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題 名 | 從16S rDNA限制片長度多型性分析臺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 |
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作 者 | 徐仕美; 林長平; | 書刊名 | 植物病理學會刊 |
卷 期 | 7:1 1998.03[民87.03] |
頁 次 | 頁43-53 |
分類號 | 433.4 |
關鍵詞 | 分類; 親緣關係; 植物菌質體; Classification; Phylogenetic relationships; Phytoplasma; |
語 文 | 中文(Chinese) |
中文摘要 | 本實驗嘗試以 16S rDNA基因之限制片段長度多型性 (RFLP) 分析七種臺灣、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)。植物藥綠體之 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。研究中使用 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、NotⅠ、RsaⅠ、 SacP、 SmaⅠ 以及 Tru9Ⅰ) 在某些或全部的植物菌質 體 16S rDNA 上有切位; 而 Alw44Ⅰ、AocⅠ、NheⅠ、PstⅠ、XbaⅠ 及 XhoⅠ 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 質體可分為六群,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:榆樹黃化病菌質體;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。 |
英文摘要 | The phylogenetic relationships of 11 phytoplasmas were established on the basis of restriction fragmet 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 Bc1Ⅰ 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 BclⅠ-digested total DNA of phytoplasma-infected plants using the primer pair (fD1 ' 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, A1luI, BamHI, BglI, ClaI, EcoRⅠ, HaeⅢ, HinfⅠ, HpaⅡ, KpnⅠ, NotⅠ, RsaⅠ, ScaⅠ, SmaⅠ, and Tru9Ⅰ digested some or all of 16S rDNAs of these 11 phytoplasmas; Alw44Ⅰ, AocⅠ, NheⅠ, PstⅠ, XbaⅠ and XhoⅠ did not have any restriction site in the 1.5 kb phytoplasma 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. |
本系統中英文摘要資訊取自各篇刊載內容。