| Genetic diversity and population genetic structure of natural population of four Lilium species from QinBa mountain area were studied at DNA level. The relationship and gene flow between these populations were discussed. Based on comprehensive analysis, the current situation of the genetic resource of Lilium was evaluated, and some suggestions were refered. The results are as follows:1. The conventional CTAB method succeedly exacted and puried the total DNA from the leaves of Lilium, its quality and the purity had met the RAPD's request.2. Using the method of single factor progressive selection to establish the RAPD program by setting different concentrations to the factors (i.e. the concentration of the factor which previous turn to the best used as fixture concentration to use in next factor secletion foundation), simultaneously, carring on the comparison to factors like annealing temperature, the number of cycle, to established the RAPD analysis system suited to the wild Lilium. The reliable RAPD analysis system was established. The reaction volume is 20μL and reaction mixture consist with 40ng DNA, 0.4μmol/L random primer, 2.25mmol/L Mg2+, 0.2mmol/L of dNTPs, 1.5U Taq polymerase and 1×buffer.RAPD program is 4 minutes at 94℃for predenaturation, then 35 cycles of 30 seconds at 94℃for denaturation, of 45 seconds at 37℃for annealing, of 90 seconds at 72℃for extension, finally extension at 72℃for 10minutes, stopped at 4℃and stored at -20℃. 3. Using the RAPD technology to analyze the genetic diversity and the genetic structure of four Lilium species form difference populations of QinBa mountain area, and then carring on the cluster analysis.The result is as follows:(1) A total of 60 DNA samples of 6 natural populations of L.lancifolium Thunb surveyed by using of 10 RAPD random primers, which generated 81 reproducible and clear amplification products with molecular weight from 350bp to 2000bp. Out of 81, 62 loci were polymorphic and accounted 76.54%. Effective number of alleles (Ne) was 1.4385, Nei's gene diversity (h) and Shannon's Information Index (I) were 0.2153 and 0.3327 in the species level respectively.Of the 6 populations, the sequence of the genetic diversity was Shannxi Chenggu (JDCG)>Shannxi Foping(JDFP)>Shannxi Nanzheng (JDNZ)>Shannxi Ningshan(JDNS) >Hubei Yunxi(JDYX)>Shannxi Pingli(JDPL). Coefficient of genetic differentiation (Gst) generated by Popgene32 was 0.3192, and in accordance with Shannon's index of phenotypic diversity showing that 31.92%of genetic diversity resided among populations. Gene flow was 1.0664. Genetic Identity and Genetic Distance between populations were estimated. Cluster analysis indicated that the 6 populations could be grouped into two clades and JDNZ, JDCG and JDPL grouped into the same group; and group second contained JDYX, JDFP and JDNZ.(2) A total of 60 DNA samples of 6 natural populations of L.leucanthum Baker surveyed by using of 10 RAPD random primers, which generated 79 reproducible and clear amplification products with molecular weight from 230bp to 2594bp. Out of 79, 65 loci were polymorphic and accounted 82.28%. Effective number of alleles (Ne) was 1.4352, Nei's gene diversity (h) and Shannon's Information Index (I) were 0.2366 and 0.3525 in the species level respectively. Of the 6 populations, the sequence of the genetic diversity was Shannxi Hanyin(YCHY)>Shannxi Zhenba(YCZB)>Henan Zhenping(YCZP)>Chongqing Wux(iYCWX) >Shannxi Taibai(YCTB)>Shannxi Foping(YCFP). Coefficient of genetic differentiation (Gst) generated by Popgene32 was 0.2469, and in accordance with Shannon's index of phenotypic diversity showing that 24.69%of genetic diversity resided among populations. Gene flow was 1.5251. Genetic Identity and Genetic Distance between populations were estimated. Cluster analysis indicated that the 6 populations could be grouped into three clades and first group contained YCTB and YCWX; the second group contained YCHY, YCZB and YCFP; and the last group contained YCZP only.(3)A total of 50 DNA samples of 5 natural populations of L.brownii F. E. Brown surveyed by using of 10 RAPD random primers, which generated 66 reproducible and clear amplification products with molecular weight from 109bp to 1741bp. Out of 66, 55 loci were polymorphic and accounted 83.33%. Effective number of alleles (Ne) was 1.6439, Nei's gene diversity (h) and Shannon's Information Index (I) were 0.2735 and 0.4385 in the species level respectively. Of the 5 populations, the sequence of the genetic diversity was Shannxi Ningqiang(YNQ)>Hubei Zhuxi(YZX)>Shannxi Chenggu(YCG)>Shannxi Nanzheng(YNZ) >Shannxi Zhashui(YZS). Coefficient of genetic differentiation (Gst) generated by Popgene32 was 0.2524, and in accordance with Shannon's index of phenotypic diversity showing that 25.24%of genetic diversity resided among populations. Gene flow was 1.4810. Genetic Identity and Genetic Distance between populations were estimated. Cluster analysis indicated that the 5 populations could be grouped into three clades and first group contained YNQ, YNZ and YCG; the second group contained YZX only; and the last group contained YZS only. (4) A total of 60 DNA samples of 6 natural populations of L.pumilum D. C. Fisch surveyed by using of 10 RAPD random primers, which generated 80 reproducible and clear amplification products with molecular weight from 184bp to 1958bp. Out of 80, 65 loci were polymorphic and accounted 81.25%. Effective number of alleles (Ne) was 1.5416, Nei's gene diversity (h) and Shannon's Information Index (I) were 0.2558 and 0.3854 in the species level respectively. Of the 6 populations, the sequence of the genetic diversity was Ganshu Wudu(SDWD)>Shannxi Shanyang(SDSY)>Shannxi Luonan(SDLN)>Shannxi Danfeng (SDDF) >Ganshu Diebu(SDDB)>Ganshu Zhouqu(SDZQ). Coefficient of genetic differentiation (Gst) generated by Popgene32 was 0.2570, and in accordance with Shannon's index of phenotypic diversity showing that 25.70% of genetic diversity resided among populations. Gene flow was 1.3095. Genetic Identity and Genetic Distance between populations were estimated. Cluster analysis indicated that the 6 populations could be grouped into three clades and first group contained SDSY and SDLN; the second group contained SDZQ, SDWD and SDDB; and the last group contained SDDF only.The study indicated that, the genetic diversity of the four Lilium wild species from the QinBa mountain area was rich, and there existed genetic differentiation between the different populations, the sequence of the genetic diversity in these four lilium species was in turn: L. brownie F. E. Brown> L. pumilum D. C. Fisch> L. leucanthum Baker>L. lancifolium Thunb.The cluster analysis demonstrated that the genetic distance and geography distance existenced certain relevance, but also there had the exception, genetic distance between different population was possibly also concerns with other factors.
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