檵木芽变生物学特性及离体培养研究
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摘要
黄檵木是继红檵木之后新发现的檵本芽变类型,除具有潜在的巨大经济价值外,且对于深入研究檵木植物遗传与变异、植物基因表达与调控、植物色素合成、代谢途径有重要意义。本文以芽变黄檵木为研究对象,探讨与红檵木、檵木母本形态特征、叶细胞超微结构、生理生化、过氧化物同工酶和DNA水平的同源性及差异性,并在此基础上建立离体培养体系,为黄檵木新种质的产业化奠定基础。
     主要研究结果如下:
     1、檵木与其芽变红檵木和黄槛檵木比较,不仅在叶色方面发生改变,而且其叶绿体超微结构也相应地发生变化。红檵木叶绿体基粒片层晶格化,一部分叶绿体解体;檵木叶绿体内存在大量同化淀粉粒,基粒片层排列紧密;黄檵木叶绿体结构介于红檵木与檵木之间。
     2、檵木、红檵木、黄檵木质体色素含量及花色素苷含量均有差异。檵木、红檵木的叶绿素a、b和类胡萝卜素含量均高于黄檵木。红檵木花色素苷含量几乎是檵木的二倍,黄檵木略低丁红檵木。芽变黄檵木叶色变化主要原因可能是由于叶绿素含量降低引起的。
     3、檵木与其芽变红檵木和黄檵木过氧化物同工酶比较,它们存在着同源性和多样性。黄檵木和红檵木具有檵木的3条酶带,但是又各有一条明显的特征谱带。
     4、檵木与黄檵木质膜透性的比较,总体看来,檵木对不良高温和低温的抗性要高于其芽变黄檵木。
     5、建立了适宜于檵木RAPD分析的最佳反应体系。研究选用了10个随机引物,共扩增74条带,其中44条为多态性条带。多态性比例为59.5%。芽变黄檵木与母本檵木遗传相似系数为0.887,芽变红檵木与母本檵木遗传相似系数为0.796。
     6、建立了黄檵木离体培养体系,并进行了快速繁殖研究,现已批量生产试管苗。
     7、经过几代离体扦插繁殖,芽变黄檵木性状稳定。笔者认为当材料繁殖达到一定数量后,像红檵木Loropetalum chinense var.rubrum一样可以把它作为檵木家族中一新变种看待,可定名为:Loropetalum chinense var.auratum。
Loropetalum chinense var . auratum(Huangjimu) is a new bud mutation from L chinense oliv(Jimu) . It is important to study it' s heredity and mutation, genetic expression and regulation and metabolizability process for it' s enormous potential economy value. This article studied on the similarity and difference of shape, chloroplast ultrastructure, physiology and biochemistry characteristics, POD and RAPD among L. chinense. L. chinense var . auratum and L. chinense var. rubrum(ftongjimu). Based on the study ,the system of culture in vitro .which is the foundation for Huangjimu' s commercial production was established.
    Following are the main results:
    1. The color and shape of leaf and the chloroplast ultrastructure of Jimu are different from Hongjimu and Huangjimu. The chloroplast of Hongjimu are crystal latticed and some disaggregated . In chloroplast, there are a lot of strained powder, and the segments were closely arrayed.
    2. The content of pigment and anthocyanin varied among the three species. The contents of chla, chlb, and chlk in Jimu and Hongjimu leaves were higher than that in Huangjimu .The content of anthocyanin in Hongjimu leaf are nearly two times higher than that in Jimu .while Huangjimu is less than Hongjimu .Thus, the main reason that leaf color changed is the decrease of chloroplast content.
    3. The POD of three species has similarity and diversity. They all have 3 common strips and also have their own special strip.
    4. The membrane permeability that is widely used on the research of plant physiologial resistance is one of the important physiological index of adversity injury. The tolerance ability of Huangjimu is stronger than that of Jimu (L. chinense) .
    5. An optimal reaction system that is suitable to the RAPD assay was established. RAPD markers were generated by 10 arbitrary primers ,44 of total 74 bands were polymorphics . The genetic similarity was 0. 887 between Jimu and sport Huangjimu, and was 0. 796 between Jimu and Hongjimu.
    
    
    6. The culture in vitro system of Huangjimu (L. chinense var . aura turn) has been established and produced hundreds of plantlet.
    7. The genetic character of Huangjimu is stable through cutting in vitro .and thus, it could be considered as a new variety of L. chinense .named L. chinense var . auratum.
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