RAPD Marker-based Mapping For Anthracnose Resistant Gene In Chinese Wild Vitis Species

By manual infection in field conditions, the resistance to anthracnose(spheceloma ampelinum de Bary) was study with interspecific hybrid cross of 88-110(83-4-96 (V.quinquangularis) × "Pink Rose" (V. vinfera)) cross. The map of RAPDmarkers for anthracnose resistant gene in Chinese wild Vitis species was establishedon the basis of the analysis of RAPD of the F1 progenies and their parents. Themarker of S353-1400 was sequenced, then the restriction-sites were analysed. Theresults are as follow: 1. The resistance of grapevines to anthracnose was study by manual infectionin field conditions with 88-110 cross F1.The results indicated all of 88-110 cross, 20cultivars were resistant to anthracnose and 24 cultivars were susceptible toanthracnose in 44 cultivars of 88-110 cross F1.The result showed 1∶1 segregation byχ2(0.2045*) analysis. The 88-110 cross F1 susceptible index of manual infection aremost between parents. There are 41 cultivars in all of 44 cross F1, the rate is about93.18%. 2. Of 444 10bp arbitrary primer, 341 primers amplified RAPD polymorphicbands when select parental DNA as templates. 341 primers were used to conductRAPD reaction on the basis of surveys of the pool DNA mixed by resistant hybridsthat showed polymorphic bands. 1∶1 and 3∶1 segregating polymorphic bands wereused for constructing disease-resistance-gene genetic maps. Anthracnose resistancegene maps consisted of 1 linkage groups spanned over 469.1cM with 24 RAPDmarkers and had an average distance of 19.55cM, a shortest distance of 5.9cM and alongest distance of 36.4cM between markers; the major gene(Unci) controlledanthracnose resistance close to marker is S353-1400(5.9cM). 3. S353-1400 fragments were reclaimed from electrophoresis gel and cloned inT-easy vector, then sequenced from one side or two sides. The DNA fragmentsS353-1400 were actually 1371bp. 4. The restriction-sites of the sequence of the S353-1400 were analyzed bysoftware "Wingene231". 141 restriction enzymes with recognition sits equal orgreater than 6 bases long had cutting sites on the sequence of S353-1400. EcoR I hada cutting site at 1147bp of the sequence of S353-1400. 5. It has been suggested that the sequence of S353-1400 are unique to thechinese wild grape (chinese wild Vitis) genome. But, these sequence were extensivelycompared by BLAST in GenBank and a small part was found to be homologous.28base pairs from the sequence of S353-1400 (from 1293-1321) matched two nucleicacid sequence code bHLH (helix-loop-helix) from Arabidopsis thaliana putativetranscription factor.