以16S-23S rDNA spacer核酸序列分析台灣植物菌質體之親緣關係

Phylogenetic Relationships of Phytoplasmas on the Basis of Sequences of 16S-23S rDNA spacers

徐仕美( S. M Hsyu) ； 林長平( C. P Lin)

《植物病理學會刊》 11卷4期 (2002/12) Pp. 199-206

https://doi.org/10.6649/PPB.200212_11(4).0005

摘要

本實驗以變異性較大的16S-23S rDNA spacer核酸序列來分析6種台灣、1種大陸及3種國外植物菌質體之親緣關係，並進行分類。利用本實驗中所設計的兩引子對：G1’/L1和fH3/rH3，以聚合酵素連鎖反應增幅16S-23S rDNA spacer區域，除了絲瓜簇葉病菌質體增幅出3條spacer DNA片段外，其餘植物菌質體均只增幅出1條spacer DNA片段。將增幅所得的spacer DNA片段進行核酸定序，結果發現在植物菌質體之spacer區域中有一tRNAI1e，但絲瓜簇葉病菌質體的其中2條spacer DNA，LWB-1及LWB-3中並無tRNA。比對排列後的植物菌質體spacer序列，發現只有tRNA區域及兩段有關rRNA processing的片段較具保守性。從spacer序列堆衍的親緣演化樹顯示這10種植物菌質體可分為5群：第一群，甘藷簇葉病、甘藷簇葉病（大陸品系）、花生簇葉病及細花牽牛簇葉病等菌質體；第二群，絲瓜簇葉病菌質體；第三群，榆樹黃化病菌質體；第四群，水稻黃萎病及甘蔗白葉病菌質體；第五群，翠菊黃化病（紐澤西及西岸品系）菌質體。

關鍵字

親緣關係 ； 植物菌質體

並列摘要

The phylogenetic relationships and classification of 10 phytoplasmas were established by analysis of 16S-23S rDNA spacers, the regions with variation in prokaryotic rrn operon. The spacer regions were amplified by polymerase chain reaction using two primer pairs, G1'/Ll and fH3/rH3. One spacer fragment was amplified for each phytoplasma, except loofah witches' broom (LWB) phytoplasma. There were three amplified spacer fragments for LWB phytoplasma. These amplified DNA were sequenced, and the results showed that the phytoplasma 16S-23S spacer region contains a single tRNAIle; however, there is no tRNA in two spacers of LWB phytoplasma, LWB-1 and LWB-3. In multiple alignment of phytoplasma spacers, only tRNAIle and two short segments, which may be involved in rRNA processing, appeared to be conserved. Based on Phylogenetic analysis of sequences from these 10 phytoplasmas using PAUP (phylogenetic analysis using parsimony) software, these phytoplasmas could be classified into five groups. These phytoplasmas groups and strains were: group 1, sweetpotato witches' broom, mainland China strain of sweetpotato witches' broom, peanut witches' broom and Ipomoea obscura witches' broom; group 2, loofah witches' broom; group 3, elm yellows; group 4, rice yellow dwarf and sugarcane white leaf disease; group 5, New Jersey strain of aster yellows and western strain of aster yellows.

並列關鍵字

phylogenetic relationships ； phytoplasma