铝诱导不同耐铝型桉树优良无性系叶片mRNA差异显示研究-林学林木遗传育种专业毕业论文.docxVIP

II II 明两个无性系在铝胁迫下发生了明显的差异表达。 （5）两个桉树无性系 BLAST 搜索同源性比对结果得出，在耐铝型无 性系 G9 中 G9_21_1 基因片段与桃金娘科桉属植物叶绿体基因同源性为 100%。G9_24_1 序列与杨树（populus）耐旱相关基因同源性为 77%。G9_9_4 序列与可可中的过氧化物酶基因有 85%的同源性。序列片段 G9_35_1 与狼 尾草（Pennisetum alopecuroides）和盐角草（Salicornia europaea）中的热休 克蛋白基因同源性分别达到 90%和 91%。序列 G9_15_2 与可可（Theobroma cacao）中的乙二醛酶基因片段同源性达 82%。这 5 个与逆境胁迫相关的基 因片段均来自经铝胁迫处理的植株，且有些片段的表达量随着胁迫时间的 延长逐渐增加。另外，G9_16_1 序列与长春花（Catharanthus roseus）的环 烯醚萜苷类葡糖基转移酶上的基因片段同源性达 85%，它也属于抗胁迫基 因但在耐铝型无性系 G9 铝胁迫植株中其表达被抑制。 铝敏感型无性系 W4 中 BLAST 搜索结果同源性最高的是 W4_10_2 序 列，该序列与可可树的动力蛋白基因同源性达 100%。W4_16_3 序列与蒺藜 状苜蓿中 GDSL 脂肪酶基因同源性为 88%。W4_19_2 序列与拟南芥 U-box 蛋白基因的同源性达 88%。以上 3 个与胁迫相关的基因在铝敏感型无性系 W4 中经铝胁迫后均抑制表达。另外，W4_22_1 序列与剥皮桉叶绿体上的基 因同源性达 100%，此抗性基因在铝敏感型无性系 W4 中经铝胁迫处理一天 诱导表达。 关键词：桉树；铝毒害；DDRT-PCR；反转录 III III ANALYSIS OF DIFFERENT ALUMINUM-INDUCED MRNA IN CLONES OF EUCALYPTUS USING DDRT-PCR METHOD ABSTRACT Eucalyptus is an important fast-growing tree, which was usually used to cultivated pure forest in southern China. Aluminum-rich soil, which was induced by acidic soil, acid rain, continuous cropping, fertilizer and other reasons, is one of the most important factors for forest recession. Analysis the molecular mechanism in response to aluminum toxicity would be useful for selective breeding resistance species of Eucalyptus. In this study, we optimized the mRNA difference display technology and screened the gene fragment with different transcript level in Eucalyptus clones G9 (insensitive to aluminium toxity) and W4 (sensitive to aluminium toxicity) under aluminum treatment. A series of differential gene fragments were sequenced analyzed. The main results were shown as follows: The RNA extraction kit was a reliable method for Total RNA extraction from Eucalyptus leaves. The purified RNA has a highly quality and quantity, which was suitable for further analysis using DDRT-PCR method. Using orthogonal experiment method, the main factors in PCR system mainly including TaqDNA polymerase, dNTP and annealing temperature, were optimi