细叶小羽藓(Haplocladium microphyllum)配子体组织培养的消毒方法及蔗糖浓度筛选

Sterilization method and sucrose concentration selection for initial culture of Haplocladium microphyllum gametophyte

以细叶小羽藓(Haplocladium microphyllum)的茎叶体为外植体,通过正交设计试验,研究消毒剂以及取材部位对配子体灭菌效果的影响;并通过芽体长度、芽体个数、原丝体直径、增殖系数等指标探讨不同蔗糖浓度对细叶小羽藓生长的影响。结果表明:细叶小羽藓配子体的最佳消毒方法为2.5%NaClO处理老枝与分枝交叉处60s;在初代培养基中加入蔗糖能促进细叶小羽藓的生长,并随蔗糖浓度的升高其促进作用增强,当蔗糖质量浓度达到30g.L-1时,芽体长度可达2.49cm,芽体个数为1.87,原丝体直径为1.14cm,增殖系数高达9.25;当蔗糖质量浓度超过30g.L-1对细叶小羽藓芽体长度及芽体数生长的促进作用开始降低,原丝体生长受到抑制,增殖系数显著低于蔗糖质量浓度为30g.L-1时细叶小羽藓配子体的增殖系数。

The effects of different disinfectants and material positions on sterlhzmg etlect ot Haplocladiurn microphylgurn gametophyte were studied hy orthogona[ design, and the effects of sucrose concentration on growth of H. rnicrophyllum gametophyte were investigated by measuring and calculating the bud length, bud number, gametophyte diameter and multiplication coefficient. The results showed that the best way of H. microphyllum gametophyte sterilization was as follows： explants gotten from intersection of branch were washed in 2.5 % NaC10 for 60 s. When 0-30 g. L 1 sucrose was added in the initial culture medium, it could promote the growth of H. rnicrophyllum gametophyte, and the effect was significant with increasing sucrose concentrations. When 30 g. L-1 sucrose was added in the initial culture medium, the bud length, bud number, gametophyte diameter and multiplication coefficient were 2.49 cm, 1.87, 1.14 cm and 9.25, respectively. But more than 30 g,L-1 sucrose was added in the initial culture medium, the branch and protonema growth of H. rnicrophyllum were decreased, and the multiplication coefficient was significantly lower than that in addition with 30 g. L 1 sucrose.