轉殖外源顫楊CAld5H基因赤桉之性狀分析

Characterization of Transgenic Eucalyptus camaldulensis Carrying the Populus tremuloides coniferyl aldehyde 5-hydroxylase gene

陳怡蒨(Yi-Chiann Chen) ； 蔡佳彬(Jia-Bin Tsai) ； 陳振榮(Zenn-Zong Chen) ； 鍾振德(Jeng-Der Chung)

《台灣林業科學》 37卷1期 (2022/03) Pp. 61-76

https://doi.org/10.7075/TJFS.202203_37(1).0004

摘要

木質素的含量與組成為製漿生產的關鍵，本研究使用農桿菌將順義式顫楊松柏醛5-羥化酶基因（coniferyl aldehyde 5-hydroxylase（PtCAld5H）gene）轉殖到赤桉，探討轉基因赤桉木材製漿性質之變化。本研究共獲取7個轉殖順義式PtCAld5H基因之赤桉轉殖系，並經由聚合酶連鎖反應及南方式雜交分析證實目標基因已經成功轉殖。H201～4與H206～7等6個品系木質素含量約在30%與對照組沒有顯著差異，而H205木質素含量32.4%，與對照組有顯著性差異。將木質素分成酸可溶與不可溶兩種形式，可發現H204～6等3個品系，其酸可溶木質素含量5.7～6.8%，極顯著（p ＜ 0.01）高於對照組之4.1%。赤桉木質素主要由紫丁香基（syringyl, S）與癒創木酚基（guaiacyl, G）木質素單元體聚合而成，其S/G比約為3，H204～6等3個品系之S/G分別為7.5, 7.5與7.3，其S木質素單元體含量顯著高於對照組。纖維素／木質素比率在轉基因品系中為2.0到2.2，而對照組為2.3。硫酸鹽法製漿分析，在活性鹼為17.0%製漿，H2O4和H2O6的得漿率提高了0.8～1.6%。

關鍵字

赤桉 ； 松柏醛5-羥化酶基因 ； 木質素單元體 ； 紫丁香基與癒創木酚基木質素單元體 ； 製漿

並列摘要

The lignin quantity and monomeric composition play important roles in wood and pulp production. In this study, the gene encoding coniferyl aldehyde 5-hydroxylase gene (CAld5H) from Populus tremuloides was transferred in a sense orientation into Eucalyptus camaldulensis by Agrobacterium-mediated transformation to assess the change in pulping properties in transgenic E. camaldulensis trees. Seven transgenic lines (H201~7) were obtained, and transformation was verified by a genomic polymerase chain reaction (PCR) and Southern hybridization. Lignin contents of transgenic lines H201~4 and H206~7 did not significantly differ compared to the control line, but that of H205 (32.4%) was significantly greater than the control (30.0%). The total lignin content was determined as the sum of acid-insoluble (AIL) and acid-soluble lignin (ASL) contents. ASL contents significantly increased in the transgenic lines H204~6 (5.7~6.8%), compared to the control line (4.1%) (p ＜ 0.01). Lignin in E. camaldulensis is polymerized from the syringyl (S) and guaiacyl (G) monolignols with an S/G ratio of ≈ 3. The S/G ratios of H204, H205, and H206 were 7.5, 7.6 and 7.3, respectively, and contents of the S monolignol were significantly higher than that of the control line. Cellulose/lignin ratios were 2.0~2.2 in the transgenic lines compared to 2.3 in the control. After Kraft pulping, results of pulp yields of H204 and H206 with 17.0% active alkali demonstrated yield increases of up to 0.8~1.6%.

並列關鍵字

Eucalyptus camaldulensis ； coniferyl aldehyde 5-hydroxylase gene ； lignin monomer ； syringyl and guaiacyl monolignols ； pulp yield