黑蒴定芽诱发增殖的初步研究

Adventitious bud induction and multiplication of Melasma arvense

关键词:  黑蒴, 定芽诱发, 组织培养, 基本培养基, 细胞分裂素, 天然有机添加物

Key words:  Melasma arvense, induction of adventitious bud, tissue culture, basic medium, cytokinin, natural organic additives

摘要:

黑蒴(Melasma arvense)为云南省民族民间珍稀药材,现已成为云南天然药物开发的热点药材种类。但目前在云南已濒临灭绝,急需对其进行资源保护和人工繁育。该研究在筛选黑蒴不同器官培养的基础上,以茎枝为外植体,依次考察了不同基本培养基、不同种类细胞分裂素和质量浓度、不同天然有机添加物对定芽(顶芽和侧芽)萌发增殖及生长的影响。结果表明:以黑蒴带节茎枝为外植体,将其剪成2～3 cm带节嫩茎枝,经严格的外植物体消毒程序处理后,接种到1/2MS+6-BA 0.5 mg·L^-1 + Pt 50 g·L^-1+ Bn 80 g·L^-1 + 25 g·L^-1蔗糖+7 g·L^-1琼脂, pH值5.8的培养基上,接种后放置在光照强度1 500～2 000 lx,光照10 h·d^-1,温度22 ℃下培养,接种后8～10 d开始萌发,40 d单茎平均增殖数达8.1,主茎40 d生长长度74.2 mm,显示出极好的诱发增殖生长效果。该研究结果初步建立了黑蒴嫰茎枝诱发定芽及增殖培养体系,为云南珍稀民族药黑蒴的资源保护、人工栽培和可持续利用提供了科学依据。

Abstract:

Melasma arvense is known as one of the precious herbs in Yunnan Province, and has the effects on leukemia, cancer, dispersing blood stasis and hemolysis. It has become a hot spot for the development of natural drugs in Yunnan. At present, because of its peculiar habitat, seed dispersal difficulty, reproductive barriers, seed germination and establishment difficulty and over-harvesting, wild plants are becoming extinctive recent years in Yunnan Province. Therefore, in order to realize resource protection and restoration, it is necessary to carry out resource conservation and artificial breeding. In order to primary establish the adventitious bud induction and multiplication system that present excellent properties of growth and multiplication, used stem shoots as tissue culture inoculation materials studied the effects of different medium type, different concentrations of cytokinin and different natural organic additives on bud multiplication and growth based on the prophase research of provenance collection, artificial propagation and screening of different organ culture of M. arvense. The results indicated that about 2-3 cm long stem with buds cultured on cultural conditions(1/2 MS with 6-BA 0.5 mg·L^-1 + Pt 50 g·L^-1 + Bn 80 g·L^-1 + 25 g·L^-1 sugar + 7 g·L^-1 agar with pH=5.8, a temperature of 22 ℃, light 10 h·d^-1)for 8-10 d, they produced numerous adventitious buds. Each explant produced 8.1 shoots and the grow length of main stem was 74.2 mm in 40 d. The experiment primarily established the adventitious bud induction and multiplication system that present excellent properties of growth and multiplication. This study provides a scientific basis for the artificial cultivation and sustainable use.