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33
2025年5月15日 星期四
农业生物技术学报  2025, Vol. 33 Issue (5): 989-998    DOI: 10.3969/j.issn.1674-7968.2025.05.004
  研究论文与报告 本期目录 | 过刊浏览 | 高级检索 |
Zl-CRK5基因克隆及其在菰茎部膨大发育中的表达分析
郑宇捷, 周雅萍, 马莹然, 王晶, 汤近天, 夏文强, 张雅芬, 叶子弘, 崔海峰*
中国计量大学 生命科学学院/国家市场监督管理总局重点实验室(微生物计量检测与生物制品质量安全),杭州 310018
Cloning and Expression Analysis of the Zl-CRK5 Gene During the Swelling and Development of Stem in Zizania latifolia
ZHENG Yu-Jie, ZHOU Ya-Ping, MA Ying-Ran, WANG Jing, TANG Jin-Tian, XIA Wen-Qiang, ZHANG Ya-Fen, YE Zi-Hong, CUI Hai-Feng*
School of Life Sciences, China Jiliang University/Key Laboratory of Microbiological Metrology, Measurement & Bio-product Quality Security, State Administration for Market Regulation, Hangzhou 310018, China
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摘要 菰黑粉菌(Ustilago esculenta)侵染菰(Zizania latifolia)植株茎部能够诱导茎部膨大发育形成可食用的肉质茎,期间菰植株茎部生长素(auxin, IAA)极性运输相关基因表达显著下调。本研究克隆获得菰钙依赖蛋白激酶相关激酶5基因(calcium-dependent protein kinase related kinase 5, Zl-CRK5),gDNA序列全长4 224 bp,可编码453个氨基酸,具有CRK激酶家族典型的STKc_CAMK保守结构域,与短花稻(Oryza brachyantha)遗传距离较近。qPCR分析显示,Zl-CRK5在菰植株茎部与叶鞘表达显著高于须根与叶片(P<0.05),并受菰黑粉菌侵染诱导调节,其在侵染7 d后的菰植株茎部表达显著下调(P<0.05);菰起始膨大发育后,植株茎部Zl-CRK5表达显著下调(P<0.05),并在随后膨大发育进程中维持较低表达(P<0.05);IAA极性运输抑制剂萘草胺(N-1-Naphthylphthalamic acid, NPA)处理分析发现,菰植株茎部Zl-CRK5与IAA极性运输相关基因PIN1 (PIN-formed)中Zl-PIN1bZl-PIN1c呈协调表达,可能参与了茎部IAA极性运输调节。本研究探讨了Zl-CRK5参与IAA极性运输相关基因Zl-PIN1bZl-PIN1c表达调节的可能作用模式,有助于深入揭示菰黑粉菌侵染诱导菰植株茎部膨大发育的调节机制。
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郑宇捷
周雅萍
马莹然
王晶
汤近天
夏文强
张雅芬
叶子弘
崔海峰
关键词 菰黑粉菌钙依赖蛋白激酶相关激酶(CRK)茎部膨大发育基因表达    
Abstract:The swollen stem was induced by the infection of Ustilago esculenta in Zizania latifolia, following with the inhibition of polar growth of plants and the down-regulation of expression of auxin (IAA) polar transport related genes. In this research, the calcium-dependent protein kinase (CDPK) related kinase 5 (CRK5) gene of Z. latifolia was cloned with the full-length gDNA sequence of 4 224 bp, encoding 453 amino acids and containing the typical STKc_CAMK conformal domain of CRK kinase family. It was closely related to Oryza brachyantha. qPCR analysis showed that Zl-CRK5 was highly expressed in stem and leaf sheath of Z. latifolia (P<0.05), but lower in root and leaf (P<0.05). The expression of Zl-CRK5 in stems was significantly down-regulated (P<0.05) after 7 d infection of U. esculenta in plantlets of Z. latifolia. It was significantly down-regulated during the initial swolling stem of normal Z. latifolia (P<0.05), and contained lower expression level following the subsequent swollen and development (P<0.05), which was similar to the reported gene of Zl-PIN1b in Z. latifolia. Further analysis showed that the Zl-CRK5 gene and IAA polar transport-related genes PIN (PIN-formed) of Zl-PIN1b and Zl-PIN1c showed a coordinated expression relationship in the stems of Z. latifolia after treatment with IAA polar transport-inhibitor of N-1-Naphthylphthalamic (NPA). It was speculated that Zl-CRK5 might be involved in the regulation of polar growth of stems by activating the expression of IAA polar transport-related genes Zl-PIN1b and Zl-PIN1c. Further research would help to reveal the regulatory mechanism of stem expansion induced by infection of U. esculenta.
Key wordsZizania latifolia    Ustilago esculenta    CDPK related kinases (CRK)    Swelling development of stem    Gene expression
收稿日期: 2024-05-13     
中图分类号: S645.2;Q946
基金资助:国家自然科学基金(U20A2043); 浙江省重点研发计划(2021C02058); 浙江省“万人计划”科技创新领军人才项目(2019R52018); 浙江省基础公益研究计划(LGN21C150012)
通讯作者: *hfcui@cjlu.edu.cn   
引用本文:   
郑宇捷, 周雅萍, 马莹然, 王晶, 汤近天, 夏文强, 张雅芬, 叶子弘, 崔海峰. Zl-CRK5基因克隆及其在菰茎部膨大发育中的表达分析[J]. 农业生物技术学报, 2025, 33(5): 989-998.
ZHENG Yu-Jie, ZHOU Ya-Ping, MA Ying-Ran, WANG Jing, TANG Jin-Tian, XIA Wen-Qiang, ZHANG Ya-Fen, YE Zi-Hong, CUI Hai-Feng. Cloning and Expression Analysis of the Zl-CRK5 Gene During the Swelling and Development of Stem in Zizania latifolia. 农业生物技术学报, 2025, 33(5): 989-998.
链接本文:  
https://journal05.magtech.org.cn/Jwk_ny/CN/10.3969/j.issn.1674-7968.2025.05.004     或     https://journal05.magtech.org.cn/Jwk_ny/CN/Y2025/V33/I5/989
 
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