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Screening for differentially expressed genes in Anoectochilus roxburghii (Orchidaceae) during symbiosis with the mycorrhizal fungus Epulorhiza sp. 被引量:7
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作者 LI Biao TANG MingJuan +2 位作者 TANG Kun ZHAO LiFang GUO ShunXing 《Science China(Life Sciences)》 SCIE CAS 2012年第2期164-171,共8页
Mycorrhizal fungi promote the growth and development of plants, including medicinal plants. The mechanisms by which this growth promotion occurs are of theoretical interest and practical importance to agriculture. Her... Mycorrhizal fungi promote the growth and development of plants, including medicinal plants. The mechanisms by which this growth promotion occurs are of theoretical interest and practical importance to agriculture. Here, an endophytic fungus (AR-18) was isolated from roots of the orchid Anoectochilus roxburghii growing in the wild, and identified as Epulorhiza sp. Tissue-cultured seedlings of A. roxburghii were inoculated with AR-l 8 and co-cultured for 60 d. Endotrophic mycorrhiza formed and the growth of A. roxburghii was markedly promoted by the fungus. To identify genes in A. roxburghii that were differentially expressed during the symbiosis with AR-18, we used the differential display reverse transcription polymerase chain reac- tion (DDRT-PCR) method to compare the transcriptomes between seedlings inoculated with the fungus and control seedlings. We amplified 52 DDRT-PCR bands using 15 primer combinations of three anchor primers and five arbitrary primers, and nine bands were re-amplified by double primers. Reverse Northern blot analyses were used to further screen the bands. Five clones were up-regulated in the symbiotic interaction, including genes encoding a uracil phosphoribosyltransferase (UPRTs; EC 2.4.2.9) and a hypothetical protein. One gene encoding an amino acid transmembrane transporter was down-regulated, and one gene encoding a tRNA-Lys (trnK) and a maturase K (matK) pseudogene were expressed only in the inoculated seedlings. The possible roles of the above genes, especially the UPRTs and marK genes, are discussed in relation to the fungal interaction. This study is the first of its type in A. roxburghii. 展开更多
关键词 anoectochilus roxburghii orchid mycorrhizal symbiosis Epulorhiza sp. differential display-PCR (DD-PCR) gene screening
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金线兰黄酮相关基因的克隆与生物信息学分析 被引量:1
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作者 陈莹 潘王韵 林思祖 《分子植物育种》 CAS CSCD 北大核心 2019年第24期8020-8028,共9页
为了对金线兰查尔酮合成酶CHS和黄酮醇合成酶FLS进行基因的克隆和生物学信息分析,本研究参照测序得到的金线兰CHS与FLS基因的序列,设计特异性引物进行扩增。利用软件,对金线兰CHS和FLS蛋白的生物信息进行预测分析。结果显示,金线兰CHS... 为了对金线兰查尔酮合成酶CHS和黄酮醇合成酶FLS进行基因的克隆和生物学信息分析,本研究参照测序得到的金线兰CHS与FLS基因的序列,设计特异性引物进行扩增。利用软件,对金线兰CHS和FLS蛋白的生物信息进行预测分析。结果显示,金线兰CHS基因的CDS总长度1 185 bp,编码394个氨基酸,金线兰FLS基因的CDS总长度1 035 bp,编码344个氨基酸。金线兰CHS蛋白与FLS蛋白皆是酸性、亲水性、无信号肽、非分泌、非跨膜蛋白,亚细胞定位在其他细胞器上,丝氨酸磷酸化位点具有调控二者构象变化的重要作用。在Blast上进行序列比对,发现金线兰CHS序列和FLS蛋白序列与一些兰科植物的相似度很高,前者序列同源性均高达85%以上,后者同源性高达82%和81%,构建的进化树显示了他们较近的亲缘关系。推测得到的CHS序列即为金线兰查尔酮合成酶的编码序列,FLS蛋白序列即为金线兰黄酮醇合成酶的编码序列。本研究为进一步研究金线兰中黄酮相关基因及其调控提供一定参考。 展开更多
关键词 金线兰(anoectochilus roxburghii) 查尔酮合成酶 黄酮醇合成酶 基因克隆 生物信息学
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