Joint Analysis Of Molecular Identification For Bamboo Germplasm

Bamboos as Bambusoideae subfamily plant in Poaceae,are the important component ofterrestrial ecosystems,That have high value of industrry and ecology. In this experiment, 10accessions different germplasms of moso bamboo (Phyllostachys edulis cv. Epruinosa , Ph.edulis cv. Huamozhu, Ph. edulis cv. Heterocycla, Ph. edulis cv. Pachyloen, Ph. edulis cv.Tubaeformis, Ph. edulis cv. Nabeshimana, Ph. edulis cv. Gimmet, Ph. edulis cv. Flexuosa and 2accessions special moso genetic resources -M,-Q) and 2 outgroups (Ph. kwangsiensis, Ph.reticulata) are taken as the subjects for the study.AFLP, ISSR, SRAP, AFLP + SRAP, ISSR +SRAP, AFLP + ISSR, AFLP + ISSR + SRAP, ITS , trnL-F, ITS + trnL-F single-species andmulti-joint identification method are respectively uesd for the genetic similarity cluster analysisand principal coordinates analysis of genetic distance.The main conclusions are as follows:(1) In the study, the locations of amplified AFLP, SRAP, ISSR are 440, 214 and 77 foreach, indicating that the provided tag information is reliable and the molecular markeridentification on the varieties is a suitable method. There are 26 variable sites and 20informative sites in ITS, the variation rate is 0.0 1.34%;there are 40 variable sites and 19informative sites in trnL-F, the variation rate is 0.32% 2.06%, which is suitable for molecularidentification on the varieties. According to amplification sites and site-specific number,theeffect of germplasm identification is: SRAP> AFLP> ISSR.(2) Based on the comprehensive analysis of support and resolution, the order of combinedidentification result is: AFLP + ISSR + SRAP> AFLP + SRAP> ISSR + SRAP> AFLP + ISSR,and the result of combined analysis is superior to the effect of a single identification. In theidentification of the sequence, the result of combined analysis by ITS and trnL-F sequence isbetter than the effect of a single sequence identification.(3) This study indicating the genetic similarity coefficient range of the 10 bamboos in thetest is 0.5997⁰.8693, the genetic distance 0.2038⁰.6764, the sequence homology97.40%⁹9.9%, the variation rate 0.0².06%. Tag information shows that Ph. edulis cv.Epruinosaan and Ph. edulis cv. Pachyloen are unique germplasm with special genetic basis. Inthe ITS sequence, there is a C nucleotide insertion in the sites 7 of Ph. edulis cv. Pachyloen andthe most differences sites with Ph. edulis-M (15).In the trnL-F sequences, there is a A-basedeletio in 869 sites of Ph. edulis cv. Pachyloen and the largest different sites between Ph.edulis cv. Tubaeformis and Ph. edulis-M (14). Ph. edulis cv. Huamozhu and Ph. edulis cv.Nabeshimana do not demonstrate a special affinity.(4) Established and optimized the bamboo germplasm SRAP-PCR reacting system for the first time,in the 50ul reacting system:20ng template DNA,2.0mmo1/L Mg²⁺,175μmol/LdNTPs,0.2μmol/L primer,1.5U Taq the DNA polymerase,5μl 10×buffer, annealing temperature52℃.In terms of Ph. kwangsiensis, the results shown that not only genetic diversity orsequence analysis together with Phyllostachys edulis germplasm, did not show symptoms ofoverseas groups, the reason should to be further explored. In addition, this study failed to makemolecular markers and gene sequence data coupled with discriminant analysis, also need thesupport of the software and methods.