Study On Genetic Diversity Of Cotton Varieties With Resistance To Fusarium Wilt And Verticillium Wilt Based On AFLP

Fusarium wilt and Verticillium wilt are main diseases which has done harm to the production of cotton in the world, and the same thing also happens in China Practice has proved that it is an effective and important method for preventing Fusarium wilt and Verticillium wilt to grow cultivars with disease resistance.In this paper, AFLP was used to study DNA fingerprinting of 30 main cotton cultivars with resistance to disease. The aim of the study is U)to establish a technical system suitable for fingerprinting study of cotton; (2)to draw DNA fingerprinting of cultivars; (3)to study genetic diversity of cotton combining with pedigree on molecular level. Some results are below:1. At first, some factors affecting establishment of AFLP technological system have been tested and analyzed again and again, and at last a most effective system was established. Some improved factors are below: the concentration of template DNA is 150ng ul; (2)in the restriction system, each of Msel and EcoRI was input 3 units, NEB buffer #2 was used and reaction time is 2 hours; (3)the most available time of ligation reaction is 10 hours;the products of ligation was diluted to 10 times for preamplification;the products of preamplification reaction was diluted to 10 times for selective amplification.2. Using a enzymes combination system including a rare cutter (EcoRl) and a frequent cutter (MseI) to perform AFLP fingerprinting research of 30 cultivars. From 100 primer combines, 25 primer pairs were selected to perform AFLP analysis. 1858 bands were amplified totally and 737 bands were polymorphismic , and 11 cultivars' DNA fingerprinting were listed. And AFLP technology is very fit for research of genetic diversity in cotton.3. AFLPs were processed by SPSS(Version 10.0). Most cultivars were classified due to their genetic relationship, and results accord to their pedigree. Results also point that two cultivars that missing their genetic origin can be classified: su-mian 4 .cultivars is similar to genetic origin of yu-mian 1; the genetic origin of lu-mian 12 is seem to Auburn623RNR's.