Studies On Genetic Diversity In Different Lilium Brownii Populations Of Chongqing

Lilium brownii belongs to Lilium of Liliaceae in plant taxonomy.It is one of commonly—used cross breeding parents.And it was provided that it has great ornamental worth and edible worth. This experiment materials come from Wushan town,Wuxi town,Chengkou town,Jiangjin town, Jinfo mountain,Sanquan town in Chongqing city,China.First,select two populations which come from Wushan town and Wuxi town as experiment materials.Establish and optimize AFLP reaction system for Lilium brownii.Then,compare the genetic diversity between the 6 populations of Lilium brownii.The results were list as following:(1)The AFLP reaction system for Lilium brownii was established and optimized.In DNA extraction process,the best ratio of CTAB and material was 4ml/g.the DNA should be extracted 3 times by the mixtion of trichlorpromazine and isoamylacohol(24:1).Incubate at 0℃for 20 minutes. Then,centrifuge in low speed(approximately 10,000rpm)for sever seconds.Discard the flow-through.The DNA was fit for AFLP.The DNA should be digested by EcorⅠand MselⅠ, every enzyme for 4 hours.The best adaptor concentration was 50 pmol/μl.The ligation products should be diluted 20 folds.1.0ul dNTP should be added into 50μl PCR reaction solution.And the pre—amplify reaction began at 72℃is better than 94℃.The pre—amplification products should be diluted 30 folds.The protocol of select-amplification was as same as pre—amplification.The products were fit for AFLP.(2)20 pairs of primer combinations were screened from 64 EcoR1/Mse1 primer combinations. They were:(1)E-AAC/M-CAC,(2)E-AAG/M-CAC,(3)E-AAG/M-CAG,(4)E-ACA/M-CAA,(5) E-ACA/M-CAG,(6)E-ACT/M-CAG,(7)E-ACC/M-CAA,(8)E-ACC/M-CAC,(9)E-ACC/M-CAG, (10)E-ACC/M-CTG,(11)E-ACG/M-CAA,(12)E-ACG/M-CAC,(13)E-ACG/M-CTG,(14)E-AGC/M-CAA, (15)E-AGC/M-CAC,(16)E-AGC/M-CAG,(17)E-AGC/M-CTC,(18)E-AGG/M-CAA,(19)E-AGG/M-CAC, (20)E-AGG/M-CAG.(3)In the AFLP experiment with 12 pairs of primer combinations,we've got 982 AFLP bands.81.83 bands per primer combination on average.In the 6 populations examineted,344 bands were obtained,28.66 bands per primer combination on average,and accounts for 35.03%in 982 total bands.638 polymorphic bands were obtained,53.17 per primer combination on average,and accounts for 64.97%.These results proved that the 6 Lilium brownii populations have the general character and genetic diversity.(4)Although every sample contain several plants.There are still some differences existing in 4 samples of the same population.The genetic distance of the same population's Lilium brownii is between 0.06-0.09.All the Lilium brownii's genetic distance were between 0.06-0.15.The results proved that the Lilium brownii show great genetic diversity form different individual.(5)Though the comparetion of genetic distance and similarity,we found that all the simple matching coefficients were more than 0.78.The 6 populations were divided into two parts.Wushan population,Wuxi population and Chengkou population belonged to the northeast part;Jiangjin population,Jinfo mountain population and Sanquan population belonged to southwest part.In the two parts,the two populations which were near in geographical position were considered as a new part first.Then,it was compared with the other population.All the results reported that the genetic distance was influenced by geography distance.