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[摘要]
目的: 建立蚬壳花椒离体再生体系并测定其内源激素量的变化。方法:以蚬壳花椒组培苗叶片为外植体, 接种在附加不同激素组合的MS培养基上, 诱导愈伤组织不定芽的分化。同时, 采用酶联免疫吸附法(ELISA)对愈伤组织分化过程中赤霉素(GA3)、生长素(IAA)、脱落酸(ABA)、玉米素核苷(ZR)4种内源激素的量进行测定。结果: 蚬壳花椒叶片不定芽诱导的最适培养基为MS+6-BA0.5mg/L+NAA0.1mg/L, 诱导率最高可达61%。内源激素测定发现, 分化过程中ZR、IAA的量相对较高, ABA量一直保持较低的水平, GA3在分化前期和中期均呈上升趋势。结论: ZR、IAA是蚬壳花椒叶片愈伤组织不定芽分化的主要因素, 培养基中适当添加GA3可能有利于提高芽的分化率; ABA可能是此过程中的负调控因子。
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[Abstract]
Objective: To establish a regenerative system in vitro and determine the endogenous hormones of Zanthoxylum dissitum. Methods: Lamina of Z. dissitum was used as the explant. Callus adventitious bud differentiation was carried out by culturing on MS with different hormones. At the same time, in the course of callus induction, four endogenous hormone Of GA3, IAA, ABA, and ZR were determined by ELISA. Results: The optimum medium for adventitious bud regeneration of Z.dissitum was MS+6-BA 0.5 mg/L+NAA 0.1 mg/L, and the highest induction rate could reach to 61%. During the process of the differentiation of explants callus, the contents of ZR and IAA were correspondingly higher than other endogenous hormones, the content of ABA was always kept in low level during this process, the content of GA3 was kept in the trend of upgrading during the earlier differentiation period.Conclusion: It could be considered that ZR and IAA should be the critical factors in the bud induction. According to the results, a proper adding of GA3 in the culture medium could improve the differentiation rate of the adventitious bud. On the other side, ABA might be the negative regulation factor.
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[基金项目]
湖南省自然科学基金项目“药用植物单面针的生物学特性研究”(07J5041);株洲市重点科技项目“药用植物单面针组培技术中试研究及组培苗栽培技术示范”;株洲市千金药业股份公司合作项目“单面针人工快繁中试项目”