| Pomegranate(Punica granatum L.)is a deciduous shrub or small tree of the genus Punica of the genus Lythraceae,which has high ornamental,medicinal and health care values,and has generated great economic,ecological and social benefits.Pomegranates have a wide range of distribution and long cultivation time.Rich and diverse varieties and types are produced with long-term natural selection and human activities.However,it is difficult to accurately identify the variety based on morphological characteristics,which limits the breeding and industrial development of pomegranates.Therefore,it is very meaningful to strengthen the research on genetic diversity analysis and fingerprint construction of pomegranate,which is important to guarantee the identification,breeding,preservation and healthy development of pomegranate varieties.In this study,we analyzed the sequence characteristics of pomegranate genome SSRs,based on the published sequence information of the whole genome of pomegranate,and developed a large number of primers.On this basis,genetic diversity analysis and fingerprint map construction of pomegranate germplasms collected in Zaozhuang City Pomegranate National Forest Germplasm Resource Bank.Based on the fingerprint information,some of the free pollinated progeny were paternally identified.The main findings are as follows:(1)SSR primer library suitable for genetic analysis of pomegranate germplasm resources was screened.719 pairs of SSR primers were developed based on the pomegranate genome sequence,and 8samples were used to analyze the polymorphism and effectiveness.22 pairs of SSR primers with good peak shape,high polymorphism and good stability were selected from them.(2)Clarified information on genetic diversity of collected pomegranate germplasm resources.The results of polymorphism analysis showed that after 179 pomegranate germplasm samples wereamplified by 22 pairs of primers,a total of 76 allele loci were detected,of which 58 were polymorphic loci,and the percentage of polymorphism was 76.3%.The average number of Na is 3.455,the average number of Ne is 2.271,the average Ho is 0.369,the average He is 0.540,the average I is 0.898,and the polymorphism information content(PIC)is between 0.326~0.690,the average value is 0.462.(3)The population genetic structure characteristics of pomegranate germplasm resources are explained.The genetic distance analysis showed that the genetic distance between 179 germplasm resources is between 0.013 and 0.831,with an average value of 0.404,indicating that the genetic diversity of pomegranate varieties is relatively rich.Cluster analysis results showed that 179 germplasm resources can be divided into 3 main clusters and can be completely distinguished by 22 pairs of primers.The results of group structure analysis showed that 179 germplasm resources are divided into four groups.AMOVA analysis results show that the mutation mainly occurs within the group.(4)Fingerprint maps of pomegranate germplasm resources were constructed.Based Specific fingerprint codes were constructed for 179 pomegranate germplasm resources and QR codes database of related data was established using 22 pairs of primers.It provides a basis for future germplasm preservation and further development and utilization.(5)Fingerprint information was used to identify 7 hybrid plants from 2 free pollinated half-sib progeny.According to the comparison of allelic configuration,1 natural hybrid of 'Qiuyan' and 6 natural hybrids of 'Huiliqingpiruanzi' were identified from 148 pomegranate progeny.The natural hybridization rates were 3.3% and 5.1%,respectively.Based on the fingerprint information,the most likely fathers of4 hybrid progeny were identified.In this study,we used the developed SSR molecular markers to conduct genetic diversity analysis and fingerprints construction on some pomegranate germplasms,which laid the foundation for further germplasms evaluation,breeding population construction and selection of new varieties. |
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