The ISSR Fingerprinting Analysis Of Verticillium Dahliae Genome Of Cotton

The molecular marking technology of ISSR is a new technology developed recently, and mainly used in the research of high mammal and plants in some fields. The fingerprinting analysis based on ISSR-PCR was carried out in 28 different Verticilliian dahliae came from different areas. The main result were as follows:1. The optimal ISSR-PCR system and program were established. In a total volume of 25ul, containing 1 XBuffer(10mM KC1, 8uM (NH4)2SO4, 10mM Tris-HCI(pH-9.0), NP-40, 2.0mM Mg2+), 200uM dNTP , 30ng template DNA, 0.2 uM ISSR primer, 1.5U Taq DNA-polymerase. Amplification performed in a AmpGene DNA Thermal Cycler 4800 was programed for 3 min at 94 "C, at annealing temperature for 60s, 90s at 72'C; 35 cyclers of 30s at 94癈, 60s at annealing temperature of each ISSR primer, 90s at 72癈; and a final extention at 72'C for 7 min after the last cycler.2. It was proved that ISSR (inter simple sequence repeat) was ubiquity in Verticillium dahilae of cotton.3. 14 ISSR primers that generated high polymorphism bands were screened from the Set# 9 bought from Canada. The results of dendrogram analysis showed that the 28 strains were classified into two ISGs (ISSR groups) under the standard value of ten . seven SISGs (Sub-ISSR groups) under the standard value of 6 respectively. The results of ISSR analysis showed that the ISSR polymorphism was related to the pathogenicity and it did not show the relation between the geographic sources, pathogenicin and consanguinity. And it indicated that genes decide the pathogenicity.