4种羊肚菌菌种提纯复壮方法比较分析

Analysis and comparison of 4 methods for purification and rejuvenation of morchella strain

羊肚菌菌种极易退化,提纯复壮是解决菌种退化的有效手段,但有关羊肚菌菌种提纯复壮技术的研究较少。为筛选出羊肚菌有效的菌种提纯复壮方法,本研究以六妹羊肚菌(M.sextelata)主栽品种‘G8’为供试菌株,采用单交配型菌株配对、子实体组织分离、多孢分离、原生质体再生4种方法对退化菌株进行提纯复壮,比较了复壮菌株的全交配型菌株比例、菌丝生长速度及生长势、菌核形成时间及数量、菌丝分枝角度、尖端菌丝细胞核数量、菌丝线粒体内活性氧含量等方面的指标。结果表明:4种方法以多孢分离效果最佳,全交配型菌株比例为100%,母种菌丝生长速度较对照快30%以上,达(15.95±0.05)mm/d;母种、原种菌核形成时间分别较对照早3和2 d,菌核浓密连成片状;菌丝一级、二级及尖端菌丝分枝角度均较小,分别为(57.63±7.99)°、(64.57±1.98)°、(62.72±10.19)°;尖端菌丝细胞核数量达(12.1±0.3)个,而对照仅为(5.4±0.3)个。

Morchella sextelata strain is prone to degradation,purification and rejuvenation technology is an effective mean to solve the strain degradation.But there are very few relevant studies.To screen out the effective strain purification and rejuvenation method on Morchella sextelata,the main cultivation variety‘G8’was used in this study.The degraded strains were purified and rejuvenated by four methods including pairing of single mating strains,fruiting body tissue isolation,multi-spore isolation,aprotoplast regeneration followed by comparisons of the proportion of fully mating strains,mycelium growth rate and growth potential,the time and number of sclerotia formation,the mycelium branching angle,the number of tip mycelium nuclei,the ROS content in the mitochondria.The results showed that the effect of the multi-spore isolation was the best among the four methods,of which the proportion of fully mating strains was 100%and the growth rate of the stock culture was up to(15.95±0.05)mm/d that was above 30%faster than the control.The sclerotia formation time of the stock culture and mother spawn was 3d and 2d earlier than the control,respectively.The sclerotia was dense and connected into pieces.The branching angles of primary,secondary and tip mycelium were as small as(57.63±7.99)°,(64.57±1.98)°and(62.72±10.19)°.The number of nucleus in apical mycelium was(12.07±0.3),while that of the control was only(5.4±0.3).