巨菌草诱导培养基的筛选及优化

Screening and optimization of Pennisetum sp.induction medium

巨菌草(Pennisetumsp.)属单子叶禾本科狼尾草属植物,可作为动物饲料和多种食用菌的培养基质,在中国及非洲广泛种植,然而不耐寒的特性限制了其推广应用。要克服其遗传缺陷,需借助转基因或体细胞杂交等技术,因此,必须建立巨菌草的组织培养体系。本研究以巨菌草幼茎为材料,研究了巨菌草外植体的消毒方式和2,4-D、IAA与NAA,细胞分裂素(6-BA和KT)的不同浓度及组合对愈伤组织诱导的影响。结果表明,用75%酒精擦拭剥离后的叶鞘,再用2%的次氯酸钠溶液消毒10 min就能达到理想的消毒目的。1.0~4.0 mg·L-1的2,4-D都可以诱导出愈伤组织,且各浓度间无显著差异(P】0.05);0.2mg·L-1 IAA对愈伤组织诱导有较好的促进作用;不同细胞分裂素对愈伤组织诱导没有明显促进作用。巨菌草愈伤组织诱导的最佳培养基配方是MS+2,4-D(3.0mg·L-1)+IAA(0.2mg·L-1)。

Pennisetumsp.is monocotyledon plant and widely planted in China and Africa as it can be used as forage and culture substrates of edible fungi.However,the lack of cold tolerance limits its popularization.Genetic technology or somatic hybridization has potential to improve the cold tolerance or quality of Pennisetumsp.which require the establishment of Pennisetumsp.tissue culture system.In the present stydy,the effects of different disinfection ways and different growth hormone such as 2,4-D,IAA,NAA and cytokinins(6-BA and KT)on callus induction were studied using caulicle of Pennisetumsp.as explants.The results showed that the ideal disinfection method was soaking in 2% sodium hypochlorite solution for10 min after wiping stripping leaf sheath with 75% alcohol.All of the medium with the different concentrations of 2,4-D ranged from 1.0mg·L-1 to 4.0mg·L-1 can induce callus without significant difference.The medium with 0.2mg·L-1 IAA can induce more callus.The medium with different cytokinins can not induce more callus.Based on these observation,the optimal callus induction medium of Pennisetumsp.was MS + 2,4-D(3.0mg·L-1)+IAA(0.2mg·L-1).