| 曹华宁, 刘博*, 刘太国, 高利, 陈万权*.小麦条锈菌hsp70基因的克隆及热胁迫下的表达特征分析[J].植物保护,2015,41(3):19-24. |
| 小麦条锈菌hsp70基因的克隆及热胁迫下的表达特征分析 |
| Cloning of a heat shock protein gene hsp70 of Puccinia striiformis f. sp. tritici and its expression in response to high-temperature stress |
| 投稿时间：2014-04-21 修订日期：2014-12-11 |
| DOI： |
| 中文关键词: 小麦条锈菌 温度敏感类型 热激蛋白70 克隆 荧光定量PCR |
| 英文关键词:wheat stripe rust temperature sensitivity HSP70 clone qRT-PCR |
| 基金项目:国家重点基础研究发展计划(“973”计划)(2013CB127704); 国家自然科学基金项目（31100110）; 国家科技支撑计划（2012BAS19B04）; 国家小麦产业技术体系岗位专家专项经费（CARS-03-04B） |
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| 中文摘要: |
| 为明确热激蛋白70在小麦条锈菌对高温适应性中的作用, 采用RACE和PCR技术扩增得到小麦条锈菌hsp70的基因全长, 并利用实时荧光定量PCR技术对热胁迫下不同温度敏感类型小麦条锈菌中hsp70的表达特征进行分析。克隆得到的小麦条锈菌hsp70基因组序列全长为2 817 bp, 包含7个内含子, cDNA序列全长为2 267 bp, 其中开放阅读框为1 917 bp, 编码639个氨基酸, 分子量为66.7 ku, 等电点为5.15。编码蛋白含3个保守的HSP70氨基酸序列。qRT-PCR试验结果表明, 28 ℃热激后, hsp70的转录水平随热激时间的延长而略有增加, 2 h达到最高值。不同温度敏感类型菌株在热激2 h后hsp70的相对表达量具有显著差异。低敏感类型菌株hsp70平均表达量为未经处理对照的7.12倍, 而高敏感类型菌株hsp70平均表达量仅为对照的1.63倍。 |
| 英文摘要: |
| To determine the response of Puccinia striiformis f. sp. tritici heat shock protein 70 to high-temperature stress, the Pst hsp70 gene was cloned via RACE and PCR, and mRNA expression of hsp70 was determined in representative isolates with low-and high-temperature sensitivity during the heat shock process. The whole genomic DNA sequence of hsp70 was 2 817 bp, including 7 introns, and the cDNA sequence was 2 267 bp, including an opening reading frame of 1 917 nucleotides. The ORF encodes a HSP70 protein of 639 amino acids, which had a theoretical pI/MW of 5.15/66.7 ku and contained three HSP70 family signature domains. Following heat shock at 28 ℃, expression of hsp70 transcripts was up-regulated in the isolates with high-and low-temperature sensitivity, and reached the maximum level 2 hours after treatment. However, the average expression level was increased by 7.12-fold upon heat shock for 2 h in the isolates with low-temperature sensitivity, but only 1.63-fold in the isolates with high-temperature sensitivity. |
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