利用裂殖酵母体系，拟南芥盐胁迫应答相关基因的克隆、鉴定及转基因分析
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摘要
盐胁迫是植物生长发育的主要限制因子之一。植物对盐胁迫的耐受性的分子生物学研究不仅对于培育耐盐农作物品种具有重要的应用价值，而且也是植物基因表达调控及信号转导等基础理论研究的重要内容。近年来，随着分子生物学技术的不断发展，对植物盐胁迫应答的分子机理研究不断深入，特别是对拟南芥在盐胁迫条件下的离子平衡和植物耐盐反应的调节途径的研究，取得了突破性的进展。尽管如此，现有数据仍不足以形成完整的盐胁迫应答的调控通路图，因此，需要继续寻找植物盐胁迫应答相关基因，以不断填补现有通路图中的缺空或未知组分。
高等植物的一些盐胁迫应答反应的分子机制被认为与酵母细胞具有相似性，许多植物基因在酵母中的表达能提高酵母细胞对盐胁迫的耐受性，或能够对酵母盐胁迫应答突变体产生功能互补，这表明真核生物的一些耐盐相关基因具有功能保守性。由于酵母是单细胞真核生物，其遗传研究远比高等植物简单，因此，利用酵母系统来分离鉴定功能保守的高等植物基因被认为是一条行之有效的快捷途径。
尽管拟南芥不属于盐生植物，但由于大量实验数据证明所有高等植物都含有耐盐基因，故拟南芥被认为也是研究耐盐机理的模式植物。本工作通过拟南芥cDNA文库在裂殖酵母细胞中的功能性表达，分离了拟南芥的编码富含甘氨酸蛋白、叶绿素a/b结合蛋白以及一些未知功能的cDNA克隆，重点对富含甘氨酸蛋白和叶绿素a/b结合蛋白这两个基因对酵母耐盐性的影响、盐胁迫和ABA对其表达的影响、在拟南芥中的组织特异性表达、转基因植株耐盐性等进行了分析。结
果表明它们可能是盐胁迫应答相关基因。
Environmental factors that impose water-deficit stress, such as salinity, drought and temperature extremes, place major limits on plant productivity, and soil salinity is a major abiotic stress in plant agriculture worldwide. Might salt stress affect plant metabolism in a complex way that disrupts homeostasis in water potential and ion distribution. To achieve salt tolerance, the damage must be prevented or alleviated. At the same time, homeostatic conditions must be re-established and plant growth must be resumed. By far, many ion/proton transporters that involves in K+/Na+ uptake and transport, establishment of proton gradients, removal and sequestration of toxic ions from the cytoplasm and organelles have been isolated. Meanwhile, many proteins have also been isolated related to osmoprotectants, reactive oxygen scavengers, stress-induced proteins, heat shock proteins, membrane fluidity, water status, signaling components, transcription control and growth regulators. However, studies of stress adaptation h
ave suggested a network of multiple signaling pathways in the course of physical adaptations and interactive molecular and cellular changes that begin after the onset of stress in plant .The most important task in the next few years remains to identify pathway components to establish their function by genetic approaches.
Despite soil salinity is the major constraint for plant agriculture, few functional determinants of salt adaptation have been identified and the significance of their involvement characterized. The fission yeast Schizosaccharomyces pombe is a single cell eukaryotic organism. Taking advantage of the genetic simplicity of S. pombe and the functional conservation of some salt tolerance-related proteins, We used this organism as a simple system to identify Arabidopsis thaliana salt tolerance proteins by screening for cDNA clones that confer salt resistance when overexpressed. An Arabidopsis thaliana cDNA library was constructed in pREPSN vector under the control of the inducible nmt-1 promoter and transformed into S. pombe. Expression of the A. thaliana sequences were induced and clones showing strong salt stress were
identified and analysed. By this 'cross-phylogenetic' screen, We have isolated some salt tolerance-related proteins such as glycine-rich protein, chlorophyll a/b-binding protein and some unknown protein. Overexpression of these proteins in the yeast cells afforded significantly increase in salt tolerance. Northern-blot analysis revealed that these genes were induced by NaCl and ABA. In addition, differentially regulated in various tissues. Transgenic Arabidopsis can develop better than wild type in salt tolerance. Taken together, these results undoubtedly suggest that these proteins are involved in the salt stress response in Arabidopsis. The AtGRP9 gene and the AtCab gene are first reported to salt stress response in Arabidopsis.
高等植物的一些盐胁迫应答反应的分子机制被认为与酵母细胞具有相似性，许多植物基因在酵母中的表达能提高酵母细胞对盐胁迫的耐受性，或能够对酵母盐胁迫应答突变体产生功能互补，这表明真核生物的一些耐盐相关基因具有功能保守性。由于酵母是单细胞真核生物，其遗传研究远比高等植物简单，因此，利用酵母系统来分离鉴定功能保守的高等植物基因被认为是一条行之有效的快捷途径。
尽管拟南芥不属于盐生植物，但由于大量实验数据证明所有高等植物都含有耐盐基因，故拟南芥被认为也是研究耐盐机理的模式植物。本工作通过拟南芥cDNA文库在裂殖酵母细胞中的功能性表达，分离了拟南芥的编码富含甘氨酸蛋白、叶绿素a/b结合蛋白以及一些未知功能的cDNA克隆，重点对富含甘氨酸蛋白和叶绿素a/b结合蛋白这两个基因对酵母耐盐性的影响、盐胁迫和ABA对其表达的影响、在拟南芥中的组织特异性表达、转基因植株耐盐性等进行了分析。结
果表明它们可能是盐胁迫应答相关基因。
Environmental factors that impose water-deficit stress, such as salinity, drought and temperature extremes, place major limits on plant productivity, and soil salinity is a major abiotic stress in plant agriculture worldwide. Might salt stress affect plant metabolism in a complex way that disrupts homeostasis in water potential and ion distribution. To achieve salt tolerance, the damage must be prevented or alleviated. At the same time, homeostatic conditions must be re-established and plant growth must be resumed. By far, many ion/proton transporters that involves in K+/Na+ uptake and transport, establishment of proton gradients, removal and sequestration of toxic ions from the cytoplasm and organelles have been isolated. Meanwhile, many proteins have also been isolated related to osmoprotectants, reactive oxygen scavengers, stress-induced proteins, heat shock proteins, membrane fluidity, water status, signaling components, transcription control and growth regulators. However, studies of stress adaptation h
ave suggested a network of multiple signaling pathways in the course of physical adaptations and interactive molecular and cellular changes that begin after the onset of stress in plant .The most important task in the next few years remains to identify pathway components to establish their function by genetic approaches.
Despite soil salinity is the major constraint for plant agriculture, few functional determinants of salt adaptation have been identified and the significance of their involvement characterized. The fission yeast Schizosaccharomyces pombe is a single cell eukaryotic organism. Taking advantage of the genetic simplicity of S. pombe and the functional conservation of some salt tolerance-related proteins, We used this organism as a simple system to identify Arabidopsis thaliana salt tolerance proteins by screening for cDNA clones that confer salt resistance when overexpressed. An Arabidopsis thaliana cDNA library was constructed in pREPSN vector under the control of the inducible nmt-1 promoter and transformed into S. pombe. Expression of the A. thaliana sequences were induced and clones showing strong salt stress were
identified and analysed. By this 'cross-phylogenetic' screen, We have isolated some salt tolerance-related proteins such as glycine-rich protein, chlorophyll a/b-binding protein and some unknown protein. Overexpression of these proteins in the yeast cells afforded significantly increase in salt tolerance. Northern-blot analysis revealed that these genes were induced by NaCl and ABA. In addition, differentially regulated in various tissues. Transgenic Arabidopsis can develop better than wild type in salt tolerance. Taken together, these results undoubtedly suggest that these proteins are involved in the salt stress response in Arabidopsis. The AtGRP9 gene and the AtCab gene are first reported to salt stress response in Arabidopsis.
引文
1 张慧，陈受宜。植物耐旱耐盐的分子生物学及基因工程策略。农业生物工程，1998：198～206
2 李子银，中科院遗传和发育生物学研究所博士论文，2000
3 沈义国，中科院遗传和发育生物学研究所博士论文，2002
4 王栋，中科院微生物研究所硕士论文，2001
5 余叔文，汤章城主编，植物生理与分子生物学(第二版)，北京：科学出版社
6 赵可夫，李法曾主编，中国盐生植物，北京：科学出版社
7 张其德，卢从明，刘丽娜，二氧化碳加富对大豆叶片光系统Ⅱ功能的影响，植物生态学报，1996，20：517～523
8 Adams P, Thomas J C, vernon D M, et al. Distinct cellular and organismic responses to salt stress. Plant Cell Physiol, 1992, 33:1215～1223
9 Alfa C, Fantes P, Hyams J, et al. Experiments with Fission Yeast: a Laboratory Course Manual, Cold Spring Harbor, NY: Cold Spring Harbor Laboratory Press 1993
10 Allakhverdiev S I, Sakamoto A, Nishiyama Y, et al. Ionic and osmotic effects of Nacl-induced inactivation of photosystems Ⅰ and Ⅱ in synechococcus sp. Plant Physiology, 2000, 123; 1047～1056
11 Allen J F, Forsberg J. Molecular recognition in thylakoid structure and function. Trends inPlant Science, 2001, 6:317～325
12 Anderson C M, Wagner T A, Perret M, et al. WAKs: cell wall-associated kinases linking the cytoplasm to the extracellular matrix. Plant Mol boil, 2001, 47:197～206
13 Apple R T, Fristrom J w. 20-Hydroxyecdysone is required for, and negatively regulates, transription of Drosophila pupal cuticle protein genes. Dev Biol, 1991, 146; 569～582
14 Apse M P, Aharon G S, Snedden W A, et al. Salt tolerance conferred by overexpression of a vacuolar Na~+/H~+ antiport in arabidopsis. Sience, 1999, 285:1256～1258
15 Apse M P, Aharon G S, Snedden W A, et al. Salt tolerance conferred by over expression of a vacuolar Na~+/H~+ antiport in Arabidopsis. Science, 1999, 285: 1256-1258
16 Balcells L, Gomez N, Cassamayor A, et al. Regulation of salt tolerance in fission yeast by a protein-phospatase-Z-like Ser/Thr protein phosphatase. Eur. J. Biochem, 1997, 250:476～483
17 Barbier-Brygoo H, Vinauger M, Colcombet J, et al. Anion channels in higher plants: functional characterization, molecular structure and physiological role. Biochinmica et Biophysica Acta, 2000, 1465; 199～218
18 Bartholoew D M, Bartley G E, Scolnik P A. Abscisic acid control of rbcS and cab transcription in tomato leaves. Plant Physiol, 1991, 96:291～296
19 Bassi R, Soen S Y, Frank G; et al. Characterization of chlorophyll apo proteins of photosystem I from chlamydomonas reinhardtii. The Journal of Biological Chemistry, 1992, 267:25714～25721
20 Bergantino Ⅰ, Dainese P, Cerovic Z, et al. A post-translational modification of the photosystem Ⅱ subunit CP29 protects maize from cold stress. The Journal of Biological Chemistry, 1995, 270:8474～8481
21 Bibby T S, Nield J, Barber J. Iron deficiency induces the formation of an antenna ring around trimeric photosystem I in cyanobacteria. Nature, 2001, 412:743～748
22 Bibby T S, Nield J, Barber J. Three-dimensional model and characterization of the iron stress-induced CP43'-photosystem Ⅰ supercomplex isolated from the cyanobacterium synechocystis PCC 6803. The Journal of Biological Chemistry, 2001, 276:43246～43252
23 Binyamin L, Falah M, Portnoy V, et al. The early light-induced protein is also produced during leaf senescence of Nicotiana tabacum. Planta, 2001, 212:591～597
24 Binzel M L, Hess F D, Bressan R A, et al. Intracellular compartmentaton of ions in salt adapted tobacco cells. Plant physiol, 1988, 86:607～614
25 Binzel M L. NaCl-induced accmulation of tonoplast and plasma membrane H~+ATPase message in tomato. Physiol Plant, 1995, 94:72～728
26 Blumwald E, Aharon G S, Apse M P. Sodium transport in plant cells. Biochimica et Biophysica Acta, 2000, 1465:140～151
27 Blumwalk E, Poole R J. Na~+/H~+ antiport in isolated tonoplast vesicles from storage tissue of Beta vulgaris. Plant Physiol, 1987, 78:163～167
28 Bonetta D, McCourt P. Genetic analysis of ABA signal transduction pathways. Trends in plant science, 1998, 3:231～235
29 Brady K P, Darvill A G; Albersheim P. Activation of a tobacco glycine-rich protein gene by a fungal glucan preparation. Plant J, 1993, 4:517～524
30 Bressan R A, Zhang C Q, Zhang H, et al. Learning from the Arabidopsis experience. Plant Physiology, 2001, 127:1354～1360
31 Cansado J, Soto T, Fernandez J,et al. Characterization of mutants devoid of neutral trehalase activity in the fission yeast schizosaccharomyces pombe: partial protection fromheat shock and high-salt stress, Journal of Bacteriology, 1998, 180:1342～1345
32 Chang Y C, Walling L L. Abscisic acid negatively regulates expression of chlorophyll a/b binding protein genes during soybean embryogeny. Plant Physiol, 1991, 97:1260～1264
33 Chang Y C, Walling L L. Chlorophyll a/b-binding protein genes are differentially expressed during soybean development. Plant Molecular Biology, 1992, 12:217～230
34 Chen T, Damaj B B, Herrera C, et al. Self-association of the single KH-domain family members Sam68, GRO33, GLD-1, and Qk1: role of the KH domain. Mol Cell Biol, 1997, 17:5707～5718
35 Cheng S H, Keller B, Condit C M. Common occurrence of homologues of petunia glycine-rich protein-1 among plants. Plant Mol Biol, 1996, 31:163～168
36 Chye M L. Arabidopsis cDNA encoding a membrane-associated protein with an acyl-CoA binding domain. Plant Molecular Biology, 1998, 38:327～838
37 Condit C M, Meagher R B. Expression of a gene encoding a glycine-rich protein in petunia. Mol Cell Biol, 1987, 7:4273～4279
38 Condit C M. Developmental expression and localization of petunia glycine-rich protein 1. Plant Cell, 1993, 5:277～288
39 Creelman R A, Mullet J E. Water deficit modulates gene expression in growing zones of soybean seedlings. Analysis of differentially expressed cDNAs, a new β-tubulin gene, and expresson of genes encoding cell wall proteins. Plant Molecular Biology, 1991, 17:591～608
40 Cruz-Alvarez M, Pellicer A. Cloning of a full-length complementary DNA for an Artemia salina glycine-rich protein. Structural relationship with RNA binding proteins. J Biol Che, 1987, 262:13377～13380
41 Delille J, Peterson E C, Johnson T, et al. A novel precursor recognition element facilitates posttranslafional binding to the signal recognition particle in chloroplasts. Proc. Natl. Acad. Sci. USA, 2000, 97:1926～1931
42 Didierjean L, Frendo P, Burkard G Stress responses in maize: sequence analysis of cDNAs encoding glycine-rich proteins. Plant Mol Biol, 1992, 18:847～849
43 Estruch F. Stress-controlled transcription factors, stress-induced genes and stress tolerance in budding yeast. FEMS Microbiology Reviews, 2000, 24:469～486
44 Fang R X, Pang Z, Gao D M, et al. cDNA sequence of a virus-inducible, glycine-rich protein gene from rice. Plant Mol Biol, 1991, 17:1255～1257
45 Feldmann K A, Marks M D. Agrobacterium-mediated transformation of germinating seeds of Arabidopsis thaliana: A non-tissue culture approach. Mol Gen Genet, 1987, 208:1-9
46 Flowers T J, Yeo A R. Solute Transport in Plants. Glasgow, Scotland: Blackie, 1992, 176pp
47 Fontes A M, Conacci M E, Monesi N, et al. The DNA puff BhB10-1 gene encodes a glycine-rich protein secreted by the late stage larval salivary glands of Bradysia hygida.
48 Freire M A, Pages M. Functional characteristics of the maize RNA-binding protein MA16. Plant Mol Biol, 1995, 29:797～807
49 Furumoto T, Hata S, Izui K. Isolation and characterization of cDNAs for differentially accumulated transcripts between mesophyll cells and bundle sheath strands of maize leaves. Plant Cell Physiol, 2000, 41:1200～1209
50 Gaxiola R A, Rao R, Sherman A, et al. The Arabidopsis thaliana proton transporters, At Nhx1 and Avp1, can function in cation detoxification in yeast. Proc Natl Acad Sci USA, 1999, 96:1480-1485
51 Geisler M, Frangne N, Gomes E, et al. The ACA4 gene of Arabidopsis encodes a vacuolar membrane calcium pump that improves salt tolerance in yeast. Plant Physiol, 2000, 124:1814～1827
52 Girault R, His I, Andeme-Onzighi C, et al. Identification and partial characterization of proteins and proteoglycans encrustion the secondary cell walls of flax fibres. Planta, 2000, 211:256～264
53 Gisbert C, Rus A M, Bolarin M C, et al. The yeast HAL1 gene improves salt tolerance of transgenic tomato. Plant Physiol, 2000, 123:393～402
54 Goddemeier M L, Wulff D, Feix G Root-specific expression of a Zea mays gene encoding a novel glycine-rich protein, zmGRP3. Plant Molecular Biology, 1998, 36: 799-802
55 Godoy J A, Pardo J M, Pintor-Toro J A. A tomato cDNA inducible by salt stress and abscisic acid: nucleotide sequence and expression pattern. Plant Molecular Biology, 1990, 15:695～705
56 Gomez J, Martinez D S, Stiefel V, et al. A gene induced by the plant hormone abscisic acid in response to water stress encodes a glycine-rich protein. Nature, 1988, 334:262～264
57 Gong Z H, Koiwa H, Cushman M A, et al. Genes that are uniquely stress regulated in salt overly sensitive (sos) mutants. Plant Physiology, 2001, 126:363～375
58 Hairier U, Ishitani M, Zhu J K. The arabidopsis sos2 protein kinase physically interacts with and is activated by the calcium-binding protein sos3. Proc. Natl. Acad. Sci. USA, 2000, 97:3735～3740
59 Haro R, Garciadeblas B, Navarro A R. A novel p-type ATPase from yeast involved in sodium transport. Federation of European Biochemical Societies, 1991, 291: 189-191
60 Harrak H, Chamberland H, Plante M, et al. A proline-, threonine-, and glycine-rich protein down-regulated by drought is localized in the cell wall of xylem elements. Plant Physiology, 1999, 121:557～564
61 Hasegawa P M, Bressan R A. Plant cellular and molecular responses to high salinity. Plant Physiol. Plant Mol. Biol, 2000, 51:463～499
62 Hechenberge M, Schwappach B, Fischer W N, et al. A family of putative chloride channels from Arabidopsis and functional complementation of a yeast strain with a CLC gene disruption. J. Biol. Chem, 1996, 271:33632～33638
63 Heddad M, Adamska I. Light stress-regulated two-helix proteins in arabidopsis thaliana related to the chlorophyll a/b-binding gene family. Proc. Nat1. Acad Sci. USA, 2000, 97:3741～3746
64 Hill D R, Hladun S L, Scherer S, et al. Water stress protein of Nostoc commune(cyanobacteria) are secreted with uv-a/b-absorbing pigments and associate with 1,4-β-D-xylanxylanohydrolase activity. The Journal of Biological Chemistry, 1994, 269:7726～7734
65 Ishitani M, Nakamura T, Hah S Y, et al. Expressio of the betaine dehydrogenase gene in barley in response to osmotic stress and abscisicc acid. Plant Molecular Biology, 1995, 27:307～315
66 Jochova J, Quaglino D, Zakeri Z, et al. Protein synthesis, DNA degradation, and morphological changes during programmed cell death in labial glands of Manduca Sexta. Dev Genet, 1997, 21:249～257
67 Kaldenhoff R, Richter G Sequence of cDNA for a novel light-induced glycine-rich protein. Nucleic Acids Res, 1989, 17:2853
68 Kanhonou R, Serrano R, Palau R R. A catalytic subunit of the sugar beet protein kinase CK2 is induced by salt stress and increases NaC1 tolerance in Saccharomyces cerevisiae. Plant Mol Biol, 2001, 47:571～579
69 Kawasaki N, Kawasaki T Yamashina I. Isolation and characterization of a mannan-binding protein from human serum. J Biochem (Tokyo), 1983, 94:937～947
70 Keller B, Sauer N Lamb C J. Glycine-rich cell wall proteins in bean: gene structure and association of the protein wit the vascular system. EMBO J, 1988, 7:3625～3633
71 Killian R, Bassi R, Schafer C. Identification and characterization of photosystem Ⅱ chlorophyll a/b binding proteins in marchantia polymorpha L. Planta, 1998, 204: 260-267
72 Klein R R, Mullet J E. Regulation of chloroplast-encoded chlorophyll-binding protein translation during higher plant chloroplast biogenesis. The Journal of Biological Chemistry, 1986, 261:11138～11145
73 Kloiger K, Weiskircher R, Kraautler B, et al. Mutational analysis and NMR spectroscopy of quail cysteine and glycine-rich protein CRP2 reveal an intrinsic segmental flexibility of LIM domains, J Mol Biol, 1999, 292:893～908
74 Knight H, Knight M R. Abiotic stress signaling pathways:specificity and cross-talk. Trends in Plant Science, 2001, 6:262～267
75 Koch A W, Holstein T W, Mala C, et al. Spinalin, a new glycine- and histidine-rich protein in spines of Hydra nematocysts. Journal of Cell Science, 1998, 111: 1545～1554
76 Kohorn B D. WAKs; cell wall associated kinases. Curr Opin Cell Biol, 2001, 13: 529-533
77 Konrat R, Krautler B, Weiskirchen R, et al. Structure of cysteine- and glycine-rich protein CRP2,J Biol Chem, 1998, 273:23233～23240
78 Kontaxis G; Konrat R, Kraurler B, et al. Structure and intramodular dynamics of the amino-terminal LIM domain from quail cysteine-and glycine-rich protein CRP2. Biochemistry, 1998, 20:7127～7134
79 Kudla J, qiang X, Klaus H, et al. Genes for calcineurin B-like proteins in Arabidopsis are differentially regulated by stress signals. Proc, Natl, Acad, Sci, USA, 1999, 96:4718～4723
80 Kumar A, Williams K R, Szer W. Purification and domain structure of core hnRNP proteins A1 and A2 and their J relationship to single-stranded DNA-binding proteins. J Biol Chem, 1986, 261:11266～11273
81 Laberge s, Castonguay Y, Vezina L P. New cold-and-drought regulated gene from medicage sativa. Plant Physiol, 1993, 101: 1411～1412
82 Lang V, Palva E T The expression of a rah-related gene, rab18, is induced by abscisic acid during the cold acclimation process of Arabidopsis thaliana (L.)Heynh. Plant Molecular Biology, 1992, 20:951～962
83 Lee J H, Van Montagu M, Verbruggen N. A highly conserved kinase is an essential component for stress tolerance in yeast and plant cells. Proc Natl Acad Sci USA, 1999, 96:5873～5877
84 Lei M, Wu R. A novel glycine-rich cell wall protein gene in rice. Plant Mol Biol, 1991, 16:187～198
85 Leonhardt K, Straus N A. An iron stress operon involved in photosynthetic electron transport in the marine cyanobacterium synechococcus sp. PCC 7002. Journal of Gereral Microbiology, 1992, 138:1613～1621
86 Leonhardt K, straus N A. Photosystem Ⅱ genes isiA, phbC in anabaena sp. PCC 7120: cloning,sequencing and the transcriptional regulation in iron-stressed and iron-repleted cells. Plant Molecular Biology, 1994, 24:63～73
87 Leung J, Merlot S, Giraudat J. The Arabidopsis ABSCISIC ACID-INSENSTIVE2(ABI2) and ABII genes encode homologous protein phosphatases 2C involved in abscisic acid signal transduction. The Plant Cell, 1997, 9:759～771
88 Li N, Parsons B L, Liu D R, et al. Accumulation of wound-inducible ACC synthase transcript in tomato fruit is inhibited by salicylic acid and polyamines. Plant Mol Biol, 1992, 18:477～487
89 Li Y Q, Ye L Z, Sugita M, et al. Tobacco nuclear gene for the 31 kd chloroplast ribonucleoprotein: genomic organization, sequence analysis and expression. Nucleic Acids Res, 1991, 19:2987～2991
90 Linthorst H j, Meuwissen R L, Kauffmann S, et al. Constitutive expressionof pathogenesis-related proteins PR-1, GRP, and PR-S in tobacco has no effect on virus infection. Plant Cell, 1989, 1:285～291
91 Linthorst H J, Memelink J, Bol J F, et al. Characterization of cDNA clones for a virs-inducible, glycine-rich protein from petunia. Plant Mol Boil, 1990, 15:521～523
92 Liu J, Zhu J K. A calcium sensor homologue required for plant salt tolerance. Science 1998, 280:1943～1945
93 Loopstra C A, Sederoff R R. Xylem-specific gene expression in loblolly pine. Plant Mol Biol, 1995, 27:277～291
94 Luan S. Protein phosphatases and signaling cascades in higher plants. Trends in plant science, 1998, 3: 271～275
95 Ludevid M D, Freire M A, Gomez J, et al. RNA binding characteristics of a 16 kDa glycine-rich protein from maize. The Plant Journal, 1992, 2:999～1003
96 Luo M, Liu H J, Mohapatra S, et al. Characterization of a gene family encoding abscisic acid- and environmental stress-inducible proteins of alfalfa. The Journal Of Biological Chemistry, 1992, 267:15367～15374
97 Matoh T, Watanabe J, Takahashi. Sodium, potassium, chloride, and betaine concentrations in isolated vacuoles from salt-grown Atriplex gmelini leaves. Plant Physiol, 1987, 84:173～177
98 Matsui M, Toyosawa I, Fukuda M. Novel N-terminal sequence of a glycine-rich protein in the aleurone layer of soybean seeds. Biosci Biotechnol Biochem, 1994, 58:1920～1922
99 Matsuyama T, Satoh H, Yamada Y, et al. A maize glycine-rich protein is synthesized in the lateral root cap and accumulates in the mucilage. Plant Physiol, 1999, 120: 665～674
100 McCue K F, Hanxon A D. Drought and salt tolerance: towards understanding and application. Biotechnology, 1990, 3:358～362
101 Mousavi A, Hiratsuka R, Takase H, et al. A novel glycine-rich protein is associated with starch grain accumulation during anther development. Plant Cell Physiol, 1999, 40:406～416
102 Mukherjee A, Cui Y, Liu Y, et al. Molecular characterization and expression of the Erwinia carotovora hrpNEccgene, which encodes an elicitor of the hypersensitive reaction. Mol Plant Microbe Interact, 1997, 10:462～471
103 Mundy J, Chua N H. Abscisic acid and water-stress induce th expression of a novel rice gene. EMBO J, 1988, 7:2279～2286
104 Munns R, Termaat A. Whole-plant responses to salinity. Aust. J. Plant Physiol, 1986, 13:143～160
105 Nadeau J A, Zhang X S, Li J, et al. Ovule development: identification of stage-specific and tissue-specific cDNAs. Plant Cell,1996,8:213～239
106 Nicolas C, Rodriguez D, Poulsen F, et al. The expressin of an abscisic acid-responsive glycine-rich protein coincides with the level of seed dormancy in Fagus sylvatica. Plant Cell Physiol, 1997, 38:1303～1310
107 Niu X, Bressan R A, Hasegawa P M, et al. Ion homeostasis in NaCl stress environments. Plant Physiol, 1995, 109:735～742
108 Nuccio M L, Rhodes D, McNeil S D, et al. Metabolic engineering of plants for osmotic stress resistance. Current Opinion in Plant Biology, 1999, 2:128～134
109 Nylander M, Heino P, Helenius E, et al. The low-temperature- and salt-induced RCI2A gene of Arabidopsis complements the sodium sensitivity caused by a deletion of the homologous yeast gene SArA1. Plant Mol Biol, 2001, 45:341～352
110 Obokata J, Ohme M, Hayashida N. Nucleotide sequence of a cDNA clone encoding a putative glycine-rich protein of 19.7 kDa in Nicotiana sylvestris. Plant Mol Biol, 1991, 17:953～955
111 Oliveira D E, Franco L 0, Simoens C, et al. Inflorescence-specific genes from Arabidopsis thaliana encoding glycine-rich proteins. Plant J, 1993:495～507
112 Oliverra D E, Seurinck J, Montagu M, et al. Differential expression of five Arabidopsis genes encoding glycine-rich protein. Plant Cell, 1990, 2:427～436
113 Park A R, Cho S K, Yun U J, et al. Interactinof the Arabidopsis receptor protein kinase Wakl with a glycine-rich protein, AtGRP-3. J Biol Chem, 2001, 276: 26688～26693
114 Park A R, Cho S K, Yun U J, et al. Interaction of the Arabidopsis receptor protein kinase wakl with a glycine-rich protein atGRP-3. The Journal of Biological Chemistry, 2001, 276:26688～26693
115 Parsons B L, Mattoo A K. A wound-repressible glycine-rich protein transcript is enriched in vascular bundles of tomato fruit and stem. Plant Cell Physiol, 1994, 35:27～35
116 Paul M H, Ray A B, Zhu J K, et al. Plant celluler and molecular responses to high salinity. Annu. Reu. Plant Physiol, 2000,51:463～499
117 Pawlowski K, Twigg P, Dobritsa S, et al. A nodule-specific gene family from Alnus glutinosa encodes glycine- and histidine-rich proteins expressed in the early stages of actinorhizal nodule development. Mol Plant Microbe Interact, 1997, 10:656～654
118 Phillips A L, huttly A K. Cloning of two gibberellin-regulated cDNAs from Arabidopsis thaliana by subtractive hybridization: expression of the tonoplast water channel, gamma-TIP, is increased by GA3. Plant Mol Biol, 1994, 24:603～615
119 Pical C, Westergren T, Dove S K, et al. Sanlinity and hyperosmotie stress induce rapid increases in phosphatidylinosital 4,5-bisphosphate, diacylglycerol pyrophosphate, and phosphatidylcholine in arabidopsis thaliana cells. The Journal of Chemistry, 1999, 274:38232～38240
120 Plana M, Irarte E, Eritja R, et al. Phosphorylation of maize RAB-17 protein by casein kinase 2. J Biol Chem, 1991, 266:22510～22514
121 Potenza C, Thomas S H, Gopalan C S. Genes induced during early response to Meloidogyne incognita in roots of resistant and susceptible alfalfa cultivars. Plant science, 2001, 161:289～299
122 Prentice H L. High efficiency transformation of schizosaccharomyces pombe by electroporation. Nucleic Acids Research, 1991, 20:621
123 Quigley F, Villiot M L, Mache R. Nucleotide sequence and expression of a novel glycine-rich protein gene from Arabidopsis thaliana. Plant Mol Biol, 1991, 17: 949～952
124 Ratajczak R, Richter J, Lutge U. Adaptation of the tonoplast V-type H~+-ATPase of Mesembryanthemum crystallinum to salt stress, C_3CAM transition and plant age. Plant Cell Enviorn, 1994, 14:1101～111
125 Reddy A S, Poovaiah B W. Accumulation of a glycine-rich protein in auxin-deprived strawberry fruits. Biochem Biophys Res Commun, 1987, 147:885～891
126 Rohde W, Rosch K, Kroger K, et al. Nucleotide sequence of a Hordeum vulgare gene encoding a glycine-rich protein with homology to vertebrate cytokeratins. Plant Mol Biol, 1990, 14:1057～1059
127 Rojik I, Huszta S, Feher O. Rapid synthesis of a glycine-rich protein in the cerebral cortex of the rat during activity. Neurobiology, 1994, 2:125～139
128 Rubio F, Gassman W, Schroeder J I. Sodium-driven potassium uptake by the plant potassium transporter HKT1 and mutations conferring salt tolerance. Science, 1995, 270:1660～1663
129 Ruiz A E, Belles J M, Serrano R, et al. Arabidopsis thaliana athal3: a flavoprotein related to salt and osmotic tolerance and plant growth. The Plant Journal, 1999, 20: 529～539
130 Ryser U, Schorderet M, Zhao G G, et al. Structural cell-wall proteins in protoxylem development: evidence for a repair process mediated by a glycine-rich protein. Plant J, 1997, 12:97～111
131 Sachetto-Martins G, Franco L O, Oliverira D E. Plant glycine-rich proteins: a family or just proteins with a common motif?. Biochimical et Biophysica Acta, 2000, 1492:1～14
132 Sakamoto A, Alia, Murata N. Metabolic engineering of rice leading to biosynthesis of glycinebetaine and tolerance to salt and cold. Plant Molecular Biology, 1998, 38: 1011-1019
133 Sakamoto A, Murata N. The use of bacterial choline oxidase, a glycinebetaine synthesizing enzyme, to create stress-resistant transgenic plants. Plant Physiol, 2001, 125:180～188
134 Sakuta C, Oda A, Yamakawa S, et al. Root-specific expression of genes for novel glycine-rich proteins cloned by use of an antiserum against xylem sap proteins of cucumber. Plant Cell Physiol, 1998, 39:1330～1336
135 Sakuta C, Satoh S. Vascular tissue-specific gene expression of xylem sap glycine-rich proteins in root and their localization in the walls of metaxylem vessels in cucumber. Plant Cell Physiol, 2000, 41:627～638
136 Sambrook J, Fritsch E F, Maniatis T Molecular Cloning: A laboratory Manual, 2nd Ed, New York, CoM spring Harbor Laboratory Press, 1989
137 Santino C G; Stanford G L, Conner T W. Developmental and transgenic analysis of two tomato fruit enhanced genes. Plant Mol Biol, 1997, 33:405～416
138 Satoh H, Akira U, Katsumi N, et al. Water-soluble chlorophyll protein in brassicaceae plants is a stress-induced chlorophyll-binding protein. Plant Cell Physiol, 2001, 42:906～911
139 Schachtman D, Liu W H. Molecular pieces to the puzzle of the interaction between potassium and sodium uptake in plants. Trends in plant science, 1991, 4:281～287
140 Schachtman D, Liu W. Molcular pieces to the puzzle of the interaction between potassium and sodium uptake in plants. Trends Plant Sci, 1999, 4:281～287
141 Shinozaki K, Yamaguchi-Shinozaki K. Molecular respones to dehydration and low temperature: differences and cross-talk between two stress signaling pathways. Current Opinion in Plant Biology, 2000, 3:217～223
142 Showalter A M, Butt A D, Kim S. Molecular details of tomato extensin and glycine-rich protein gene expression. Plant Molecular Biology, 1992, 19:205～215
143 Showalter A M, Zhou J, Rumeau D, et al. Tomato extensin and extensin-like cDNAs: structure and expression in response to wounding. Plant Mol Biol, 1991, 16: 547～565
144 Skriver K, Mundy J. Gene expression in response to abscisic acid and osmotic stress. The Plant Cell, 1990, 2:503～512
145 Smart L B, Camerson K D, Bennett A B. Isolation of genes predominantly expressed in guard cells and epidermal cells of Nicotianz glauca. Plant Mol Biol., 2000, 42:857～869
146 Staiger D, Heintzen C. The circadian system of Arabidopsis thaliana: forward and reverse genetic approaches. Chronobiol Int, 1999, 16:1～16
147 Sto S, Toya T, Kawahara R, et al. Isolation of a carrot gene expressed specifically during early-stage somatic embryogenesis. Plant Mol Biol, 1995, 28:39～46
148 Taipalensuu J, Falk A, Ek B, et al. Myrosinase-binding proteins are derived from a large wound-inducible and repetitive transcription. Eur J Biochem, 1997, 243:605～611
149 Trossat C, Nolte K D, Hanson A D. Evidence that the pathway of dimethylsufoniopropionate biosynthesis begins in the cytosol and ends in the chlorolart. Plant Physiol, 1996, 111: 965～973
150 Van de Rhee M D, Van Kan J A, Gonzalez-Jaen M T, et al. Analysis of regulatory elements involved in the induction of two tobacco genes by salicylate treatment and virus infection. Plant Cell, 1990, 2:357～366
151 Vilardell J, Goday A, Freire M A, et al. Gene sequence, developmental expression, and protein phosphorylation of RAB-17 in maize. Plant Mol Biol, 1990, 14:423～432
152 Wadsworth G J, Redinbaugh M G, Scandalios J G. A procedure for small-scale isolation of plant RNA suitable for RNA blot analysis. Anal Biochem, 1988, 172: 279～283
153 Weretilnyk E A, Hanson A D. Molecular cloning of a plant betaine aldehydedehydrogenase, an enzyme implicated in adaptation to salinity and drought. Proc Natl Acad Sci USA, 1990, 87:2745～2749
154 Wong G, Muller O, Clark R, et al. Molecular cloning and nucleic acid binding properties of the GAP-associated tyrosine phosphoprotein p62. Cell, 1992, 69:551～558
155 Wu J, Saupe S J, Glass N L. Evidence for balancing selectin operating at the het-c heterokaryon incompatibility locus in a group of filamentous fungi. Proc Nat1 Acad Sci USA, 1998, 95: 12398～123403
156 Wyatt R E, Nagao R T, Key J L. Patterns of soybean proline-rich protein gene expression. Plant Cell, 1992, 4:99～110
157 Yasuda E, Ebinuma H, Wabiko H. Anovel glycine-rich/hydrophobic 16 kDa polypeptide gene from tobacco: similarity to proline-rich protein genes and its wound-inducible and developmentally regulated expression. Plant Mol Biol, 1997, 33: 667～678
158 Ye G N, Stone D, Pang S Z, et al. Arabidopsis ovule is the target for agrobacterium in planta vacuum infiltration transformation. The Plant Journal, 1999, 19:249～257
159 Yu L X, Chamberland H, Lafontaine J G, et al. Negative regulation of gene expression of a novel proline-, threonine-, and glycine-rich protein by water stress in Lycopersicon chilense. Genome, 1996, 39:1185～1193
160 Zhao Z Y, Tagliani L, Cai T, et al. Agrobacterum-mediated sorghum transformation. Plant Molecular Biology, 2000, 44:789～798
161 Zhu J K, Hasegawa P M, Bressan R A. Molecular aspects of osmotic stess in plants. CRC Crit Rev Plant Sci, 1997,16:253～277
162 Zhu J K, Liu J, Xiong L. Genetic analysis of salt tolerance in Arabidopsis thaliana: evidence of a critical role for potassium nutrition. Plant Cell, 1998,10:1181～1192
163 Zhu J K. Genetic analysis of plant salt tolerance using Arabidopsis. Plant Physiology, 2000, 124:941～948
164 Zhu J K. Plant salt tolerance. Trends in Plant Science, 2001, 6:66～71
165 Zhu W, MaGbanua M M, White F F. Identification of two novel hrp-associated genes in the hrp helle cluster of Xanthomonas oryzae pv. Oryzae. J Bacteriol, 2000, 182: 1844～1853
2 李子银，中科院遗传和发育生物学研究所博士论文，2000
3 沈义国，中科院遗传和发育生物学研究所博士论文，2002
4 王栋，中科院微生物研究所硕士论文，2001
5 余叔文，汤章城主编，植物生理与分子生物学(第二版)，北京：科学出版社
6 赵可夫，李法曾主编，中国盐生植物，北京：科学出版社
7 张其德，卢从明，刘丽娜，二氧化碳加富对大豆叶片光系统Ⅱ功能的影响，植物生态学报，1996，20：517～523
8 Adams P, Thomas J C, vernon D M, et al. Distinct cellular and organismic responses to salt stress. Plant Cell Physiol, 1992, 33:1215～1223
9 Alfa C, Fantes P, Hyams J, et al. Experiments with Fission Yeast: a Laboratory Course Manual, Cold Spring Harbor, NY: Cold Spring Harbor Laboratory Press 1993
10 Allakhverdiev S I, Sakamoto A, Nishiyama Y, et al. Ionic and osmotic effects of Nacl-induced inactivation of photosystems Ⅰ and Ⅱ in synechococcus sp. Plant Physiology, 2000, 123; 1047～1056
11 Allen J F, Forsberg J. Molecular recognition in thylakoid structure and function. Trends inPlant Science, 2001, 6:317～325
12 Anderson C M, Wagner T A, Perret M, et al. WAKs: cell wall-associated kinases linking the cytoplasm to the extracellular matrix. Plant Mol boil, 2001, 47:197～206
13 Apple R T, Fristrom J w. 20-Hydroxyecdysone is required for, and negatively regulates, transription of Drosophila pupal cuticle protein genes. Dev Biol, 1991, 146; 569～582
14 Apse M P, Aharon G S, Snedden W A, et al. Salt tolerance conferred by overexpression of a vacuolar Na~+/H~+ antiport in arabidopsis. Sience, 1999, 285:1256～1258
15 Apse M P, Aharon G S, Snedden W A, et al. Salt tolerance conferred by over expression of a vacuolar Na~+/H~+ antiport in Arabidopsis. Science, 1999, 285: 1256-1258
16 Balcells L, Gomez N, Cassamayor A, et al. Regulation of salt tolerance in fission yeast by a protein-phospatase-Z-like Ser/Thr protein phosphatase. Eur. J. Biochem, 1997, 250:476～483
17 Barbier-Brygoo H, Vinauger M, Colcombet J, et al. Anion channels in higher plants: functional characterization, molecular structure and physiological role. Biochinmica et Biophysica Acta, 2000, 1465; 199～218
18 Bartholoew D M, Bartley G E, Scolnik P A. Abscisic acid control of rbcS and cab transcription in tomato leaves. Plant Physiol, 1991, 96:291～296
19 Bassi R, Soen S Y, Frank G; et al. Characterization of chlorophyll apo proteins of photosystem I from chlamydomonas reinhardtii. The Journal of Biological Chemistry, 1992, 267:25714～25721
20 Bergantino Ⅰ, Dainese P, Cerovic Z, et al. A post-translational modification of the photosystem Ⅱ subunit CP29 protects maize from cold stress. The Journal of Biological Chemistry, 1995, 270:8474～8481
21 Bibby T S, Nield J, Barber J. Iron deficiency induces the formation of an antenna ring around trimeric photosystem I in cyanobacteria. Nature, 2001, 412:743～748
22 Bibby T S, Nield J, Barber J. Three-dimensional model and characterization of the iron stress-induced CP43'-photosystem Ⅰ supercomplex isolated from the cyanobacterium synechocystis PCC 6803. The Journal of Biological Chemistry, 2001, 276:43246～43252
23 Binyamin L, Falah M, Portnoy V, et al. The early light-induced protein is also produced during leaf senescence of Nicotiana tabacum. Planta, 2001, 212:591～597
24 Binzel M L, Hess F D, Bressan R A, et al. Intracellular compartmentaton of ions in salt adapted tobacco cells. Plant physiol, 1988, 86:607～614
25 Binzel M L. NaCl-induced accmulation of tonoplast and plasma membrane H~+ATPase message in tomato. Physiol Plant, 1995, 94:72～728
26 Blumwald E, Aharon G S, Apse M P. Sodium transport in plant cells. Biochimica et Biophysica Acta, 2000, 1465:140～151
27 Blumwalk E, Poole R J. Na~+/H~+ antiport in isolated tonoplast vesicles from storage tissue of Beta vulgaris. Plant Physiol, 1987, 78:163～167
28 Bonetta D, McCourt P. Genetic analysis of ABA signal transduction pathways. Trends in plant science, 1998, 3:231～235
29 Brady K P, Darvill A G; Albersheim P. Activation of a tobacco glycine-rich protein gene by a fungal glucan preparation. Plant J, 1993, 4:517～524
30 Bressan R A, Zhang C Q, Zhang H, et al. Learning from the Arabidopsis experience. Plant Physiology, 2001, 127:1354～1360
31 Cansado J, Soto T, Fernandez J,et al. Characterization of mutants devoid of neutral trehalase activity in the fission yeast schizosaccharomyces pombe: partial protection fromheat shock and high-salt stress, Journal of Bacteriology, 1998, 180:1342～1345
32 Chang Y C, Walling L L. Abscisic acid negatively regulates expression of chlorophyll a/b binding protein genes during soybean embryogeny. Plant Physiol, 1991, 97:1260～1264
33 Chang Y C, Walling L L. Chlorophyll a/b-binding protein genes are differentially expressed during soybean development. Plant Molecular Biology, 1992, 12:217～230
34 Chen T, Damaj B B, Herrera C, et al. Self-association of the single KH-domain family members Sam68, GRO33, GLD-1, and Qk1: role of the KH domain. Mol Cell Biol, 1997, 17:5707～5718
35 Cheng S H, Keller B, Condit C M. Common occurrence of homologues of petunia glycine-rich protein-1 among plants. Plant Mol Biol, 1996, 31:163～168
36 Chye M L. Arabidopsis cDNA encoding a membrane-associated protein with an acyl-CoA binding domain. Plant Molecular Biology, 1998, 38:327～838
37 Condit C M, Meagher R B. Expression of a gene encoding a glycine-rich protein in petunia. Mol Cell Biol, 1987, 7:4273～4279
38 Condit C M. Developmental expression and localization of petunia glycine-rich protein 1. Plant Cell, 1993, 5:277～288
39 Creelman R A, Mullet J E. Water deficit modulates gene expression in growing zones of soybean seedlings. Analysis of differentially expressed cDNAs, a new β-tubulin gene, and expresson of genes encoding cell wall proteins. Plant Molecular Biology, 1991, 17:591～608
40 Cruz-Alvarez M, Pellicer A. Cloning of a full-length complementary DNA for an Artemia salina glycine-rich protein. Structural relationship with RNA binding proteins. J Biol Che, 1987, 262:13377～13380
41 Delille J, Peterson E C, Johnson T, et al. A novel precursor recognition element facilitates posttranslafional binding to the signal recognition particle in chloroplasts. Proc. Natl. Acad. Sci. USA, 2000, 97:1926～1931
42 Didierjean L, Frendo P, Burkard G Stress responses in maize: sequence analysis of cDNAs encoding glycine-rich proteins. Plant Mol Biol, 1992, 18:847～849
43 Estruch F. Stress-controlled transcription factors, stress-induced genes and stress tolerance in budding yeast. FEMS Microbiology Reviews, 2000, 24:469～486
44 Fang R X, Pang Z, Gao D M, et al. cDNA sequence of a virus-inducible, glycine-rich protein gene from rice. Plant Mol Biol, 1991, 17:1255～1257
45 Feldmann K A, Marks M D. Agrobacterium-mediated transformation of germinating seeds of Arabidopsis thaliana: A non-tissue culture approach. Mol Gen Genet, 1987, 208:1-9
46 Flowers T J, Yeo A R. Solute Transport in Plants. Glasgow, Scotland: Blackie, 1992, 176pp
47 Fontes A M, Conacci M E, Monesi N, et al. The DNA puff BhB10-1 gene encodes a glycine-rich protein secreted by the late stage larval salivary glands of Bradysia hygida.
48 Freire M A, Pages M. Functional characteristics of the maize RNA-binding protein MA16. Plant Mol Biol, 1995, 29:797～807
49 Furumoto T, Hata S, Izui K. Isolation and characterization of cDNAs for differentially accumulated transcripts between mesophyll cells and bundle sheath strands of maize leaves. Plant Cell Physiol, 2000, 41:1200～1209
50 Gaxiola R A, Rao R, Sherman A, et al. The Arabidopsis thaliana proton transporters, At Nhx1 and Avp1, can function in cation detoxification in yeast. Proc Natl Acad Sci USA, 1999, 96:1480-1485
51 Geisler M, Frangne N, Gomes E, et al. The ACA4 gene of Arabidopsis encodes a vacuolar membrane calcium pump that improves salt tolerance in yeast. Plant Physiol, 2000, 124:1814～1827
52 Girault R, His I, Andeme-Onzighi C, et al. Identification and partial characterization of proteins and proteoglycans encrustion the secondary cell walls of flax fibres. Planta, 2000, 211:256～264
53 Gisbert C, Rus A M, Bolarin M C, et al. The yeast HAL1 gene improves salt tolerance of transgenic tomato. Plant Physiol, 2000, 123:393～402
54 Goddemeier M L, Wulff D, Feix G Root-specific expression of a Zea mays gene encoding a novel glycine-rich protein, zmGRP3. Plant Molecular Biology, 1998, 36: 799-802
55 Godoy J A, Pardo J M, Pintor-Toro J A. A tomato cDNA inducible by salt stress and abscisic acid: nucleotide sequence and expression pattern. Plant Molecular Biology, 1990, 15:695～705
56 Gomez J, Martinez D S, Stiefel V, et al. A gene induced by the plant hormone abscisic acid in response to water stress encodes a glycine-rich protein. Nature, 1988, 334:262～264
57 Gong Z H, Koiwa H, Cushman M A, et al. Genes that are uniquely stress regulated in salt overly sensitive (sos) mutants. Plant Physiology, 2001, 126:363～375
58 Hairier U, Ishitani M, Zhu J K. The arabidopsis sos2 protein kinase physically interacts with and is activated by the calcium-binding protein sos3. Proc. Natl. Acad. Sci. USA, 2000, 97:3735～3740
59 Haro R, Garciadeblas B, Navarro A R. A novel p-type ATPase from yeast involved in sodium transport. Federation of European Biochemical Societies, 1991, 291: 189-191
60 Harrak H, Chamberland H, Plante M, et al. A proline-, threonine-, and glycine-rich protein down-regulated by drought is localized in the cell wall of xylem elements. Plant Physiology, 1999, 121:557～564
61 Hasegawa P M, Bressan R A. Plant cellular and molecular responses to high salinity. Plant Physiol. Plant Mol. Biol, 2000, 51:463～499
62 Hechenberge M, Schwappach B, Fischer W N, et al. A family of putative chloride channels from Arabidopsis and functional complementation of a yeast strain with a CLC gene disruption. J. Biol. Chem, 1996, 271:33632～33638
63 Heddad M, Adamska I. Light stress-regulated two-helix proteins in arabidopsis thaliana related to the chlorophyll a/b-binding gene family. Proc. Nat1. Acad Sci. USA, 2000, 97:3741～3746
64 Hill D R, Hladun S L, Scherer S, et al. Water stress protein of Nostoc commune(cyanobacteria) are secreted with uv-a/b-absorbing pigments and associate with 1,4-β-D-xylanxylanohydrolase activity. The Journal of Biological Chemistry, 1994, 269:7726～7734
65 Ishitani M, Nakamura T, Hah S Y, et al. Expressio of the betaine dehydrogenase gene in barley in response to osmotic stress and abscisicc acid. Plant Molecular Biology, 1995, 27:307～315
66 Jochova J, Quaglino D, Zakeri Z, et al. Protein synthesis, DNA degradation, and morphological changes during programmed cell death in labial glands of Manduca Sexta. Dev Genet, 1997, 21:249～257
67 Kaldenhoff R, Richter G Sequence of cDNA for a novel light-induced glycine-rich protein. Nucleic Acids Res, 1989, 17:2853
68 Kanhonou R, Serrano R, Palau R R. A catalytic subunit of the sugar beet protein kinase CK2 is induced by salt stress and increases NaC1 tolerance in Saccharomyces cerevisiae. Plant Mol Biol, 2001, 47:571～579
69 Kawasaki N, Kawasaki T Yamashina I. Isolation and characterization of a mannan-binding protein from human serum. J Biochem (Tokyo), 1983, 94:937～947
70 Keller B, Sauer N Lamb C J. Glycine-rich cell wall proteins in bean: gene structure and association of the protein wit the vascular system. EMBO J, 1988, 7:3625～3633
71 Killian R, Bassi R, Schafer C. Identification and characterization of photosystem Ⅱ chlorophyll a/b binding proteins in marchantia polymorpha L. Planta, 1998, 204: 260-267
72 Klein R R, Mullet J E. Regulation of chloroplast-encoded chlorophyll-binding protein translation during higher plant chloroplast biogenesis. The Journal of Biological Chemistry, 1986, 261:11138～11145
73 Kloiger K, Weiskircher R, Kraautler B, et al. Mutational analysis and NMR spectroscopy of quail cysteine and glycine-rich protein CRP2 reveal an intrinsic segmental flexibility of LIM domains, J Mol Biol, 1999, 292:893～908
74 Knight H, Knight M R. Abiotic stress signaling pathways:specificity and cross-talk. Trends in Plant Science, 2001, 6:262～267
75 Koch A W, Holstein T W, Mala C, et al. Spinalin, a new glycine- and histidine-rich protein in spines of Hydra nematocysts. Journal of Cell Science, 1998, 111: 1545～1554
76 Kohorn B D. WAKs; cell wall associated kinases. Curr Opin Cell Biol, 2001, 13: 529-533
77 Konrat R, Krautler B, Weiskirchen R, et al. Structure of cysteine- and glycine-rich protein CRP2,J Biol Chem, 1998, 273:23233～23240
78 Kontaxis G; Konrat R, Kraurler B, et al. Structure and intramodular dynamics of the amino-terminal LIM domain from quail cysteine-and glycine-rich protein CRP2. Biochemistry, 1998, 20:7127～7134
79 Kudla J, qiang X, Klaus H, et al. Genes for calcineurin B-like proteins in Arabidopsis are differentially regulated by stress signals. Proc, Natl, Acad, Sci, USA, 1999, 96:4718～4723
80 Kumar A, Williams K R, Szer W. Purification and domain structure of core hnRNP proteins A1 and A2 and their J relationship to single-stranded DNA-binding proteins. J Biol Chem, 1986, 261:11266～11273
81 Laberge s, Castonguay Y, Vezina L P. New cold-and-drought regulated gene from medicage sativa. Plant Physiol, 1993, 101: 1411～1412
82 Lang V, Palva E T The expression of a rah-related gene, rab18, is induced by abscisic acid during the cold acclimation process of Arabidopsis thaliana (L.)Heynh. Plant Molecular Biology, 1992, 20:951～962
83 Lee J H, Van Montagu M, Verbruggen N. A highly conserved kinase is an essential component for stress tolerance in yeast and plant cells. Proc Natl Acad Sci USA, 1999, 96:5873～5877
84 Lei M, Wu R. A novel glycine-rich cell wall protein gene in rice. Plant Mol Biol, 1991, 16:187～198
85 Leonhardt K, Straus N A. An iron stress operon involved in photosynthetic electron transport in the marine cyanobacterium synechococcus sp. PCC 7002. Journal of Gereral Microbiology, 1992, 138:1613～1621
86 Leonhardt K, straus N A. Photosystem Ⅱ genes isiA, phbC in anabaena sp. PCC 7120: cloning,sequencing and the transcriptional regulation in iron-stressed and iron-repleted cells. Plant Molecular Biology, 1994, 24:63～73
87 Leung J, Merlot S, Giraudat J. The Arabidopsis ABSCISIC ACID-INSENSTIVE2(ABI2) and ABII genes encode homologous protein phosphatases 2C involved in abscisic acid signal transduction. The Plant Cell, 1997, 9:759～771
88 Li N, Parsons B L, Liu D R, et al. Accumulation of wound-inducible ACC synthase transcript in tomato fruit is inhibited by salicylic acid and polyamines. Plant Mol Biol, 1992, 18:477～487
89 Li Y Q, Ye L Z, Sugita M, et al. Tobacco nuclear gene for the 31 kd chloroplast ribonucleoprotein: genomic organization, sequence analysis and expression. Nucleic Acids Res, 1991, 19:2987～2991
90 Linthorst H j, Meuwissen R L, Kauffmann S, et al. Constitutive expressionof pathogenesis-related proteins PR-1, GRP, and PR-S in tobacco has no effect on virus infection. Plant Cell, 1989, 1:285～291
91 Linthorst H J, Memelink J, Bol J F, et al. Characterization of cDNA clones for a virs-inducible, glycine-rich protein from petunia. Plant Mol Boil, 1990, 15:521～523
92 Liu J, Zhu J K. A calcium sensor homologue required for plant salt tolerance. Science 1998, 280:1943～1945
93 Loopstra C A, Sederoff R R. Xylem-specific gene expression in loblolly pine. Plant Mol Biol, 1995, 27:277～291
94 Luan S. Protein phosphatases and signaling cascades in higher plants. Trends in plant science, 1998, 3: 271～275
95 Ludevid M D, Freire M A, Gomez J, et al. RNA binding characteristics of a 16 kDa glycine-rich protein from maize. The Plant Journal, 1992, 2:999～1003
96 Luo M, Liu H J, Mohapatra S, et al. Characterization of a gene family encoding abscisic acid- and environmental stress-inducible proteins of alfalfa. The Journal Of Biological Chemistry, 1992, 267:15367～15374
97 Matoh T, Watanabe J, Takahashi. Sodium, potassium, chloride, and betaine concentrations in isolated vacuoles from salt-grown Atriplex gmelini leaves. Plant Physiol, 1987, 84:173～177
98 Matsui M, Toyosawa I, Fukuda M. Novel N-terminal sequence of a glycine-rich protein in the aleurone layer of soybean seeds. Biosci Biotechnol Biochem, 1994, 58:1920～1922
99 Matsuyama T, Satoh H, Yamada Y, et al. A maize glycine-rich protein is synthesized in the lateral root cap and accumulates in the mucilage. Plant Physiol, 1999, 120: 665～674
100 McCue K F, Hanxon A D. Drought and salt tolerance: towards understanding and application. Biotechnology, 1990, 3:358～362
101 Mousavi A, Hiratsuka R, Takase H, et al. A novel glycine-rich protein is associated with starch grain accumulation during anther development. Plant Cell Physiol, 1999, 40:406～416
102 Mukherjee A, Cui Y, Liu Y, et al. Molecular characterization and expression of the Erwinia carotovora hrpNEccgene, which encodes an elicitor of the hypersensitive reaction. Mol Plant Microbe Interact, 1997, 10:462～471
103 Mundy J, Chua N H. Abscisic acid and water-stress induce th expression of a novel rice gene. EMBO J, 1988, 7:2279～2286
104 Munns R, Termaat A. Whole-plant responses to salinity. Aust. J. Plant Physiol, 1986, 13:143～160
105 Nadeau J A, Zhang X S, Li J, et al. Ovule development: identification of stage-specific and tissue-specific cDNAs. Plant Cell,1996,8:213～239
106 Nicolas C, Rodriguez D, Poulsen F, et al. The expressin of an abscisic acid-responsive glycine-rich protein coincides with the level of seed dormancy in Fagus sylvatica. Plant Cell Physiol, 1997, 38:1303～1310
107 Niu X, Bressan R A, Hasegawa P M, et al. Ion homeostasis in NaCl stress environments. Plant Physiol, 1995, 109:735～742
108 Nuccio M L, Rhodes D, McNeil S D, et al. Metabolic engineering of plants for osmotic stress resistance. Current Opinion in Plant Biology, 1999, 2:128～134
109 Nylander M, Heino P, Helenius E, et al. The low-temperature- and salt-induced RCI2A gene of Arabidopsis complements the sodium sensitivity caused by a deletion of the homologous yeast gene SArA1. Plant Mol Biol, 2001, 45:341～352
110 Obokata J, Ohme M, Hayashida N. Nucleotide sequence of a cDNA clone encoding a putative glycine-rich protein of 19.7 kDa in Nicotiana sylvestris. Plant Mol Biol, 1991, 17:953～955
111 Oliveira D E, Franco L 0, Simoens C, et al. Inflorescence-specific genes from Arabidopsis thaliana encoding glycine-rich proteins. Plant J, 1993:495～507
112 Oliverra D E, Seurinck J, Montagu M, et al. Differential expression of five Arabidopsis genes encoding glycine-rich protein. Plant Cell, 1990, 2:427～436
113 Park A R, Cho S K, Yun U J, et al. Interactinof the Arabidopsis receptor protein kinase Wakl with a glycine-rich protein, AtGRP-3. J Biol Chem, 2001, 276: 26688～26693
114 Park A R, Cho S K, Yun U J, et al. Interaction of the Arabidopsis receptor protein kinase wakl with a glycine-rich protein atGRP-3. The Journal of Biological Chemistry, 2001, 276:26688～26693
115 Parsons B L, Mattoo A K. A wound-repressible glycine-rich protein transcript is enriched in vascular bundles of tomato fruit and stem. Plant Cell Physiol, 1994, 35:27～35
116 Paul M H, Ray A B, Zhu J K, et al. Plant celluler and molecular responses to high salinity. Annu. Reu. Plant Physiol, 2000,51:463～499
117 Pawlowski K, Twigg P, Dobritsa S, et al. A nodule-specific gene family from Alnus glutinosa encodes glycine- and histidine-rich proteins expressed in the early stages of actinorhizal nodule development. Mol Plant Microbe Interact, 1997, 10:656～654
118 Phillips A L, huttly A K. Cloning of two gibberellin-regulated cDNAs from Arabidopsis thaliana by subtractive hybridization: expression of the tonoplast water channel, gamma-TIP, is increased by GA3. Plant Mol Biol, 1994, 24:603～615
119 Pical C, Westergren T, Dove S K, et al. Sanlinity and hyperosmotie stress induce rapid increases in phosphatidylinosital 4,5-bisphosphate, diacylglycerol pyrophosphate, and phosphatidylcholine in arabidopsis thaliana cells. The Journal of Chemistry, 1999, 274:38232～38240
120 Plana M, Irarte E, Eritja R, et al. Phosphorylation of maize RAB-17 protein by casein kinase 2. J Biol Chem, 1991, 266:22510～22514
121 Potenza C, Thomas S H, Gopalan C S. Genes induced during early response to Meloidogyne incognita in roots of resistant and susceptible alfalfa cultivars. Plant science, 2001, 161:289～299
122 Prentice H L. High efficiency transformation of schizosaccharomyces pombe by electroporation. Nucleic Acids Research, 1991, 20:621
123 Quigley F, Villiot M L, Mache R. Nucleotide sequence and expression of a novel glycine-rich protein gene from Arabidopsis thaliana. Plant Mol Biol, 1991, 17: 949～952
124 Ratajczak R, Richter J, Lutge U. Adaptation of the tonoplast V-type H~+-ATPase of Mesembryanthemum crystallinum to salt stress, C_3CAM transition and plant age. Plant Cell Enviorn, 1994, 14:1101～111
125 Reddy A S, Poovaiah B W. Accumulation of a glycine-rich protein in auxin-deprived strawberry fruits. Biochem Biophys Res Commun, 1987, 147:885～891
126 Rohde W, Rosch K, Kroger K, et al. Nucleotide sequence of a Hordeum vulgare gene encoding a glycine-rich protein with homology to vertebrate cytokeratins. Plant Mol Biol, 1990, 14:1057～1059
127 Rojik I, Huszta S, Feher O. Rapid synthesis of a glycine-rich protein in the cerebral cortex of the rat during activity. Neurobiology, 1994, 2:125～139
128 Rubio F, Gassman W, Schroeder J I. Sodium-driven potassium uptake by the plant potassium transporter HKT1 and mutations conferring salt tolerance. Science, 1995, 270:1660～1663
129 Ruiz A E, Belles J M, Serrano R, et al. Arabidopsis thaliana athal3: a flavoprotein related to salt and osmotic tolerance and plant growth. The Plant Journal, 1999, 20: 529～539
130 Ryser U, Schorderet M, Zhao G G, et al. Structural cell-wall proteins in protoxylem development: evidence for a repair process mediated by a glycine-rich protein. Plant J, 1997, 12:97～111
131 Sachetto-Martins G, Franco L O, Oliverira D E. Plant glycine-rich proteins: a family or just proteins with a common motif?. Biochimical et Biophysica Acta, 2000, 1492:1～14
132 Sakamoto A, Alia, Murata N. Metabolic engineering of rice leading to biosynthesis of glycinebetaine and tolerance to salt and cold. Plant Molecular Biology, 1998, 38: 1011-1019
133 Sakamoto A, Murata N. The use of bacterial choline oxidase, a glycinebetaine synthesizing enzyme, to create stress-resistant transgenic plants. Plant Physiol, 2001, 125:180～188
134 Sakuta C, Oda A, Yamakawa S, et al. Root-specific expression of genes for novel glycine-rich proteins cloned by use of an antiserum against xylem sap proteins of cucumber. Plant Cell Physiol, 1998, 39:1330～1336
135 Sakuta C, Satoh S. Vascular tissue-specific gene expression of xylem sap glycine-rich proteins in root and their localization in the walls of metaxylem vessels in cucumber. Plant Cell Physiol, 2000, 41:627～638
136 Sambrook J, Fritsch E F, Maniatis T Molecular Cloning: A laboratory Manual, 2nd Ed, New York, CoM spring Harbor Laboratory Press, 1989
137 Santino C G; Stanford G L, Conner T W. Developmental and transgenic analysis of two tomato fruit enhanced genes. Plant Mol Biol, 1997, 33:405～416
138 Satoh H, Akira U, Katsumi N, et al. Water-soluble chlorophyll protein in brassicaceae plants is a stress-induced chlorophyll-binding protein. Plant Cell Physiol, 2001, 42:906～911
139 Schachtman D, Liu W H. Molecular pieces to the puzzle of the interaction between potassium and sodium uptake in plants. Trends in plant science, 1991, 4:281～287
140 Schachtman D, Liu W. Molcular pieces to the puzzle of the interaction between potassium and sodium uptake in plants. Trends Plant Sci, 1999, 4:281～287
141 Shinozaki K, Yamaguchi-Shinozaki K. Molecular respones to dehydration and low temperature: differences and cross-talk between two stress signaling pathways. Current Opinion in Plant Biology, 2000, 3:217～223
142 Showalter A M, Butt A D, Kim S. Molecular details of tomato extensin and glycine-rich protein gene expression. Plant Molecular Biology, 1992, 19:205～215
143 Showalter A M, Zhou J, Rumeau D, et al. Tomato extensin and extensin-like cDNAs: structure and expression in response to wounding. Plant Mol Biol, 1991, 16: 547～565
144 Skriver K, Mundy J. Gene expression in response to abscisic acid and osmotic stress. The Plant Cell, 1990, 2:503～512
145 Smart L B, Camerson K D, Bennett A B. Isolation of genes predominantly expressed in guard cells and epidermal cells of Nicotianz glauca. Plant Mol Biol., 2000, 42:857～869
146 Staiger D, Heintzen C. The circadian system of Arabidopsis thaliana: forward and reverse genetic approaches. Chronobiol Int, 1999, 16:1～16
147 Sto S, Toya T, Kawahara R, et al. Isolation of a carrot gene expressed specifically during early-stage somatic embryogenesis. Plant Mol Biol, 1995, 28:39～46
148 Taipalensuu J, Falk A, Ek B, et al. Myrosinase-binding proteins are derived from a large wound-inducible and repetitive transcription. Eur J Biochem, 1997, 243:605～611
149 Trossat C, Nolte K D, Hanson A D. Evidence that the pathway of dimethylsufoniopropionate biosynthesis begins in the cytosol and ends in the chlorolart. Plant Physiol, 1996, 111: 965～973
150 Van de Rhee M D, Van Kan J A, Gonzalez-Jaen M T, et al. Analysis of regulatory elements involved in the induction of two tobacco genes by salicylate treatment and virus infection. Plant Cell, 1990, 2:357～366
151 Vilardell J, Goday A, Freire M A, et al. Gene sequence, developmental expression, and protein phosphorylation of RAB-17 in maize. Plant Mol Biol, 1990, 14:423～432
152 Wadsworth G J, Redinbaugh M G, Scandalios J G. A procedure for small-scale isolation of plant RNA suitable for RNA blot analysis. Anal Biochem, 1988, 172: 279～283
153 Weretilnyk E A, Hanson A D. Molecular cloning of a plant betaine aldehydedehydrogenase, an enzyme implicated in adaptation to salinity and drought. Proc Natl Acad Sci USA, 1990, 87:2745～2749
154 Wong G, Muller O, Clark R, et al. Molecular cloning and nucleic acid binding properties of the GAP-associated tyrosine phosphoprotein p62. Cell, 1992, 69:551～558
155 Wu J, Saupe S J, Glass N L. Evidence for balancing selectin operating at the het-c heterokaryon incompatibility locus in a group of filamentous fungi. Proc Nat1 Acad Sci USA, 1998, 95: 12398～123403
156 Wyatt R E, Nagao R T, Key J L. Patterns of soybean proline-rich protein gene expression. Plant Cell, 1992, 4:99～110
157 Yasuda E, Ebinuma H, Wabiko H. Anovel glycine-rich/hydrophobic 16 kDa polypeptide gene from tobacco: similarity to proline-rich protein genes and its wound-inducible and developmentally regulated expression. Plant Mol Biol, 1997, 33: 667～678
158 Ye G N, Stone D, Pang S Z, et al. Arabidopsis ovule is the target for agrobacterium in planta vacuum infiltration transformation. The Plant Journal, 1999, 19:249～257
159 Yu L X, Chamberland H, Lafontaine J G, et al. Negative regulation of gene expression of a novel proline-, threonine-, and glycine-rich protein by water stress in Lycopersicon chilense. Genome, 1996, 39:1185～1193
160 Zhao Z Y, Tagliani L, Cai T, et al. Agrobacterum-mediated sorghum transformation. Plant Molecular Biology, 2000, 44:789～798
161 Zhu J K, Hasegawa P M, Bressan R A. Molecular aspects of osmotic stess in plants. CRC Crit Rev Plant Sci, 1997,16:253～277
162 Zhu J K, Liu J, Xiong L. Genetic analysis of salt tolerance in Arabidopsis thaliana: evidence of a critical role for potassium nutrition. Plant Cell, 1998,10:1181～1192
163 Zhu J K. Genetic analysis of plant salt tolerance using Arabidopsis. Plant Physiology, 2000, 124:941～948
164 Zhu J K. Plant salt tolerance. Trends in Plant Science, 2001, 6:66～71
165 Zhu W, MaGbanua M M, White F F. Identification of two novel hrp-associated genes in the hrp helle cluster of Xanthomonas oryzae pv. Oryzae. J Bacteriol, 2000, 182: 1844～1853