摘要
多聚半乳糖醛酸酶抑制蛋白(poly-galacturonase inhibiting proteins,PGIPs)能与真菌分泌的内切多聚半乳糖醛酸酶(endo-polygalacturonase,PG)可逆专一性的结合,具有抑制真菌对植物细胞壁降解,激活植物的防卫反应,减缓真菌的繁殖速度等作用,因此被认为是一种植物抗病性相关防卫蛋白。本文以高山离子芥(Chorispora bungeana Fisch,et.Mey,Chorispora bungean)为实验材料,克隆到离子芥PGIP1(CbPGIP1)基因。通过免疫荧光分析了CbPGIP1在植株不同空间的分布。通过荧光定量PCR和Western blotting分析了生物胁迫和非生物胁迫下CbPGIP1基因的变化。此外,一氧化氮(nitric oxide,NO)在植物和病原菌互作中具有重要的重要。本文以拟南芥野生型(wild type)和一氧化氮合成酶突变体(Atnos1)为材料,研究真菌病原体胁迫下NO对植物抗病性以及AtPGIP1表达的调节机理。内容概括如下:
1.从高山离子芥中克隆了CbPGIP1,此基因全长1203bp,编码332个氨基酸的蛋白质,分子量为36.9KD,等电点约为6.93。氨基酸序列分析表明,CbPGIP1蛋白中含有具有典型的LRR(leucine-rich repeat,富亮氨酸重复序列)特征。CbPGIP1与拟南芥AtPGIP1的同源性最高。因此命名为CbPGIP1。
2.CbPGIP1和GFP融合蛋白通过基因枪转入洋葱表皮细胞的实验证实,CbPGIP1在细胞中没有特定的定位,均匀分布在细胞质中。
3.荧光定量PCR和蛋白杂交证明茄匐柄霉,水杨酸(salicylic acid,SA)能够强烈的诱导CbPGIP1的表达,说明CbPGIP1在植物抗病中具有重要的作用。
4.CbPGIP1的荧光免疫定位显示,在叶片中的CbPGIP1均匀分布在这个组织中,而茎和根中的CbPGIP1主要分布在表皮细胞和维管束中。进一步的实验显示CbPGIP1在叶中的含量高于在茎和根中的含量。这些结果显示CbPGIP1的空间分布做为一道天然的物理屏障抵御病源物的入侵。
5.4℃,-4℃和NO均能诱导CbPGIP1基因的表达,CbPGIP1可能在非生物胁迫中具有重要作用。因为低温胁迫就像其他的胁迫条件一样能够降低植物的的抗病性。由此可见低温诱导CbPGIP的表达是为了增强植物的抗病性。NO诱导CbPGIP1基因表达说明了NO参与并调控了该基因的表达。
6.荧光免疫定位显示,在拟南芥叶片中的AtPGIP1均匀分布在整个叶片组织中,而叶柄、茎和根中的AtPGIP1主要分布在表皮细胞和维管束中。这和离子芥的研究一致。AtPGIP1空间分布做为一道天然的物理屏障抵御病源物的入侵。
7.NO增加植物对茄匐柄霉的抗性。T-DNA插入Atnos1基因的拟南芥突变体中,一氧化氮合成酶(nitirc oxide systhase,NOS)活性下降,NO释放减少。和拟南芥野生型相比,Atnos1突变体对于病原菌的生物胁迫更敏感。用NO供体SNP可以减缓Atnos1突变体对于病原菌的生物胁迫的敏感性。在植株水平上,Atnos1突变体在病原菌茄柄霉菌侵染后几乎不形成过敏性病斑。在组织水平上,病原菌茄匐柄霉在Atnos1突变体中繁殖更快。在细胞水平上,NO可以调控植物防卫反应中抗氧化酶含量。在信号分子水平上,突变体中游离态的SA含量比野生株的低。在基因水平上,突变体的一些抗病基因—AtPGIP1的表达量都比野生株的低。由此可见,NO作为信号分子,参与了对植物抗病胁迫的正调控。
总之,病原菌,低温,SA和NO对离子芥PGIP诱导表明PGIP在植物的抗性反应中具有重要作用。同时,在植物的抗病防卫反应中,NO作为信号分子增强植物的抗病性并调控AtPGIP1基因的表达。
Polygalacturonase-inhibiting proteins(PGIPs)are plant proteins believed to play an important role in the defence against plant pathogen fungals.PGIPs specifically inhibit fungal PGs,which reduce the hydrolytic activity of polygalacturonases(PGs),limit the growth of plant pathogens,and also elicit defence responses in plant.In our paper,PGIP gene was cloned from Chorispora bungeana(Chorispora bungeana Fisch,et.Mey,Chorispora bungean)(CbPGIP1). Immunofluorescence localizes the spatial distribution.Quantitative real-time PCR and western blotting characterized CbPGIP1 expression profile in response to biotic and abiotic stresses.In addition,Nitric oxide(NO)has emerged as a key molecule involved in the interaction between plant and pathogens.In our paper,the signaling functions of ON in plant fungal pathogen stress and the regulation of AtPGIP1 was studied using Arabidopsis thaliana wild type and Atnos1 mutant plants.The results were showed as following:
1.The cloned full-length CbPGIP1 cDNA from Chorispora bungeana was 1203 bp.The cDNA encoded a protein of 332 amino acids with a calculated molecular weight of about 36.9 KD and with an isoeleetric point(pI)of 6.93. Prediction of amino acid analysis shows that the full-length cDNA of CbPGIP1 contains tepical LRR(leucine-rich repeat)domain.The protein exhibits closest homology to a subgroup of AtPGIP1 and we thus refer to it as CbPGIP1.
2.CbPGI1P-GFP fusion protein was expressed transiently in epidermal cells of onions by microprojectile bombardment.The result showed that the intracellular distribution of fusion protein was located in cytosol,indicating no specific cellular localization of CbPGIP1 to any cellular compartment.
3.Quantitative real-time PCR and western blotting showed that Stemphylium solani,SA(salicylic acid)notably induced the CbPGIP1 gene expression,indicating that CbPGIP1 plays key role in plant disease resistance.
4.The results of immunofluorescence suggested that the CbPGIP1 in leaf was evenly localized in all tissues but the CbPGIP1 in stem and root was mainly localized in the epidermis and vascular bundle.Further study suggests that CbPGIP1 has higher levels in leaves and stems than in roots.Perhaps such impediments of the spatial distribution can be major factors as host resistance that places severe restrictions on the host range for a pathogen.
5.4℃,-4℃and NO all can induce the CbPGIP1 gene expression,surgesting that CbPGIP1 would play important role in abiotic stresses.Low temperature like other stresses can increase susceptibility to diseases,so cold induction of CbPGIP1 might provide protection from infections.In addition,NO induced the expression of CbPGIP1,indicating that NO may take part in CbPGIP1 expression in Chorispora bungeana.
6.The results of immunofluorescence showed that the AtPGIP1 in leaf was evenly localized in all tissues but the AtPGIP1 in petiole,stem and root was mainly localized in the epidermis and vascular bundle.This is consistent with the result of Chorispora bungeana.Those results indicate that the spatial distribution of AtPGIP1 as a physical barrier against the attack of fungal pathogen.
7.NO increased the plant resistance to Stemphylium solani.Atnos1 mutant was homozygous mutant line with a T-DNA insertion in the first exon of Atnos1 gene.NOS activity in Atnos1 was about 25%of wild type under normal condition. Atnos1 was identified to have impaired NO production.Compared with wild type plants,Atnos1 mutant plants were more sensitive to disease stress.Treatment of Atnos1 mutant plants with SNP(a NO donor)decreased the sensitive to disease stress.There were not hypersensitive response-like lesions in Atnos1 mutant plants infected by Stemphylium solani SS31.The spread of Stemphylium solani was more quickly in Atnos1 mutant plants than wild type plants.In addition,NO can regulate the antioxidative enzymes in plant defence.Compared with wild type plants,the extent of SA and AtPGIP1 were both lower in the mutant infected by Stemphylium solani.In a word,as a singal molecular,NO plays an important role in plant diease resistance.
In a word,the induction of PGIP by infection,low temperature,SA and NO indicates that PGIP has a role in defence of subnival plant Chorispora bungeana.In addition,NO is a signal molecule involed in the enhancement of disease resistance and regulation of AtPGIP1 expression.
引文
Abu-Goukh AA, Labavitch JM (1983) The in vivo role of 'Bartlett' pear fruit polygalacturonase inhibitors.Physiol Plant Pathol 23:123 - 135
Aguero C, Dandekar A, and Meredith C (2003) Evaluation of Tolerance to Pierce's Disease and Botrytis in Transgenic Plants of Vitis vinifera L. Expressing the Pear pgip Gene. ASEV 54th Annual Meeting Reno, Nevada
Ahsan, N., Yoon, H., S., Jo, J (2005) Molecular cloning of a BcPGIP cDNA from Brassica campestris and its expression to several stresses. Plant Sci. 169: 1081 -1089
Albersheim P, Anderson AJ (1971) Proteins from plant cell walls inhibit polygalacturonases secreted by plant patho- gens. Proc Natl Acad Sci USA. 68: 1815-1819
An LZ, Liu YH, Feng GN, Feng HY, Chen G (2000b) Studies on characteristics of communities in the source region of Urumqi River. Acta Bot. Boreal-Occident. Sinicb 20:80-86
An LZ, Liu YH, Feng HY, Feng GN, Chen GD (2000a) Studies on characteristics of element contents of altifrigetic subnival vegetation at the source area of Urumqi River. Acta Bot. Boreal-Occident.Sinica 20:80-86
An, L., Zh., Liu, Y.,H., Feng, G.,N., Feng, H.,Y., Feng, G.,N., Chen, G.,D(2000) Studies on the characteristics of element contents of altifrigetic subnival vegetation at the source area of Urumchi river. Acta Bot. Boreal.-Occident. Sin. 20:1063-1069
Ayitu, R., Dunyan, T. Zhi J., Li, Y (1998) The relationship between the structure of vegetative organs in Chorispora bungeana and its environment. J.Xinjiang Agric. Univ. 21: 273 -277
Beligni M V, Lamattina L (2000) Nitric oxide stimulates seed germination and de -etiolation, and inhibits hypocotyl elongation, three light inducible responses in plants.Planta 210:215-221
Berger DK, Oelofse D, Arendse MS, Du Plessis E, Dubery IA (2000) Bean polygalacturonase inhibiting protein-1 (PGIP-1) inhibits polygalacturonases from Stenocarpella maydis. Physiol. Mol. Plant P 57: 5 - 14
Bergmann C, Ito Y, Singer D, Albersheim P, Darvill AG, Benhamou N, Nuss L, Salvi G, Cervone F, De Lorenzo G (1994) Polygalacturonase-inhibiting protein accumulates in Phaseolus vulgaris L. in response to wounding, elicitors and fungal infection. Plant J 5: 625-634
Bézier A, Lambert B, Baillieul F (2002) Study of defense-related gene expression in grapevine leaves and berries infected with Botrytis cinerea. Eur. J. Plant Pathol 108: 111-120
Boccara, M., Mills, C. E., Zeier, J., Anzi, C., Lamb, C., Poole, R. K. and Dellednne, M (2005) Flavohaemoglobin HmpX from Erwinia chrysanthemi confers nitrosative stress tolerance and affects the plant hypersensitive reaction by intercepting nitric oxide produced by the host. Plant J. 43: 226 - 237
Bonivento, D., Pontiggia, D., DiMatteo, A., Fernandez-Recio,J., Salvi,G., Tsernoglou, D., Cervone,F., DeLorenzo, G., and Federici, L (2008) Crystal structure of the endopolygalacturonase from the phytopathogenic fungusColletotrichum lupini and its interaction with polygalacturonase-inhibiting proteins. Proteins: Structure, Function, and Bioinformatics 70: 294 - 299
Bowler C, Montagu MV, lrge D (1992) Superoxide disnimtase and stress tolerance. Annv Rev Plant Physiol Plant Mol Biol.43:83- 116
Bowler C., Alliote T., De Loose M., Van Montagu M., Inzé D (1989) The induction of manganese Superoxide dismutase in reponse to stress in Nicotiana plumbaginifolia. EMBO J 8:31-38
Bradford, M.M (1976) A rapid and sensitive method for quantitative of microgram quantities of protein using the principle of protein-dye binding. Anal. Biochem.72:248 - 254
Buza NL, Krinitsyna AA, Protsenko MA, and Vartapetyan VV (2004) Role of the Polygalacturonidase Inhibitor Protein in the Ripening of Apples and Their Resistance To Monilia fructigena,a Causative Agent of Fruit Rot. Applied Biochemistry and Microbiology 40: 89 - 92
Cervone, F., De Lorenzo, G., Pressey, R., Darvill, A., G., and Albersheim, P (1990) Can phaseolus PGIP inhibit pectic enzymes from microbes and plants? Phytochemistry 29:447 - 449
Chang, J., F., Fu, X., Y., An, L.,Zh., Xu, Sh.J.,Wang, J.,H.,Zhang,M.,X., Feng, H.,Y., Chen, T (2006) Properties of cellular ubiquinone and stress-resistance in suspension-cultured cells of Chorispora bungeana during early chilling. Environ. Exp. Bot. 57:116 - 122
Chen Z, Silva H, Klessig D F (1993) Active oxygen species in the induction of plant systemic acquired resistance by salicylic acid.Science 262:883-1886
Clarke A, Desikan R, Hurst R D , Hancock J T, Neill S J (2000) NO way back : nitric oxide and programmed cell death in Arabidopsis thaliana suspension cultures. Plant J 24:667-677
D'Ovidio R, Mattei B, Roberti S, Bellincampi D (2004) Polygalacturonases, polygalacturonase-inhibiting proteins and pectic oligomers in plant-pathogen interactions. Biochimica et Biophysica Acta 1696:237 - 244
De Lorenzo G, and Ferrari S (2002) Polygalacturonase-inhibiting proteins in defense against phytopathogenic fungi. Curr. Opin. in Plant Biol 5: 295 - 299
Delaney T P , Uknes S , Vernooij B , et al (1994) A central role of salicylic acid in plant disease resistance. Science 266: 1247-1250
Delledonne M, Xia YJ, Dixon R A, Lamb C (1998) Nitric oxide functions as a signal in plant disease resistance. Nature 394: 585-588
Delledonne, M., Zeier, J., Marocco, A. and Lamb, C (2001) Signal interaction between nitric oxide and reactive oxygen intermediates in the plant hypersensitive disease resistance response. Proc. Natl Acad. Sci.USA. 98: 13454-13459
Desiderio A, Aracri B, Leckie F, Mattei B, Salvi G, Tigelaar H, Van Roekel JS, Baulcombe DC, Melchers LS, De lorenzo G, and Cervone F (1997) Polygalacturonase-inhibiting proteins (PGIPS) with different specificities are expressed in Phaseolus vulgaris. Mol. Plant- Microbe Interact 10: 852 - 860
Devoto, A., Leckie, F., Lupotto, E., Cervone, F., and De Lorenzo, G (1998) The promoter of a gene encoding a polygalacturonase-inhibiting protein of Phaseolus vulgaris L. is activated bywounding but not by elicitors or pathogen infection. Planta 205:165 - 174
Di C., X., Zhang, M., X., Xu, Sh., J., Cheng, T., An, L., Zh (2006) Role of Poly-galacturonase Inhibiting Protein in Plant Defense. Crit. rev. microbial 32: 91 -100
Di Matteo, A., Federici, L., Salvi, G., Johnson, K. A., Savino, C., De Lorenzo, G., Tsernoglou, D., and Cervone, F (2003) The crystal structure of polygalacturonase-inhibiting protein (PGIP), a leucine-rich repeat protein involved in plant defense. Proc. Natl. Acad. Sci. U.S.A. 100:10124 - 10128
Di Matteo,A., Bonivento, D., Tsernoglou, D., Federici, L., Cervone, F (2006) Polygalacturonase-inhibiting protein (PGIP) in plant defence: a structural view. Phytochemistry 67:528 - 533
D'Ovidio R, Raiola A, Capodicasa C., Devoto A, Pontiggia D, Roberti S, Galletti R, Conti E, O'Sullivan DM and De Lorenzo G (2004) Characterization of the complex locus of Phaseolus vulgaris encoding polygalacturonase-inhibiting proteins (PGIPs) reveals sub-functionalization for defense against fungi and insects. Plant Physiol 135:2424 - 2435
Durner J , Wendehenne D , Klessig D F (1998) Defense gene induction in tobacco by nitric oxide , cyclic GMP , and cyclic ADP- ribose. Proc Natl Acad Sci USA 95: 10328-10333
Durner J, Klessig D F (1999) Nitric oxide as a signal in plants. Current Opinion in Plant Biology 2:369-374
Elad Y, Evensen K (1995) Physiological aspects of resistance to Botrytis cinerea.Phytopathology 85:637 - 643
Favaron F, Destro T and D'Ovidio R (2000) Transcript Accumulation of Polygalacturonase Inhibiting Protein (PGIP) Following Pathogen Infections in Soybean. Journal of Plant Pathol 82: 103-109
Favaron, F., D'ovidio, R., Porceddu, E., Alghisi, P(1994) Purification and molecular characterization of a soybean polygalacturonase-inhibiting protein. Planta 195: 80 - 87
Federici L, Di Matteo A, Fernandez-Recio J, Tsernoglou D, and Cervone F (2006) Polygalacturonase inhibiting proteins: players in plant innate immunity? Trends Plant Sci 11: 65 - 70
Ferrari S, Galletti R, Vairo D, Cervone F and De Lorenzo G(2006) Antisense Expressionof the Arabidopsis thaliana AtPGIPl Gene Reduces Polygalacturonase-inhibiting Protein Accumulation and Enhances Susceptibility to Botrytis cinerea. Mol. Plant- Microbe Interact 19: 931 - 936
Ferrari S, Vairo D, Ausubel FM, Cervone F, and De Lorenzo G (2003) Tandemly Duplicated Arabidopsis Genes That Encode Polygalacturonase-inhibiting Proteins Are Regulated Coordinately by Different Signal Transduction Pathwaysin Response to Fungal Infection. Plant Cell 15:93 -106
Fish WW (2005) Polygalacturonase-inhibiting protein activity in cantaloupe fruit as a function of fruit maturation and tissue origin. European Journal of Plant Pathology 111: 67-76
Fridovich I (1995) Superoxide radical and superoxide dismutases. Annu. Rev. Biochem 64: 97-112
Fu XY, Chang JF, An LZ, Zhang MX, Xu SJ,Chen T, Liu YH, Xin H,
Wang JH (2006) Association of the cold-hardiness of Chorispora bungeana with the distribution and accumulation of calcium in the cells and tissues. Environmental and Experimental Botany 55: 282-293
Gadre RV, Van Driessche G, Van Beeumen J, Bhat MK (2003) Purification, characterisation and mode of action of an endo-polygalacturonase from the psychrophilic fungus Mucor .avus. Enzyme Microb Technol 32:321-30
Gaffney T , Friedrich L , Vernooij B , et a 1 (1993) Requirement of salicylic acid for the induction of systemic acquired resistance. Science 261: 754-756
Gardes M, Bruns T D(1993) ITS primer with enhanced specificity for basidiomycetes:Application to the identification of mycorrhizae and rusts. MolEcol2:113-118
Gazendam I, Oelofse D, Berger DK (2004) High-level expression of apple PGIP1 is not sufficient to protect transgenic potato against Verticillium dahliae. Physiological and Molecular Plant Pathology 65: 145-155
Gomathi V and Gnanamanickam SS (2004) Polygalacturonase-inhibiting proteins in plant defense. Current Science 87: 1211-1217
Guo FQ, Okamoto M, and Crawford NM (2006) Identication of a Plant Nitric Oxide Synthase Gene Involved in Hormonal Signaling. Science 302: 100-103
Guo FX, Zhang MX, Chen Y, Zhang WH, Xu SJ, Wang JH,An LZ (2006) Relation of several antioxidant enzymes to rapid freezing resistance in suspension cultured cells from alpine Chorispora bungeana. Cryobiology 52: 241-250
Hadfield KA and Bennett AB (1998) Polygalacturonases: many genes in search of a function. Plant physiol 117: 337 - 343
Hahn MG, Bucheli P, Cervone F, Doares SH, O'Neill RA, Dan ill RA and Albersheim P (1989) The role of cell wall constituents in plant-pathogen interactions. Plant-Microbe Interact 13:131-181
Hammond-Kosack KE, Jones JDG (1997) Plant disease resistance genes. Annu. Rev. Plant Physiol. Plant Mol. Biol 48: 575 - 607
He Y K, Tang R H, Hao Y, et al (2004) Nitric oxide represses the Arabidopsis floral transition. Science 305:1968-1971
Henrik U. Stotz, Ann 1. T. Powell, Susan E. Damon, 1. Carl Greve, Alan B. Bennett, and John M (1993) Labavitch.Molecular Characterization of a Polygalacturonase Inhibitor from Pyrus communis 1. cv Bartlett'. Plant Physiol 102:133-138
Hoffman RM and Turner JG (1982) Partial purification of proteins from Pea leaflets that inhibit Ascophyta Pisi endopolygalaturonase. Physiol Plant Pathol 20: 173-187
Hoffman RM and Turner JG (1984) Occurence and specificity of an endopolygalacturonase inhibitor in Pisum sativum. Physiol Plant Pathol 24: 49 -59
James JT, Dubery IA (2001) Inhibition of polygalacturonase from Verticillium dahliae by a polygalacturonase inhibiting protein from cotton .Phytochemistry 57:149-156
Jang S, Lee B, Kim C, Kim SJ, Yim J, Han JJ, Lee S, Kim SR, An G.(2003) The OsFORl gene encodes a polygalacturonase-inhibiting protein (PGIP) that regulates floral organ number in rice. Plant Mol. Biol. 53: 357 - 369
Janni M, Sella L, Favaron F, Blechl AE, De Lorenzo G, and D'Ovidio R (2008) The Expression of a Bean PGIP in Transgenic Wheat Confers Increased Resistance to the Fungal Pathogen Bipolaris sorokiniana. Mol. Plant- Microbe Interact 21: 171-177
Jones TM, Anderson AJ, albersheim P (1972) Host-pathogen interactions.IV. Studies on the polysaccharide-degrading enzymes secreted by Fusarium oxysporum f.Sp.lycopersici.Physiol.plant pathol 2:153-166
Jones DA, and Jones JDG (1997) The role of leucine-rich repeat proteins in plant defense. Adv Bot Res. 24: 89-166
Jones J D (2001) Putting knowledge of plant disease resistance genes to work. Curr. Opin. Plant Biol. 4:281-287
Joubert DA, Kars I, Wagemakers L, Bergmann C, Kemp G, Vivier MA, van Kan JA(2007) A polygalacturonase-inhibiting protein from grapevine reduces the symptoms of the endopolygalacturonase BcPG2 from Botrytis cinerea in Nicotiana benthamiana leaves without any evidence for in vitro interaction. Mol. Plant- Microbe Interact 20: 392 - 402
Karr ALL and Albersheim P (1970) Polysacride-degrading enzymes are unable to attack plant cell walls without prior action by a "wall-modifying enzymes". Plant Physiol 46: 69 - 80
Kemp G, Bergmann CW, Clay R, Van der Westhuizen AJ, Pretorius ZA (2003) Isolation of polygalacturonase-inhibitingprotein (PGIP) from wheat. Mol. Plant-Microb. Interact 16: 955-961
Kessmann H, Ward E, Ryals J (1994) A central role of salicylic acid in plant-disease resistance. Science 266:1247-1250
Klepper L (1975) Evolution of nitrogen oxide gases from herbicidet reated plant tissues. WSSA Abstracts 184:70
Klepper L (1979) Nitric oxide (NO) and nitrogen dioxide ( NO_2) emissions from herbicide-treated soybean plants. Atmos Environ 13: 537 - 542
Klepper L (1991) NOx evolution by soybean leaves treated with salicylic acid and selected derivatives. Pestic Biochem Phys39: 43 -48
Klessig D F , Durner J , Noad R , Navarre D A , Wendehenne D , Kumar D , Zhou J M, Shah J , Zhang S Q , Kachroo P , Trifa Y, Pontier D , Lam E , Silva H (2000) Nitric oxide and salicylic acid signaling in plant defense. Proc Natl Acad Sci USA 97:8849-8855
Kornfeld R, Kornfeld S (1985) Assembly of asparagine-linked oligosaccharides.Ann Rev Biochem 54: 631-64
Kumar S, Tamura K, Nei M (2004) MEGA 3: Integrated software for molecular evolutionary genetics analysis and sequence alignment. Brief Bioinform 5: 150 -163
Leckie F, Mattei B, Capodicasa C, Hemmings A, Nuss L, Aracri B, De Lorenzo G., and Cervone F (1999) The specificity of polygalacturonase-inhibiting proteins (PGIP): A single amino-acid substitution in the solvent-exposed β-strand/β-turn region of the leucine-rich repeats (LRRs) confers a new recognition capability. EMBO J. 18: 2352 - 2363
Li R, Rimmer R, Yu M, Sharpe AG, Seguin-Swartz G, Lydiate D, Hegedus DD (2003) Two Brassica napus polygalacturonase inhibitory protein genes are expressed at di.erent levels in response to biotic and abiotic stresses. Planta 217: 299 - 308
Liang FS, Zhang KCH, Zhou ChJ, Kong Fn, Lia J, Wang B (2005) Cloning,characterization and expression of the gene encoding polygalacturonase-inhibiting proteins (PGIPs) of peach [prunus persica (L.) Batch]. Plant Sci 168: 481 -486
Liochev SI and Fridovich I (1994) Paraquat diaphorases in Escherichia coli, Free Radical Biology and Medicine 16:555-559
Loake G, Grant M (2007) Salicylic acid in plant defence-the players and protagonists. Curr Opin Plant Biol. 10: 466 - 72
Machinandiarena MF, Olivieri FP, Daleo GR, Oliva C (2001) Isolation and characterization of a polygalacturonase inhibiting protein from potato leaves: accumulation in response to salicylic acid, wounding and infection. Plant Physio. Bioch. 39:129-136
Magalhaes J R, Monte D C , Durzan D (2000) Nitric oxide and ethylene emission in Arabidopsis thaliana. Physiol Mol Biol Plant 6:117- 127
Mattei B, Galletti R, Manfredini C, Pontiggia D, Salvi G, Spadoni S, Caprari C, Ferrari S, Bellincampi D, Cervone F, and De Lorenzo G (2005) Recognition and signalling in the cell wall: The case of endopolygalacturonase,PGIP and oligogalacturonides.Plant Biosystems 139: 24 - 27
Mehli L, Schaart JG, Kjellsen TD, Hong Tran D, Salentijn EMJ, Schouten HJ, Iversen T-H (2004) A gene encoding a polygalacturonase- inhibiting protein (PGIP) shows developmental regulation and pathogen-induced expression in strawberry. New Phytol 163: 99 - 110
Milagros F. Machinandiarena, Florencia P. Olivieri, Gustavo R. Daleo, Claudia R. Oliva (2001) Isolation and characterization of a polygalacturonase-inhibiting protein from potato leaves. Accumulation in response to salicylic acid, wounding and infection. Plant Physiol. Biochem 39:129-136
Millar A H, Day D A (1996) Nitric oxide inhibits the cytochrome oxidase but not the alternative oxidase of p lantmitochondria. FEBS Lett 398: 155-158
Navarre D A , Wendehenne D , Durner J , Noad R , Klessig D F (2000) Nitric oxide modulates the activity of tobacco aconitase. Plant Physiol 122:573-582
Neill S J, Desikan R, Clarke A, Hancock J T (2002) Nitric oxide is a novel component of abscisic acid signaling in stomatal guard cells. Plant Physio 1128:13-16
Neill SJ, Desikan R, Hancock JT (2003) Nitric oxide signalling in plants. New Phytol. 159:11-35
O'Donnell PJ, Jones JB, Antoine FR, Ciardi J and Klee H.J (2001) Ethylene-dependent salicylic acid regulates an expanded cell death response to a plant pathogen. Plant J 25: 315 - 323
Okkes A.and Barbaros N (2003 ) Antifreeze proteins in higher plants. Phytochemistry 64: 1187-1196
Powell AL, van Kan J, ten Have A, Visser J, Greve LC, Bennett AB and Labavitch JM (2000) Transgenic Expression of Pear PGIP in Tomato Limits Fungal Colonization. Mol. Plant-Microbe Interact 13: 942 - 950
Raskin I (1992) Salicylic acid, A new plant hormone.Plant Physiol 99: 799-803
Richter A,de Kathen A,de Lorenzo G,Briviba K,Hain R,Ramsay Jacobsen GH,Kiesecker H(2006)Transgenic peas(Pisum sativum)expressing polygalacturonase inhibiting protein from raspberry(Rubus idaeus)and stilbene synthase from grape(Vitis vinifera).Plant Cell Rep.25:1166-1173
Ruan Q P,ZHOU L,LIU Y(2000)小麦在与水杨酸诱导的应答过程中PGIP的积累[J].Chin J Appl Environ Biol(应用与环境生物学报)6:313-316
Ruan Q P,ZHOU L,LIU Y(2000)小麦在损伤、真菌感染应答过程中内源PGIP 积累的研究初报.ACTA PHYTO PA THOLO G ICA S IN ICA(植物病理学报)30:372-373
Ruan Q P,ZHOU L,ZHENG Y Q((2000)PGIP在植物抗病方面的研究进展[J].Chinese Bulletin of Botany(植物学通报)17:60-63
Schaart JG.,Mehli L,Schouten HJ(2005)Quantification of allele-specific expression of a gene encoding strawberry polygalacturonase-inhibiting protein (PGIP)using Pyrosequencing(TM).Plant J.41:493-500
Shackel KA,de la Paz Celorio-Mancera M,Ahmadi H,Greve LC,Tenber LR,Backus EA,Labavitchl JM(2005)Micro-Injection of Lygus Salivary Gland Proteins to Simulate Feeding Damage in Alfalfa and Cotton Flowers.Archives of Insect Biochemistry and Physiology 58:69-83
Shanmugam V(2005)Role of extracytoplasmic leucine rich repeat proteins in plant defence mechanisms.Microbio.Res 160:83-94
Shirasu K,Nakajima H,Rajasekhar VK,Dixon RA and Lamb C(1997)Salicylic acid potentiates an agonistdependent gain control that amplifies pathogen signals in the activation of defense mechanisms.Plant Cell 9:261-270
Stotz HU,Contos JJ,Powell AL,Bennett AB,Labavitch JM(1994)Structure and expression of an inhibitor of fungal polygalacturonases from tomato.Plant Mol Biol 25:607-617
Szankowski I,Briviba K,Fleschhut J,Schonherr J,Jacobsen H J,Kiesecker H (2003) Transformation of apple (Malus domestica Borkh.)with the stilbene synthase gene from grapevine (Vitis vinifera L.)And a PGIP gene from kiwi(Actinidia deliciosa). Plant Cell Rep 22:141-149
Tamura M. Gao M, Tao R, Labavitch JM, Dandekar AM (2004) Transformation of persimmon with a pear fruit polygalacturonase inhibiting protein (PGIP) gene . Scientia Horticulturae 103: 19-30
Taylor RJ, Secor, G A (1988) An improved diffusion assay for quantifying the polygalacturonase content of Erwinia culture filtrates. Phytopathology 78: 1101 -1103
Thompson JD, Higgins DG, Gibson TJ (1994) CLUSTAL W: improving the sensitivity of progressive multiple sequence alignment through sequence weighting, position specific gap penalties and weight matrix choice. Nucleic Acids Res. 22:4673 - 4680
Toubart P, Desiderio A, Salvi G, Cervone F, Daroda L, De Lorenzo G, Bergmann C, Darvill AG, and Albersheim P (1992) Cloning and characterization of the gene encoding the endopolygalacturonase-inhibiting protein (PGIP) of Phaseolus vulgaris L. Plant J. 2: 367-373
Turner JG and Hoffman RM (1985) Effect of the PGIP from Pea on the hydrolysis of Pea cell walls by the endo-PG from Ascochyta pisi. Plant Pathol 34: 54 - 60
Tzeng D D, Devay J E (1993) Role of oxygen radicals in p lant disease development. Adv. Plant PathologylO: 1 - 34
White RF (1979) Acetyl salicylic acid (aspirin) induces resistance to tobacco mosaic virus in tobacco. Virology 99:410 -412
Worrall D, Elias L, Ashford D, Smallwood M, Sidebottom C, Lillford P, Telford J, Holt C, and Bowles D (1998) Acarrot leucine-rich-repeat protein that inhibits ice recrystallization. Science 282:115 - 117
Yang CH L,CHEN ZH L (1997) 高等植物的LRR蛋白结构与功能.生物工程进展17:43-47
Yao CL,William S.Conway,Ren RH,David Smithl,Gavin S.Ross and Carl E.Sams(1999)Gene encoding polygalacturonase inhibitor in apple fruit is developmentally regulated and activated by wounding and fungal infection.Plant Molecular Biology 39:1231-1241
Yuan YB,Liu CL,Ju ZG(1997)水杨酸对苹果叶片中过氧化氢的调节及其机制.园艺学报24:20-24
Zhang XY(2000)水杨酸诱导植物抗病性机制的研究进展.河北林果研究3:288-291
Zhang TG,Liu YB,Xue LG,Xu SJ,Chen T,Yang TW,Zhang LJ,An LZ(2006)Molecular cloning and characterization of a novel MAP kinase gene in Chorispora bungeana.Plant Physio.Bioch.44:78-84
Zhao Z,Chen G,Zhang C(2001)Interaction between reactive oxygen species and nitric oxide in drought-induced abscisic acid synthesis in root tips of wheat seedlings.Aust J Plant Physiol 28:1055-1061
Zheng Y Q,YANG Z J,LI J W,ZHOU L,WU W L(1994)小麦内切多聚半乳糖醛酸酶抑制蛋白的分离纯化研究.ACTA BIOCHIMICA etBIOPGYSIOA SINICA(生物化学与生物物理学报)26:555-558
Zhou L,LI J W,ZHENG Y Q,LU Y Q(1996)还原糖法测定多聚半乳糖醛酸酶抑制蛋白活性的研究[J].Journal of Sichuan University(四川大学学报)33:311-315
ZHOU L,LI J W,ZHENG Y Q,YU L(1997)Endo-PG诱导培养小麦细胞PGIP 产生的研究.Chinese Biochemical Journal(生物化学杂志)13:365-367
Zhou L,Lui Y,Li J W,Yang X H,CAO Y(1998)小麦多聚半乳糖醛酸酶抑制蛋白对几种病原真菌抑制作用的研究.ACTA PHYTO PA THOLO GICA S IN ICA(植物病理学报)28:107-112
Zhou L,RUAN Q P,LI J W,ZHANG J(2000)小麦PGIP的免疫组织细胞化学定位.Journal of Sichuan University(Natural Science Edition)(四川大学学报(自然科学版))37:916-923