| 陈博雯，肖玉菲，李军集，张 烨，覃子海，张晓宁，刘海龙.EuCAD 基因小片段 RNAi对烟草木质素合成的影响[J].广东农业科学,2020,47(6):1-7 |
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EuCAD 基因小片段 RNAi对烟草木质素合成的影响 |
| Effect of RNA Interference Triggered by Small Fragment of EuCAD Gene on Lignin Synthesis in Tobacco |
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| DOI：10.16768/j.issn.1004-874X.2020.06.001 |
| 中文关键词: 肉桂酰乙醇脱氢酶 RNA 干扰片段 木质素合成 烟草转化 |
| 英文关键词: cinnamoyl alcohol dehydrogenase RNA interference fragment lignin synthesis tobacco transformation |
| 基金项目:广西林业科技项目（桂林科字〔2016〕第 11 号）；广西科技基地和人才专项（桂科 AD18281086） |
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| 中文摘要: |
| 【目的】深入研究尾叶桉（Eucalyptus urophylla）肉桂酰乙醇脱氢酶（CAD）基因序列中的关键片段并利用其进行木质素合成调控。【方法】采用生物信息学工具对 EuCAD 基因进行序列分析，根据编码蛋白保守结构域的分布情况进行筛选，获得一段 44 bp 关键序列用于构建 RNA 干扰载体并转化烟草，获得转基因植株并对基因表达水平、木质素含量等表型进行检测。【结果】获得 28 个转基因株系，转基因植株正常生长发育，与野生型无异，随机挑选 3 个株系进行检测，发现 CAD 基因表达水平均较野生型显著降低，以 R1 株系降幅最大，降至野生型的 12.51%。对 R1 株系进一步检测发现，其茎部直径与野生型无异，但木质部厚度较野生型显著降低，其中第 4 节处降低 11.75%，第 6 节处降低 10.06%，茎部整体木质素含量较野生型降低 15.26%。表明该 RNAi 片段显著抑制烟草中 CAD 基因表达并影响烟草木质素合成。【结论】通过序列分析筛选出的 EuCAD 基因小片段，可通过 RNA 干扰显著影响烟草木质素合成，为尾叶桉木质素合成调控研究提供了技术参考。 |
| 英文摘要: |
| 【Objective】The study aimed to explore the key sequence of cinnamoyl alcohol dehydrogenase（CAD）gene of Eucalyptus urophylla and use them to regulate lignin synthesis.【Method】Sequence analysis of EuCAD gene was carried out by bioinformatics method. According to the distribution of conservative domain of coding protein, a 44 bp key sequence was obtained to construct RNA interference vector and transformed to tobacco. Transgenic plants were obtained and phenotypes such as gene expression and lignin content were detected.【Result】Twenty-eight transgenic lines were obtained, which grew normally as same as the wild type. Three transgenic lines were randomly selected for detection. The results showed that the expression level of CAD gene was significantly lower than that of wild type. The largest decline was observed in line R1 which decreased to 12.51% of the wild type. It was found that the stem diameter of line R1 was the same as that of the wild type, but the xylem thickness was significantly lower than that of the wild type, which decreased by 11.75% at the 4th node and 10.06% at the 6th node. The lignin content of the stem in line R1 decreased by 15.26% compared with that of the wild type. The results showed that the RNAi fragment significantly inhibited the expression of CAD gene in tobacco and affected the lignin synthesis of tobacco.【Conclusion】The small fragment of EuCAD gene screened by sequence analysis can significantly affect lignin synthesis in tobacco by triggering RNA interference, which provides a technical reference for the study on the regulation of lignin synthesis in E. urophylla. |
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