广西野生灵芝生物学特性及遗传多样性分析

Biological characteristics and genetic diversity of wild Ganoderma lucidum in Guangxi

  • 摘要: 【目的】对广西灵芝菌株资源进行生物学特性和遗传多样性分析,为广西野生灵芝种质资源的开发利用及新品种选育提供理论依据。【方法】以广西区内收集到的19个野生灵芝菌株和区外引进的5个灵芝菌株为试验材料,比较供试菌株菌丝体在不同温度、pH、碳源和氮源上的生长情况,筛选出灵芝最适生长条件,并利用SSR分子标记对供试菌株进行遗传多样性分析和聚类分析。【结果】 24个灵芝菌株菌丝10℃时均不生长; 15~30℃均能生长,15℃菌丝生长速度最慢,均低于1.26 mm/d; 20和25℃菌丝生长速度较快,分别为1.93~3.93 mm/d和4.02~5.92 mm/d; 30℃菌丝生长速度最快,均高于6.17 mm/d,35℃只有2个菌株可萌发。8个菌株最适生长的pH≤5.0; 16个菌株最适生长的pH≥8.0。13个菌株菌丝的最适碳源是果糖,6个菌株菌丝的最适碳源是蔗糖,3个菌株的最适碳源是葡萄糖; 2个菌株菌丝生长的最适碳源有2种,为果糖和蔗糖;乳糖、麦芽糖和淀粉均不适合作供试灵芝菌株的碳源。以酵母粉、硝酸铵、硫酸铵、牛肉膏、硝酸钾和蛋白胨为最佳氮源的分别有15、5、2、1、2和1个菌株,其中RSLZ-1901的最适氮源有3种,分别是酵母粉、硝酸铵和硫酸铵; CLZ-119的最适氮源为酵母粉和蛋白胨,尿素不适合作为灵芝的氮源。24个灵芝菌株间遗传相似性系数为0.49~0.93,在遗传相似性系数0.68时分为5个类群,表明各菌株间存在较大的遗传差异。【结论】灵芝菌株菌丝的生长速度和长势均与温度呈正相关,最适生长温度为30℃; 8个菌株耐酸性较强; 16个菌株耐碱性较强;最适碳源有果糖、蔗糖和葡萄糖;最适氮源有酵母粉、硝酸铵、硫酸铵、牛肉膏、硝酸钾和蛋白胨。24个供试菌株的生物学特性及遗传多样性存在明显差异,表明其遗传多样性丰富,可用于广西灵芝种质创新及品种选育。

     

    Abstract: 【Objective】In order to lay a theoretical foundation for the selection of wild Ganoderma lucidum resources development,utilization and breeding of new varieties in Guangxi,the biological characteristics and genetic diversity of G. lucidum strains collected in Guangxi were analyzed.【Method】 The optimum growth conditions of 19 wild G. lucidum strains collected in Guangxi and 5 cultivated G. lucidum strains collected from out of Guangxi were determined by comparing the mycelial growth rate at different growth temperatures,pH values, carbon sources and nitrogen sources. Genetic diversity analysis of each stain was carried out using SSR molecular markers.【Result】 The mycelium of the 24 collected G. lucidum strains could grow at 15-30℃,but stopped growth at 10℃. The mycelial growth rate was the slowest at 15℃ for all strains(below 1.26 mm/d);the growth rate increased at 20℃ and 25℃,ranging from 1.93 to 3.93 mm/d and 4.02 to 5.92 mm/d respectively;the mycelial growth rate was the fastest at 30℃(higher than 6.17 mm/d for all strains). Only 2 out of the 24 strains could survive at 35℃. The optimal growth pH for 8 strains were ≤ 5.0,and ≥ 8.0 for the rest 16 strains. Fructose was the optimal carbon source for 13 strains,sucrose for 6 strains,glucose for 3 strains,and fructose and sucrose for the last two strains. Lactose,maltose and starch were not suitable carbon sources for the tested G. lucidum strains. Yeast powder was the optimal nitrogen source for 15 strains,ammonium for 5 strains,ammonium nitrate for 2 strains,ammonium sulfate for 1 strain,beef extract for 2 strains,and potassium nitrate and peptone for 1 strain. The optimal nitrogen sources for RSLZ-1901 were yeast powder,ammonium nitrate and ammonium sulfate. The optimal nitrogen sources for CLZ-119 were yeast powder and peptone. Urea was not suitable nitrogen source for G. lucidum. The level of genetic similarities among the 24 strains were 0.49-0.93. At a genetic similarity level of 0.68,the 24 G. lucidum strains could be divided into 5 groups, indicating great genetic diversities among them.【Conclusion】 The growth rate and vigor of mycelium of the selected G. lucidum strains are positively correlated with temperature,and the optimal growth temperature is 30℃. Eight strains have strong acid resistance, and 16 strains have strong alkali resistance. The most suitable carbon sources include fructose,sucrose,and glucose;the best nitrogen sources are yeast powder,ammonium nitrate,ammonium sulfate,beef extract,potassium nitrate and peptone. These 24 selected G. lucidum strains from Guangxi show great genetic diversities and significantly different biological characteristics. The results indicate that the genetic diversity of G. lucidum strains is rich and can be used for germplasm innovation and variety breeding in Guangxi.

     

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