利用逢機複製多型性DNA分析測定溶鈣磷細菌之遺傳歧異度

Genetic Diversity of Calcium Phosphate-Solubilizing Bacteria Determined by Random Amplified Polymorphic DNA Analysis

楊秋忠(C.C.Young) ； 陳鴻森(H. S. Chen)

《土壤與環境》 2卷2期 (1999/06) Pp. 147-158

https://doi.org/10.30058/SE.199906.0005

摘要

本研究利用六株溶鈣磷細菌，2株Bacillus pumillu Ca-48與Ca-49菌系，2株pseudomonas cepacia Al-72與PSB2菌系，及2株Rhizobium fredii SB+PSB與SB818菌系，進行限制指紋分析及逢機複制多型性DNA分析。限制指紋分析結果顯示，同屬的菌種之基因相似度較高，而SB+PSB和SB818雖然同屬於Rhizobium fredii，但其限制指紋分析的結果卻有明顯差異。本研究以40條逢機引子進行六株溶磷菌的基因組指紋分析，其中的19條逢機引子的反應效果不佳，因此，以另外的21條逢機引子進行實驗所得的電泳圖中之254條複制產物，結果顯示逢機複製多型性DNA分析的結果與限制指紋分析結果是相同的。經RAPDistance軟體計算各菌株之間的相似度，發現6株溶磷菌的相似度在0.256～0.863之間，顯示菌種間具有相當高的歧異度，其中Bacillus Ca-48與Ca-49及Pseudomonas Al-74與PSB2，二組內的相似度很高，分別為0.812及0.863，而SB+PSB與SB818的相似度只有0.314。若以相似度0.65為準，則6株溶磷菌可分成兩個群叢（cluster）-Bacillus Ca-48與Ca-49、Pseudomonas Al-74與PSB2，這兩群叢彼此的親緣差異很大，相似度約只有0.4。溶磷的大豆根瘤菌Phizobium SB+PSB、SB818與另兩群叢的親緣是相當疏遠的，其中SB+PSB與其他溶磷菌的親緣相差最遠。本研究證明RAPD指紋分析法可有效分析鑑別溶鈣磷菌之同菌種中菌系間之遺傳之差異。

關鍵字

溶鈣磷菌 ； 枯草菌屬 ； 擬球菌屬 ； 根瘤菌屬 ； 遺傳歧異度 ； 逢機複製多型性DNA分析 ； DNA指紋

並列摘要

The genetic diversity among six strains of calcium phosphate-solubilizing bacteria, including 2 strains of Bacillus pumillu Ca-48 and Ca-49, 2 strains of Pseudomonas cepacia Al-74 and PSB 2, and 2 strains of Rhizobium fredii SB+PSB and SB818, was determined by using the restriction fingerprint (RF) analysis and the random amplified polymorphic DNA (RAPD) techniques. In the restriction fingerprint analysis, two strains in the same species were closely related with the similarities among DNA samples, but had less similarity between Rhizobium SB+PSB and SB818. In the study, 40 arbitrary 10mer primers were employed for RAPD analysis, and 19 of 40 primers were not duplicated on the synthetic reaction. Twenty one primers produced 254 amplification products. The study showed similar results in the RF analysis and the RAPD analysis. In the RAPD istance software analysis, the coefficient similarity of six strains was ranged from 0.259 to 0.863 with the great genetic diversity. The coefficient similarity of Bacillus Ca-48 and Ca-49, Pseudomonas Ai-74 and PSB2 and Rhizobium SB+PSB and SB818 were 0.812, 0.863 and 0.314, respectively. The results showed that six strains divided into two clusters-Bacillus and Pseudomonas with the coefficient similarity above 0.65. It was about 0.4 similarity between these two clusters. Two Rhizobium, SB+PSB and SB818, had less similarity from other two genus. The molecular methods employed RAPD analysis should be reasonably representative the difference of genome between strains in same species of calcium phosphate-solubilizing becteria.

並列關鍵字

Calcium phosphate-solubilizing bacteria ； Bacillus ； Pseudomonas ； Rhizobium ； Genetic diversity ； Random amplified polymorphic DNA analysis (RAPD) ； DNA fingerprint