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斑茅是甘蔗近缘野生种之一,具有较强的抗逆性,研究并开发利用斑茅的强抗逆基因,对甘蔗育种具有重要意义。本研究以高粱Cu/Zn-SOD(登录号:XM 002445626.1)cDNA序列为探针,对甘蔗属EST数据库进行检索、比对、拼接,通过设计特异引物,PCR扩增和序列分析验证,克隆得到2个Cu/Zn-SOD(SaSOD-1a和SaSOD-1b,登录号:KJ001795和KJ001796)基因全序列,其中SaSOD-1a基因组序列全长2 473 bp,SaSOD-1b基因组序列全长2 228 bp,均有8个外显子,7个内含子,其cDNA序列长度均为692 bp,编码206个氨基酸。序列比较分析表明,SaSOD-1a和SaSOD-1b序列相似性为98.6%,有8个单核苷酸多态性位点,蛋白质相似性为99.0%,2个氨基酸变异位点。用推导出的氨基酸序列与其它植物做同源进化分析,与高粱、玉米、谷子等比较均表现出较高的保守性。研究结果可为进一步了解超氧化物歧化酶与斑茅耐旱性的关系奠定基础。
Umbelliferae is one of the wild relatives of sugarcane and has a strong resistance to stress. Studying and developing the strong resistance gene of Umbelliferae is of great significance to the breeding of sugarcane. In this study, the cDNA sequence of sorghum Cu / Zn-SOD (accession number: XM 002445626.1) was used as a probe to search, align, and splicing the EST database of sugarcane, and the specific primers were designed and verified by PCR amplification and sequence analysis. The full-length sequence of SaSOD-1a gene was 2 473 bp in length and 2 228 bp in full length in SaSOD-1b genomic sequence. The sequence of SaSOD-1a and SaSOD-1b was identical to SaSOD-1a and SaSOD-1b, There are 8 exons and 7 introns, both of which have a 692 bp cDNA sequence encoding 206 amino acids. Sequence analysis showed that the sequence similarity of SaSOD-1a and SaSOD-1b was 98.6%. There were 8 single nucleotide polymorphisms (SNPs) with 99.0% similarity and 2 amino acid mutations. Using the deduced amino acid sequence to do homologous evolution analysis with other plants, it showed high conservation compared with sorghum, maize and millet. The results may lay a foundation for further understanding of the relationship between superoxide dismutase and drought tolerance of