台灣原住民部落土芭樂之分子多樣性與系統演化分析   全文下載

Molecular Identification and Analysis of Psidium guajava L. from Indigenous Tribes of Taiwan

Psidium guajava L.為生長於熱帶及亞熱帶常綠灌木果樹，近年來的研究報告中指出P. guajava具抑菌、止腹痛、抗發炎及退熱等療效，多被原住民用來治療腹痛、腸痛等。本實驗利用分子標誌方式作親緣鑑定，如土芭樂葉rDNA基因序列之18S及internal transcribe spacer（ITS）區域，以及葉綠體DNA基因上的trnL intron與trnL-trnF intergenic spacer（IGS），對幾個不同地區的土芭樂進行分析比對，但這些區域之保守性高，對於親源鑑定無法得到明確的分析結果。故利用RAPD的方法，使用隨機引子進行對土芭樂DNA進行逢機擴增，經電泳分析結果顯示，OPB 17 、OPG 6 、OPY 15及OPY 18四條引子擴增出共82個性狀特徵條帶，32個樣品總共記錄1,476個條帶，根據cluster analysis繪製之樹狀圖，明確地將經濟品種及土芭樂區分為兩群，紅心土芭樂均被歸在G1-II，推測比G1-I與G1-II的分群為紅心土芭樂與非紅心土芭樂的分群。"

Psidium guajava L. is a perennial fruit tree in subtropical and tropical areas. In Taiwan, P. guajava has been used as anthro-pharmacological plants by aboriginal people to treat acute diarrhea, cough and intestinal spasmodic diseases. The classification and functional identification of P. guajava remains unsolved these days. In this study, molecular markers 18S rRNA, internal transcribe spacer (ITS) region of ribosomal DNA, trnL intron and trnL-trnF intergenic spacer (IGS) of chloroplast DNA (cpDNA), and random amplified polymorphic DNA (RAPD) were used for the molecular identification of 18 P. guajava samples from different indigenous tribes, 2 from non-indigenous tribe, and 12 commercial cultivars from markets in Taiwan. Molecular methods restricted fragment length polymorphism (RFLP) and denatured gradient gel electrophoresis (DGGE) are found timeconsuming and less efficient as compared to RAPD, thus are not suitable for samples of high homology. In this study, 18S rDNA, ITS and cpDNA trnL intron and trnL-trnK intergenic spacer were also tested molecular marker; however, results analyzed by molecular algorithm UPGMA, Neighbor-Joining, Parsimony or Maximum likelihood showed no discriminations (data not shown). On the other side, ten 10-mer oligonucleotide primers were used in RAPD to amplify the specific genes from 32 guava samples. Four primers, OPB 17, OPG 6, OPY 15 and OPY 18, were able to direct the amplification and yielded a total of 82 polymorphic RAPD patterns. Thirty-two genotypes on the dendrogram were identified and were divided into two major groups, the uncultivated and commercial cultivars. Based on the cluster analysis, the red-flesh Psidium samples that were believed to have high medical function were grouped independently. The results suggest that RAPD is useful for the discrimination of uncultivated, cultivars and potential Psidium of high economy.