| Lycium barbarum L. is one of the most important plant resources in our country,which belong to the family of Solanaceae and the category of Lycium L.. However,based on current development of Lycium barbarum L., there exist some obviousproblems, like standardized production has not been in norm, and synonym andhomonym happened occasionally because parts of the cultivars name are based on thename of place or sequencing, etc.and given by the person who plants them, which hasnot only brought pretty much inconvenience in production process, but also createdobstacles for genetic breeding and academic communication of Lycium barbarum L..In order to clearly understand the species, family relationship and the genetic basis ofbreeding of Lycium barbarum L., a sort of feature plant resources in western area ofChina, we set10species of Lycium barbarum L. as subject, and made use ofmolecular biological method to systematic study the genetic relationship among them,then got the final results.The experimental material in this research came from two area, the first areainclude nine Lycium barbarum L. cultivated varieties, Ningqi1, Ningqi3, Ningqi4,Ningqi5, Ningqi7, Mengqi1, Flat fruit medlar,07-2and0901, which all came formLycium barbarum L. demonstration garden of Yujie agricultural trade limitedcompany, located in Jingtai county, Gansu province; the other species was wildLycium ruthenicum Murray (LMNQH), which grow in Nuomu Hong area, Qinghaiprovince. We tried to construct evolutionary tree about phylogeny by setting rDNAinternal transcribed spacer (nrDNA-ITS) sequence as object, and using method ofCTAB to extracted genomic DNA, aimed to establish its phylogenetic tree, and thenfigure out the system evolutionary relationship about the plants of Lycium L., thegenetic background of main cultivated Lycium barbarum L., and the most effectiveway in saving fresh leaves, which would provide theoretical basis for parent selectingin crossbreeding and heterosis utilization of Lycium barbarum L.. The mainconclusions we got are as follows:(1) We used two method in this study, turn colors silica gel drying way and liquid nitrogen, to keep fresh leaves of Lycium barbarum L.. After making use ofA260/A280and agarose gel electrophoresis to identify the selective DNA under twodifferent saving ways, the tests results showed that there was no obvious differenceappeared in extracted DNA. Since turn colors silica gel has much advantages likecheaper, easier and much convince in saving process than liquid nitrogen, so wesuggested turn colors silica gel drying way is more adoptable to keep fresh leaves.(2) The length characteristic of ITS sequence demonstrated that there were noobvious difference between the length of ITS sequences, ITS1sequences and ITS2sequences in0901, Ningqi3, Ningqi5, Ningqi7Mengqi1and Lycium barbarum L.,and the length of them were all582bp、215bp、213bp, and they clustered into the samecategory because of the short length. The total ITS length of07-2, Ningqi1, Ningqi4,and Lycium ruthenicum Murray were655bp、653bp、656bp、627bp respectively, andthey clustered into the same category because of the relatively long length; the ITS1length of07-2, Ningqi1, Ningqi4were all269bp, while the Lycium ruthenicumMurray was242bp; the ITS1length of07-2, Ningqi1, Ningqi4and Lyciumruthenicum Murray were232bp,230bp,233bp,231bp severally.(3) The G+C content of ITS sequence in0901, Mengqi1, Lycium barbarum L.were all55.3%, Ningqi3, Ningqi5, Ningqi7were all55.1%, and those6species hada rather close relationship with each other due to the similar and low G+C content.The G+C content in07-2, Ningqi1, Ningqi4and Lycium ruthenicum Murray variedfrom54.0to69.6, and they had a close relationship with each other because of thehigher content of G+C.(4) The analyzed results of genetic distance inflected the genetic differentiationdistance of10Lycium L. species was from0.000to0.158. Ningqi3, Ningqi7, Lyciumbarbarum L.,0901, Mengqi1and Ningqi5had a pretty close relationship with eachother and the genetic distance of them were from0.000to0.005; Ningqi4, Ningqi1,07-2and Lycium ruthenicum Murray had a rather close relationship with each otherand the genetic distance of them were from0.002to0.048; the phylogeneticrelationship between Ningqi4, Ningqi1,07-2, Lycium ruthenicum Murray and Ningqi3, Ningqi7, Lycium barbarum L.,0901, Mengqi1and Ningqi5were relatively further, and their genetic distance were range from0.126to0.158.(5)The phylogenetic tree showed that the10Lycium L. species were classifiedinto2stable components: one component was formed by Ningqi3, Ningqi7, Lyciumbarbarum L.,0901, Mengqi1and Ningqi5, in which Lycium barbarum L.,0901,Mengqi1and Ningqi5are clustered into a separate branch and Ningqi3, Ningqi7clustered into another one; the other component was formed by Lycium ruthenicumMurray,07-2, Ningqi4and Ningqi1, in which Lycium ruthenicum Murray and07-2clustered into one category and Ningqi4and Ningqi1clustered into the other one. |
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