Tissue culture of Sinningia speciosa and analysis of the in vitro-generated tricussate whorled phyllotaxis (twp) variant
Tissue culture of Sinningia speciosa and analysis of the in vitro-generated tricussate whorled phyllotaxis (twp) variant
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DOI：
10.1007/s11627-009-9257-z
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发表时间：
2009-10
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影响因子：
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通讯作者：
Quan Xu;Zhe Hu;Chun-yuan Li;Xin-yu Wang;Chongying Wang
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文献类型：
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作者：
Quan Xu;Zhe Hu;Chun-yuan Li;Xin-yu Wang;Chongying Wang
以大岩桐叶片为外植体，通过两种途径建立了大岩桐叶片高效再生体系。第一种方法是在含有2.0 mg l−16-苄基氨基嘌呤（BA）和0.2 mg l−1α-萘乙酸（NAA）的MS培养基中形成愈伤组织和芽，然后进行根的生长，再生效率为99.0%。第二种方法涉及产生愈伤组织和根，随后在添加1.0-5.0 mg l− 1 NAA的MS培养基中形成芽，并且导致再生效率为90.4%。我们的实验表明，根优先途径导致较低的植株再生效率。通过芽先法连续5代，最后一代共获得215株再生植株，其中8株表现为三簇轮生叶序（twp）。6个再生的twp变异植株保持其三对生轮生叶序表型，没有表现出其他异常，而一个在开花前恢复为野生型，另一个形成两轮萼片。生理分析表明，两株植株对外源NAA和2，3，5-三碘苯甲酸（TIBA）的反应与野生型不同，而高效液相色谱（HPLC）分析表明，两株植株内源吲哚乙酸（IAA）和赤霉素（GA）含量低于野生型。这些结果表明，该突变体的形成可能与植物激素有关，该突变体可能是研究植物叶序格局形成的分子机制的重要材料。
Two protocols were developed for the efficient regeneration ofSinningia speciosafrom leaf explants via two developmental pathways. The first method involved formation of callus and buds, followed by subsequent root growth, in Murashige and Skoog medium (MS) containing 2.0 mg l−16-benzylaminopurine (BA) and 0.2 mg l−1α-naphthalene acetic acid (NAA), with a regeneration efficiency of 99.0%. The second method involved producing callus and roots, followed by subsequent formation of buds, in MS medium supplemented with 1.0–5.0 mg l−1NAA, and resulted in a regeneration efficiency of 90.4%. Our experiments indicate that the root-first pathway resulted in a lower plant regeneration efficiency. Through five continual generations using the buds-first method, a total of 215 regenerated plants were obtained in the last generation, and eight exhibited a phenotype we namedtricussate whorled phyllotaxis(twp). Six of the regeneratedtwpvariant plants maintained their tricussate whorled phyllotaxis phenotype, showing no other abnormalities, while one reverted to a wild type before flowering and another formed two rounds of sepals. Physiological analysis revealed that thetwpplants responded differently than wild type to exogenous NAA and 2,3,5-triiodobenzoic acid (TIBA), while high-performance liquid chromatography (HPLC) analysis showed that the levels of endogenous indole-3-acetic acid (IAA) and gibberellin (GA) were lower intwpthan wild-type plants. These results suggest that the formation of thetwpmutant may be related to phytohormones and that thetwpvariant could be an important material for investigating the molecular mechanism of plant phyllotaxis patterning.