番茄谷氧还蛋白基因SlGRX1的克隆与功能分析
|
摘要
活性氧(Reactive oxygen species, ROS)是植物体内正常代谢的信号小分子,在植物生长发育和抗逆反应的信号转导中具有重要作用。但是,过多的活性氧积累会严重影响植物的生长发育,造成氧化胁迫。谷氧还蛋白是一类小分子的氧化还原蛋白,能够调节蛋白质的氧化还原状态,从而维持蛋白质的活性,在植物抗氧化胁迫及生长发育反应中具有重要作用。近年来,越来越多的谷氧还蛋白基因被克隆,其分子功能被阐明。本研究从番茄中克隆得到了一个新的CGFS类谷氧还蛋白基因,并在番茄中利用病毒诱导的基因沉默技术以及在拟南芥中利用超量表达技术研究了其在植物抗氧化、干旱及高盐胁迫中的作用。
根据已发表的水稻和拟南芥的CGFS类谷氧还蛋白基因的序列信息,采用同源克隆和电子克隆的方法,从番茄中分离得到了一个新的CGFS类谷氧还蛋白基因,命名为S1GRX1。该基因是从番茄中分离得到的第一个谷氧还蛋白基因,编码的蛋白质有292个氨基酸,其结构具有谷氧还蛋白基因家族保守的结构域和典型的CGFS催化位点。
通过Southern杂交分析表明,SlGRX1在番茄在基因组中以单拷贝的形式存在。采用Real time PCR的方法分析了SlGRX1基因在番茄中的表达模式,发现该基因能够在番茄的根、茎、叶片和花瓣中表达,其中叶片中的表达量最高,根和茎中的表达量相对较低,同时也证明该基因能够受外界逆境条件如氧化、干旱和高盐的诱导表达。
构建了SlGRX1和GFP的融合植物表达载体,将SlGRX1融合到GFP蛋白的C端以及N端,采用农杆菌浸润法,在本氏烟叶片的表皮细胞瞬时表达重组蛋白,通过激光共聚焦显微镜观察SlGRX1蛋白的亚细胞定位,结果表明,SlGRX1能够定位在植物细胞的细胞核和细胞质中。
构建了基于双生病毒卫星DNAβ的SlGRX1的沉默载体,并将该载体导入农杆菌菌株EHA105中,以中国番茄黄化曲叶病毒(Tomato yellow leaf curl China virus, TYLCCNV)为辅助病毒,在番茄中沉默了SlGRX1基因,以接种TYLCCNV和空载体的番茄作为对照。SlGRX1基因沉默番茄与对照相比对氧化和盐胁迫更为敏感,主要表现为叶绿素降解速度加快；在干旱胁迫下,SlGRX1基因沉默番茄与对照相比则更容易出现黄化和萎蔫的症状,叶片内的相对含水量有明显的下降。这些结果表明,SlGRX1基因在植物的抗氧化、干旱和高盐反应中具有重要的调控作用。
将SlGRX1基因构建到植物表达载体pBin438中,采用农杆菌介导的花序浸蘸法对模式植物拟南芥进行遗传转化,获得了2个单拷贝的纯合T3代转基因株系Y1、Y2,同时将pBin438空载体也转化到拟南芥中作为研究的对照。通过分析株系Y1、Y2与对照在氧化、干旱和高盐胁迫下的表型,进一步明确了SlGRX1基因的功能,无论拟南芥在离体生长条件下还是土壤生长条件下,SlGRX1基因超量表达的株系Y1、Y2对氧化、干旱和高盐的抗性都明显高于对照,说明超量表达SlGRX1基因能够提高植物对上述逆境的抗性。
为了进一步分析了SlGRX1基因在提高植物抗逆性可能的作用机理,利用Real time PCR技术分析了拟南芥抗氧化、干旱和高盐反应中的Marker基因在Y1、Y2和对照中的表达差异。结果显示,拟南芥的Apx2、Apx6和RD22的表达水平有明显的上调,表明SlGRX1基因有可能通过提Apx2、Apx6和RD22的表达水平实现其功能。
Reactive oxygen species (ROS) are redox signals essential to many physiological processes in plants. But high concentration of ROS can cause damage to macromoleces thus disrupt normal signaling in plant. Glutaredoxins (Grxs) are ubiquitous small heat-stable disulfide oxidoreductases that play a crucial role in plant response to oxidative stress. Recently, studies have extended our knowledge on the physiological and molecular functions of Grxs in plant. In the present study, a novel CGFS type Grx gene was isolated from tomato and its functions were validated using loss-of-function and gain-of-function approaches by virus induced gene silencing (VIGS) of the gene in tomato and over-expression of the gene in Arabidopsis.
Based on the sequence information of CGFS type Grxs from Arabidopsis and rice, a cDNA encoding a protein containing the consensus Grx family domain with a CGFS active site was isolated using homologue cloning and in silio cloning methods. Quantitative real-time RT-PCR (qRT-PCR) analysis revealed that SlGRX1 is expressed ubiquitously in tomato including leaf, root, stem and flower and can be induced by oxidative, drought and salt stresses.
To investigate the subcellular localization of SlGRX1 protein in plant cells, the SlGRX1 protein was fused to both C-terminus and N-terminus of GFP and the construct was transiently expressed in Nicotiana benthamiana leaf epidermal cells then monitored by capturing GFP fluorescence. Our results suggested that SlGRX1 protein was localized in both nucleus and cytoplasm.
SlGRX1 gene silencing vector DNAmβ-SlGRX1 was constructed based on the geminivirus satellite DNAmβinduced silencing system and then introduced into Agrobacterium tumefaciens strain EHA105. Agrobecterium harboring recombinant silencing vector DNAmβ-SlGRX1 and TYLCCNV (Tomato yellow leaf curl China virus) infectious clone were co-agroinoculated into tomato plants. Meanwhile, tomato plants co-agroinoculated with TYLCCNV and empty silencing vector DNAmP was used as the control. VIGS mediated silencing of SlGRX1 in tomato led to increased sensitivity to oxidative and salt stresses with decreased relative chlorophyll content, and reduced tolerance to drought stress with decreased relative water content. These results suggested that SlGRX1 was required for plant resistance to oxidative, drought and salt stresses.
The coding sequence of SlGRX1 was then cloned into the binary vector pBin438 to produce pBin438-SlGRX1 in which the SlGRX1 gene was expressed under the control of the cauliflower mosaic virus (CaMV) 35S promoter. The plasmid pBin438-SlGRX1 was then introduced into A. tumefaciens strain EHA105. Transformation of Arabidopsis was performed using the floral dip method. The empty vector pBin438 was also transformed into Arabidopsis as the control. Two independent homozygous lines (Y1, Y2) with a single copy of SlGRX1 were allowed to grow for 3 generations for further studies. We found over-expression of SlGRX1 in Arabidopsis plants enhanced lines Yl and Y2 the tolerance to oxidative, drought and salt stresses in both in vitro and in soil-grown conditions.
Expression levels of some oxidative related ascorbate peroxidase (Apx) genes Apx2, Apx3, Apx4, Apx5, Apx6, and well-known drought/salt stress marker genes RD29A, RD22 and RAB18 in Y1,Y2 and the control plants were analyzed by qRT-PCR to explore the possible mechanism of the enhanced abiotic stress tolerance in SlGRX1 over-expression plants. We found that over-expression of the SlGRX1 gene in Arabidopsis can activate the expression of the Arabidopsis Apx2 and Apx6, indicating that the SlGRX1 gene may positively regulate the Apx2 and Apx6 genes to improve plants resistance to oxidative stress.
根据已发表的水稻和拟南芥的CGFS类谷氧还蛋白基因的序列信息,采用同源克隆和电子克隆的方法,从番茄中分离得到了一个新的CGFS类谷氧还蛋白基因,命名为S1GRX1。该基因是从番茄中分离得到的第一个谷氧还蛋白基因,编码的蛋白质有292个氨基酸,其结构具有谷氧还蛋白基因家族保守的结构域和典型的CGFS催化位点。
通过Southern杂交分析表明,SlGRX1在番茄在基因组中以单拷贝的形式存在。采用Real time PCR的方法分析了SlGRX1基因在番茄中的表达模式,发现该基因能够在番茄的根、茎、叶片和花瓣中表达,其中叶片中的表达量最高,根和茎中的表达量相对较低,同时也证明该基因能够受外界逆境条件如氧化、干旱和高盐的诱导表达。
构建了SlGRX1和GFP的融合植物表达载体,将SlGRX1融合到GFP蛋白的C端以及N端,采用农杆菌浸润法,在本氏烟叶片的表皮细胞瞬时表达重组蛋白,通过激光共聚焦显微镜观察SlGRX1蛋白的亚细胞定位,结果表明,SlGRX1能够定位在植物细胞的细胞核和细胞质中。
构建了基于双生病毒卫星DNAβ的SlGRX1的沉默载体,并将该载体导入农杆菌菌株EHA105中,以中国番茄黄化曲叶病毒(Tomato yellow leaf curl China virus, TYLCCNV)为辅助病毒,在番茄中沉默了SlGRX1基因,以接种TYLCCNV和空载体的番茄作为对照。SlGRX1基因沉默番茄与对照相比对氧化和盐胁迫更为敏感,主要表现为叶绿素降解速度加快；在干旱胁迫下,SlGRX1基因沉默番茄与对照相比则更容易出现黄化和萎蔫的症状,叶片内的相对含水量有明显的下降。这些结果表明,SlGRX1基因在植物的抗氧化、干旱和高盐反应中具有重要的调控作用。
将SlGRX1基因构建到植物表达载体pBin438中,采用农杆菌介导的花序浸蘸法对模式植物拟南芥进行遗传转化,获得了2个单拷贝的纯合T3代转基因株系Y1、Y2,同时将pBin438空载体也转化到拟南芥中作为研究的对照。通过分析株系Y1、Y2与对照在氧化、干旱和高盐胁迫下的表型,进一步明确了SlGRX1基因的功能,无论拟南芥在离体生长条件下还是土壤生长条件下,SlGRX1基因超量表达的株系Y1、Y2对氧化、干旱和高盐的抗性都明显高于对照,说明超量表达SlGRX1基因能够提高植物对上述逆境的抗性。
为了进一步分析了SlGRX1基因在提高植物抗逆性可能的作用机理,利用Real time PCR技术分析了拟南芥抗氧化、干旱和高盐反应中的Marker基因在Y1、Y2和对照中的表达差异。结果显示,拟南芥的Apx2、Apx6和RD22的表达水平有明显的上调,表明SlGRX1基因有可能通过提Apx2、Apx6和RD22的表达水平实现其功能。
Reactive oxygen species (ROS) are redox signals essential to many physiological processes in plants. But high concentration of ROS can cause damage to macromoleces thus disrupt normal signaling in plant. Glutaredoxins (Grxs) are ubiquitous small heat-stable disulfide oxidoreductases that play a crucial role in plant response to oxidative stress. Recently, studies have extended our knowledge on the physiological and molecular functions of Grxs in plant. In the present study, a novel CGFS type Grx gene was isolated from tomato and its functions were validated using loss-of-function and gain-of-function approaches by virus induced gene silencing (VIGS) of the gene in tomato and over-expression of the gene in Arabidopsis.
Based on the sequence information of CGFS type Grxs from Arabidopsis and rice, a cDNA encoding a protein containing the consensus Grx family domain with a CGFS active site was isolated using homologue cloning and in silio cloning methods. Quantitative real-time RT-PCR (qRT-PCR) analysis revealed that SlGRX1 is expressed ubiquitously in tomato including leaf, root, stem and flower and can be induced by oxidative, drought and salt stresses.
To investigate the subcellular localization of SlGRX1 protein in plant cells, the SlGRX1 protein was fused to both C-terminus and N-terminus of GFP and the construct was transiently expressed in Nicotiana benthamiana leaf epidermal cells then monitored by capturing GFP fluorescence. Our results suggested that SlGRX1 protein was localized in both nucleus and cytoplasm.
SlGRX1 gene silencing vector DNAmβ-SlGRX1 was constructed based on the geminivirus satellite DNAmβinduced silencing system and then introduced into Agrobacterium tumefaciens strain EHA105. Agrobecterium harboring recombinant silencing vector DNAmβ-SlGRX1 and TYLCCNV (Tomato yellow leaf curl China virus) infectious clone were co-agroinoculated into tomato plants. Meanwhile, tomato plants co-agroinoculated with TYLCCNV and empty silencing vector DNAmP was used as the control. VIGS mediated silencing of SlGRX1 in tomato led to increased sensitivity to oxidative and salt stresses with decreased relative chlorophyll content, and reduced tolerance to drought stress with decreased relative water content. These results suggested that SlGRX1 was required for plant resistance to oxidative, drought and salt stresses.
The coding sequence of SlGRX1 was then cloned into the binary vector pBin438 to produce pBin438-SlGRX1 in which the SlGRX1 gene was expressed under the control of the cauliflower mosaic virus (CaMV) 35S promoter. The plasmid pBin438-SlGRX1 was then introduced into A. tumefaciens strain EHA105. Transformation of Arabidopsis was performed using the floral dip method. The empty vector pBin438 was also transformed into Arabidopsis as the control. Two independent homozygous lines (Y1, Y2) with a single copy of SlGRX1 were allowed to grow for 3 generations for further studies. We found over-expression of SlGRX1 in Arabidopsis plants enhanced lines Yl and Y2 the tolerance to oxidative, drought and salt stresses in both in vitro and in soil-grown conditions.
Expression levels of some oxidative related ascorbate peroxidase (Apx) genes Apx2, Apx3, Apx4, Apx5, Apx6, and well-known drought/salt stress marker genes RD29A, RD22 and RAB18 in Y1,Y2 and the control plants were analyzed by qRT-PCR to explore the possible mechanism of the enhanced abiotic stress tolerance in SlGRX1 over-expression plants. We found that over-expression of the SlGRX1 gene in Arabidopsis can activate the expression of the Arabidopsis Apx2 and Apx6, indicating that the SlGRX1 gene may positively regulate the Apx2 and Apx6 genes to improve plants resistance to oxidative stress.
引文
1. Alscher, R.G., Erturk, N., and Heath, L.S. (2002). Role of superoxide dismutases (SODs) in controlling oxidative stress in plants. J Exp Bot 53, 1331-1341.
2. Apel, K., and Hirt, H. (2004). Reactive oxygen species: metabolism, oxidative stress, and signal transduction. Annu Rev Plant Biol 55,373-399.
3. Ardie, S.W., Xie, L.N., Takahashi, R., Liu, S.K., and Takano, T. (2009). Cloning of a high-affinity K+ transporter gene PutHKT2;1 from Puccinellia tenuiflora and its functional comparison with OsHKT2;1 from rice in yeast and Arabidopsis. J Exp Bot 60,3491-3502.
4. Aroca, R., Irigoyen, J.J., and Sanchez-Diaz, M. (2003). Drought enhances maize chilling tolerance. Ⅱ. Photosynthetic traits and protective mechanisms against oxidative stress. Physiol Plant 117,540-549.
5. Arrabaca, J.D., and Loureito Dias, M.C. (1982). Cyanide-resistant respiration in a respiration-deficient mutant of Saccharomyces cerevisae. ZAllg Mikrobiol 22, 437-442.
6. Asada, K. (1999). The water-water cycle in chloroplasts:Dissipation of Excess Photons. Annu Rev Plant Physiol Plant Mol Biol 50,601-639.
7. Bandyopadhyay, S., Gama, F., Molina-Navarro, M.M., Gualberto, J.M., Claxton, R., Naik, S.G., Huynh, B.H., Herrero, E., Jacquot, J.P., Johnson, M.K., and Rouhier, N. (2008). Chloroplast monothiol glutaredoxins as scaffold proteins for the assembly and delivery of [2Fe-2S] clusters. EMBO J 27, 1122-1133.
8. Bolwell, G.P. (1999). Role of active oxygen species and NO in plant defence responses. Curr Opin Plant Biol 2,287-294.
9. Buchanan, B.B., and Balmer, Y. (2005). Redox regulation:a broadening horizon. Annu Rev Plant Biol 56,187-220.
10. Bushweller, J.H., Aslund, F., Wuthrich, K., and Holmgren, A. (1992). Structural and functional characterization of the mutant Escherichia coli glutaredoxin (C14----S) and its mixed disulfide with glutathione. Biochemistry 31, 9288-9293.
11. Cai, X.Z., Wang, C.C., Xu, Y.P., Xu, Q.F., Zheng, Z., and Zhou, X.P. (2007). Efficient gene silencing induction in tomato by a viral satellite DNA vector. Virus Res 125,169-175.
12. Chamnongpol, S., Willekens, H., Moeder, W., Langebartels, C., Sandermann,
H., Jr., Van Montagu, M., Inze, D., and Van Camp, W. (1998). Defense activation and enhanced pathogen tolerance induced by H2O2 in transgenic tobacco. Proc Natl Acad Sci USA 95,5818-5823.
13. Chen, Z., Ricigliano, J.W., and Klessig, D.F. (1993). Purification and characterization of a soluble salicylic acid-binding protein from tobacco. Proc Natl Acad Sci USA 90,9533-9537.
14. Cheng, N.H. (2008). AtGRX4, an Arabidopsis chloroplastic monothiol glutaredoxin, is able to suppress yeast grx5 mutant phenotypes and respond to oxidative stress. FEBS Lett 582,848-854.
15. Cheng, N.H., Liu, J.Z., Brock, A., Nelsono, R.S., and Hirschi, K.D. (2006). AtGRXcp, an Arabidopsis chloroplastic glutaredoxin, is critical for protection against protein oxidative damage. J. Biol. Chem 281,26280-26288.
16. Chuang, C.F., and Meyerowitz, E.M. (2000). Specific and heritable genetic interference by double-stranded RNA in Arabidopsis thaliana. Proc Natl Acad Sci USA 97,4985-4990.
17. Clough, S.J., and Bent, A.F. (1998). Floral dip:a simplified method for Agrobacterium-mediated transformation of Arabidopsis thaliana. Plant J 16, 735-743.
18. Cross, J.V., and Templeton, D.J. (2004). Oxidative stress inhibits MEKK1 by site-specific glutathionylation in the ATP-binding domain. Biochem J 381, 675-683.
19. Cui, X., Li, G., Wang, D., Hu, D., and Zhou, X. (2005). A Begomovirus DNAbeta-encoded protein binds DNA, functions as a suppressor of RNA silencing, and targets the cell nucleus. J Virol 79,10764-10775.
20. Dalle-Donne, I., Rossi, R., Giustarini, D., Colombo, R., and Milzani, A. (2007). S-glutathionylation in protein redox regulation. Free Radic Biol Med 43, 883-898.
21. Dalle-Donne, I., Rossi, R., Colombo, G., Giustarini, D., and Milzani, A. (2009). Protein S-glutathionylation:a regulatory device from bacteria to humans. Trends Biochem Sci 34,85-96.
22. Ding, X.S., Schneider, W.L., Chaluvadi, S.R., Mian, M.A.R., and Nelson, R.S. (2006). Characterization of a Brome mosaic virus strain and its use as a vector for gene silencing in monocotyledonous hosts. Mol Plant Microbe Interact 19, 1229-1239.
23. Dutilleul, C., Garmier, M., Noctor, G., Mathieu, C., Chetrit, P., Foyer, C.H.,
and de Paepe, R. (2003). Leaf mitochondria modulate whole cell redox homeostasis, set antioxidant capacity, and determine stress resistance through altered signaling and diurnal regulation. Plant Cell 15,1212-1226.
24. Fire, A., Xu, S., Montgomery, M.K., Kostas, S.A., Driver, S.E., and Mello, C.C. (1998). Potent and specific genetic interference by double-stranded RNA in Caenorhabditis elegans. Nature 391,806-811.
25. Foyer, C.H., and Noctor, G. (2005). Redox homeostasis and antioxidant signaling:a metabolic interface between stress perception and physiological responses. Plant Cell 17,1866-1875.
26. Fryer, M.J., Ball, L., Oxborough, K., Karpinski, S., Mullineaux, P.M., and Baker, N.R. (2003). Control of Ascorbate Peroxidase 2 expression by hydrogen peroxide and leaf water status during excess light stress reveals a functional organisation of Arabidopsis leaves. Plant J 33,691-705.
27. Ghezzi, P. (2005). Oxidoreduction of protein thiols in redox regulation. Biochem Soc Trans 33,1378-1381.
28. Gill, R.W., and Sanseau, P. (2000). Rapid in silico cloning of genes using expressed sequence tags (ESTs). Biotechnol Annu Rev 5,25-44.
29. Godge, M.R., Purkayastha, A., Dasgupta, I., and Kumar, P.P. (2008). Virus-induced gene silencing for functional analysis of selected genes. Plant. Cell Rep 27,209-219.
30. Gossele, V., Fache, I., Meulewaeter, F., Cornelissen, M., and Metzlaff, M. (2002). SVISS-a novel transient gene silencing system for gene function discovery and validation in tobacco plants. Plant J 32,859-866.
31. Green, R., and Fluhr, R. (1995). UV-B-Induced PR-1 Accumulation Is Mediated by Active Oxygen Species. Plant Cell 7,203-212.
32. Gupte, S.A., and Wolin, M.S. (2008). Oxidant and redox signaling in vascular oxygen sensing:implications for systemic and pulmonary hypertension. Antioxid Redox Signal 10,1137-1152.
33. Hamilton, A.J., and Baulcombe, D.C. (1999). A species of small antisense RNA in posttranscriptional gene silencing in plants. Science 286,950-952.
34. He, X.X., Jin, C.W., Li, G.X., You, G.Y., Zhou, X.P., and Zheng, S.J. (2008). Use of the modified viral satellite DNA vector to silence mineral nutrition-related genes in plants:silencing of the tomato ferric chelate reductase gene, FRO1, as an example. Sci China Ser C 51,402-409.
35. Hill, I.M., Beswick, P.H., and Donaldson, K. (1996). Enhancement of the
macrophage oxidative burst by immunoglobulin coating of respirable fibers: fiber-specific differences between asbestos and man-made fibers. Exp Lung Res 22,133-148.
36. Ho, T., Wang, H., Pallett, D., and Dalmay, T. (2007). Evidence for targeting common siRNA hotspots and GC preference by plant Dicer-like proteins. FEBS Lett 581,3267-3272.
37. Holmgren, A. (1979). Glutathione-dependent synthesis of deoxyribonucleotides-purification and characterization of glutaredoxin from Escherichia coli. J. Biol. Chem 254,3664-3671.
38. Huang, C.J., Xie, Y., and Zhou, X.P. (2009). Efficient virus-induced gene silencing in plants using a modified geminivirus DNA1 component. Plant Biotechnol J 7,254-265.
39. Jami, S.K., Clark, G.B., Turlapati, S.A., Handley, C., Roux, S.J., and Kirti, P.B. (2008). Ectopic expression of an annexin from Brassica juncea confers tolerance to abiotic and biotic stress treatments in transgenic tobacco. Plant Physiol Biochem 46,1019-1030.
40. Jones, L., Hamilton, A.J., Voinnet, O., Thomas, C.L., Maule, A.J., and Baulcombe, D.C. (1999). RNA-DNA interactions and DNA methylation in post-transcriptional gene silencing. Plant Cell 11,2291-2301.
41. Kalamaki, M.S., Alexandrou, D., Lazari, D., Merkouropoulos, G., Fotopoulos, V., Pateraki, I., Aggelis, A., Carrillo-Lopez, A., Rubio-Cabetas, M.J., and Kanellis, A.K. (2009). Over-expression of a tomato N-acetyl-L-glutamate synthase gene (SINAGS1) in Arabidopsis thaliana results in high ornithine levels and increased tolerance in salt and drought stresses. J Exp Bot 60,1859-1871.
42. Kang, J.Y., Choi, H.I., Im, M.Y., and Kim, S.Y. (2002). Arabidopsis basic leucine zipper proteins that mediate stress-responsive abscisic acid signaling. Plant Cell 14,343-357.
43. Karaffa, L., Vaczy, K., Sandor, E., Biro, S., Szentirmai, A., and Pocsi, I. (2001). Cyanide-resistant alternative respiration is strictly correlated to intracellular peroxide levels in Acremonium chrysogenum. Free Radic Res 34, 405-416.
44. Karyotou, K., and Donaldson, R.P. (2005). Ascorbate peroxidase, a scavenger of hydrogen peroxide in glyoxysomal membranes. Arch Biochem Biophys 434, 248-257.
45. Kjemtrup, S., Sampson, K.S., Peele, C.G., Nguyen, L.V., Conkling, M.A.,
Thompson, W.F., and Robertson, D. (1998). Gene silencing from plant DNA carried by a Geminivirus. Plant J 14,91-100.
46. Kumagai, M.H., Donson, J., della-Cioppa, G., Harvey, D., Hanley, K., and Grill, L.K. (1995). Cytoplasmic inhibition of carotenoid biosynthesis with virus-derived RNA. Proc Natl Acad Sci USA 92,1679-1683.
47. Lam, E., Kato, N., and Lawton, M. (2001). Programmed cell death, mitochondria and the plant hypersensitive response. Nature 411,848-853.
48. Lemaire, S.D. (2004). The glutaredoxin family in oxygenic photosynthetic organisms. Photosynth. Res 79,305-318.
49. Li, S., Lauri, A., Ziemann, M., Busch, A., Bhave, M., and Zachgo, S. (2009). Nuclear activity of ROXY1, a glutaredoxin interacting with TGA Factors, is required for petal development in Arabidopsis thaliana. Plant Cell 21,429-441.
50. Liu, Y.L., Schiff, M., and Dinesh-Kumar, S.P. (2002a). Virus-induced gene silencing in tomato. Plant J 31,777-786.
51. Liu, Y.L., Schiff, M., Marathe, R., and Dinesh-Kumar, S.P. (2002b). Tobacco Rarl, EDS1 and NPR1/NIM1 like genes are required for N-mediated resistance to tobacco mosaic virus. Plant J 30,415-429.
52. Livak, K.J., and Schmittgen, T.D. (2001). Analysis of relative gene expression data using real-time quantitative PCR and the 2-△△CT method. Methods 25, 402-408.
53. Lu, R., Martin-Hernandez, A.M., Peart, J.R., Malcuit, I., and Baulcombe, D.C. (2003). Virus-induced gene silencing in plants. Methods 30,296-303.
54. Marchler-Bauer, A., Anderson, J.B., Derbyshire, et al. (2007). CDD:a conserved domain database for interactive domain family analysis. Nucleic Acids Res 35, D237-240.
55. Matthew, L. (2004). RNAi for Plant Functional Genomics. Comp Funct Genomics 5,240-244.
56. Mikuni, T., Tatsuta, M., and Kamachi, M. (1987). Scavenging effect of butylated hydroxytoluene on the production of free radicals by the reaction of hydrogen peroxide with N-methyl-N'-nitro-N-nitrosoguanidine. J Natl Cancer Inst 79,281-283.
57. Miller, G., Suzuki, N., Ciftci-Yilmaz, S., and Mittler, R. (2009). Reactive oxygen species homeostasis and signalling during drought and salinity stresses. Plant Cell & Environ.
58. Mittler, R. (2002). Oxidative stress, antioxidants and stress tolerance. Trends
Plant Sci 7,405-410.
59. Miwa, N., Yamazaki, H., Nagaoka, Y., Kageyama, K., Onoyama, Y., Matsui-Yuasa, I., Otani, S., and Morisawa, S. (1988). Altered production of the active oxygen species is involved in enhanced cytotoxic action of acylated derivatives of ascorbate to tumor cells. Biochim Biophys Acta 972,144-151.
60. Miyake, C., and Yokota, A. (2000). Determination of the rate of photoreduction of O2 in the water-water cycle in watermelon leaves and enhancement of the rate by limitation of photosynthesis. Plant Cell Physiol 41,335-343.
61. Mlotshwa, S., Pruss, G.J., and Vance, V. (2008). Small RNAs in viral infection and host defense. Trends Plant Sci 13,375-382.
62. Moller, I.M. (2001). Plant Mitochondria and oxidative stredd:Electron transport, NADPH turnover, and metabolism of reactive oxygen species. Annu Rev Plant Physiol Plant Mol Biol 52,561-591.
63. Narendra, S., Venkataramani, S., Shen, G., Wang, J., Pasapula, V., Lin, Y., Kornyeyev, D., Holaday, A.S., and Zhang, H. (2006a). The Arabidopsis ascorbate peroxidase 3 is a peroxisomal membrane-bound antioxidant enzyme and is dispensable for Arabidopsis growth and development. J Exp Bot 57, 3033-3042.
64. Narendra, S., Venkataramani, S., Shen, G.X., Wang, J., Pasapula, V, Lin, Y., Kornyeyev, D., Holaday, A.S., and Zhang, H. (2006b). The Arabidopsis ascorbate peroxidase 3 is a peroxisomal membrane-bound antioxidant enzyme and is dispensable for Arabidopsis growth and development. J Exp Bot 57, 3033-3042.
65. Ndamukong, I., Al Abdallat, A., Thurow, C., Fode, B., Zander, M., Weigel, R., and Gatz, C. (2007). SA-inducible Arabidopsis glutaredoxin interacts with TGA factors and suppresses JA-responsive PDF1.2 transcription. Plant J 50,128-139.
66. Nicolas, R. (2010). Plant glutaredoxins:pivotal players in redox biology and iron-sulphur centre assembly centre assembly. New Phytol 186,365-372.
67. Noctor, G., and Foyer, C.H. (1998). Ascorbate and glutathione:keeping active oxygen under control. Annu Rev Plant Physiol Plant Mol Biol 49,249-279.
68. Nordstrand, K., slund, F., Holmgren, A., Otting, G., and Berndt, K.D. (1999). NMR structure of Escherichia coli glutaredoxin 3-glutathione mixed disulfide complex:implications for the enzymatic mechanism. J Mol Biol 286,541-552.
69. Overmyer, K., Brosche, M., Pellinen, R., Kuittinen, T., Tuominen, H., Ahlfors, R., Keinanen, M., Saarma, M., Scheel, D., and Kangasjarvi, J. (2005).
Ozone-induced programmed cell death in the Arabidopsis radical-induced cell deathl mutant. Plant Physiol 137,1092-1104.
70. Panchuk, II, Zentgraf, U., and Volkov, R.A. (2005). Expression of the Apx gene family during leaf senescence of Arabidopsis thaliana. Planta 222,926-932.
71. Pasqualini, S., Paolocci, F., Borgogni, A., Morettini, R., and Ederli, L. (2007). The overexpression of an alternative oxidase gene triggers ozone sensitivity in tobacco plants. Plant Cell & Environ 30,1545-1556.
72. Pasqualini, S., Piccioni, C., Reale, L., Ederli, L., Della Torre, G., and Ferranti, F. (2003). Ozone-induced cell death in tobacco cultivar Bel W3 plants. The role of programmed cell death in lesion formation. Plant Physiol 133,1122-1134.
73. Peele, C., Jordan, C.V., Muangsan, N., Turnage, M., Egelkrout, E., Eagle, P., Hanley-Bowdoin, L., and Robertson, D. (2001). Silencing of a meristematic gene using geminivirus-derived vectors. Plant J 27,357-366.
74. Pitzschke, A., Forzani, C., and Hirt, H. (2006). Reactive oxygen species signaling in plants. Antioxid Redox Signal 8,1757-1764.
75. Rasmusson, A.G, Heiser, V., Zabaleta, E., Brennicke, A., and Grohmann, L. (1998). Physiological, biochemical and molecular aspects of mitochondrial complex I in plants. Biochimica et Biophysica Acta (BBA)-Bioenergetics 1364, 101-111.
76. Ratcliff, F., Martin-Hernandez, A.M., and Baulcombe, D.C. (2001). Tobacco rattle virus as a vector for analysis of gene function by silencing. Plant J 25, 237-245.
77. Rinna, A., Torres, M., and Forman, H.J. (2006). Stimulation of the alveolar macrophage respiratory burst by ADP causes selective glutathionylation of protein tyrosine phosphatase 1B. Free Radic Biol Med 41,86-91.
78. Rizhsky, L., Liang, H., and Mittler, R. (2003). The water-water cycle is essential for chloroplast protection in the absence of stress. J Biol Chem 278, 38921-38925.
79. Robertson, D. (2004). VIGS vectors for gene silencing:Many targets, many tools. Annu Rev of Plant Biol 55,495-519.
80. Robson, C.A., and Vanlerberghe, G.C. (2002). Transgenic plant cells lacking mitochondrial alternative oxidase have increased susceptibility to mitochondria-dependent and-independent pathways of programmed cell death. Plant Physiol 129,1908-1920.
81. Rodriguez-Manzaneque, M.T., Ros, J., Cabiscol, E., Sorribas, A., and
Herrero, E. (1999). Grx5 glutaredoxin plays a central role in protection against protein oxidative damage in Saccharomyces cerevisiae. Mol Cell Biol 19, 8180-8190.
82. Rossel, J.B., Walter, P.B., Hendrickson, L., Chow, W.S., Poole, A., Mullineaux, P.M., and Pogson, B.J. (2006). A mutation affecting ascorbate peroxidase 2 gene expression reveals a link between responses to high light and drought tolerance. Plant Cell & Environ 29,269-281.
83. Rouhier, N., Gelhaye, E., and Jacquot, J.P. (2004a). Plant glutaredoxins:still mysterious reducing systems. Cell Mol Life Sci 61,1266-1277.
84. Rouhier, N., Couturier, J., and Jacquot, J.P. (2006). Genome-wide analysis of plant glutaredoxin systems. J Exp Bot 57,1685-1696.
85. Rouhier, N., Lemaire, S.D., and Jacquot, J.P. (2008). The role of glutathione in photosynthetic organisms:Emerging functions for glutaredoxins and glutathionylation. Annu Rev of Plant Biol 59,143-166.
86. Rouhier, N., Unno, H., Bandyopadhyay, S., Masip, L., et al. (2007). Functional, structural, and spectroscopic characterization of a glutathione-ligated [2Fe-2S] cluster in poplar glutaredoxin C1. Proc Natl Acad Sci USA 104, 7379-7384.
87. Ruiz, M.T., Voinnet, O., and Baulcombe, D.C. (1998). Initiation and maintenance of virus-induced gene silencing. Plant Cell 10,937-946.
88. Rusanen, H., Majamaa, K., and Hassinen, I.E. (2000). Increased activities of antioxidant enzymes and decreased ATP concentration in cultured myoblasts with the 3243A->G mutation in mitochondrial DNA. Biochim Biophys Acta 1500, 10-16.
89. Scandalios, J.G. (2002). The rise of ROS. Trends Biochem Sci 27,483-486.
90. Senthil-Kumar, M., Gowda, H.V.R., Hema, R., Mysore, K.S., and Udayakumar, M. (2008). Virus-induced gene silencing and its application in characterizing genes involved in water-deficit-stress tolerance. J. Plant Physiol 165,1404-1421.
91. Shigeoka, S., Ishikawa, T., Tamoi, M., Miyagawa, Y., Takeda, T., Yabuta, Y., and Yoshimura, K. (2002). Regulation and function of ascorbate peroxidase isoenzymes. J Exp Bot 53,1305-1319.
92. Smirnoff, N. (2000). Ascorbic acid:metabolism and functions of a multi-facetted molecule. Curr Opin Plant Biol 3,229-235.
93. Smith, N.A., Singh, S.P., Wang, M.B., Stoutjesdijk, P.A., Green, A.G., and Waterhouse, P.M. (2000). Total silencing by intron-spliced hairpin RNAs. Nature 407,319-320.
94. Tada, Y., Spoel, S.H., Pajerowska-Mukhtar, K., Mou, Z., Song, J., Wang, C., Zuo, J., and Dong, X. (2008). Plant immunity requires conformational changes of NPR1 via S-nitrosylation and thioredoxins. Science 321,952-956.
95. Tao, X.R., and Zhou, X.P. (2004). A modified viral satellite DNA that suppresses gene expression in plants. Plant J 38,850-860.
96. Tao, X.R., Qian, Y.J., and Zhou, X.P. (2006). Modification of a viral satellite DNA-based gene silencing vector and its application to leaf or flower color change in Petunia hybrida. Chinese Sci Bull 51,2208-2213.
97. Turner, N.C. (1981). Techniques and Experimental Approaches for the Measurement of Plant Water Status. Plant & Soil 58,339-366.
98. Unver, T., and Budak, H. (2009). Virus-induced gene silencing, a post transcriptional gene silencing method. Int J Plant Genomics 2009, 198680-198687.
99. Voinnet, O. (2001). RNA silencing as a plant immune system against viruses. Trends Genet 17,449-459.
100.Voinnet, O. (2003). RNA silencing bridging the gaps in wheat extracts. Trends Plant Sci 8,307-309.
101.Vranova, E., Inze, D., and Van Breusegem, F. (2002). Signal transduction during oxidative stress. J Exp Bot 53,1227-1236.
102.Wang, Z., Xing, S.P., Birkenbihl, R.P., and Zachgo, S. (2009). Conserved functions of Arabidopsis and rice CC-type glutaredoxins in flower development and pathogen response. Molecular Plant 2,323-335.
103.Waterhouse, P.M., Wang, M.B., and Lough, T. (2001). Gene silencing as an adaptive defence against viruses. Nature 411,834-842.
104.Willekens, H., Langebartels, C., Tire, C., Van Montagu, M., Inze, D., and Van Camp, W. (1994). Differential expression of catalase genes in Nicotiana plumbaginifolia (L.). Proc Natl Acad Sci USA 91,10450-10454.
105.Wu, G., Shortt, B.J., Lawrence, E.B., Leon, J., Fitzsimmons, K.C., Levine, E.B., Raskin, I., and Shah, D.M. (1997). Activation of host defense mechanisms by elevated production of H2O2 in transgenic plants. Plant Physiol 115,427-435.
106.Xing, S., and Zachgo, S. (2008). ROXY1 and ROXY2, two Arabidopsis glutaredoxin genes, are required for anther development. Plant J 53,790-801.
107.Xing, S.P., Rosso, M.G., and Zachgo, S. (2005). ROXY1, a member of the plant glutaredoxin family, is required for petal development in Arabidopsis thaliana. Development 132,1555-1565.
108.Xiong, R.Y., Wu, J.X., Zhou, Y.J., and Zhou, X.P. (2009). Characterization and subcellular localization of an RNA silencing suppressor encoded by Rice stripe tenuivirus. Virology 387,29-40.
109.Yamaguchi, K., Mori, H., and Nishimura, M. (1995). A novel isoenzyme of ascorbate peroxidase localized on glyoxysomal and leaf peroxisomal membranes in pumpkin. Plant Cell Physiol 36,1157-1162.
110.Yoshimura, K., Yabuta, Y., Ishikawa, T., and Shigeoka, S. (2000). Expression of spinach ascorbate peroxidase isoenzymes in response to oxidative stresses. Plant Physiol 123,223-234.
111.Zhu, J.K. (2002). Salt and drought stress signal transduction in plants. Annu Rev Plant Biol 53,247-273.
112.Ziemann, M., Bhave, M., and Zachgo, S. (2009). Origin and diversification of land plant CC-Type glutaredoxins. Genome Biol Evol 2009,265-277.
2. Apel, K., and Hirt, H. (2004). Reactive oxygen species: metabolism, oxidative stress, and signal transduction. Annu Rev Plant Biol 55,373-399.
3. Ardie, S.W., Xie, L.N., Takahashi, R., Liu, S.K., and Takano, T. (2009). Cloning of a high-affinity K+ transporter gene PutHKT2;1 from Puccinellia tenuiflora and its functional comparison with OsHKT2;1 from rice in yeast and Arabidopsis. J Exp Bot 60,3491-3502.
4. Aroca, R., Irigoyen, J.J., and Sanchez-Diaz, M. (2003). Drought enhances maize chilling tolerance. Ⅱ. Photosynthetic traits and protective mechanisms against oxidative stress. Physiol Plant 117,540-549.
5. Arrabaca, J.D., and Loureito Dias, M.C. (1982). Cyanide-resistant respiration in a respiration-deficient mutant of Saccharomyces cerevisae. ZAllg Mikrobiol 22, 437-442.
6. Asada, K. (1999). The water-water cycle in chloroplasts:Dissipation of Excess Photons. Annu Rev Plant Physiol Plant Mol Biol 50,601-639.
7. Bandyopadhyay, S., Gama, F., Molina-Navarro, M.M., Gualberto, J.M., Claxton, R., Naik, S.G., Huynh, B.H., Herrero, E., Jacquot, J.P., Johnson, M.K., and Rouhier, N. (2008). Chloroplast monothiol glutaredoxins as scaffold proteins for the assembly and delivery of [2Fe-2S] clusters. EMBO J 27, 1122-1133.
8. Bolwell, G.P. (1999). Role of active oxygen species and NO in plant defence responses. Curr Opin Plant Biol 2,287-294.
9. Buchanan, B.B., and Balmer, Y. (2005). Redox regulation:a broadening horizon. Annu Rev Plant Biol 56,187-220.
10. Bushweller, J.H., Aslund, F., Wuthrich, K., and Holmgren, A. (1992). Structural and functional characterization of the mutant Escherichia coli glutaredoxin (C14----S) and its mixed disulfide with glutathione. Biochemistry 31, 9288-9293.
11. Cai, X.Z., Wang, C.C., Xu, Y.P., Xu, Q.F., Zheng, Z., and Zhou, X.P. (2007). Efficient gene silencing induction in tomato by a viral satellite DNA vector. Virus Res 125,169-175.
12. Chamnongpol, S., Willekens, H., Moeder, W., Langebartels, C., Sandermann,
H., Jr., Van Montagu, M., Inze, D., and Van Camp, W. (1998). Defense activation and enhanced pathogen tolerance induced by H2O2 in transgenic tobacco. Proc Natl Acad Sci USA 95,5818-5823.
13. Chen, Z., Ricigliano, J.W., and Klessig, D.F. (1993). Purification and characterization of a soluble salicylic acid-binding protein from tobacco. Proc Natl Acad Sci USA 90,9533-9537.
14. Cheng, N.H. (2008). AtGRX4, an Arabidopsis chloroplastic monothiol glutaredoxin, is able to suppress yeast grx5 mutant phenotypes and respond to oxidative stress. FEBS Lett 582,848-854.
15. Cheng, N.H., Liu, J.Z., Brock, A., Nelsono, R.S., and Hirschi, K.D. (2006). AtGRXcp, an Arabidopsis chloroplastic glutaredoxin, is critical for protection against protein oxidative damage. J. Biol. Chem 281,26280-26288.
16. Chuang, C.F., and Meyerowitz, E.M. (2000). Specific and heritable genetic interference by double-stranded RNA in Arabidopsis thaliana. Proc Natl Acad Sci USA 97,4985-4990.
17. Clough, S.J., and Bent, A.F. (1998). Floral dip:a simplified method for Agrobacterium-mediated transformation of Arabidopsis thaliana. Plant J 16, 735-743.
18. Cross, J.V., and Templeton, D.J. (2004). Oxidative stress inhibits MEKK1 by site-specific glutathionylation in the ATP-binding domain. Biochem J 381, 675-683.
19. Cui, X., Li, G., Wang, D., Hu, D., and Zhou, X. (2005). A Begomovirus DNAbeta-encoded protein binds DNA, functions as a suppressor of RNA silencing, and targets the cell nucleus. J Virol 79,10764-10775.
20. Dalle-Donne, I., Rossi, R., Giustarini, D., Colombo, R., and Milzani, A. (2007). S-glutathionylation in protein redox regulation. Free Radic Biol Med 43, 883-898.
21. Dalle-Donne, I., Rossi, R., Colombo, G., Giustarini, D., and Milzani, A. (2009). Protein S-glutathionylation:a regulatory device from bacteria to humans. Trends Biochem Sci 34,85-96.
22. Ding, X.S., Schneider, W.L., Chaluvadi, S.R., Mian, M.A.R., and Nelson, R.S. (2006). Characterization of a Brome mosaic virus strain and its use as a vector for gene silencing in monocotyledonous hosts. Mol Plant Microbe Interact 19, 1229-1239.
23. Dutilleul, C., Garmier, M., Noctor, G., Mathieu, C., Chetrit, P., Foyer, C.H.,
and de Paepe, R. (2003). Leaf mitochondria modulate whole cell redox homeostasis, set antioxidant capacity, and determine stress resistance through altered signaling and diurnal regulation. Plant Cell 15,1212-1226.
24. Fire, A., Xu, S., Montgomery, M.K., Kostas, S.A., Driver, S.E., and Mello, C.C. (1998). Potent and specific genetic interference by double-stranded RNA in Caenorhabditis elegans. Nature 391,806-811.
25. Foyer, C.H., and Noctor, G. (2005). Redox homeostasis and antioxidant signaling:a metabolic interface between stress perception and physiological responses. Plant Cell 17,1866-1875.
26. Fryer, M.J., Ball, L., Oxborough, K., Karpinski, S., Mullineaux, P.M., and Baker, N.R. (2003). Control of Ascorbate Peroxidase 2 expression by hydrogen peroxide and leaf water status during excess light stress reveals a functional organisation of Arabidopsis leaves. Plant J 33,691-705.
27. Ghezzi, P. (2005). Oxidoreduction of protein thiols in redox regulation. Biochem Soc Trans 33,1378-1381.
28. Gill, R.W., and Sanseau, P. (2000). Rapid in silico cloning of genes using expressed sequence tags (ESTs). Biotechnol Annu Rev 5,25-44.
29. Godge, M.R., Purkayastha, A., Dasgupta, I., and Kumar, P.P. (2008). Virus-induced gene silencing for functional analysis of selected genes. Plant. Cell Rep 27,209-219.
30. Gossele, V., Fache, I., Meulewaeter, F., Cornelissen, M., and Metzlaff, M. (2002). SVISS-a novel transient gene silencing system for gene function discovery and validation in tobacco plants. Plant J 32,859-866.
31. Green, R., and Fluhr, R. (1995). UV-B-Induced PR-1 Accumulation Is Mediated by Active Oxygen Species. Plant Cell 7,203-212.
32. Gupte, S.A., and Wolin, M.S. (2008). Oxidant and redox signaling in vascular oxygen sensing:implications for systemic and pulmonary hypertension. Antioxid Redox Signal 10,1137-1152.
33. Hamilton, A.J., and Baulcombe, D.C. (1999). A species of small antisense RNA in posttranscriptional gene silencing in plants. Science 286,950-952.
34. He, X.X., Jin, C.W., Li, G.X., You, G.Y., Zhou, X.P., and Zheng, S.J. (2008). Use of the modified viral satellite DNA vector to silence mineral nutrition-related genes in plants:silencing of the tomato ferric chelate reductase gene, FRO1, as an example. Sci China Ser C 51,402-409.
35. Hill, I.M., Beswick, P.H., and Donaldson, K. (1996). Enhancement of the
macrophage oxidative burst by immunoglobulin coating of respirable fibers: fiber-specific differences between asbestos and man-made fibers. Exp Lung Res 22,133-148.
36. Ho, T., Wang, H., Pallett, D., and Dalmay, T. (2007). Evidence for targeting common siRNA hotspots and GC preference by plant Dicer-like proteins. FEBS Lett 581,3267-3272.
37. Holmgren, A. (1979). Glutathione-dependent synthesis of deoxyribonucleotides-purification and characterization of glutaredoxin from Escherichia coli. J. Biol. Chem 254,3664-3671.
38. Huang, C.J., Xie, Y., and Zhou, X.P. (2009). Efficient virus-induced gene silencing in plants using a modified geminivirus DNA1 component. Plant Biotechnol J 7,254-265.
39. Jami, S.K., Clark, G.B., Turlapati, S.A., Handley, C., Roux, S.J., and Kirti, P.B. (2008). Ectopic expression of an annexin from Brassica juncea confers tolerance to abiotic and biotic stress treatments in transgenic tobacco. Plant Physiol Biochem 46,1019-1030.
40. Jones, L., Hamilton, A.J., Voinnet, O., Thomas, C.L., Maule, A.J., and Baulcombe, D.C. (1999). RNA-DNA interactions and DNA methylation in post-transcriptional gene silencing. Plant Cell 11,2291-2301.
41. Kalamaki, M.S., Alexandrou, D., Lazari, D., Merkouropoulos, G., Fotopoulos, V., Pateraki, I., Aggelis, A., Carrillo-Lopez, A., Rubio-Cabetas, M.J., and Kanellis, A.K. (2009). Over-expression of a tomato N-acetyl-L-glutamate synthase gene (SINAGS1) in Arabidopsis thaliana results in high ornithine levels and increased tolerance in salt and drought stresses. J Exp Bot 60,1859-1871.
42. Kang, J.Y., Choi, H.I., Im, M.Y., and Kim, S.Y. (2002). Arabidopsis basic leucine zipper proteins that mediate stress-responsive abscisic acid signaling. Plant Cell 14,343-357.
43. Karaffa, L., Vaczy, K., Sandor, E., Biro, S., Szentirmai, A., and Pocsi, I. (2001). Cyanide-resistant alternative respiration is strictly correlated to intracellular peroxide levels in Acremonium chrysogenum. Free Radic Res 34, 405-416.
44. Karyotou, K., and Donaldson, R.P. (2005). Ascorbate peroxidase, a scavenger of hydrogen peroxide in glyoxysomal membranes. Arch Biochem Biophys 434, 248-257.
45. Kjemtrup, S., Sampson, K.S., Peele, C.G., Nguyen, L.V., Conkling, M.A.,
Thompson, W.F., and Robertson, D. (1998). Gene silencing from plant DNA carried by a Geminivirus. Plant J 14,91-100.
46. Kumagai, M.H., Donson, J., della-Cioppa, G., Harvey, D., Hanley, K., and Grill, L.K. (1995). Cytoplasmic inhibition of carotenoid biosynthesis with virus-derived RNA. Proc Natl Acad Sci USA 92,1679-1683.
47. Lam, E., Kato, N., and Lawton, M. (2001). Programmed cell death, mitochondria and the plant hypersensitive response. Nature 411,848-853.
48. Lemaire, S.D. (2004). The glutaredoxin family in oxygenic photosynthetic organisms. Photosynth. Res 79,305-318.
49. Li, S., Lauri, A., Ziemann, M., Busch, A., Bhave, M., and Zachgo, S. (2009). Nuclear activity of ROXY1, a glutaredoxin interacting with TGA Factors, is required for petal development in Arabidopsis thaliana. Plant Cell 21,429-441.
50. Liu, Y.L., Schiff, M., and Dinesh-Kumar, S.P. (2002a). Virus-induced gene silencing in tomato. Plant J 31,777-786.
51. Liu, Y.L., Schiff, M., Marathe, R., and Dinesh-Kumar, S.P. (2002b). Tobacco Rarl, EDS1 and NPR1/NIM1 like genes are required for N-mediated resistance to tobacco mosaic virus. Plant J 30,415-429.
52. Livak, K.J., and Schmittgen, T.D. (2001). Analysis of relative gene expression data using real-time quantitative PCR and the 2-△△CT method. Methods 25, 402-408.
53. Lu, R., Martin-Hernandez, A.M., Peart, J.R., Malcuit, I., and Baulcombe, D.C. (2003). Virus-induced gene silencing in plants. Methods 30,296-303.
54. Marchler-Bauer, A., Anderson, J.B., Derbyshire, et al. (2007). CDD:a conserved domain database for interactive domain family analysis. Nucleic Acids Res 35, D237-240.
55. Matthew, L. (2004). RNAi for Plant Functional Genomics. Comp Funct Genomics 5,240-244.
56. Mikuni, T., Tatsuta, M., and Kamachi, M. (1987). Scavenging effect of butylated hydroxytoluene on the production of free radicals by the reaction of hydrogen peroxide with N-methyl-N'-nitro-N-nitrosoguanidine. J Natl Cancer Inst 79,281-283.
57. Miller, G., Suzuki, N., Ciftci-Yilmaz, S., and Mittler, R. (2009). Reactive oxygen species homeostasis and signalling during drought and salinity stresses. Plant Cell & Environ.
58. Mittler, R. (2002). Oxidative stress, antioxidants and stress tolerance. Trends
Plant Sci 7,405-410.
59. Miwa, N., Yamazaki, H., Nagaoka, Y., Kageyama, K., Onoyama, Y., Matsui-Yuasa, I., Otani, S., and Morisawa, S. (1988). Altered production of the active oxygen species is involved in enhanced cytotoxic action of acylated derivatives of ascorbate to tumor cells. Biochim Biophys Acta 972,144-151.
60. Miyake, C., and Yokota, A. (2000). Determination of the rate of photoreduction of O2 in the water-water cycle in watermelon leaves and enhancement of the rate by limitation of photosynthesis. Plant Cell Physiol 41,335-343.
61. Mlotshwa, S., Pruss, G.J., and Vance, V. (2008). Small RNAs in viral infection and host defense. Trends Plant Sci 13,375-382.
62. Moller, I.M. (2001). Plant Mitochondria and oxidative stredd:Electron transport, NADPH turnover, and metabolism of reactive oxygen species. Annu Rev Plant Physiol Plant Mol Biol 52,561-591.
63. Narendra, S., Venkataramani, S., Shen, G., Wang, J., Pasapula, V., Lin, Y., Kornyeyev, D., Holaday, A.S., and Zhang, H. (2006a). The Arabidopsis ascorbate peroxidase 3 is a peroxisomal membrane-bound antioxidant enzyme and is dispensable for Arabidopsis growth and development. J Exp Bot 57, 3033-3042.
64. Narendra, S., Venkataramani, S., Shen, G.X., Wang, J., Pasapula, V, Lin, Y., Kornyeyev, D., Holaday, A.S., and Zhang, H. (2006b). The Arabidopsis ascorbate peroxidase 3 is a peroxisomal membrane-bound antioxidant enzyme and is dispensable for Arabidopsis growth and development. J Exp Bot 57, 3033-3042.
65. Ndamukong, I., Al Abdallat, A., Thurow, C., Fode, B., Zander, M., Weigel, R., and Gatz, C. (2007). SA-inducible Arabidopsis glutaredoxin interacts with TGA factors and suppresses JA-responsive PDF1.2 transcription. Plant J 50,128-139.
66. Nicolas, R. (2010). Plant glutaredoxins:pivotal players in redox biology and iron-sulphur centre assembly centre assembly. New Phytol 186,365-372.
67. Noctor, G., and Foyer, C.H. (1998). Ascorbate and glutathione:keeping active oxygen under control. Annu Rev Plant Physiol Plant Mol Biol 49,249-279.
68. Nordstrand, K., slund, F., Holmgren, A., Otting, G., and Berndt, K.D. (1999). NMR structure of Escherichia coli glutaredoxin 3-glutathione mixed disulfide complex:implications for the enzymatic mechanism. J Mol Biol 286,541-552.
69. Overmyer, K., Brosche, M., Pellinen, R., Kuittinen, T., Tuominen, H., Ahlfors, R., Keinanen, M., Saarma, M., Scheel, D., and Kangasjarvi, J. (2005).
Ozone-induced programmed cell death in the Arabidopsis radical-induced cell deathl mutant. Plant Physiol 137,1092-1104.
70. Panchuk, II, Zentgraf, U., and Volkov, R.A. (2005). Expression of the Apx gene family during leaf senescence of Arabidopsis thaliana. Planta 222,926-932.
71. Pasqualini, S., Paolocci, F., Borgogni, A., Morettini, R., and Ederli, L. (2007). The overexpression of an alternative oxidase gene triggers ozone sensitivity in tobacco plants. Plant Cell & Environ 30,1545-1556.
72. Pasqualini, S., Piccioni, C., Reale, L., Ederli, L., Della Torre, G., and Ferranti, F. (2003). Ozone-induced cell death in tobacco cultivar Bel W3 plants. The role of programmed cell death in lesion formation. Plant Physiol 133,1122-1134.
73. Peele, C., Jordan, C.V., Muangsan, N., Turnage, M., Egelkrout, E., Eagle, P., Hanley-Bowdoin, L., and Robertson, D. (2001). Silencing of a meristematic gene using geminivirus-derived vectors. Plant J 27,357-366.
74. Pitzschke, A., Forzani, C., and Hirt, H. (2006). Reactive oxygen species signaling in plants. Antioxid Redox Signal 8,1757-1764.
75. Rasmusson, A.G, Heiser, V., Zabaleta, E., Brennicke, A., and Grohmann, L. (1998). Physiological, biochemical and molecular aspects of mitochondrial complex I in plants. Biochimica et Biophysica Acta (BBA)-Bioenergetics 1364, 101-111.
76. Ratcliff, F., Martin-Hernandez, A.M., and Baulcombe, D.C. (2001). Tobacco rattle virus as a vector for analysis of gene function by silencing. Plant J 25, 237-245.
77. Rinna, A., Torres, M., and Forman, H.J. (2006). Stimulation of the alveolar macrophage respiratory burst by ADP causes selective glutathionylation of protein tyrosine phosphatase 1B. Free Radic Biol Med 41,86-91.
78. Rizhsky, L., Liang, H., and Mittler, R. (2003). The water-water cycle is essential for chloroplast protection in the absence of stress. J Biol Chem 278, 38921-38925.
79. Robertson, D. (2004). VIGS vectors for gene silencing:Many targets, many tools. Annu Rev of Plant Biol 55,495-519.
80. Robson, C.A., and Vanlerberghe, G.C. (2002). Transgenic plant cells lacking mitochondrial alternative oxidase have increased susceptibility to mitochondria-dependent and-independent pathways of programmed cell death. Plant Physiol 129,1908-1920.
81. Rodriguez-Manzaneque, M.T., Ros, J., Cabiscol, E., Sorribas, A., and
Herrero, E. (1999). Grx5 glutaredoxin plays a central role in protection against protein oxidative damage in Saccharomyces cerevisiae. Mol Cell Biol 19, 8180-8190.
82. Rossel, J.B., Walter, P.B., Hendrickson, L., Chow, W.S., Poole, A., Mullineaux, P.M., and Pogson, B.J. (2006). A mutation affecting ascorbate peroxidase 2 gene expression reveals a link between responses to high light and drought tolerance. Plant Cell & Environ 29,269-281.
83. Rouhier, N., Gelhaye, E., and Jacquot, J.P. (2004a). Plant glutaredoxins:still mysterious reducing systems. Cell Mol Life Sci 61,1266-1277.
84. Rouhier, N., Couturier, J., and Jacquot, J.P. (2006). Genome-wide analysis of plant glutaredoxin systems. J Exp Bot 57,1685-1696.
85. Rouhier, N., Lemaire, S.D., and Jacquot, J.P. (2008). The role of glutathione in photosynthetic organisms:Emerging functions for glutaredoxins and glutathionylation. Annu Rev of Plant Biol 59,143-166.
86. Rouhier, N., Unno, H., Bandyopadhyay, S., Masip, L., et al. (2007). Functional, structural, and spectroscopic characterization of a glutathione-ligated [2Fe-2S] cluster in poplar glutaredoxin C1. Proc Natl Acad Sci USA 104, 7379-7384.
87. Ruiz, M.T., Voinnet, O., and Baulcombe, D.C. (1998). Initiation and maintenance of virus-induced gene silencing. Plant Cell 10,937-946.
88. Rusanen, H., Majamaa, K., and Hassinen, I.E. (2000). Increased activities of antioxidant enzymes and decreased ATP concentration in cultured myoblasts with the 3243A->G mutation in mitochondrial DNA. Biochim Biophys Acta 1500, 10-16.
89. Scandalios, J.G. (2002). The rise of ROS. Trends Biochem Sci 27,483-486.
90. Senthil-Kumar, M., Gowda, H.V.R., Hema, R., Mysore, K.S., and Udayakumar, M. (2008). Virus-induced gene silencing and its application in characterizing genes involved in water-deficit-stress tolerance. J. Plant Physiol 165,1404-1421.
91. Shigeoka, S., Ishikawa, T., Tamoi, M., Miyagawa, Y., Takeda, T., Yabuta, Y., and Yoshimura, K. (2002). Regulation and function of ascorbate peroxidase isoenzymes. J Exp Bot 53,1305-1319.
92. Smirnoff, N. (2000). Ascorbic acid:metabolism and functions of a multi-facetted molecule. Curr Opin Plant Biol 3,229-235.
93. Smith, N.A., Singh, S.P., Wang, M.B., Stoutjesdijk, P.A., Green, A.G., and Waterhouse, P.M. (2000). Total silencing by intron-spliced hairpin RNAs. Nature 407,319-320.
94. Tada, Y., Spoel, S.H., Pajerowska-Mukhtar, K., Mou, Z., Song, J., Wang, C., Zuo, J., and Dong, X. (2008). Plant immunity requires conformational changes of NPR1 via S-nitrosylation and thioredoxins. Science 321,952-956.
95. Tao, X.R., and Zhou, X.P. (2004). A modified viral satellite DNA that suppresses gene expression in plants. Plant J 38,850-860.
96. Tao, X.R., Qian, Y.J., and Zhou, X.P. (2006). Modification of a viral satellite DNA-based gene silencing vector and its application to leaf or flower color change in Petunia hybrida. Chinese Sci Bull 51,2208-2213.
97. Turner, N.C. (1981). Techniques and Experimental Approaches for the Measurement of Plant Water Status. Plant & Soil 58,339-366.
98. Unver, T., and Budak, H. (2009). Virus-induced gene silencing, a post transcriptional gene silencing method. Int J Plant Genomics 2009, 198680-198687.
99. Voinnet, O. (2001). RNA silencing as a plant immune system against viruses. Trends Genet 17,449-459.
100.Voinnet, O. (2003). RNA silencing bridging the gaps in wheat extracts. Trends Plant Sci 8,307-309.
101.Vranova, E., Inze, D., and Van Breusegem, F. (2002). Signal transduction during oxidative stress. J Exp Bot 53,1227-1236.
102.Wang, Z., Xing, S.P., Birkenbihl, R.P., and Zachgo, S. (2009). Conserved functions of Arabidopsis and rice CC-type glutaredoxins in flower development and pathogen response. Molecular Plant 2,323-335.
103.Waterhouse, P.M., Wang, M.B., and Lough, T. (2001). Gene silencing as an adaptive defence against viruses. Nature 411,834-842.
104.Willekens, H., Langebartels, C., Tire, C., Van Montagu, M., Inze, D., and Van Camp, W. (1994). Differential expression of catalase genes in Nicotiana plumbaginifolia (L.). Proc Natl Acad Sci USA 91,10450-10454.
105.Wu, G., Shortt, B.J., Lawrence, E.B., Leon, J., Fitzsimmons, K.C., Levine, E.B., Raskin, I., and Shah, D.M. (1997). Activation of host defense mechanisms by elevated production of H2O2 in transgenic plants. Plant Physiol 115,427-435.
106.Xing, S., and Zachgo, S. (2008). ROXY1 and ROXY2, two Arabidopsis glutaredoxin genes, are required for anther development. Plant J 53,790-801.
107.Xing, S.P., Rosso, M.G., and Zachgo, S. (2005). ROXY1, a member of the plant glutaredoxin family, is required for petal development in Arabidopsis thaliana. Development 132,1555-1565.
108.Xiong, R.Y., Wu, J.X., Zhou, Y.J., and Zhou, X.P. (2009). Characterization and subcellular localization of an RNA silencing suppressor encoded by Rice stripe tenuivirus. Virology 387,29-40.
109.Yamaguchi, K., Mori, H., and Nishimura, M. (1995). A novel isoenzyme of ascorbate peroxidase localized on glyoxysomal and leaf peroxisomal membranes in pumpkin. Plant Cell Physiol 36,1157-1162.
110.Yoshimura, K., Yabuta, Y., Ishikawa, T., and Shigeoka, S. (2000). Expression of spinach ascorbate peroxidase isoenzymes in response to oxidative stresses. Plant Physiol 123,223-234.
111.Zhu, J.K. (2002). Salt and drought stress signal transduction in plants. Annu Rev Plant Biol 53,247-273.
112.Ziemann, M., Bhave, M., and Zachgo, S. (2009). Origin and diversification of land plant CC-Type glutaredoxins. Genome Biol Evol 2009,265-277.