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水稻磷酸核酮糖激酶基因OsPRK克隆、亚细胞定位与诱导表达分析
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引用本文:陈林,吕静,李晨羊,周书行,娄永根.水稻磷酸核酮糖激酶基因OsPRK克隆、亚细胞定位与诱导表达分析.植物保护学报,2020,47(2):283-291
DOI:10.13802/j.cnki.zwbhxb.2020.2019113
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作者单位E-mail
陈林 浙江大学昆虫科学研究所, 杭州 310058  
吕静 浙江大学昆虫科学研究所, 杭州 310058  
李晨羊 浙江大学生物技术研究所, 杭州 310058  
周书行 浙江大学昆虫科学研究所, 杭州 310058  
娄永根 浙江大学昆虫科学研究所, 杭州 310058 yglou@zju.edu.cn 
中文摘要:为探究水稻磷酸核酮糖激酶基因OsPRK在水稻诱导抗虫反应中的功能,以水稻秀水110为材料克隆OsPRK基因的全长,通过生物信息学软件分析其序列特征,并应用实时荧光定量PCR技术分析OsPRK基因在水稻不同组织中的分布情况以及在虫害诱导、激素和机械损伤处理水稻中的表达特征。结果显示,水稻OsPRK基因序列全长为1 212 bp,编码403个氨基酸,分子量为44.86 kD,具有1个磷酸核酮糖激酶保守结构域。OsPRK蛋白亚细胞定位结果显示其定位于叶绿体。OsPRK基因在水稻中的表达具有组织特异性,其在内叶、外叶、内叶鞘、外叶鞘和根系这5个组织中相对于内参基因ACTIN的表达量分别为35.83、20.53、6.25、3.21和0.03。与对照相比,二化螟Chilo suppressalis为害能够强烈抑制水稻茎秆中OsPRK基因的表达;褐飞虱Nilaparvata lugens怀卵雌成虫为害1.5、24 h、白背飞虱Sogatella furcifera怀卵雌成虫为害1、8、24 h以及机械损伤处理3、6、24 h均能显著诱导水稻茎秆中OsPRK基因的表达;而OsPRK基因的表达量在茉莉酸处理6、12 h时以及水杨酸处理0.5、1.5 h时被显著抑制,在茉莉酸处理48 h和水杨酸处理24 h时被显著诱导。表明OsPRK基因可能参与了水稻对害虫的诱导防御反应。
中文关键词:水稻  磷酸核酮糖激酶  亚细胞定位  防御反应  诱导表达分析
 
Cloning, subcellular localization and expression patterns of the phosphoribulokinase gene OsPRK in the rice plant
Author NameAffiliationE-mail
CHEN Lin Institute of Insect Sciences, Zhejiang University, Hangzhou 310058, Zhejiang Province, China  
Lü Jing Institute of Insect Sciences, Zhejiang University, Hangzhou 310058, Zhejiang Province, China  
LI Chenyang Institute of Biotechnology, Zhejiang University, Hangzhou 310058, Zhejiang Province, China  
ZHOU Shuxing Institute of Insect Sciences, Zhejiang University, Hangzhou 310058, Zhejiang Province, China  
LOU Yonggen Institute of Insect Sciences, Zhejiang University, Hangzhou 310058, Zhejiang Province, China yglou@zju.edu.cn 
Abstract:To investigate the role of phosphoribulokinase gene OsPRK in the herbivore-induced defenses of rice plants, the full-length cDNA sequence of OsPRK from rice variety XS110 was cloned and its characteristics were analyzed with bioinformatics tools. The expression patterns of OsPRK in different tissues of rice and in leaf sheaths of rice in response to different treatments were detected by quantitative real-time PCR (qPCR). The results showed that the full-length cDNA sequence of OsPRK was 1 212 bp, encoding 403 amino acid residues with a molecular weight of 44.86 kD, which possesses a conserved phosphoribulokinase (PRK) domain. Subcellular localization showed that OsPRK was localized in the chloroplast. The qPCR results revealed that OsPRK was expressed in a tissue-specific manner:transcript levels of OsPRK in inner leaves, outer leaves, inner leaf sheaths, outer leaf sheaths and roots of rice plants, relative to the reference gene ACTIN, were 35.83, 20.53, 6.25, 3.21 and 0.03, respectively. The relative transcript level of OsPRK was strongly repressed after infestation of striped stem borer Chilo suppressalis, whereas brown planthopper Nilaparvata lugens infestation for 1.5 h and 24 h, whitebacked planthopper Sogatella furcifera infestation for 1, 8 and 24 h, and mechanical wounding for 3, 6 and 24 h induced the expression of OsPRK in rice sheaths. The expression levels of OsPRK were suppressed after jasmonic acid (JA) treatment for 6 h and 12 h, and salicylic acid (SA) treatment for 0.5 h and 1.5 h, whereas OsPRK expression was induced after JA treatment for 48 h and SA treatment for 24 h. These results indicated that OsPRK might be involved in defense responses in rice against insect pests.
keywords:rice  phosphoribulokinase  subcellular localization  plant defense  induced expression pattern
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