| Lilium pumilum DC. is important genetic resources for lily breeding and breed improvement. Studying on genetic variation of population under the different ecological factors, and exploring the genetic relationship among L. pumilum and other wild and cultivate Lilies. It has important significance for protection and innovation of germplasm resource, and lily breeding. This paper used ISSR molecular marker technology, researched nine geographical populations, such as Baishishan, Xiaowutai, Wulingshan, Huairou, Kaiyuan, Chifeng, Liupanshan, Weifang, Yulin. This paper inquired into the genetic relationship among L.pumilum, L.davidii var leichtlinii, L.concolor var. buschianum Baker, L.tsingtauense, L.lancifolium,‘Turino’,‘Elite’,‘Sorbonne’,‘Prito’. The correlationship between the population variation and differentiation and the effecting biotic factors and abiotic factors was analyzed with the use of SPSS17.0. The adaptability of L. pumilum DC. to different environmental factors is discussed. The main results are as follows:(1)The orthogonal design method was used to optimize ISSR-PCR amplification system on L. pumilum in 4 levels of 5 factors, respectively. The suitable reaction system was established as follows: Template DNA 50～60 ng; 15 mmol/L Mg2+ 2.5μL; 2.5 mmol/L dNTP 4μL; Primer 0.4μmol/L; EsTaq DNA polymerase 1.5 U. The result provided a standardizing ISSR-PCR program for L. pumilum: predenaturation 5 min at 94℃; followed by 40 cycles of denaturation for 50 s at 94℃, anneal 45 s at 49℃, extension 75 s at 72℃,then elongated 8 min at 72℃, and preserved at 4℃.(2)Selected 8 arbitrary primers which had rich polymorphic loci from 14 primers, and used them to ISSR-PCR amplify. There was 78 loci detected. The percentage of polymorphic loci of 100%, the Nei’s diversity index (H) was 0.2977, the Shannon’s information index (I) was 0.4611. It was proved that L. pumilum populations had very rich genetic diversity. The genetic variation among the nine L. pumilum populations were mainly origined among the populations. The genetic differentiation coefficient (Gst) was 0.6254, the gene flow (Nm) was 0.2995. A significant genetic differentiation was in the the L. pumilum germplasm of different origins.(3) Based on the results showed that the correlation between genetic distance and geographic distance between them was no significant correlation. This paper analysised the correlation from the genetic diversity, soil, climate and other factors of L. pumilum population. The result showed that the genetic diversity and nitrogen content of soil available was significantly negatively correlated. Other soil and climatic factors, annual average temperature, annual precipitation The amount, altitude, P, K content in soil had no significant correlation relationship with the genetic diversity.(4) Samples from 9 wild Lilies and cultivated Lilies were amplified with 8 primers. The percentage of polymorphic loci (P) was 91.41%, Nei’s diversity index (H) was 0.1933, Shannon information index (I) was 0.3134. The genetic diversity of wild lily was higher than the genetic diversity of cultivated lily. These lilies were clustered into five categories:‘Torino’,‘Jingcui’, L.concolor var. buschianum Baker, L. pumilum clustered into one class; L.davidii var leichtlinii and L.lancifolium clustered into one class; L.tsingtauense clustered into one class;‘Sorbonne’clustered a class;‘Torino’clustered into one group. This result was showed that L. pumilum and Asian lily had close genetic relationship. L. pumilum was excellent lily breeding. |
PDF Full Text Request