甘菊下胚轴离体再生体系的建立
Establishing an in vitro regeneration system from hypocotyls of Chrysanthemum lavandulifolium.
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摘要: 以甘菊下胚轴为外植体，通过器官发生途径诱导不定芽分化，建立了甘菊下胚轴高效再生体系，为甘菊遗传转化体系的建立奠定了基础。结果表明：将甘菊种子播种于MS培养基上，置于黑暗条件下培养7 d可以迅速获得大量下胚轴；下胚轴在MS+1.0 mg/L 2, 4-D+0.5 mg/L 6-BA的培养基上培养15 d后，愈伤组织形成率最高为8666%，愈伤组织呈淡黄色，大小一致；将愈伤组织转接到MS培养基中培养30 d左右即可分化不定芽，不定芽分化率达25.50%，平均每个外植体产生不定芽数目为4.25个；不定芽在添加0.1 mg/L NAA的1/2 MS培养基中培养15 d后，生根率达到100%。生根试管苗出瓶后，经过适宜培养，均可以获得健壮的开花植株。Abstract: An efficient regeneration protocol had been established for plantlet regeneration from hypocotylinduced calli of Chrysanthemum lavandulifolium, which laid the foundation for genetic transformation of C. lavandulifolium. When long hypocotyls from the seeds that germinated in the dark for 7 days were cultured on the Murashige and Skoogs (MS) medium containing 1.0 mg/L 2, 4-dichlorophenoxyacetic acid (2, 4-D) and 0.5 mg/L 6-benzyl aminopurine(6-BA) for 15 days, the calli inducing rate could reach the maximum of 86.66%. The calli were light yellow with almost uniform size. Following approximately 30 days of culture on the MS medium, adventitious buds were differentiated. The highest adventitious bud rate (25.50%) with an average of about 4.25 adventitious buds per explant was obtained from calli cultured on a MS medium without 6-BA. The rooting rate was 100% when the adventitious buds were cultured on the 1/2 MS medium supplemented with 0.1 mg/L naphthaleneacetic acid (NAA) for 15 days. After acclimatized to greenhouse conditions, normal growth and blossom C. lavandulifolium plants were obtained.
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Keywords:
- Chrysanthemum lavandulifolium /
- hypocotyl /
- regeneration system
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