Department of Pharmaceutical Technology, Division of Microbiology and Biotechnology, Jadavpur University, Kolkata, 700032, India.
Division of Plant Biology, Bose Institute, P-1/12,C.I.T. Scheme VII M, Kolkata, 700054, India.
World J Microbiol Biotechnol. 2021 Mar 22;37(4):69. doi: 10.1007/s11274-021-03032-3.
Two edible mushrooms Calocybe indica and Pleurotus sajor-caju were chosen as parent strains in this study to approach the concept of hybridization through the protoplast fusion technique. Protoplast fusion in presence of polyethylene glycol (PEG) was conducted between the parent strains and by further double selection screening method, six somatic hybrid lines were developed. Those fruit bodies of the hybrid lines showed phenotypic resemblance with Pleurotus sajor-caju when grown on paddy straw under favorable conditions. The hybridity of the newly developed somatic hybrid strains was established by barrage reaction, morphological traits, fruitbody parameter and, inter single sequence repeat (ISSR) profiling. One-way analysis of variance (ANOVA) was used for the analysis of phenotypic data of hybrid lines and parents. Five ISSR primers were used to generate 51 amplified DNA fragments ranged between 250 and 3000 bp in size in six hybrids and two parents with 90.19% polymorphism. Some of the hybrids contain some non-parental bands which indicate that recombination might happen in the hybrid genome hence confirming the hybridity of newly developed strains. The dendrogram was created using the Average Linkage (Between Groups) method based on ISSR profiling and genetic distance between parent-hybrids and hybrid-hybrid was analyzed by Jaccard's proximity matrix. A definite improvement in nutritional properties and biological activity was observed in the study. Due to ease in their cultivation, it can play a significant role in the rural economic development.
在这项研究中,选择了两种可食用的蘑菇——草菇和姬松茸作为亲本菌株,通过原生质体融合技术来探讨杂交的概念。在聚乙二醇(PEG)存在的情况下,对亲本菌株进行原生质体融合,并通过进一步的双重选择筛选方法,开发了 6 株体细胞杂种。在有利条件下,这些杂种在稻草上生长时,其子实体形态与姬松茸相似。通过阻断反应、形态特征、子实体参数和单核苷酸重复序列(ISSR)分析,确立了新开发的体细胞杂种菌株的杂种特性。采用单向方差分析(ANOVA)对杂种系和亲本的表型数据进行分析。使用 5 个 ISSR 引物在 6 个杂种和 2 个亲本中产生了 51 个扩增的 DNA 片段,大小在 250 到 3000 bp 之间,多态性为 90.19%。一些杂种含有一些非亲本带,这表明重组可能发生在杂种基因组中,从而证实了新开发菌株的杂种特性。基于 ISSR 分析,采用平均连锁(组间)方法创建了系统发育树,并通过 Jaccard 相似性矩阵分析了亲本-杂种和杂种-杂种之间的遗传距离。研究中观察到营养特性和生物活性的明显改善。由于其易于栽培,可以在农村经济发展中发挥重要作用。