摘要

一般植物本身會產生小分子量熱休克蛋白質來克服熱逆境，而台灣黃菀屬植物大多分布於海拔400-3950公尺以上山區，其生育環境為向陽坡地，易受環境逆境的直接衝擊，為研究小分子量熱休克蛋白質基因家族之最佳素材。本研究之目的是研究黃菀屬植物內小分子量熱休克基因家族之遺傳變異程度、演化關係和生理功能。親緣分析結果，黃菀屬植物內第一型和葉綠體型的小分子量熱休克蛋白質在親緣樹狀圖上，皆顯示物種之間無顯著分群，彼此混雜，證明黃菀屬物種早在種化之前就經由基因複製（gene duplication）的機制產生基因多樣性。天擇測驗的結果，第一型小分子量熱休克蛋白質基因家族之Tajima’s D值呈現顯著負值、Ka/Ks值普遍小於1，顯示此基因受到強烈負向天擇作用，偏向同義置換來保留重要功能性基因，因此胺基酸序列呈現高度保守，其中C端胺基酸序列相似度高於N端區域。而葉綠體型小分子量熱休克蛋白質基因家族在天擇選汰測驗下，其Tajima’s D呈現負值、Ka/Ks值普遍小於1，顯示此基因受到負向天擇作用，但不顯著，其胺基酸序列變異程度偏低，並擁有甲硫氨酸區域（Met-rich）。在生理實驗中顯示黃菀屬植物的確在受到熱逆境時會產生第一型小分子量熱休克蛋白質的表現，顯示此基因是具有分子伴隨子（molecular chaperone）的保護功能，至於表現調控機制未來仍需深入探究。另外在黃菀屬植物內第一型小分子熱休克蛋白質基因家族內發現有7個偽基因（pseudogene）存在，雖然尚不清楚其真正的生物特性，希望在未來能提供學者進一步研究。

Abstract

In plants, small heat-shock proteins (sHSPs) are the predominant stress proteins in response heat stress. Senecio (Asteraceae) plants grow at habitats of elevations of 400-3950m with highly fluctuating temperatures that they are good samples to study sHSPs. The main purpose of this project is to study the genetic diversity, evolution and gene functions in small heat shock protein gene family in the Senecio. Phylogeny tree reveals that the gene duplications of sHSPs were prior to the speciation events in the genus Senecio. Results of neutral selection tests, the significant negative values of Tajima’s D test and Ka/Ks are less than 1 and the cytosolic class I sHSPs are under the negative selection. The cytosolic class I sHSPs prefer have synomous changes to nonsynomous changes indicates the gene function is important and unchangeable. They are more conserved in the C-terminal domain than in the N-terminal domain in the cytosolic class I sHSPs amino sequences. The negative values of Tajima’s D test and Ka/Ks are less than 1 reveal that the CP class sHSPs is also under negative selection. There is Met-rich region in the conserved of amino acid sequence in the CP class sHSPs. Under the heat treatment, cytosolic class I sHSPs was produced proteins as molecular chaperone to protect Senecio. The regulation and function of sHSPs genes are needed to study in the future. Finally, 7 pseudogenes were found in the cytosolic class I sHSP gene family. Although much effort has been devoted to understanding the function of pseudogenes, their biological roles remain largely unknown. Hope that they will provide information for the future research.

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