Isolation and expression analysis of cellulose synthase 3 (Ces3) genes from sugarcane (Saccharum officinarum L.)

IF 5.2 2区 农林科学 Q1 AGRICULTURE, MULTIDISCIPLINARY
Cheng-Mei Huang, Kai-Chao Wu, Krishan K. Verma, Hai-Bin Luo, Zhi-Nian Deng, Guo-Ying Shi, Hui-Qing Cao, Xing-Jian Wu, Li-Ping Ye, Yuan-Wen Wei, Lin Xu
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Abstract

Plant cellulose synthase is one of the important glycosyltransferases, which catalyzes the synthesis of the paracrystalline of H-bonded-β-(1,4)-glucose chains. This study isolated the cellulose synthase 3 (Ces3) sequence from sugarcane (Saccharum officinarum L.) leaves. SoCes3 (GenBank accession No. MG324347) has a full-length cDNA sequence of 3625 bp. It contains an open reading frame (3225 bp), encoding 1074 amino acids with a molecular weight of about 120.89 kDa and isoelectric point of 6.26. SoCes3 protein showed high activity with other plant cellulose synthases. The recombinant protein contains plant cellulose synthase (Ces) protein conservative domains. In subcellular localization experiments, the fusion protein of SoCes3 with green fluorescent protein (GFP) was specifically localized in the cell membrane. The gene expression of SoCes3 was found in the leaf, leaf sheath, and internodes of the sugarcane stem. The highest expression level was found in the internode, especially with the highest expression level in the 5th internode and lowest in the leaves, and the gene expression level of SoCes3 was upregulated by PP333 and not in gibberellic acid-treated plants. It was conducted in tobacco plants to understand the biotechnological potential of SoCes3. The contents of cellulose and lignin were increased in SoCes3-overexpressing tobacco. Transcriptomic analysis showed that the transgenic tobacco induced different genes associated with different biological regulatory processes. Differentially expressed genes (DEGs) mediated plant hormone signal transduction, starch and sucrose metabolism signaling pathways were widely induced and mostly upregulated. The transcription levels in SoCes3-overexpressing transgenic lines were higher than wild-type.

Graphical Abstract

甘蔗(Saccharum officinarum L.)纤维素合成酶 3(Ces3)基因的分离和表达分析
植物纤维素合成酶是重要的糖基转移酶之一,可催化H键-β-(1,4)-葡萄糖链旁晶的合成。本研究从甘蔗(Saccharum officinarum L.)叶片中分离出纤维素合成酶 3(Ces3)序列。SoCes3(GenBank登录号:MG324347)的全长cDNA序列为3625 bp。它包含一个开放阅读框(3225 bp),编码 1074 个氨基酸,分子量约为 120.89 kDa,等电点为 6.26。与其他植物纤维素合成酶相比,SoCes3 蛋白具有较高的活性。重组蛋白含有植物纤维素合成酶(Ces)蛋白保守结构域。在亚细胞定位实验中,SoCes3 与绿色荧光蛋白(GFP)的融合蛋白特异性地定位在细胞膜上。SoCes3基因在甘蔗茎的叶片、叶鞘和节间均有表达。SoCes3的基因表达水平受PP333的调控而不受赤霉素的调控。为了了解 SoCes3 的生物技术潜力,研究人员对烟草植株进行了研究。在SoCes3过表达的烟草中,纤维素和木质素的含量都有所增加。转录组分析表明,转基因烟草诱导了与不同生物调控过程相关的不同基因。介导植物激素信号转导、淀粉和蔗糖代谢信号通路的差异表达基因(DEGs)被广泛诱导且大部分上调。SoCes3高表达转基因品系的转录水平高于野生型。
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来源期刊
Chemical and Biological Technologies in Agriculture
Chemical and Biological Technologies in Agriculture Biochemistry, Genetics and Molecular Biology-Biotechnology
CiteScore
6.80
自引率
3.00%
发文量
83
审稿时长
15 weeks
期刊介绍: Chemical and Biological Technologies in Agriculture is an international, interdisciplinary, peer-reviewed forum for the advancement and application to all fields of agriculture of modern chemical, biochemical and molecular technologies. The scope of this journal includes chemical and biochemical processes aimed to increase sustainable agricultural and food production, the evaluation of quality and origin of raw primary products and their transformation into foods and chemicals, as well as environmental monitoring and remediation. Of special interest are the effects of chemical and biochemical technologies, also at the nano and supramolecular scale, on the relationships between soil, plants, microorganisms and their environment, with the help of modern bioinformatics. Another special focus is the use of modern bioorganic and biological chemistry to develop new technologies for plant nutrition and bio-stimulation, advancement of biorefineries from biomasses, safe and traceable food products, carbon storage in soil and plants and restoration of contaminated soils to agriculture. This journal presents the first opportunity to bring together researchers from a wide number of disciplines within the agricultural chemical and biological sciences, from both industry and academia. The principle aim of Chemical and Biological Technologies in Agriculture is to allow the exchange of the most advanced chemical and biochemical knowledge to develop technologies which address one of the most pressing challenges of our times - sustaining a growing world population. Chemical and Biological Technologies in Agriculture publishes original research articles, short letters and invited reviews. Articles from scientists in industry, academia as well as private research institutes, non-governmental and environmental organizations are encouraged.
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