CAJ | 학술논문

SUMO化修饰与植物抗病防御、信号转导和耐旱等有着直接的关系。本文以抑制差减杂交（sup—pression subtractive hybridization．SSH）技术获得SUMO的EST为信息探针，对棉花EST数据库进行同源搜索和电子克隆，获得了全长为396bp的SUMO基因编码区cDNA全长，我们将该基因命名为GhSUMO，推测该基因编码95个氨基酸。分别以抗黄萎病陆地棉品种豫棉21号的cDNA和DNA为模板，对该基因进行了PCR扩增验证，测序结果表明，GhSUMO基因序列与电子克隆序列一致，且没有内含子。蛋白序列分析表明，该蛋白具有保守泛素结构域和C端双Gly的断裂／连接位点，以及保守的疏水表面和Ulpl—Smt3互作位点。系统进化分析表明，该蛋白与蓖麻的同源序列表现了最高的相似性，与其它双子叶植物同源序列次之，而与单子叶植物的相似性较低。棉苗接菌后实时定量PCR结果显示，该基因表达量在接黄萎病菌的量48h后明显上调，96h达到未接菌对照的5倍以上。GhsSUMO基因可受黄萎病菌诱导表达，表明该基因在陆地棉抗黄萎病的机制中可能发挥重要作用。
SUMO화수식여식물항병방어、신호전도화내한등유착직접적관계。본문이억제차감잡교（sup—pression subtractive hybridization．SSH）기술획득SUMO적EST위신식탐침，대면화EST수거고진행동원수색화전자극륭，획득료전장위396bp적SUMO기인편마구cDNA전장，아문장해기인명명위GhSUMO，추측해기인편마95개안기산。분별이항황위병륙지면품충예면21호적cDNA화DNA위모판，대해기인진행료PCR확증험증，측서결과표명，GhSUMO기인서렬여전자극륭서렬일치，차몰유내함자。단백서렬분석표명，해단백구유보수범소결구역화C단쌍Gly적단렬／련접위점，이급보수적소수표면화Ulpl—Smt3호작위점。계통진화분석표명，해단백여비마적동원서렬표현료최고적상사성，여기타쌍자협식물동원서렬차지，이여단자협식물적상사성교저。면묘접균후실시정량PCR결과현시，해기인표체량재접황위병균적량48h후명현상조，96h체도미접균대조적5배이상。GhsSUMO기인가수황위병균유도표체，표명해기인재륙지면항황위병적궤제중가능발휘중요작용。
As an important post-translational modification, sumoylation SUMO participates in a number of biological processes, including plant disease resistance, signal transduction and drought tolerance et al. Using an EST of a small ubiquitin-related modifier gene as the query probe to blast cotton EST database, a SUMO gene, with a complete open reading fragment （ORF） 396 bp encoding a protein of 95 amino acids, was successfully obtained in silico cloning and was named GhSUMO. The GhSUMO gene was verified by PCR amplifications using the cDNA and DNA templates from the root of upland cotton Yumian 21, which was of resistance to Verticillium wilt, and sequencing results showed that the GhSUMO had no intron. Sequence analysis showed that the protein has a conserved ubiqitin domain, the C-terminal double Gly fracture/Connection site, a conservative hydrophobic surface and an Ulpl-Smt3 interaction sites. Phylogenetic analysis showed the GhSUMO was highly identical with its ortholog from Ricinus communis, and the next most similar orthologs coming from other dicotyledons, but far away from monocotyledons. Quantitative PCR analysis showed that GhSUMO expression was significantly uprelated at the 48 h past innoculation with Verticillium dahliae, and the fold of relative gene expression at 96 h reached, more than 5 times. The induced expression of GhSUMO suggested that the gene may play important roles in the mechanism ofresitance to Verticillium wilt of upland cotton.