作物学报 ›› 2014, Vol. 40 ›› Issue (10): 1756-1766.
刘荣榜1,2,陈明2,*,郭萌萌2,司青林2,高世庆3,徐兆师2,李连城2,马有志2,尹钧1,*
LIU Rong-Bang1,2,CHEN Ming2,*,GUO Meng-Meng2,SI Qing-Lin2,GAO Shi-Qing3,XU Zhao-Shi2,LI Lian-Cheng2,MA You-Zhi2,YIN Jun1,*
摘要:
用拟南芥AVP1为诱饵采用膜蛋白酵母双杂交系统筛选拟南芥cDNA文库, 获得一个与AVP1互作的小GTP结合蛋白AtRAB。酵母互作试验表明, AVP1和AtRAB存在互作; 双分子荧光互补试验(BiFC)证明, AVP1和AtRAB能在质膜和细胞核上发生相互作用。野生型拟南芥(WT)、AtRAB和AVP1的拟南芥突变体rab和avp1在高盐胁迫条件下, 随着NaCl浓度的加大, 突变体rab和avp1的表型变化相似, 相对于WT都表现为根长缩短, 侧根数减少; 在低磷处理条件下, 随着磷浓度的逐渐降低, 2个突变体的表型相似, 相对于WT同样表现为根长缩短, 总根面积减少, 侧根数减少; 在相同磷浓度条件下, 突变体rab对胁迫的反应比avp1强; 在低钾处理条件下, 随着钾浓度的降低, 2个突变体的表型与在低磷胁迫处理条件下的表型相似。结果说明, AVP1和AtRAB可以在细胞膜和细胞核上相互作用, AtRAB能够影响植物对离子的吸收, AVP1和AtRAB的突变体在高盐、低磷和低钾等胁迫条件下表型变化相似, 证明了2个基因都正向调控植物对高盐、低磷和低钾胁迫的反应, 它们属于同一信号途径。
[1]Rea P A, Poole R J. Vacuolar H+-translocating pyrophosphatase. Ann Rev Plant Physiol, 1993, 44: 157–180[2]Nakanishi Y, Matsuda N, Aizawa K, Kashiyama T, Yamamoto K, Mimura T, Ikeda M, Maeshima M. Molecular cloning and sequencing of the cDNA for vacuolar H+-pyrophosphatase from Chara coralline. Biochim Biophys Acta, 1999, 1418: 245–250[3]Baykov A A, Bakuleva N P, Rea P A. Steady-state kinetics of substrate hydrolysis by vacuolar H+-pyrophosphatase. A simple three-state model. Eur J Biochem, 1993, 217: 755–762[4]Long A R, Williams L E, Nelson S J, Hall J L. Localization of membrane pyrophosphatase activity in Ricinus communis seedlings. J Plant Physiol, 1995, 146: 629–638[5]Robinson D G, Haschke H P, Hinz G, Hoh B, Maeshima M, Marty F. Immunological detection of tonoplast polypeptides in the plasma membrane of pea cotyledons. Planta, 1996, 198: 95–103[6]Robinson D G, Hoppenrath M, Oberbeck K, Luykx P, Ratajczak R. Localization of pyrophosphatase and V-ATPase in Chlamydomonas reinhardtii. Bot Acta, 1998, 111: 108–122[7]Lerchl J, Geigenberger P, Stitt M, Sonnewald U. Impaired photoassimilate partitioning caused by phloem-specific removal of pyrophosphate can be complemented by a phloem-specific cytosolic yeast-derived invertase in transgenic plants. Plant Cell, 1995, 7: 259–270[8]Jose R, Castineira P, Hernandez A, Drake R, Serrano. A plant proton-pumping inorganic pyrophosphatase functionally complements the vacuolar ATPase transport activity and confers bafilomycin resistance in yeast. Biochem J, 2011, 437: 269–278[9]Blumwald E. Tonopast vesicles for the study of ion transport in plant vacuoles. Plant, 1987, 69: 731–734[10]Hedrich R, Schroeder J I. The physiology of ion channels and electrogenic pumps in higher plants. Ann Rev Plant Physiol, 1989, 40: 539–569[11]Guo S L, Yin H B, Zhang X, Zhao F Y, Li P H, Chen S H, Zhao Y X, Zhang H. Molecular cloning and characterization of a vacuolar H+-pyrophosphatase gene, SsVP, from the halophyte Suaeda salsa and its overexpression increases salt and drought tolerance of Arabidopsis. Plant Mol Biol, 2006, 60: 41–50[12]Sun Q H, Gao F, Zhao L, Li K P, Zhang J R. Identification of a new 130 bp cis-acting element in the TsVP1 promoter involved in the salt stress response from Thellungiella halophila. BMC Plant Biol, 2010, 10: 1471–2290[13]Liu L, Wang Y, Wang N, Dong Y Y, Fan X D, Liu X M, Yang J, Li H Y. Cloning of a Vacuolar H+-pyrophosphatase gene from the halophyte Suaeda corniculata whose heterogonous overexpression improves salt, saline-alkali and drought tolerance in Arabidopsis. J Integr Plant Biol, 2011, 53: 731–742[14]Bhaskaran S, Savithramma D L. Co-expression of Pennisetum glaucum vacuolar Na+/H+ antiporter and Arabidopsis H+-pyrophosphatase enhances salt tolerance in transgenic tomato. J Exp Bot, 2011, 62: 5561–5570[15]Park S H, Li J S, Pittman J K, Berkowitz G A, Yang H B, Undurraga S, Morris J, Hirschi K D, Gaxiola R A. Up-regulation of a H+-pyrophosphatase (H+-PPase) as a strategy to engineer drought-resistant crop plants. PNAS, 2005, 102: 18830–18835[16]Zhang H, Shen G X, Kuppu S, Gaxiola R, Payton P. Creating drought- and salt-tolerant cotton by overexpressing a vacuolar pyrophosphatase gene. Plant Signal Behav, 2011, 6:6: 861–863[17]Pasapula V, Shen G X, Kuppu S, Paez-Valencia J, Mendoza M, Hou P, Chen J, Qiu X Y, Zhu L F, Zhang X L, Auld D, Blumwald E, Zhang H, Gaxiola R, Payton P. Expression of an Arabidopsis vacuolar H+-pyrophosphatase gene (AVP1) in cotton improves drought-and salt tolerance and increases fibre yield in the field conditions. Plant Biotechnol J, 2011, 9: 88–99[18]Dong Q L, Liu D D, An X H, Hu D G, Yao Y X, Hao Y J. MdVHP1 encodes an apple vacuolar H+-PPase and enhances stress tolerance in transgenic apple callus and tomato. J Plant Physiol, 2011, 168: 2124–2133[19]Zhang J, Li J Q, Wang X C, Chen J. OVP1, a Vacuolar H+-translocating inorganic pyrophosphatase (V-PPase), overexpression improved rice cold tolerance. Plant Physiology and Biochemistry, 2011, 49: 33–38[20]Kabala K, Janicka-Russak M, Klobus G. Different responses of tonoplast proton pumps in cucumber roots to cadmium and copper. J Plant Physiol, 2010, 167: 1328–1335[21]Migocka M, Papierniak A, Kosatka E, Klobus G. Comparative study of the active cadmium efflux systems operating at the plasma membrane and tonoplast of cucumber root cells. J Exp Bot, 2011, 62: 4903–4916[22]Khoudi H, Maatar Y, Gouiaa S, Masmoudi K. Transgenic tobacco plants expressing ectopically wheat H+-pyrophosphatase (H+-PPase) gene TaVP1 show enhanced accumulation and tolerance to cadmium. J Plant Physiol, 2012, 169: 98–103[23]Li J S, Yang H B, Peer W A, Richter G, Blakeslee J, Bandyopadhyay A, Titapiwantakun B, Undurraga S, Khodakovskaya M, Richards E L, Krizek B, Murphy A S, Gilroy S, Gaxiola R. Arabidopsis H+-PPase AVP1 regulates Auxin-mediated organ development. Science, 2005, 310: 121–125[24]Yao Y X, Dong Q L, You C X, Zhai H, Hao Y J. Expression analysis and functional characterization of apple MdVHP1 gene reveals its involvement in Na+, malate and soluble sugar accumulation. Plant Physiol Biochem, 2011, 49: 1201–1208[25]Krebs M, Beyhl D, Gorlich E, Al-Rasheid K A S, Marten I, Stierhof Y D, Hedrich R, Schumacher K. Arabidopsis V-ATPase activity at the tonoplast is required for efficient nutrient storage but not for sodium accumulation. Proc Natl Acad Sci USA, 2010, 107: 3251–3256[26]Yang H B, Knapp J, Koirala P, Rajagopal D, Peer W A, Silbart L K, Murphy A, Gaxiola R A. Enhanced phosphorus nutrition in monocots and dicots over-expressing a phosphorus-responsive type I H+-pyrophosphatase. Plant Biotechnol J, 2007, 5: 735–745[27]Stagljar I, Korostensky C, Johnsson N, te Heesen S. A genetic system based on split-ubiquitin for the analysis of interactions between membrane proteins in vivo. Proc Natl Acad Sci USA, 1998, 95: 5187–5192[28]李敏, 杨双, 阮燕晔, 樊金娟, 张立军. 拟南芥T DNA插入突变体atsuc3的PCR鉴定. 植物生理学通讯, 2006, 42: 91–94Li M, Yang S, Ruan Y Y, Fan J J, Zhang L J. Identification of atsuc3 with T-DNA Insertion by PCR. Plant Physiol Commun, 2006, 42: 91–94 (in Chinese)[29]Waizenegger I, Lukowitz W, Assaad F, Schwarz H, Jurgens G, Mayer U. The Arabidopsis KNOLLE and KEULE genes interact to promote vesicle fusion during cytokinesis. Curr Biol, 2000, 10: 1371–1374[30]Geldner N, Friml J, Stierhof Y D, Jurgens G, Palme K. Auxin transport inhibitors block PIN1 cycling and vesicle trafficking. Nature, 2001, 413: 425–428[31]Surpin M, Raikhel N. Traffic jams affect plant development and signal transduction. Nat Rev Mol Cell Biol, 2004, 5: 100–109[32]Molendijk A J, Ruperti B, Palme K. Small GTPases in vesicle trafficking. Curr Opin Plant Biol, 2004, 7: 694–700[33]Vernoud V, Horton A C, Yang Z B, Nielsen E. Analysis of the small GTPase gene superfamily of Arabidopsis. Plant Physiol, 2003, 131: 1191–1208[34]Peng J L, Ilarslan H, Wurtele E S, Bassham D C. AtRabD2b and AtRabD2c have overlapping functions in pollen development and pollen tube growth. BMC Plant Biol, 2011, 11: 25[35]Mazel A, Leshem Y, Tiwari B S, Levine A. induction of salt and osmotic stress tolerance by overexpression of an intracellular vesicle trafficking protein AtRab7 (AtRabG3e). Plant Physiol, 2004, 134: 118–128[36]White P J, Marshall J, Smith J A C. Substrate kinetics of the tonoplast H+-translocating inorganic pyrophosphatase and its activation by free Mg2+. Plant Physiol, 1990, 93: 1063–1070[37]Parvanova D, Ivanov S, Konstantinova T, Karanovc E, Atanassov A, Tsvetkov T, Alexieva V, Djilianov D. Transgenic tobacco plants accumulating osmolytes show reduced oxidative damage under freezing stress. Plant Physiol Biochem, 2004, 42: 57–63[38]Boguski M S, McCormick F. Proteins regulating Ras and its relatives. Nature, 1993, 366: 643–654[39]Takai Y, Sasaki T, Matozaki T: Small GTP-binding proteins. Physiol Rev, 2001, 81: 153–208[40]Preuss M L, Serna J, Falbel T G, Bednarek S Y, Nielsen E. The Arabidopsis Rab GTPase RabA4b localizes to the tips of growing root hair cells. Plant Cell, 2004, 16: 1589–1603 |
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