Subcellular Localization Analysis of Rice Protein Kinase Gene <i>OsCIPK</i>5 and Generation of Its RNAi Transgenic Plants

XIONG Gui-hong; LIU Xiao-juan; YANG Liang; WU Zu-jian

doi:10.19303/j.issn.1008-0384.2017.04.001

Volume 32 Issue 4

May 2017

Turn off MathJax

Article Contents

Article Navigation > Fujian Journal of Agricultural Sciences > 2017 > 32(4): 353-358

XIONG Gui-hong, LIU Xiao-juan, YANG Liang, WU Zu-jian. Subcellular Localization Analysis of Rice Protein Kinase Gene OsCIPK5 and Generation of Its RNAi Transgenic Plants[J]. Fujian Journal of Agricultural Sciences, 2017, 32(4): 353-358. doi: 10.19303/j.issn.1008-0384.2017.04.001

Citation:

XIONG Gui-hong, LIU Xiao-juan, YANG Liang, WU Zu-jian. Subcellular Localization Analysis of Rice Protein Kinase Gene OsCIPK5 and Generation of Its RNAi Transgenic Plants[J]. Fujian Journal of Agricultural Sciences, 2017, 32(4): 353-358. doi: 10.19303/j.issn.1008-0384.2017.04.001

Citation:

PDF( 2797 KB)

Subcellular Localization Analysis of Rice Protein Kinase Gene OsCIPK5 and Generation of Its RNAi Transgenic Plants

doi: 10.19303/j.issn.1008-0384.2017.04.001

Institute of Plant Virology/Key Laboratory of Plant Virology of Fujian Province, Fujian Agriculture and Forestry University, Fuzhou, Fujian 350002, China

Received Date: 2016-12-20
Rev Recd Date: 2017-03-07
Publish Date: 2017-04-28

Abstract

Abstract

Plant CBL-interacting protein kinases (CIPKs) play an important role in stress signaling transduction and enhancing plant stress tolerance. However, the functions of OsCIPK5 in the rice (Oryza sativa) have not been studied. In order to study the functions of OsCIPK5, RT-PCR method was used to clone OsCIPK5 gene from rice leaf (Oryza sativa Japonica Group cultivar Nipponbare).The bioinformatics analysis showed that OsCIPK5 contained two functional domains, N-terminal kinase activation loop domain and C-terminal NAF domain, which were the same as other CIPK that had been reported.OsCIPK5 was highly homologous in amino acid with other five plant species, especially with Oryza brachyantha, which was about 94%. By Real-time PCR approach the expression pattern of OsCIPK5 was detected in the rice seedlings responses to potassium. The result showed that the expression of OsCIPK5 in root was up-regulated under low potassium treatment, but not in leaf. The recombinant expression plasmid pEarleyGate101-OsCIPK5 contained yellow fluorescent protein (YFP) was transformed into GV3101 and then the positive clones were infiltrated into the epidermal leaves of Nicotiana benthamiana. The results revealed that the OsCIPK5 protein localized on the nucleus, cytolemma and cytoplasm. The RNAi vector containing OsCIPK5 specific gene was constructed and transferred into EHA105 (Agrobacterium tumefaciens), then rice transformation was conducted through agrobacterium-mediated system. T₀ RNAi transgenic seedlings of OsCIPK5 were obtained and 26 plants were identified to be positive.The expression of OsCIPK5 in transgenic plants was obviously reduced compared with that in wild type by Real-time PCR analysis.These results including bioinformatics analysis, expression pattern, subcellular localization analysis and transgenic rice may be useful in study on the important role of OsCIPK5 in the process of plant growth.
- OsCIPK5,
- bioinformatic analysis,
- expression pattern,
- subcellular localization,
- RNAi

FullText(HTML)

References(28)

References

[1]	FUJITA M, FUJITA Y, NOUTOSHI Y, et al. Crosstalk between abiotic and biotic stress responses: a current view from the points of convergence in the stress signaling networks [J]. Current Opinion in Plant Biology, 2006, 9(4):436-442. doi: 10.1016/j.pbi.2006.05.014
[2]	XIONG L M, SCHUMAKER K S, ZHU J K. Cell Signaling during Cold, Drought, and Salt Stress [J]. Plant Cell, 2001, 14 (S1):165-183.
[3]	DEFALCO T A, BENDER K W, SNEDDEN W A. Breaking the code: Ca sensors in plant signaling [J]. Biochemical Journal, 2010, 425(1):27-40. doi: 10.1042/BJ20091147
[4]	YANG T B, POOVAIAH B W. Calcium/calmodulin-mediated signal network in plants [J]. Trends in Plant Science, 2003, 8(10):505-512. doi: 10.1016/j.tplants.2003.09.004
[5]	HARMON A C, GRIBSKOV M, HARPER J F. CDPKs-a kinase for every Ca²⁺ signal? [J]. Trends in Plant Science, 2000, 5(4):154-159. doi: 10.1016/S1360-1385(00)01577-6
[6]	LUAN S, KUDLA J, RODRIGUEZ-CONCEPCION M, et al. Calmodulins and Calcineurin B-like Proteins: Calcium Sensors for Specific Signal Response Coupling in Plants [J]. Plant Cell, 2002, 14(1):S389-S400.
[7]	SHI J R, KIM K N, RITZ O. Novel protein kinases associated with calcineurin B-like calcium sensors in Arabidopsis [J]. Plant Cell, 1999, 11(12):2393-2405. doi: 10.1105/tpc.11.12.2393
[8]	LIU J K, ISHITANI M, HALFTER U, et al. The Arabidopsis thaliana SOS2 gene encodes a protein kinase that is required for salt tolerance [J]. Proceedings of the National Academy of Sciences of the United States of America, 2000, 97(7):3730-3734. doi: 10.1073/pnas.97.7.3730
[9]	GUO Y, HALFTER U, ISHITANL M, et al. Molecular characterization of functional domains in the protein kinase SOS2 that is required for plant salt tolerance [J]. Plant Cell, 2001, 13(6):1383-1400. doi: 10.1105/tpc.13.6.1383
[10]	HALFTER U, ISHITANI M, ZHU J K. The Arabidopsis SOS2 Protein Kinase Physically Interacts with and Is Activated by the Calcium-Binding Protein SOS3 [J]. Proceedings of the National Academy of Sciences of the United States of America, 2000, 97(7):3735-3740. doi: 10.1073/pnas.97.7.3735
[11]	BATISTIC O, KUDLA J. Integration and channeling of calcium signaling through the CBL calcium sensor/CIPK protein kinase network[J]. Planta, 2004, 219(6):915-924. doi: 10.1007/s00425-004-1333-3
[12]	WEINL S, KUDLA J. The CBL CIPK Ca²⁺ -decoding signaling network: function and perspectives [J]. New Phytologist, 2009, 184(3):517-528. doi: 10.1111/nph.2009.184.issue-3
[13]	YU Y, XIA X, YIN W, et al. Comparative genomic analysis of CIPK gene family in Arabidopsis and Populus[J]. Plant Growth Regulation, 2007, 52(2):101-110. doi: 10.1007/s10725-007-9165-3
[14]	TAI F J, YUAN Z H, LI S P, et al. ZmCIPK8, a CBL-interacting protein kinase, regulates maize response to drought stress [J]. Plant Cell, Tissue and Organ Culture (PCTOC), 2016, 124(3):459-469. doi: 10.1007/s11240-015-0906-0
[15]	凌秋平, 曾巧英, 胡斐, 等.甘蔗CIPK23基因克隆及对低钾、干旱、盐胁迫的响应[J].分子植物育种, 2015, (6):1329-1335. http://www.cnki.com.cn/Article/CJFDTOTAL-FZZW201506029.htm
[16]	LV F L, ZHANG H C, XIA X L, et al. Expression profiling and functional characterization of a CBL-interacting protein kinase gene from Populus euphratica[J]. Plant Cell Reports, 2014, 33(5):807. doi: 10.1007/s00299-013-1557-4
[17]	刘实, 赵小英, 唐冬英, 等.拟南芥CIPK1基因的功能初步分析[J].西北植物学报, 2007, 27(6):1091-1095. http://www.cnki.com.cn/Article/CJFDTOTAL-DNYX200706003.htm
[18]	CHEN L, WANG Q Q, ZHOU L, et al. Arabidopsis CBL-interacting protein kinase (CIPK6) is involved in plant response to salt/osmotic stress and ABA [J]. Molecular Biology Reports, 2013, 40(8):4759-4767. doi: 10.1007/s11033-013-2572-9
[19]	XU J, LI H D, CHEN L Q, et al. A Protein Kinase, Interacting with Two Calcineurin B-like Proteins, Regulates K + Transporter AKT1 in Arabidopsis[J]. Cell, 2006, 125(7):1347-1360. doi: 10.1016/j.cell.2006.06.011
[20]	YONG H C, PANDEY G K, GRANT J J, et al. Two calcineurin B-like calcium sensors, interacting with protein kinase CIPK23, regulate leaf transpiration and root potassium uptake in Arabidopsis [J]. Plant Journal, 2007, 52(2):223-239. doi: 10.1111/tpj.2007.52.issue-2
[21]	XIANG Y, HUANG Y, XIONG L. Characterization of stress-responsive CIPK genes in rice for stress tolerance improvement[J].Plant Physiology, 2007, 144(3):1416-1428. doi: 10.1104/pp.107.101295
[22]	LI J, LONG Y, QI G N, et al. The OsAKT1 channel is critical for K⁺ uptake in rice roots and is modulated by the rice CBL1-CIPK23 complex [J]. Plant Cell, 2014, 26(8):3387-3402. doi: 10.1105/tpc.114.123455
[23]	XIONG G H, LIU X J, QIU P, et al. Rice grassy stunt virus p5 interacts with two protein components of the plant-specific CBL-CIPK Ca²⁺ signaling network of rice[J].Virus Gene, 2017:1-8.
[24]	KOLUKISAOGLU U, WEINL S, BLAZEVIC D, et al. Calcium Sensors and Their Interacting Protein Kinases: Genomics of the Arabidopsis and Rice CBL-CIPK Signaling Networks[J]. Plant Physiology, 2004, 134(1):43-58. doi: 10.1104/pp.103.033068
[25]	MIKI D, ITOH R, SHIMAMOTO K. RNA Silencing of Single and Multiple Members in a Gene Family of Rice[J]. Plant Physiology, 2005, 138(4):1903-1913. doi: 10.1104/pp.105.063933
[26]	WANG P, LIANG Z F, ZENG J, et al. Generation of tobacco lines with widely different reduction in nicotine levels via RNA silencing approaches [J]. Journal of Biosciences, 2008, 33(2):177-184. doi: 10.1007/s12038-008-0035-6
[27]	SHIMIZU T, YOSHII M, WEI T Y, et al. Silencing by RNAi of the gene for Pns12, a viroplasm matrix protein of Rice dwarf virus, results in strong resistance of transgenic rice plants to the virus[J]. Plant Biotechnology Journal, 2009, 7(1):24-32. doi: 10.1111/pbi.2009.7.issue-1
[28]	陈析丰, 顾志敏, 刘峰, 等.生物与非生物胁迫下水稻CIPK基因的鉴定分析[J].中国水稻科学, 2010, 24(6):567-574. http://www.cnki.com.cn/Article/CJFDTOTAL-ZGSK201006003.htm