| Paulownia(Paulownia Sieb.et Zucc),as the only deciduous trees of Scrophulariaceae,is a kind of improtant fast-growing trees of economicvalue,its natural distribution and cultivat area covers areas in China.Paulownia is not only cross-pollination,between each kind of all can cross,and produce fertile offspring,but also they can clone and easy to form and retain trantional types.After a long interspecific hybridization and repeated and the natural selection,it makes the Paulownia belong to the difficulty in botany classification that combination and infiltration with each kind of genes.The focus of the traditional morpholoay base on the phenotypic differences put Paulownia into Scrophularaceae or Bignoniaceae,but with the development of pollen morphology,they put Paulownia into Scrophularaceae because of the significant differences in pollen morphology.There has not yet been reported about the relationship between Paulownia and its allied genera.In order to analyze the relatives within families and systematics of paulownia taxa relationship and clear the classification between the relationship with the different species of genus Paulownia,the study adopts two pairs of primer combination method like the chlolplast genome sequence psb E-pet L and trn D-trn T,and provide a theoretical basis and reference data for the reasonable use of plants of the genus Paulownia and genetic breeding,providing important theoretical guidance for seed breeding,suitable trees,creating high productivity of artificial forest.The main results are as follows:(1)Two kinds of methods with CTAB method and urea protocol and modified from genome DNA,and detect the quality and concentration of DNA,compared to the two methods,results show that the extracted by improved CTAB method of DNA of higher quality,more suitable for the demand of the chloroplast genome amplification.(2)This study through the optimization of PCR reaction system,for the establishment of Paulownia Plants and related system of plants,as 25μL system containe 12.5μLTap PCR Master Mix,50 ng DNA,each 0.40 mmol · L-1reverse primer and 5.5μL dd H2 O.By improving PCR amplification conditions,select the appropriate concentration of DNA templates,primers and the annealing temperature,so that the PCR products sequencing complete.From 10 clear difference in chloroplast genome sequence of genetic screening than2 for primer sequence: psb E-pet L,trn D-trn T,and use the corresponding software specific to construct phylogenetic tree analysis sequence.(3)Through constructing model results showed that the genetic distance aboutPaulownia genus and its relatives,the conclusion are as follows: the genetic distance between P.fortunei,P.tomentosa and P.fargesii is larger,the genetic distance between P.elongata,P.australis,P.catalpifolia,P.lamprophylla is smaller and these nodes results in line with the morphological studies.(4)By 2 of the primer sequences and sequences with data collection system construction of phylogenetic tree shows that Paulownia Plants of the genus were to 100%bootstrap support coalescence rate for a group,the out group Bignonia and Digitalis is clustered into a group,Paulownia species at the molecular level that do not belong to the Bignoniaceae,neither to Digitalis.The comparison of the 3 data sets shows that the highest proportion of simple information in trn D-trn T sequence is the highest,and the rate of variation is also the highest.Construction of a phylogenetic tree of P.australis and P.kawakamii,P.tomentosa and P.lamprophylla,P.elongata and P.elongata f.alba elongata total poly a group that they evolved independently. |
