云耳原生质体制备及再生的研究

Study on Protoplast Preparation and Regeneration of Auricularia herimuer

云耳是广西特色的食用菌品种,长期以来,云耳的生产方式仍处于野生或半野生状态,加上缺少适合广西栽培的优良品种,广西云耳产量日益萎缩,失去原有的市场优势。因此,利用原生质体技术对云耳进行育种,改良品种以及恢复菌种的活力显得尤为重要。本试验选用‘桂云3号’为实验材料,研究了原生质体制备和再生条件,包括菌龄、酶解时间、溶壁酶浓度以及再生培养基种类,以期获得高质量、高产量的原生质,为后续云耳遗传育种、优良性状筛选和培育新品种提供良好的试验材料。结果表明:云耳‘桂云3号’菌株制备的适宜条件是菌丝培养时间为7 d,溶壁酶浓度为3%,浓度为0.6 mol·L^(-1)的甘露醇做稳渗剂,30℃酶解3 h,可获得大量原生质体,对优化条件下得到的原生质体再生,获得最佳再生培养条件为PTA培养基。这些研究成果将为未来的云耳遗传育种、优良性状筛选和培育新品种提供重要的实验基础和支持。

Auricularia herimuer is a characteristic edible fungus of Guangxi,but for a long time,the production mode of A.herimuer is still in wild or semi-wild state,In additon to lack of excellent varieties suitable for cultivation in Guangxi,the yield of A.herimuer in Guangxi is shrinking day by day,and the original market advantage is lost.Therefore,it is very important to use protoplast technology to breed,improve the varieties and restore the vitality of the strains.In this experiment,the‘GuiYun 3’was used as material,the conditions of plasma preparation and regeneration were studied,including fungus age,digestion time,concentration of lysing enzymes and regeneration medium types,in order to obtain high quality,high yield of protoplasm for subsequent A.herimuer genetic breeding,excellent traits screening and breeding new varieties to provide good experimental material.The results showed that:the suitable conditions for the preparation of A.herimuer strain were as follows:mycelium culture time of 7 d,lysozyme concentration of 3%,mannitol concentration of 0.6 mol·L^(-1) as infiltration stabilizing agent,enzymatic hydrolysis at 30℃for 3 h,a large number of protoplasts could be obtained.The protoplasts obtained under optimized conditions were regenerated,and the best regeneration culture conditions were PTA medium.These research results will provide important experimental basis and support for the future genetic breeding,selection of good traits and breeding of new varieties.