g/L.The for adventitious buds from callus of Chinese wild Vitis pseudoreticulata Baihe-35-1"Baihe-35-1"was B5 +3.0 mg/LTDZ+0.2 mg/LNAA+CH 500mg/L+1g /lAC, ra t i o n of adventitious buds 6.67%; The medium for adventitious buds from callus of"Red Globe"was MS+2.0 mg/LTDZ+ 0.1mg/LNAA+CH500mg/L+1g/lAC.Ration of adventitious buds is 10%.The callus from Hunan-1 grow better and longer in mediumB5+2.0mg/LTDZ+0.5mg/L 2.4-D+CH500mg/L +1g/lPVP.