Genetic diversity of Phalaris arundinacea Linn germplasm detected by SRAP markers

摘要

Sequence-related amplified polymorphism (SRAP) molecular marker was used to detect the genetic diversity of 25 accessions of Phalaris arundinacea Linn that collected from the USA, Russia, Kazakhstan, Canada, LanZhou and HuBei. The following results were obtained: 1) Sixteen primer pairs produced 131 polymorphic bands, with an average of 8.19 bands per primer pair. The percentage of polymorphic bands on average was 89.25%. The polymorphism information content(PIC) ranged from 0.784 to 0.9069, with an average of 0.8696. 2) The Nei's genetic similarity coefficient of the tested accessions ranged from 0.5959 to 0.8425, and the average of Nei's coefficient was 0.7422. These results suggested that there was rich genetic diversity among the resources of Phalaris arundinacea Linn. 3) Twenty-five accessions were clustered into five groups. Moreover, the accessions from the same or similar origin frequently clustered into one group. The findings implied that the correlation among the resources, geographical and ecological environment. Sequence Related Amplified Polymorphism (SRAP) is a relatively simple marker system, it bases on PCR technique and aims for the amplification of open reading frames(ORFs). It is based on two-primer amplification. The primers are 17 or 18 nucleotides long and consist of the following elements. Core sequences, which are 13 to 14 bases long, where the first 10 or 11 bases starting at the 5' end, are sequences of no specific constitution ( "filler" sequences), followed by the sequence CCGG in the forward primer and AATT in the reverse primer. The core is followed by three selective nucleotides at the 3' end. The filler sequences of the forward and reverse primers must be different from each other and can be 10 or 11 bases long. SRAP technique is simple, convenient and there is no need to know in advance the sequence information of the species, therefor, it is widely used in analysis of genetic diversity(Zhang et al. 2010), germplasm appraisal (Zhang et al. 2005), genetic linkage mapping(Huang et al. 2009), gene location(Li and Quiros et al. 2001) and comparative genomics(Barrett B et al. 2004). SRAP technique has been used for enetic analysis in a number of species, including Brassica oleracea L(Li and Quiros et al. 2001), Cucurbita(Ferriol M et al. 2003) and Paeonia L(Han X Y et al.2008), Medicago sativa(Vandemark G J et al. 2006), Buchloe dactyloides(Budak H et al. 2004), and Zoysia japonica(Chen et al. 2009; Guo et al. 2009) . Phalaris arundinacea Linn belongs to phalaris of Gramineae and includes 22 species(Sahramaa M et al. 2004), they are always grow in North America, Northern Europe and Asian. In China, it is mainly grown in the Northeast, the north, northwest and central section. Phalaris arundinacea Linn lives in wet area and often lives with Phragmites australis. Phalaris arundinacea Linn is a perennial rhizomatous grass.