金邊也門鐵花葉性狀差異表達基因的cDNA-AFLP分析

劉和平 何業華 林劍波 張俊麗

摘要：【目的】篩選金邊也門鐵花葉性狀相關差異表達基因，為深入探索也門鐵葉色形成的分子調控機制打下基礎。【方法】基于cDNA擴增片段長度多態性分析技術（cDNA-AFLP），利用64對EcoR I和Mse I選擇性擴增引物組合分析金邊也門鐵葉片綠色與黃色部分組織cDNA，經篩選、回收、克隆、測序和BLAST比對分析獲得差異表達基因。【結果】利用64對引物組合共擴增出1726條可分辨的條帶，并獲得58條差異表達的轉錄衍生片段（TDFs），其中12條TDFs為特異性表達。12條特異性TDFs中，5條TDFs具有同源序列，另外7條TDFs無同源序列，為未知功能基因。在葉片黃化部位特異表達同源性為96%的TDFs序列M8E3B-2和M8E3B-3與大麗花花葉病毒Holland包涵體蛋白基因、紫莖澤蘭明脈病毒基因、洋丁香黃化卷葉病毒基因等花椰菜花葉病毒科病毒基因具有較高的同源性。【結論】M8E3B-2和M8E3B-3兩條TDFs序列或來源于一種花椰菜花葉病毒科病毒基因，且有可能參與了金邊也門鐵花葉葉色性狀的形成過程。

關鍵詞： 也門鐵;花葉性狀;cDNA-AFLP;差異表達基因

中圖分類號： S687.9? ? ? ? ? ? ? ? ? ? ? ? 文獻標志碼：A 文章編號：2095-1191（2018）11-2129-07

cDNA-AFLP analysis on differentially expressed genes related to floral leaf characters of Dracaena fragrans cv. lindenii

LIU He-ping1，2，3， HE Ye-hua2*， LIN Jian-bo1， ZHANG Jun-li1

（1Yangjiang Polytechnic，Yangjiang， Guangdong 529500，China; 2College of Horticulture， South China Agricultural University， Guangzhou? 510642， China; 3Institute of Fruit Science in Maoming， Maoming， Guangdong? 525000， China）

Abstract：【Objective】Differentially expressed genes related to floral leaf characters of Dracaena fragrans cv. lindenii were selected to provide reference for further? research into the molecular regulation mechanisms in the formation of leaf color of D. fragrans（L.） Ker-Gaw. 【Method】Based on the cDNA-amplified fragment length polymorphism（cDNA-AFLP）， 64 pairs of EcoR I and Mse I selective amplification primer groups were used to investigate the cDNA of green and yellow parts of D. fragrans cv. lindenii leaves using cDNA-AFLP analysis. Then the differentially expressed genes were obtained via screening， recovery， cloning， sequencing and Blast alignment. 【Result】Sixty-four pairs of primer combinations amplified 1726 distinguishable bands. Of which， 58 differentially expressed transcript-derived fragments（TDFs） were identified. Out of the 58 TDFs， 12 specifically expressed TDFs were obtained. Among the twelve TDFs， five shared homologous sequence， and the other seven had no homologous sequence and were unknown functional genes. Two TDFs sequence（M8E3B-2 and M8E3B-3） whose specific expression homology in yellow part of leaves was 96%? shared high homology with cauliflower mosaic virus genes such as Dahlia mosaic virus Holland inclusion body，? Eupatorium vein clearing virus gene and Cestrum yellow leaf curling virus. 【Conclusion】These results suggest that M8E3B-2 and M8E3B-3 TDFs sequences may both derive from one kind of cauliflower mosaic virus gene， and may be involved in the formation process of floral leaf color characters in D. fragrans cv. lindenii.

Key words： Dracaena fragrans（L.） Ker-Gaw.; floral leaf character; cDNA-AFLP; differentially expressed gene

0 引言

【研究意義】也門鐵[Dracaena fragrans（L.） Ker-Gaw.]又稱香龍血樹，是龍舌蘭科（Agavaceae）龍血樹屬（Dracaena）常綠喬木，原產于幾內亞和埃塞俄比亞，我國引種栽培后已發展成為重要的盆栽觀葉植物，市場上主要有金心也門鐵（D. fragrans cv. massangeana）、金邊也門鐵（D. fragrans cv. lindenii）和普通也門鐵（D. fragrans cv.）3個品種。其中，金心也門鐵和金邊也門鐵是普通也門鐵的葉色嵌合體變異種，其葉片上的黃色和綠色條紋相間排列，色彩明亮、觀賞周期長，具有較高的觀賞價值和特殊園林用途，已成為園林造景的優選物種（何業華等，2008）。……