Down-Regulated Expression of RACK1 Gene by RNA Interference Enhances Drought Tolerance in Rice

doi:10.1016/S1672-6308(08)60051-7

摘要/Abstract

摘要： The receptor for activated C-kinase 1 (RACK1) is a highly conserved scaffold protein with versatile functions, and plays important roles in the regulation of plant growth and development. Transgenic rice plants, in which the expression of RACK1 gene was inhibited by RNA interference (RNAi), were studied to elucidate the possible functions of RACK1 in responses to drought stress in rice. Real-time PCR analysis showed that the expression of RACK1 in transgenic rice plants was inhibited by more than 50%. The tolerance to drought stress of the transgenic rice plants was higher as compared with the non-transgenic rice plants. The peroxidation of membrane and the production of malondialdehyde were significantly lower, and the superoxide dismutase activity in transgenic rice plants was significantly higher than those in non-trangenic rice plants. It is suggested that RACK1 negatively regulated the redox system-related tolerance to drought stress of rice plants.

关键词: Oryza sativa, receptor for activated C-kinase 1 gene, RNA interference, transgenic plant, drought stress, real-time quantitative RT-PCR, gene expression

Li Da-hong, Liu Hui, Yang Yan-li, Zhen Ping-ping, Liang Jian-sheng. Down-Regulated Expression of RACK1 Gene by RNA Interference Enhances Drought Tolerance in Rice[J]. RICE SCIENCE, 2009, 16(1): 14-20 .

参考文献

1 Fire A, Xu S, Montgomery M K, Kostas S A, Driver S E, Mello C C. Potent and specific genetic interference by double -stranded RNA in Caenorhabditis elegans. Nature, 1998, 391: 806–811.
2 Fire A. RNA-triggered gene silencing. Trends Genet, 1999, 15: 358–363.
3 Zhu N F, Zhang X N. RNAi and its application in plant genetic improvement. J Huazhong Agric Univ, 2004, 23(4): 472–477. (in Chinese with English abstract)
4 Ashique A M, Kharazia V, Yaka R, Phamluong K, Peterson A S, Ron D. Localization of the scaffolding protein RACK1 in the developing and adult mouse brain. Brain Res, 2006, 1069: 31–38.
5 Chen J G, Ullah H, Temple B, Liang J, Guo J, Alonso J M, Ecker J R, Jones A M. RACK1 mediates multiple hormone responsiveness and developmental processes in Arabidopsis. J Exp Bot, 2006, 57(11): 2697–2708.
6 Yarwood S J, Steele M R, Scotland G, Houslay M D, Bolger G B. The RACK1 signaling scaffold protein selectively interacts with the cAMP-specific phosphodiesterase PDE4D5 isoform. J Biol Chem, 1999, 274: 14909–14917.
7 Steele M R, McCahill A, Thompson D S, MacKenzie C, Isaacs N W, Houslay M D, Bolger G B. Identification of a surface on the beta-propeller protein RACK1 that interacts with the cAMP-specific phosphodiesterase PDE4D5. Cell Signaling, 2001, 13: 507–513.
8 Mamidipudi V, Zhang J, Lee K C, Cartwright C A. RACK1 regulates G1/S progression by suppressing Src kinase activity. Mol Cell Biol, 2004, 24: 6788–6798.
9 Patterson R L, Rossum D B, Barrow R K, Snyder S H. RACK1 binds to inositol 1,4,5-trisphosphate receptors and mediates Ca2+ release. Proc Natl Acad Sci USA, 2004, 101: 2328–2332.
10 Link A J, Eng J, Schieltz D M, Carmack E, Mize G J, Morris D R, Garvik B M, Yates J R. Direct analysis of protein complexes using mass spectrometry. Nat Biotechnol, 1999, 17: 676–682.
11 Ceci M, Gaviraghi C, Gorrini C, Sala L A, Offenhauser N, Marchisio P C, Biffo S. Release of eIF6 (p27BBP) from the 60S subunit allows 80S ribosome assembly. Nature, 2003, 426: 579–584.
12 Sengupta J, Nilsson J, Gursky R, Spahn C M, Nissen P, Frank J. Identification of the versatile scaffold protein RACK1 on the eukaryotic ribosome by cryo-EM. Nat Struc Mol Biol, 2004, 11: 957–962.
13 Chang I F, Szick-Miranda K, Pan S, Bailey-Serres J. Proteomic characterization of evolutionarily conserved and variable proteins of Arabidopsis cytosolic ribosomes. Plant Physiol, 2005, 137: 848–862.
14 Giavalisco P, Wilson D, Kreitler T, Lehrach H, Klose J, Gobom J, Fucini P. High heterogeneity within the ribosomal proteins of the Arabidopsis thaliana 80S ribosome. Plant Mol Biol, 2005, 57: 577–591.
15 Liu Q Q, Zhang J L, Wang Z Y, Hong M M, Gu M H. A highly efficient transformation system mediated by Agrobacterium tumefaciens in rice (Oryza sativa L.). Acta Phytophysiol Sin, 1998, 24(3): 259–271. (in Chinese with English abstract)
16 Jefferson R A, Kavangh T A, Bevan M W. GUS fusions: β-glucuronidase as a sensitive and versatile gene fusion marker in higher plants. EMBO J, 1987, 13: 3901–3907.
17 Murray M G, Thompson W F. Rapid isolation of high molecular weight plant DNA. Nucl Acids Res, 1980, 8: 4321–4325.
18 Sambrook J, Russell D W. Molecular Cloning: A Laboratory Manual. 3rd ed. Translated by Huang P T. Beijing: Science Press, 2002: 487–513. (in Chinese)
19 Chen J X. Plant Physiology Experimental Guidance. Guangzhou: South China University of Technology Press, 2002. (in Chinese)
20 Chuang C F, Meyerowitz E M. Specific and heritable genetic interference by double-stranded RNA in Arabidopsis thaliana. Proc Natl Acad Sci USA, 2000, 97(9): 4985–4990.

[1]	WANG Wei-xia, LAI Feng-xiang, LI Kai-long, FU Qiang. Selection of Reference Genes for Gene Expression Analysis in Nilaparvata lugens with Different Levels of Virulence on Rice by Quantitative Real-Time PCR[J]. RICE SCIENCE, 2014, 21(6): 305-311.
[2]	AI Li-ping, SHEN Ao, GAO Zhi-chao, LI Zheng-long, SUN Qiong-lin, LI Ying-ying, LUAN Wei-jiang. Reverse Genetic Analysis of Transcription Factor OsHox9, a Member of Homeobox Family, in Rice[J]. RICE SCIENCE, 2014, 21(6): 312-317.
[3]	ZHENG Xiao-guo, CHEN Liang, LOU Qiao-jun, XIA Hui, LI Ming-shou, LUO Li-jun. Changes in DNA Methylation Pattern at Two Seedling Stages in Water Saving and Drought-Resistant Rice Variety after Drought Stress Domestication[J]. RICE SCIENCE, 2014, 21(5): 262-270.
[4]	Md. Nasim ALI, Bhaswati GHOSH, Saikat GANTAIT, Somsubhra CHAKRABORTY. Selection of Rice Genotypes for Salinity Tolerance Through Morpho-Biochemical Assessment[J]. RICE SCIENCE, 2014, 21(5): 288-298.
[5]	Bidhan ROY, Sanjib BHADRA. Effects of Toxic Levels of Aluminium on Seedling Parameters of Rice under Hydroponic Culture[J]. RICE SCIENCE, 2014, 21(4): 217-223.
[6]	B SAILAJA, SR VOLETI, D SUBRAHMANYAM, N SARLA, VISHNU PRASANTH V, VP BHADANA，Satendra K. MANGRAUTHIA. Prediction and Expression Analysis of miRNAs Associated with Heat Stress in Oryza sativa[J]. RICE SCIENCE, 2014, 21(1): 3-12.
[7]	P. MANIMARAN, G. RAVI KUMAR, M. RAGHURAMI REDDY, S. JAIN, T. BHASKAR RAO, S. K. MANGRAUTHIA, R. M. SUNDARAM, S. RAVICHANDRAN, S. M. BALACHANDRAN. Infection of Early and Young Callus Tissues of Indica Rice BPT 5204 Enhances Regeneration and Transformation Efficiency[J]. RICE SCIENCE, 2013, 20(6): 415-426.
[8]	Zahra MARDANI, Babak RABIEI, Hossein SABOURI, Atefeh SABOURI. Mapping of QTLs for Germination Characteristics under Non-stress and Drought Stress in Rice[J]. RICE SCIENCE, 2013, 20(6): 391-399.
[9]	YU Shun-wu, LUO Hua-cheng, LI Jia-jia, YU Xin-qiao. Molecular Variation and Application from Aerospace Mutagenesis in Upland Rice Huhan 3 and Huhan 7[J]. RICE SCIENCE, 2013, 20(4): 249-258.
[10]	PAN Xiao-wu, LI Yong-chao, LI Xiao-xiang, LIU Wen-qiang, MING Jun, LU Ting-ting, TAN Jiang, SHENG Xin-nian. Differential Regulatory Mechanisms of CBF Regulon Between Nipponbare (Japonica) and 93-11 (Indica) During Cold Acclimation[J]. RICE SCIENCE, 2013, 20(3): 165-172.
[11]	LU Chao1, LIU Jian1, JIANG Jian-hua1, Caleb Manamik BRERIA1, TAN He-lin1, Masahiko ICHII2, HONG De-lin1. Development and Application of SCAR Markers for Discriminating Cytoplasmic Male Sterile Lines from Their Cognate Maintainer Lines in Indica Rice[J]. RICE SCIENCE, 2013, 20(3): 191-199.
[12]	GUO Si-bin1, 2, WEI Yu1, LI Xiao-qiong1, LIU Kai-qiang1, HUANG Feng-kuan2, 3, CHEN Cai-hong1, 2, GAO Guo-qing1, 2. Development and Identification of Introgression Lines from Cross of Oryza sativa and Oryza minuta[J]. RICE SCIENCE, 2013, 20(2): 95-102.
[13]	WANG Cai-hong, XU Qun, YU Ping, YUAN Xiao-ping, YU Han-yong, WANG Yi-ping, TANG Sheng-xiang, WEI Xing-hua. Comparison of Cheng’s Index- and SSR Marker-based Classification of Asian Cultivated Rice[J]. RICE SCIENCE, 2013, 20(2): 103-110.
[14]	FENG Bao-hua1, YANG Yang1, 2, SHI Yong-feng1, LIN Lu3, CHEN Jie1, WEI Yan-lin1, 2, Hei LEUNG4, WU Jian-li1. Genetic Analysis and Gene Mapping of Light Brown Spotted Leaf Mutant in Rice[J]. RICE SCIENCE, 2013, 20(1): 13-18.
[15]	ZHANG Qi-jun, LI Cong, LIU Shao-kui, LAI Dong, QI Qing-ming, LU Chuan-gen. Breeding and Identification of Insect-Resistant Rice by Transferring Two Insecticidal Genes, sbk and sck[J]. RICE SCIENCE, 2013, 20(1): 19-24.