东方杉的组织培养繁殖技术研究

Research on propagation of×Taxodiomeria peizhongii (Taxodium mucronatum×Cryptomeria fortunei) by tissue culture

以东方杉大树基部当年生的幼嫩萌枝为外植体,进行了组织培养技术研究。结果表明,外植体的最佳消毒方法为75%酒精消毒30～60 s后用升汞0.2%浓度消毒10～15 min。采用茎段微型扦插和诱导丛生芽的方法来增殖扩繁,适宜东方杉芽增殖培养基为:MHJ+BA 0.5 mg/L+NAA 0.05 mg/L+L-谷氨酰胺0.5 g/L+活性碳5 g/L+蔗糖2%,茎段腋芽萌发率可达90%。芽伸长生长的培养基为:MHJ+L-谷氨酰胺0.5 g/L+活性碳5 g/L+蔗糖2%,35 d平均增高4.2 cm。试管苗的生根培养基为:1/2 MHJ+L-谷氨酰胺0.5 g/L+活性碳5 g/L+蔗糖1%,最高生根率达75%,以沙为培养基质的效果比琼脂好。生根的组培苗先在沙床上进行炼苗,炼苗成活率达87%,然后移栽到带混合基质的营养钵中育苗,移栽成活率可达90%,育苗1～2个月后移栽大田。

The shoots newly sprouted within one year from the basal part of mature tree of x Taxodioraeria peizhongii （Taxodium mucronatum x Cryptomeria fortunei） were used as explants, and the technology of propa- gation by tissue culture was studied. The results were as follows： the optimal disinfection of explants was to disinfect them with 75% ethanol for 30～60s, and then with 0.2% mercuric chloride for l0～15 min; the stem cuttings were used to induce regenerated shoots and adventitious shoots ; the appropriate medium of shoot multiplication was MHJ ＋ 0.5 mg/L BA ＋ 0.05 mg/L NAA ＋ 2 g/L glutamine ＋ 5 g/L activated carbon ＋ 2% sucrose, and the germination percentage of stem＇s axillary buds could reach 90% ; the elongation medium was MHJ ＋ 2 g/L glutamine ＋ 5 g/L activated carbon ＋ 2% sucrose, and the shoot height growth was averagely 4.2 cm in 35 days ; the rooting medium for test-tube plantlets was 1/2 MHJ ＋ 2 g/L glutamine ＋ 5 g/L activated carbon ＋ 1% sucrose, and the highest rooting frequency was 75%; as culture substance, sand was better than agar; the rooted plantlets were firstly transferred to sand bed in greenhouse for 4～6 weeks of hardening off, in which the survival rate was 87%, and then into nutrition pot with a survival rate of 90% ; after one or two months the regenerated plantlets were transplanted into field.