| Lentinula edodes is the second biggest commercial fungi in terms of total world production,following Agaricus bisporus.China has been the largest producer.consumer and exporter of Lentinula edodes in the world.Fine strains of Lentinula edodes is the basic of sustainable development of Lentinula edodes industry.Mushroom cultivation in our country is transformed from small-scale production parttern to large-scale professional production,gradually,which require accurate,high-quality strains.However,Chinese mushroom markert has been short of reasonable mechanism,which leads to strains confusion.This study aims to establish a quick?simple?accurate identification method of Lentinula edodes by SSR markers based on whole genome sequencing.SSR analysis,based on genetic difference,can solve the problems of Lentinula edodes strains disorder,pseudo strains.As a result,intellectual property rights of Lentinula edodes strain can be protected.Sustainable development of Lentinula edodes can be assured.The genome of strain L135 was sequenced using the combined Solexa and 454 method by the Shenzhen Genomics Institute.340 SSR primer pairs were designed by Primer Premier 5.0 software and synthesized by Shanghai Sangon Co.Ltd.Generally,DNA library method?database searching method of ISSR-Inhibition PCR method are used to develop SSR markers.But,these three methods are inferior to genome sequencing method in terms of the quality and quantity of SSR markers.Whereas,genome sequencing method require a lot of money.That's why SSR markers based on genome sequencing are not widely used in Lentinula edodes study.It has not been reported that such a lot of genome sequencing SSR markers were used to analyse the genentic background of Lentinula edodes strains.Thirty-nine cultivated strains of Xianggu(Lentinula edodes)recently in use collected from Chinese main producing region,two wild strains isolated from Sichuan and Shaanxi province,and two commercial strains from Japan were analyzed using 92 SSR primer pairs.A fingerprinting system of was constructed based on the genetic analysis.According to the genotyping analysis,a total of 445 bands were detected of which 433 bands(97.6%)were polymorphic between two or more strains.The number of alleles of each SSR primer pair ranged from 1 to 14 with an average value of 4.83.The PIC value of SSR primer pairs is varied from 0.1862 to 0.7541,the mean value is 0.4712.The pairwise genetic similarity values were calculated using SM coefficients and cluster analysis was conducted.The genetic similarity values varied from 0.561 to 1.000,and the average genetic similarity value is 0.756.At the similarity of 0.60,experimental population can be divided into two groups,Cultivars and wild strains.Sixteen strains can be distinguished from all tested strain.The rest strains were grouped into six Communites,containing 2 to 9 strains.Lentinula edodes strains could not be differentiate within groups.Clustering analysis of SSR markers is in line with the strains' Pedigree and cultivation experiment.In conclusion,SSR markers based on genome sequencing is a fine and promising method to evaluate the genetic background of Lentinula edodes strains and to identify strains... |
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