黑老虎带芽茎段的组织培养及快速增殖

Tissue culture and rapid propagation of bud and stem segments of Kadsura coccinea

【目的】为黑老虎的工厂化育苗及遗传转化研究提供参考。【方法】以大田培养和温室盆栽的黑老虎带芽茎段为外植体,比较不同消毒方式对黑老虎带芽茎段消毒效果的影响,研究不同植物生长调节剂浓度配比下外植体的腋芽诱导和增殖效果,找到适合培养黑老虎带芽茎段的最佳培养基。【结果】各处理中大田黑老虎带芽茎段的消毒效果均较差,腋芽成活率均为0。剪掉大田黑老虎地上部分的茎段,然后移栽到花盆中,放在温室内培养,待新梢长出后,取半木质化的带芽茎段作为组织培养的外植体,这样可以解决黑老虎外植体消毒难的问题。适宜的黑老虎带芽茎段消毒方法为75%酒精浸泡2 min+0.1%HgCl_(2)浸泡处理45 min,采用该方法消毒后污染率可以控制在18.5%之内,腋芽诱导率可达59.8%。适宜的黑老虎带芽茎段初代培养基配方为1/2 MS+2.0 mg/L6-BA+0.2 mg/L IAA,在该培养基上腋芽诱导率可达46.2%;最佳继代增殖的培养基配方为1/2 MS+3.0 mg/L6-BA+0.1 mg/L 3-IBA,在该培养基上腋芽增殖系数可达8.5。【结论】以温室盆栽植株的带芽茎段作为外植体,可显著降低污染率,提高腋芽萌发率;消毒后的带芽茎段经过初代培养及继代增殖培养后,可以得到生长健壮的黑老虎无菌苗。

【Objective】To provide reference for industrial breeding and genetic transformation of Kadsura coccinea.【Method】The stem segments with buds cultured in field and potted in greenhouse were used as explants to compare the effects of different disinfection methods on the disinfection effect of stem segments with buds of K. coccinea, and to study the axillary bud induction and proliferation effect of explants under different plant growth regulators concentration ratios,so as to find the optimal medium for cultivating stem segments with buds of K. coccinea.【Result】The disinfection effect of stem segments with bud of K. coccinea in field was poor, and the survival rate of axillary bud was 0. The stem segments of the aboveground part of the K. coccinea in the field were cut off, and then transplanted into the flowerpot,and cultured in the greenhouse. After the new shoots grew out, the semi-lignified stem segments with buds were taken as explants for tissue culture, as to solve the problem of difficult disinfection of K. coccinea explants. The appropriate disinfection method for stem segments with buds of K. coccinea was soaking in 75% ethanol for 2 min + 0.1% HgCl_(2) for 45 min. After disinfection, the contamination rate could be controlled within 18.5%, and the axillary bud induction rate could reach 59.8%. The suitable medium for the primary generation of stem segments with buds of K. coccinea was 1/2 MS + 2.0 mg/L 6-BA + 0.2 mg/L IAA, and the axillary bud induction rate could reach 46.2% on this medium.The optimal medium for subculture was 1/2 MS + 3.0 mg/L 6-BA + 0.1 mg/L 3-IBA, and the axillary bud proliferation coefficient could reach 8.5 on this medium.【Conclusion】Taking the stem segments with buds of potted plants in greenhouse as explants can significantly reduce pollution rate and improve axillary bud germination rate. After primary culture and subculture, the sterilized stem segments with buds could obtain robust aseptic seedlings of K. coccinea.