以分子標誌檢視綠竹及莿竹屬物種之遺傳相似度

Abstract

本研究以逢機擴增多型性DNA (RAPD)與簡單重複序列(ISSR)兩種分子標誌分析綠竹(Bambusa oldhamii)及莿竹屬(Bambusa spp.)竹種的遺傳相關性。試驗分為兩部分，自台灣南部及北部綠竹產區所收集的96個綠竹材料，先依地區分開檢視區內的遺傳相似性，並進行RAPD引子的篩選，再將材料合併檢視，在73個能擴增清楚、並具再現性條帶之引子，都只得到單型性條帶，而少數差異係來自混雜的烏腳綠竹(Bambusa edulis)樣本及來源為橫山、三灣的開花綠竹，其餘所採集之綠竹樣品間沒有遺傳差異。莿竹屬內遺傳相關性以 23個RAPD引子在莿竹屬 15參試樣品中，共產生 182個多型性marker，平均每引子可產生7.91 個。材料間相似度介於0.11-0.92，以綠竹(橫山)與綠竹相似度最高，簕竹與烏葉竹相似度最低。UPGMA之親緣關係樹狀圖中，簕竹遺傳距離最遠，其餘材料分成兩群，第一群包括綠竹、烏腳綠竹及麻竹三個竹種，第二群中有六個莿竹屬物種及種內之兩栽培種加上’潭子樣本’共九個材料； ISSR使用 13個引子，在莿竹屬15個參試樣品共產生135 個多型性marker，平均每引子可產生10.38 個，樣品間相似度介於0.15-0.88之間，以綠竹(橫山)與綠竹相似度最高，簕竹與硬頭黃竹相似度最低。UPGMA親緣關係樹狀圖中，與RAPD結果同樣分成兩群，但綠竹在第一群中與烏腳綠竹及麻竹相似性較低，在RAPD分析結果中，綠竹與烏腳綠竹關係較近。硬頭黃竹在第二群中與其他竹種都較遠，其餘分群結果則與RAPD結果相同。Mantel’s test 顯示兩種分子標誌所得的相似度矩陣有極高相關性。’潭子樣本’所稱’四季竹’係農民所取，經分子標誌分析，其分類認定上與’竹變’有極高相似性。

Both random amplified polymorphic DNA (RAPD) and inter simple sequence repeat markers (ISSR) assays were applied to detect the molecular variation among green bamboo (Bambusa oldhamii) accessions grown in cultivation and among Bambusa spp. collection from a park. A total of 96 B. oldhamii accessions were tested with 73 decamer oligonucleotide primers and only monomorphic fragments were generated. Accessions from northern and from southern Taiwan were assessed separately and assessed together in different tests and only primers with distinct and reproducible bands were selected. No polymorphism was observed among B. oldhamii accessions.Different band was only amplified from B. edulis and flowered B. oldhamii. One hundred and eighty two polymorphic RAPD markers were amphified among 15 Bambusa collections by 23 primers with an average of 7.91 markers per primer. The genetic similarity ranges between 0.11-0.92 with the highest similarity between B. oldhamii and flowered B. oldhamii and lowest between B. dissimulator and B. utilis. In UPGMA dendrogram, all Bambusa collections were grouped in 2 clusters, i.e. B. oldhamii, B. edulis and Dendrocalamus latiflorus in one group, six Bambusa species and two varieties and ‘accession of Tanzih’ in anther group. The same 15 Bambusa clones were tested by 13 microsatellite-primers and 135 polymorphic bands were recorded. The genetic similarity ranges between 0.15-0.88 and the lowest similarity was between B. dissimulator and B. fecunda. The UPGMA dendrogram separates 15 clones similarly as in RAPD analysis into 2 clusters with minor differentiation. Mantel’s test showed very high correlation between RAPD and ISSR similarity matrix. The molecular marker assay indicated that ‘accession of Tanzih’ bamoo belong to B. becheyana var. pubescens.