元宝草高频植株再生体系的建立  

Establishment of high-frequency plant regeneration system in Hypericum sampsonii

作　　者：杨鸿淋 何金玉 张欣钰 候英英 吉能么友喝 邹利娟[1] YANG Honglin;HE Jinyu;ZHANG Xinyu;HOU Yingying;JI Nengmeyouhe;ZOU Lijuan(Ecological Security and Protection Key Laboratory of Sichuan Province,Mianyang Teachers1 College,Mianyang,Sichuan 621000,China)

机构地区：[1]绵阳师范学院生态安全与保护四川省重点实验室,四川绵阳621000

出　　处：《植物生理学报》2021年第12期2379-2386,共8页Plant Physiology Journal

基　　金：国家自然科学基金(31972258);绵阳师范学院科研启动项目(QD2019A12);四川省科技厅应用基础项目(2018JY0192)。

摘　　要：以元宝草(Hypericum sampsonii)无菌苗顶芽和茎段为实验材料,以MS为基本培养基,研究不同植物生长调节物质种类及浓度对愈伤组织诱导、从生芽发生及植株再生的影响。结果表明:顶芽通过间接器官发生途径,在MS+0.3 mg·L^(-1)IAA+2.0 mg·L^(-1)TDZ中, 15 d左右诱导出再生能力极强的愈伤组织,诱导率100%;30 d后愈伤组织在此培养基上分化出绿色丛生芽,平均每个外植体形成的再生芽数25.5个,分化率100%。茎段通过直接器官发生途径,在MS+0.3 mg·L^(-1)IAA+3.0 mg·L^(-1)TDZ中培养25 d,平均每个外植体诱导的从生芽数为32.4个。2种培养获得的试管苗矮小瘦弱,不利于生根培养,丛生芽在MS+3.0 mg·L^(-1)6-BA+0.3 mg·L^(-1)IAA+0.1 mg·L^(-1)GA_(3)+0.5 g·L^(-1)VC中培养20 d后可获得健壮的试管苗,增殖系数可达21.3,平均株高达6.2 cm;在1/2MS+1.5 mg·L^(-1)IBA培养基中进行生根培养, 20 d即可获得健康的再生植株,生根率可达83.5%,平均生根数为34.5,平均根长为3.5 cm。组培苗炼苗5 d后移栽到基质中,成活率可达100%。本实验建立了元宝草无性快速繁殖体系,为优质种苗繁育及工厂化生产提供了有效途径,并为其遗传转化研究奠定了基础。The experiment explored the effects of different types and concentrations of plant growth regulators on callus induction, adventitious shoot formation and plant regeneration, using apical buds and stem segments of aseptic seedlings of Hypericum sampsonii as explants. The results showed that MS medium supplemented with 0.3 mg·L^(-1)IAA and 2.0 mg·L^(-1)TDZ was best for the apical bud callus induction with a strong capacity by the indirect organogenesis, and the callus induction was up to 100%. The callus started to generate green multiple shoots and yield the average number of shoots(25.5) with 100% after 30 d. By direct organogenesis way, stem segments yielded the highest and number of regenerated shoots(32.4) supplemented with a combination of MS, 0.3 mg·L^(-1)IAA and 3.0 mg·L^(-1)TDZ after 25 d. The plantlet derived from two explants was too weak for rooting, the multiplication coefficient(21.3) and average height of the shoot(6.2 cm) were obtained on MS medium supplemented with 3.0 mg·L^(-1)6-BA, 0.3 mg·L^(-1)IAA, 0.1 mg·L^(-1)GA_(3)and 0.5 g·L^(-1)VC to rejuvenation for 20 d. The regenerated shoots developed roots in 1/2 MS containing 1.5 mg L^(-1)IBA, healthy regenerated plants were obtained with 83.5% rooting rate, the average number of roots was 34.5, and the average root length was 3.5 cm after 20 days. Successful acclimatization of rooted plantlets in a greenhouse was achieved with 100% survival rate. Our protocol offers, an efficient, scalable propagation procedure for H. sampsonii useful of large-scale cultivation with the purposes of conservation and genetic manipulation.

关 键 词：元宝草 愈伤组织 丛生芽 无性快速繁殖 再生体系 

分 类 号：S567.239[农业科学—中草药栽培]