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香根草茎节腋芽离体再生成苗研究

Study on Seedling Regeneration from Axillary Buds of Stem Nodes of Vetiveria zizanioides in Vitro
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摘要 香根草是一种重要的生态保护用草,本研究预建立一种以香根草茎节腋芽为外植体的再生成苗体系,提高香根草的繁殖速度。为探究最优再生体系,本试验取香根草茎节腋芽为外植体,首先设计不同升汞浓度与时间梯度处理外植体,选取最佳消毒处理,进而在各浓度多菌灵MS培养基上,选取最佳多菌灵添加浓度;然后设计12种NAA与6-BA激素组合的外植体展芽培养基探究最佳展芽配方,再取展芽接在15种6-BA与2,4-D组合的芽增殖培养基上优化增殖配方;最后取增殖芽到12种IBA与NAA组合的生根培养基上,筛选最佳生更激素配比。结果表明,适宜的外植体消毒方法是75%酒精浸泡30 s,0.6%升汞处理10 min,同时在MS中加入60 mg/L多菌灵,真菌污染率仅为3.87%;适宜的展芽培养基为MS+3 mg/L 6-BA+0.2 mg/LNAA,展芽率高达93.3%;适宜不定芽增殖培养基为MS+0.5 mg/L 2,4-D+2 mg/L 6-BA,增殖新芽数达34个;适宜不定芽生根培养基为MS+0.2 mg/L NAA+0.5 mg/L IBA,生根率达99.78%。应用本研究建立的香根草离体再生体系,单个香根草茎节腋芽可获得30株幼苗,显著提高了香根草的繁殖效率,同时为香根草转基因方面的研究打下坚实基础。 Vetiveria zizanioides is an important grass used in ecological protection.In this study,a seedling regeneration system was established using the axillary buds of stem segments of Vetiveria zizanioides as explants.In order to explore the optimal regeneration system,this experiment took the axillary bud of the stem node of Vetiveria zizanioides as explant.First,different concentration of mercury and time gradient were designed to treat 30 explants,and the optimal disinfection treatment was selected,and then the optimal carbendazim addition concentration was selected on the MS medium of each concentration of carbendazim.Then,explant budding culture medium of 12 kinds of NAA and 6-BA hormone combination was designed to explore the optimal sprout formula,and then the spreading bud was optimized on the bud multiplication medium of 15 6-BA and 2,4-D combination.Finally,the proliferation buds were taken to rooting medium of 12 kinds of IBA and NAA combination,each treatment was 10 bottles and 3 plants per bottle.The results showed that,the suitable disinfection method for explant was soaking them in 75%alcohol for 30 s,treating them in 0.6%mercuric chloride for 10 min,and adding 60 mg/L carbendazim to MS,which could effectively inhibit fungi,and the fungal contamination rate was only 3.87%;the suitable budding medium was MS+3 mg/L 6-BA+0.2 mg/L NAA,and the budding rate was 93.3%,and the suitable adventitious bud multiplication medium was MS+0.5 mg/L 2,4-D+2 mg/L 6-BA,the number of new shoots increased to 34;suitable adventitious bud rooting medium was MS+0.2 mg/L NAA+0.5 mg/L IBA,rooting rate reached 99.78%.Using the In vitro regeneration system established in this study,30 seedlings could be obtained from single axillary buds of stem segments of Vetiveria zizanioides.This study would not only enhance the reproductive efficiency of Vetiveria zizanioides,but also lay a solid foundation for the research of Vetiveria zizanioides transgene.
作者 王栋 曹苑楠 马乐 陈秀文 张菲 陈吉宝 Wang Dong;Cao Yuanan;Ma Le;Chen Xiuwen;Zhang Fei;Chen Jibao(Key Laboratory of Ecological Security for Water Source Region of Mid-line of South-to-North Water Diversion Project of Henan Province,Nanyang Normal University,Nanyang,473061)
出处 《分子植物育种》 CAS CSCD 北大核心 2019年第4期1313-1319,共7页 Molecular Plant Breeding
基金 河南省科技攻关项目(152102110099)资助。
关键词 香根草 腋芽 组织培养 Vetiveria zizanioides Axillary bud Tissue culture
作者简介 通信作者:陈吉宝,chenjibao2012@163.com。
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