红三叶新品系种质鉴定及无性繁殖特性的分析-identification of new red clover germplasm and analysis of its asexual reproduction characteristics.docx

clover was 2n=2x=14=8m+6sm(2SAT). The asymmetry of interchromosomal is greater than intrachromosomal, and was relatively high in the red clover populations. According to the the Langlet’s chromosome morphology criteria for the classification, the new line of red clover chromosome type is the T-type, primitive relatively.4、The analysis of SSR marker showed that using 8 pairs of primers in 32 materialsobtain 219 polymorphic bands totally. The polymorphism percentage was 100%, and the number of effective alleles was 1.1636. The Nei’ diversity index, shannon information index, total variance, variance between the materials, coefficient of genetic differentiation and gene flow were 0.1229, 0.2241, 0.1218, 0.0387, 0.3179 respectively. 31.79% genetic variation of the red clover exists in external populations, 68.21% was in internal populations. The fingerprints of the red clover new line was obtained, which can effectively distinguish from two registered varieties of red clover in China. The Nei diversity index of red clover new line was 0.0665, and Shannons information index was 0.1065, lower than the average population diversity level. SSR genetic distance showed that the new line of red clover was similar to the number of 60, 22 ,53, which were from Russia, Spain and Russia respectively.5、The best ramets per plant was 6~7 in division propagation, and the highestproliferation coefficient was 6.38; The highest proliferation coefficient of cutting can up to6.40 using entire branches in branching period.The study of tissue culture showed: the best method of seeds sterilization for red clover is the 75% ethyl alcohol treatment followed by disinfection with 0.1% HgCl2 for 10 min. 1/4MS medium with 1.5% sucrose without hormones was most effective for seed culture. Cotyledons, hypocotyl, petiole, leaf and root callus were easy to be induced in medium of MS containing 0.5 mg·L-1 6-BA and 1~2 mg·L-1 2,4-D. The best callus subculture medium was MS containing 0.5 mg·L-