【作者】 洑香香；

【导师】 施季森；

【作者基本信息】 南京林业大学 ， 林木遗传育种， 2006， 博士

【摘要】 利用差异显示法(DDRT-PCR)研究杉木的2个自然变异类型(句容0号及独干杉)皮层(包含周皮、韧皮部、形成层及初生木质部)基因表达差异，共获得29个差异片段，测序后进行Blast比对分析。结果表明： 1)Blastn比对(分值＞60)分析表明共有9个片段在GeneBank中找到了相似的功能。分别与核糖体蛋白基因、信号传导功能、泛素蛋白基因、抗性功能、组氨酸磷酸转移蛋白、细胞发生功能及能量代谢相关。 2)Blastx比对分析有12个序列可进行功能推测。其中7个片段和细菌、动物的功能相似，涉及膜结构、细胞因子、代谢过程中酶的活性及一些未知蛋白。 3)还有8个差异片段在数据库中未发现匹配信息。 对句容0号杉茎段皮层cDNA文库随机测序。分析表明有效EST序列1，119条，聚类分析和拼接后获得354个UniGenes。Unigene比对结果表明： 1)高度相似和中度相似的序列分别为58个和70个，低度相似和无匹配序列合计达63.8％。 2)可以推测基因功能的高丰度、中丰度及低丰度表达序列分别为5条、25条和98条。 3)基因功能：功能注释中表达的基因主要有：一类参与光合作用相关的相关基因，包括多种光合系统的组分、叶绿体a／b结合蛋白、氧捕获增强蛋白和能量传递等；另一类是与受外界胁迫表达的抗逆性相关基因。除此，表达频率较高的ARP(Auxin-RepressedProtein)可能与杉木分枝及茎的伸长有关。更多还原

【Abstract】 Differential display genes derived from the bark of two natural mutants of Cunninghamia lanceolata var. "dugan" and Cunninghamia lanceolata var. "jurong 0" were analyzed using mRNA differential display reverse transcriptase polymerase chain reaction (DDRT-PCR). The results showed: 1) Twenty-nine differential expressed fragments were screened and sequenced. Nine differential display transcrips matche to sequences of known or unknown function in GeneBank by Blastn (score>60). Functions involve: ribosomal protein, signal transferring, polyubiquitin, resistant protein, histidine-containing phosphotransfer protein and ATP synthase respectively.2) Blastx analysis showed that putative function were assigned 12 DD fragments: seven fragments were similar to that of bacteria and animals, function of them involved in membrane structure, cellular component, enzyme activity of metabolizing process and unknown protein; function forecast of 5 DD fragments came from plants which function were related to zinc finger protein、cf2-like protein、Glycerophosapokiester phosphodiesterase/kinase and catalase CAT-2.3) there were still eight fragments with no hit in the GenBank.Clones from cDNA library of Cunninghamia lanceolata var. jurong 0 bark tissue were selected randomly and sequenced from 5’ end. 1,119 effective ESTs were grouped into 354 Unigenes. Blastx showed:1) the number of sequences with high and moderate similarity were 58 and 70 respectively, total of low similarity and no hit with GenBank was up to 63.8%;2) putative gene function were assigned to 5 Unigenes consisted of high expressed frequency ESTs, 25 consisted of moderate expressed frequency ESTs and 98 Unigenes consisted of low expressed frequency ESTs.3) gene function: annotated gene expressed with higher frequency in the bark library of Chinese fir involved: one group was genes of photosynthesis process including components of photosystem pathway, chlorophyll a/b-binding protein, oxygen evolving enhancer protein and energy transferring etc.; another group was resistant protein induced by environmental stress, such as Cu/Zu-superoxide dismutase, bZIP transcription factor, heat stock protein, cytochrome etc.. Besides, ARP (Auxin-Repressed Protein) with high expressed frequency may be related with branching and elongate of stem in Chinese fir.更多还原