從16S rDNA限制片長度多型性分析台灣植物菌質體之親緣關係

Phylogenetic relationships of phytoplasmas on the basis of restriction fragment length polymorphisms of 16S rDNAs

徐仕美(S. M. Hsyu) ； 林長平(C. P. Lin)

《植物病理學會刊》 7卷1期 (1998/03) Pp. 43-53

https://doi.org/10.6649/PPB.199803_7(1).0006

摘要

本實驗嘗試以16S rDNA基因之限制片段長度多型性(RFLP)分析七種台灣、一種大陸及三種外國植物菌質體之親緣關係。參照Schneider等人發展之方法，以聚合酵素連鎖反應(PCR)可增幅植物菌質體幾近全長之16S rDNAs (25)。植物葉綠體之16s rDNA含有一BclI切位，而植物菌質體之16s rDNA則無；利用此差別，並配合fD1與rP1，引子對，可從BclI酵解後之罹病植物全DNA中以聚合酵素連鎖反應增幅出1.5 kb大小之植物菌質體16S rDNA。硏究中使用21種內限制酵素進行16S rDNA之RFLP分析，結果有15種內限制酵素(AluI 、Bam HI、BglI、ClaI、EcoRI、HaeIII、HindIII、HinfI、HpaII、KpnI、NotI、Rsal、SacI、SmaI以及Tru9I)在某些或全部的植物菌質體16S rDNA上有切位；而Alw 44I、AocI、Nhel、PstI、XbaI及XtoI等六種內限制酵素在1.5 kb片段上無切位。從RFLP結果推衍之親緣演化樹顯示這11種植物菌質體可分爲六群，第一群：甘藷簇葉病、甘藷簇葉病(大陸品系)花生簇葉病及細花野牽牛簇葉病等菌質體；第二群：絲瓜簇葉病菌質體；第三群：水稻黃萎病及甘蔗白葉病菌質體；第四群：楡樹黃化病菌質體；第五群：泡桐簇葉病菌質體；第六群：翠菊黃化病(紐澤西及西岸品系)菌質體。

關鍵字

分類 ； 親緣關係 ； 植物菌質體

並列摘要

The phylogenetic relationships of 11 phytoplasmas were established on the basis of restriction fragment length polymorphism (RFLP) patterns of 16s ribosomal RNA genes. Approximately full length of 16s rDNAs of 11 phytoplasma strains were amplified by polymerase chain reaction (PCR) using the method developed by Schneider et al. (25). The method, depending on the presence of a BclI restriction site in the 16s rDNA of the chloroplast but not in that of the phytoplasma, permits the specific amplification of the 1.5 kb 16S rDNA of phytoplasmas from BcIT-digested total DNA of phytoplasma-infected plants using the primer pair (fDi' and rP1') in conserved regions of this gene. A total of 21 restriction enzymes were used in RFLP analyses of the 1.5 kb phytoplasma 16S rDNA fragments. Among these restriction enzymes, Alul, BamHI, Bgl, Clal, EcoRI, HaelII, HindIII, Hinfl, HpalI, Kpnl, Notl, Rsa I, Sacl, smal and Tru9I digested some or all of 16s rDNAs of these 11 phytoplasmas; Alw 44I. AocI, Nhel, PstI. Xbal and xhol did not have any restriction site in the 1.5 kb phyto plasma 16S rDNA fragment. Based on phylogenetic analysis of RFLP patterns from these 11 phytoplasmas using PAUP (phylogenetic analysis using parsimony) software, these phytoplasmas could be classified into six groups These phytoplasmas groups and strains were as follows: 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, rice yellow dwarf and sugarcane white leaf disease; group 4, elm yellows; group 5, paulownia witches' broom: group 6, New Jersey strain of aster yellows and western strain of aster yellows.

並列關鍵字

Classification ； phylogenetic relationships ； phytoplasma