啤酒花不同外植体筛选及再生体系构建

doi:10.11686/cyxb2019160

摘要/Abstract

摘要： 为研究不同外植体、不同激素配比对啤酒花愈伤组织诱导、分化和植株再生的影响,建立稳定高效的啤酒花再生体系,选用5种不同基因型啤酒花半木质化枝条,通过扦插生根发芽筛选最适外植体供体。以啤酒花根尖、腋芽、叶片为外植体,研究不同外源激素配比对其愈伤组织诱导、不定芽分化及生根的影响。结果表明:1)PJ274材料在相同生长条件下生根率最高为85%,且在日本园试配方的水培液中发芽率高达40%,是最理想的啤酒花外植体供试母体材料;2)对比根尖、腋芽和叶片为外植体诱导愈伤组织情况,腋芽为最适外植体,诱导率为51.11%;3)啤酒花腋芽愈伤组织诱导的最适激素配比是6-BA(6-苄氨基嘌呤)0.1 mg·L^-1+IAA(吲哚乙酸)1.0 mg·L^-1+2,4-D(2,4-二氯苯氧乙酸)2.0 mg·L^-1,愈伤组织诱导率可达86.67%;4)啤酒花腋芽愈伤组织芽的分化率在6-BA 1.0 mg·L^-1+NAA(萘乙酸)0.2 mg·L^-1+2,4-D 2.0 mg·L^-1的条件下最高,为66.67%;5)啤酒花腋芽愈伤组织根分化的最佳激素调控模式为6-BA 0.1 mg·L^-1+IBA(乙哚丁酸)1.0 mg·L^-1,最高生根率为66.67%。试管苗经炼苗后移栽,成活率达80%。最终成功构建了以腋芽为外植体的啤酒花高频再生体系,为啤酒花种质资源保存、组培快繁和基因工程育种研究奠定基础。

关键词: 啤酒花, 外植体, 愈伤组织, 再生体系, 激素

Abstract: The aim of this research was to study the effects of different explants and hormone combinations on callus induction, differentiation and regeneration of hops, and finally to establish a stable and efficient hop plant regeneration system. Five different hop genotypes with semi-lignified shoots were selected, and the optimal explant donors were screened by cutting rooting and germination. The root tip, axillary bud and leaves of the hop genotypes were used as explants to study the effects of different exogenous hormone combinations on callus induction, adventitious bud differentiation and rooting. It was found that a genotype ‘PJ274’ had a rooting rate of up to 85% under the test growth conditions, and the germination rate of PJ274 material in hydroponic liquid of the ‘Japanese garden’ formula was as high as 40%, making PJ274 the most suitable among the tested genotypes for producing hop explants. Comparing the root tip, axillary bud and leaf as tissues for explant-induced callus, we found that axillary bud to be optimal with an induction rate of 51.11 %. The optimum culture conditions for callus induction using hop axillary buds were: 6-Benzylaminopurine (6-BA) 0.1 mg·L^-1+IAA (indole-3-acetic acid) 1.0 mg·L^-1+2,4-dichlorophenoxyacetic acid (2,4-D) 2.0 mg·L^-1, and with this formulation callus induction rate reached 86.67%. The differentiation rate of axillary bud callus was achieved using 6-BA 1.0 mg·L^-1+1-naphthaleneacetic acid) 0.2 mg·L^-1+2,4-D 2.0 mg·L^-1, with up to 66.67% success. The optimal hormoned regime for promotion of callus root differentiation of hops was 6-BA 0.1 mg·L^-1+indole-3-butytric acid 1.0 mg·L^-1, with a maximum rooting rate of 66.67% achieved. The test tube seedlings were transplanted after refining, and the survival rate was 80%. With these protocols, a high-output regeneration system for hops, using axillary buds as explants, was successfully established, so laying a foundation for the preservation of hop germplasm resources, tissue culture and rapid propagation, and genetically engineered breeding.

Key words: Humulus lupulus, explant, callus, regeneration system, hormone

杨轲, 陈一酉, 汪军成, 姚立蓉, 孟亚雄, 马小乐, 李葆春, 司二静, 王化俊. 啤酒花不同外植体筛选及再生体系构建[J]. 草业学报, 2019, 28(8): 209-217.

YANG Ke, CHEN Yi-you, WANG Jun-cheng, YAO Li-rong, MENG Ya-xiong, MA Xiao-le, LI Bao-chun, SI Er-jing, WANG Hua-jun. Selection of different explants from hops and establishment of a regeneration system[J]. Acta Prataculturae Sinica, 2019, 28(8): 209-217.

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