柑橘屬植物葉綠體trnL intron與trnL-trnF IGS區域及檸檬苦素葡萄糖轉移酶基因序列親緣性分析

Phylogenetic analysis using sequences of trnL intron and trnL-trnF IGS of chloroplast genome and limonoid glucosyltransferase gene in Taiwan citrus

吳思節(Sz-Jie Wu) ； 徐源泰(Yuan-Tay Shyu)

《台灣農學會報》 17卷2期 (2016/06) Pp. 158-176

https://doi.org/10.6730/JAAT.201606_17(2).0003

摘要

臺灣栽培多種遺傳多樣性豐富的柑橘類果樹，廣泛應用於鮮食和加工。由於種源接近，在許多加工應用上難以被分類鑑定，加上近年來因果物生產過剩，導致農產廢棄物的增加，間接帶來許多不必要的農業和環境成本。為求在加工上因應不同柑橘的特性，作有效率和務實的利用，本研究開發了新的分子標誌，以柑橘之葉綠體trnL intron 及trnL-trnF IGS 區域與檸檬苦素葡萄糖轉移酶limonoid glucosyltransferase (LGTase) 作為標誌，分析鑑定臺灣12 種常見柑橘的分子親緣性。由遺傳演化分析結果，柑橘使用葉綠體DNA 的trnL intron 與trnL-trnF區域，柚類、甜橙、葡萄柚和檸檬較難區別出來。但使用LGTase 為標誌，可將12 種柑橘屬植物分為3 大群，第一群包括椪柑、桶柑、葡萄柚、金桔與血橙，第二群包括蜂蜜柚、蜜柚、斗柚、文旦、石頭柚與雞蛋丁，第三群為檸檬。此結果更能鑑別原先葉綠體DNA 難以區分的葡萄柚與檸檬。同時，也可利用此段序列將柚類與其他柑橘屬植物鑑別之。本研究發現柑橘屬植物中之檸檬苦素葡萄醣轉移酶有兩種型態，此外可發現柑橘屬植物中兩種型態的LGTase 基因的遺傳演化是分別進行的，後續研究將進行檸檬苦素葡萄醣轉移酶基因存在與表現的研究，期利用分子技術進一步對此兩型基因進行探討。

關鍵字

柑橘屬 ； 分類 ； 分子標誌 ； 檸檬苦素葡萄醣轉移酶

並列摘要

Taiwan is rich with wide varieties and cultivars of citrus for fresh fruit and processing uses. In juice processing, bitterness is the main factor determining the juice quality. Owing to their close genetic relationships, the identification and classification of citrus fruits from the genetic perspective is rather inconsistent. To accurately identify citrus varieties for quality control of juice processing, this study used molecular markers chloroplast trnL intron trnL-trnF IGS region and limonoid glucosyltransferase gene (LGTase) to identify 12 citrus fruits commonly found in Taiwan. LGTase gene was found to be suitable for differentiating grapefruit, lemon and other citrus plants. LGT-GF/LGT-GR primers amplify a 1620 bp open reading frame containing a sequence of 40 amino acid UDP-Glycosyltransferase signatures. Results also revealed that trnL intron and trnL-trnF regions could not differentiate pomelo, sweet orange, grapefruit and lemon. However, when using LGTase as the molecular marker, 12 citrus fruits were grouped into three clades. This marker was found especially useful for distinguishing pomelo from other citrus fruits. Further study should focus on the expression of limonoid glucosyltransferase and examining their roles in bitterness development in citrus fruits.

並列關鍵字

Citrus spp. ； classification ； molecular marker ； limonoid glucosyltransferase