| Pomegranate(Punica granatum L.)is deciduous shrub or small trees which belongs to Punicaceae,Punica L.It is only 1 genus and 2 species,one for the pomegranate,another for wild species without cultivation value only distributed in the western Indian Ocean on the island of Socotra.It originated in Iran,Afghanistan and other places,pomegranate has formed different variation resources in the long-term transmission and cultivation process due to natural hybridization,gene mutation,seedling stand,grafting,artificial selection and other reasons.In this study,35 species and types of pomegranate germplasms preserved in Shandong Zaozhuang Pomegranate Resources were studied.ISSR molecular markers were used to analyze the genetic diversity and genetic relationship of these resources.The origin and evolution of pomegranate germplasm resources were discussed,to protect,use pomegranate germplasm and breeding new varieties to provide scientific basis.The main results are as follows:1.The origin of 35 pomegranate germplasm,color,flower pattern and peel color and oth er preliminary statistics.From the origin to classification can be divided into five categories,f rom the color classification can be divided into four categories,from the flower type can be di vided into two categories,from the peel color classification can be divided into five categorie s,from origin,color,Peel color are the same can be divided into five categories,the test of 35 pomegranate germ rose pomegranate and six germplasm,peony pomegranate and five germpl asm,agate pomegranate and three germplasm,sour pomegranate and 8 germplasm,sweet po megranate and three germplasm.The phenotype of some pomegranate germplasm is described in detail.2.PCR amplification of 35 pomegranate germplasms was performed with the eight selec ted ISSR random primers.A total of 168 DNA loci were amplified,with an average of 21 DN A loci from each primer,of which 158 were polymorphic loci,with the percentage of polymo rphic loci(P)=94.05 %.The number of DNA loci amplified by eight ISSR primers was betw een 16 and 24,and P was between 86.96 % and 100 %.Among eight ISSR primers,UBC811 with the lowest P among the 35 germplasms was 86.96 %,while UBC816 amplified the 16 D NA loci,with the least number of DNA loci,but with the highest P,up to 100 %.The results s howed that the primers had good polymorphism,and the genetic diversity of 35 pomegranate germplasm was rich.Each primer amplification generated an average of 21 DNA loci and 19.75 polymorphic loci.The length of the amplified DNA fragment was between 101 and 2000 b p.The results showed that the number of amplified loci was great different,but the difference between the percentage of polymorphic loci was not significant with different ISSR primers t o amplify the pomegranate germplasms.3.The genetic diversity parameters of 35 pomegranate germplasms were statistically ana lyzed by Popgene 1.32 software.The results showed that the average observed number of alle les(Na)is 1.9405,the average effective number of alleles(Ne)is1.6785,and the average Nei's gene diversity index(He)is 0.3799,and the average Shannon's information index(I)is 0.5530.The results showed that the genetic variation among 35 pomegranate germplasms was larg e,and there was abundant genetic diversity among varieties.4.The genetic similarity coefficient(GS)among 35 germplasms was calculated by NTS YS2.10 software.The UPGMA phylogenetic map of 35 germplasms was constructed accordi ng to the GS value.Eight ISSR primers could distinguish 35 pomegranate germplasms.35 po megranate germplasms tested were divided into 6 groups at Gs 0.668.The genetic similarities coefficient(Gs)between germplasms ranged from 0.4167 to 0.9226,of which Gs between N o.9 and No.26 was the smallest,suggesting that their genetic relationship was relatively dista nt,and between No.28 and No.30 was the largest,suggesting that their genetic relationship was the closest.35 pomegranate germplasms with rich genetic diversity was further confirme d by ISSR. |
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