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热带作物学报 ›› 2020, Vol. 41 ›› Issue (5): 971-977.doi: 10.3969/j.issn.1000-2561.2020.05.017

• 生物技术与组织培养 • 上一篇    下一篇

低磷胁迫柱花草差异表达基因分析研究

文亦芾1,韩蓉蓉2,单贵莲1,史亮涛3,罗富成1,赵小雪1,曾黎琼4   

  1. 1. 云南农业大学动物科学技术学院,云南昆明 650201
    2. 西南大学动物科学技术学院/重庆市高校草食动物工程中心,重庆 400715
    3. 云南省农业科学院热区生态农业研究所,云南元谋 651300
    4. 云南省农业科学院生物技术与种质资源研究所,云南昆明 650223
  • 收稿日期:2019-06-03 修回日期:2019-09-07 出版日期:2020-05-25 发布日期:2020-06-15
  • 作者简介:文亦芾(1972—),女,博士,教授,研究方向:草种质资源与育种。E-mail: 1301814839@qq.com
  • 基金资助:
    国家自然科学基金(31560665);国家自然科学基金(31160481)

Analysis of Differentially Expressed Genes in Low Phosphorus Stress of Stylosanthes Guianensis

WEN Yifu1,HAN Rongrong2,SHAN Guilian1,SHI Liangtao3,LUO Fucheng1,ZHAO Xiaoxue1,ZENG Liqiong4   

  1. 1. College of Animal Science and Technology, Yunnan Agricultural University, Kunming, Yunnan 650201, China
    2. College of Animal Science and Technology, Southwest University / College Herbivore Engineering Center of Chongqing, Chongqing 400715, China
    3. Research Institute of Tropical Eco-agriculture Sciences, Yunnan Academy of Agricultural Sciences, Yuanmou, Yunnan 651300, China
    4. Biotechnology and Germplasm Resources Institute, Yunnan Academy of Agricultural Sciences, Kunming, Yunnan 650223, China
  • Received:2019-06-03 Revised:2019-09-07 Online:2020-05-25 Published:2020-06-15

摘要:

研究柱花草在低磷条件下生长的分子机理和反应机制。采用cDNA-SRAP分子标记技术分离柱花草茎和叶在低磷胁迫下相关基因的差异表达片段,并对其进行生物信息学分析。结果表明,8对引物组合共扩增出差异片段195条,其中抑制型表达片段56条,诱导型表达片段92条,上调表达型差异片段36条,下调表达型片段11条,大小均在50~1000 bp之间。二次扩增后将特异性条带回收后进行测序,通过BLAST比对,共比对出14条差异片段的同源序列,其中8个差异表达的核苷酸序列与已知功能基因(JCVI-FLLj-1K4未知mRNA、NBS-LRR型抗病蛋白、ATP合成酶、醛固酮还原酶、叶绿体RF2)有较高同源性,5个序列与假定基因或预测基因(UPF0051蛋白、蛋白酶启动子、SCEI结合酶、吲哚-3-丙酮酸盐单氧酶)同源性较高。本研究为筛选柱花草响应低磷胁迫的差异基因提供参考。

关键词: 柱花草, 磷胁迫, 同源性, cDNA-SRAP, 差异表达

Abstract:

The cDNA-SRAP molecular marker technique was used to isolate the differentially expressed genes of the stems and leaves of Stylosanthes under low phosphorus stress, and bioinformatics analysis was carried out to study the molecular mechanism and reaction mechanism of Stylosanthes growing under low phosphorus condition. A total of 195 differential fragments were amplified from 8 pairs of primer combinations. Among them, there were 56 inhibitory expression fragments, 92 inducible expression fragments, 36 upregulated expression fragments and 11 down-regulated expression fragments, and the sizes ranged from 50 to 1000 bp. After the second amplification, the specific bands were recovered and sequenced. Through blast alignment, the homologous sequences of 14 different fragments were compared, and the 8 differentially expressed nucleotide sequences had high homology with known functional genes (JCVI-FLLj-1K4 unknown mRNA, NBS-LRR type disease resistance protein, ATP synthase, aldosterone reductase, chloroplast RF2), five sequences were highly homologous to the putative or predicted genes (UPF0051 protein, protease promoter, SCEI binding enzyme, indole-3-pyruvate monooxygenase). This study has laid a foundation for screening differential genes in response to low phosphorus stress.

Key words: Stylosanthes, phosphorus deficiency, homolog, cDNA-SRAP, differential expression

中图分类号: 

  • S541