甘草愈伤组织诱导及带芽茎段丛生芽增殖条件优化

李伟　张东向　刘丽杰　吕晴　任艳波

摘 要：为筛选适于甘草愈伤组织诱导及带芽茎段丛生芽增殖的最佳激素配比，选取萌发能力最强的甘草种子作为试验材料，获得无菌苗，取其下胚轴和带芽茎段为外植体，在不同激素配比下诱导愈伤组织，统计愈伤组织的生长状况、颜色状态、诱导率和出愈率等指标，并研究不同激素对甘草带芽茎段丛生芽增殖的影响，统计带芽茎段丛生芽株高、节间距、真叶数、增殖倍数等指标。结果表明：MS+6BA 1.0 mg·L-1+2，4D 2.0 mg·L-1和MS+6BA 1.0 mg·L-1+NAA 2.0 mg·L-1有利于下胚軸愈伤组织的诱导，MS+6BA 0.2～0.6 mg·L-1有利于带芽茎段愈伤组织的形成;MS+6BA 1.8 mg·L-1对带芽茎段的生长最为有利，MS+6BA 1.0～2.2 mg·L-1有利于带芽茎段丛生芽的增殖，随着6BA浓度的升高，甘草带芽茎段丛生芽的增殖倍数也随之增加。

关键词：甘草;愈伤组织;带芽茎段;丛生芽;激素

中图分类号：S 567.23 文献标志码：A 文章编号：0253-2301（2021）10-0027-07

DOI： 10.13651/j.cnki.fjnykj.2021.10.006

Optimization on the Callus Induction and Proliferation of Cluster Budsfrom Stem Segments with Buds of Glycyrrhiza Uralensis

LI Wei， ZHANG Dong-xiang*， LIU Li-jie， LYU Qing， REN Yan-bo

（College of Life Sciences and Agriculture and Forestry， Qiqihar University/Heilongjiang Key Laboratory of

Resistance Gene Engineering and Biodiversity Protection in Cold Areas， Qiqihar， Heilongjiang 161006， China）

Abstract： In order to screen the best hormone ratio for callus induction and the proliferation of cluster buds from the stem segments with buds of Glycyrrhiza uralensis， the seeds of Glycyrrhiza uralensis with the strongest germination ability were selected as the experimental materials to obtain the aseptic seedlings. The hypocotyls and stem segments with buds were taken as the explants to induce the callus under different hormone ratios， and the growth status， color status， induction rate and healing rate of callus were counted. Then， the effects of different hormones on the proliferation of cluster buds from the stem segments with buds of Glycyrrhiza uralensis were studied. And the indicators such as plant height， node spacing， number of true leaves and proliferation multiple of cluster buds from the stem segments with buds were counted. The results showed that MS+6BA 1.0 mg·L-1+2，4D 2.0 mg·L-1 and MS+6BA 1.0 mg·L-1+NAA 2.0 mg·L-1 were conducive to the induction of hypocotyl callus， and MS+6BA 0.2-0.6 mg·L-1 was conducive to the callus formation of the stem segments with buds. MS+6BA 1.8 mg·L-1 was the most beneficial to the growth of the stem segments with buds， and MS+6BA 1.0-2.2 mg·L-1 was conducive to the proliferation of cluster buds from the stem segments with buds. With the increase of 6BA concentration， the proliferation multiple of cluster buds from the stem segments with buds of Glycyrrhiza uralensis also increased.

Key words： Glycyrrhiza uralensis; Callus; Stem segments with buds; Cluster buds; Hormone

甘草Glycyrrhiza uralensis Fisch为豆科Leguminosae甘草属Glycyrrihiza多年生草本植物，是中国传统医学中最古老和最常用的草药之一。它含有20多种三萜和300多种黄酮类化合物。近年来，研究表明从甘草中分离的活性化合物具有抗肿瘤、抗菌、抗病毒、抗炎、免疫调节功能，同时具有恢复和保护神经、消化、呼吸、内分泌和心血管系统的功效[1]。《中国药典》2020年版规定甘草有3个来源，即乌拉尔甘草G.Uralensis Fisch.、光果甘草G.glabra Linn. 及脹果甘草G.inflata Batalin.[2]。甘草具有多种药用价值[3]，由于过度和不可持续的采挖，甘草的野生资源减少得非常迅速，现在虽然甘草在中国西北部地区广泛种植[4]。但人工栽培易受外界环境影响，因此无性系育苗是保持亲本优良特性的首选途径，特别是植物组织培养方法具有生长周期短、外植体遗传背景一致性高、生长条件可控制等优势，可快速、高效的繁育种苗[5]。……