橙黄硬皮马勃菌种的分离纯化及其菌根的人工合成

Isolation and purification of Scleroderma citrinum and its artificial synthesis

【目的】为分离纯化菌根食用菌橙黄硬皮马勃的菌种,促进云南松无菌苗菌根的人工合成,实现云南松菌根化苗木的定向培育和菌根食用菌的人工驯化栽培。【方法】采用子实体组织分离法和综合PDA培养基对橙黄硬皮马勃子实体进行菌种分离,对其子实体、组织分离菌株和菌根菌的核糖体转录间隔区片段(ITS)进行分子鉴定,研究不同栽培基质条件下的接种处理对云南松菌根合成及其幼苗生长的影响。【结果】1)从邻接法构建的系统发育树上可以看出,采集的子实体、组织分离菌株和菌根菌确认为橙黄硬皮马勃。2)接种处理后的云南松幼苗生长指标显著高于未接种处理,而且在不同基质处理间差异显著,其中在蛭石和珍珠岩和泥炭土(质量比10:1:5)的混合基质中均表现最优。3)橙黄硬皮马勃在此混合基质中的菌根侵染率、菌根依赖性和菌根贡献率均最高,分别为92.66%、292.86%和65.85%,显著高于其他两种基质处理,可能是橙黄硬皮马勃菌丝更适宜生长在该混合基质,进而利于云南松无菌苗的菌根合成,也间接促进了云南松幼苗的生长。【结论】橙黄硬皮马勃菌种的分离纯化和菌根合成基质的筛选是云南松无菌苗菌根合成的关键。

【Objective】This work aimed to isolate and purify the edible mycorrhizal fungi(EMF)of Scleroderma citrinum,and promote the artificial synthesis of ectomycorrhizal of Pinus yunnanensis seedlings,to provide a theoretical basis for the directional cultivation of mycorrhizal seedlings of P.yunnanensis and the artificial domestication and cultivation of edible mycorrhizal fungi.【Method】The fruiting bodies of S.citrinum were isolated by tissue isolation method using the comprehensive PDA medium.Then the internal transcribed spacers(ITS)of fruiting bodies,tissue isolated strains and ectomycorrhizal fungi were identified.Also,the effects of inoculation treatment on the mycorrhizal synthesis and seedling growth of P.yunnanensis were studied by using different cultivation substrates.【Result】1)Based on the phylogenetic tree constructed by the neighbor-joining(NJ)method,the collected fruiting bodies,tissue isolated strains and ectomycorrhizal fungi were confirmed to be S.citrinum.2)The growth indexes of P.yunnanensis seedlings under the inoculation treatment were significantly higher than those without inoculation treatment.Meanwhile,there were significant differences among different substrate treatments,and the seedlings showed the best growth in the mixed substrate of vermiculite,perlite and peat soil(mass ratio 10:1:5).3)The mycorrhizal infection rate,mycorrhizal dependence and mycorrhizal contribution rate in this mixed substrate were 92.66%,292.86%and 65.85%respectively,which were significantly higher than those of the other two substrate treatments.The results suggested that S.citrinum mycelium was more suitable to grow in this mixed substrate,which was also conducive to the mycorrhizal synthesis of P.yunnanensis seedlings,and indirectly promoted the growth of P.yunnanensis seedlings.【Conclusion】The isolation and purification of S.citrinum and the screening of mycorrhizal synthesis substrates are the keys to the mycorrhizal synthesis of P.yunnanensis seedlings.