Abstract
In Arabidopsis, it has been clarified that AGO4 protein is implicated in a phenomenon termed RNA-directed DNA methylation (RdDM). Previously, four orthologs of AtAGO4 were cloned in tomato, designated as SlAGO4A–SlAGO4D. Here, we studied the role of the SlAGO4A gene in regulating salt and drought tolerance in tomato. SlAGO4A-down-regulating (AS) transgenic tomato plants showed enhanced tolerance to salt and drought stress compared to wild-type (WT) and SlAGO4A-overexpressing (OE) transgenic plants, as assessed by physiological parameters such as seed germination rate, primary root length, chlorophyll/proline/MDA/soluble sugar/RWC content, and survival rate. Moreover, several genes involved in ROS scavenging and plant defense, including CAT, SOD, GST, POD, APX, LOX, and PR1, were up- or down-regulated consistently under salt and drought stress. Notably, expression levels of some DNA methyltransferase genes and RNAi pathway genes were significantly lower in AS plants than in WT. Taken together, our results suggest that SlAGO4A gene plays a negative role under salt and drought stress in tomato probably through the modulation of DNA methylation as well as the classical RNAi pathway. Hence, it may serve as a useful biotechnological tool for the genetic improvement of stress tolerance in crops.





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Agorio A, Vera P (2007) ARGONAUTE4 is required for resistance to Pseudomonas syringae in Arabidopsis. Plant Cell 19:3778–3790
Bai M, Yang GS, Chen WT, Mao ZC, Kang HX, Chen GH, Yang YH, Xie BY (2012) Genome-wide identification of Dicer-like, Argonaute and RNA-dependent RNA polymerase gene families and their expression analyses in response to viral infection and abiotic stresses in Solanum lycopersicum. Gene 501:52–62
Banu MSK, Huda KMK, Sahoo RK, Garg B, Tula S, Islam SMS, Tuteja R, Tuteja N (2015) Pea p68 imparts salinity stress tolerance in rice by scavenging of ROS-mediated H2O2 and interacts with Argonaute. Plant Mol Biol Rep 33:221–238
Bates L, Waldren R, Teare I (1973) Rapid determination of free proline for water-stress studies. Plant Soil 39:205–207
Bilichak A, Ilnystkyy Y, Hollunder J, Kovalchuk I (2012) The progeny of Arabidopsis thaliana plants exposed to salt exhibit changes in DNA methylation, histone modifications and gene expression. PLoS One 7:e30515
Boyko A, Blevins T, YaoY Golubov A, Bilichak A, Ilnytskyy Y, Hollander J, Meins F, Kovalchuk I (2010) Transgenerational adaptation of Arabidopsis to stress requires DNA methylation and the function of Dicer-like proteins. PLoS One 5:e9514
Brodersen P, Vioinnet O (2006) The diversity of RNA silencing pathways in plants. Trends Genet 22:268–280
Chan SWL, Zilberman D, Xie ZX, Johansen LK, Carrington JC, Jacobsen SE (2004) RNA silencing genes control de novo DNA methylation. Science 303:1336
Chandar BY, Mehanathan M, Pandey Garima, Manoj P (2015) Identification, characterization and expression profiling of Dicer-Like, Argonaute and RNA-dependent RNA polymerase gene families in foxtail millet. Plant Mol Boil Rep 33:43–55
Choi CS, Sano H (2007) Abiotic-stress induces demethylation and transcriptional activation of a gene encoding a glycerophosphodiesterase-like protein in tobacco plants. Mol Genet Genomics 277:589–600
Choudhury S, Panda P, Sahoo L, Panda SK (2013) Reactive oxygen species signaling in plants under abiotic stress. Plant Signal Behav 8:e23681
Dat J, Vandenabeele S, Vranova E, Van Montagu M, Inze D, Van Breusegem F (2000) Dual action of the active oxygen species during plant stress responses. Cell Mol Life Sci 57:779–795
Fukao T, Xu K, Ronald PC, Bailey-Serres J (2006) A variable cluster of ethylene response factor-like genes regulates metabolic and developmental acclimation responses to submergence in rice. Plant Cell 18:2021–2034
Gill SS, Tuteja N (2010) Reactive oxygen species and antioxidant machinery in abiotic stress tolerance in crop plants. Plant Physiol Biochem 48:909–939
Heath RL, Packer L (1968) Photoperoxidation in isolated chloroplasts. I. Kinetics and stoichiometry of fatty acid peroxidation. Arch Biochem Biophys 125:189–198
Higo K, Ugawa Y, Iwamoto M, Korenaga T (1999) Plant cis-acting regulatory DNA elements (PLACE) database. Nucleic Acids Res 27:297–300
Hoagland DR, Arnon DI (1950) The water-culture method for growing plants without soil. Circular. California Agricultural Experiment Station 347 (2nd edit)
Hutvagner G, Simard MJ (2008) Argonaute proteins: key players in RNA silencing. Nat Rev Mol Cell Biol 9:22–32
Ito H, Gaubert H, Bucher E, Mirouze M, Vaillant I, Paszkowski J (2011) An siRNA pathway prevents transgenerational retrotransposition in plants subjected to stress. Nature 472:115–119
Kapoor M, Arora R, Lama T, Nijhawan A, Khurana JP, Tyagi AK, Kapoor S (2008) Genome-wide identification, organization and phylogenetic analysis of Dicer-like, Argonaute and RNA-dependent RNA polymerase gene families and their expression analysis during reproductive development and stress in rice. BMC Genom 9:451
Karan R, DeLeon T, Biradar H, Subudhi PK (2012) Salt stress induced variation in DNA methylation pattern and its influence on gene expression in contrasting rice genotypes. PLoS One 7:e40203
Khraiwesh B, Zhu JK, Zhu J (2012) Role of miRNAs and siRNAs in biotic and abiotic stress responses of plants. Biochim Biophys Acta 1819:137–148
Kimura S, Sinha N (2008) Tomato (Solanum lycopersicum): a model fruit-bearing crop. Cold Spring Harb Protoc 11:1–9
Li CH, Chiang CP, Yang JY, Ma CJ, Chen YC, Yen HE (2014) RING-type ubiquitin ligase McCPN1 catalyzes UBC8-dependent protein ubiquitination and interacts with Argonaute 4 in halophyte ice plant. Plant Physiol Biochem 80:211–219
Lichtenthaler HK (1987) Chlorophylls and carotenoids: pigments of photosynthetic biomembranes. Methods Enzymol 148:350–382
Mallory A, Vaucheret H (2010) Form, function, and regulation of Argonaute proteins. Plant cell 22:3879–3889
Matzke M, Kanno T, Daxinger L, Huettel B, Matzke AJ (2009) RNA mediated chromatin-based silencing in plants. Curr Opin Cell Biol 21:367–376
Meng FR, Jia HY, Ling N, Xue YL, Liu H, Wang KT, Yin J, Li YC (2013) Cloning and characterization of two Argonaute genes in wheat (Triticum aestivum L.). BMC Plant Biol 13:18
Mittler R (2002) Oxidative stress, antioxidants and stress tolerance. Trends Plant Sci 7:405–410
Mittler R, Vanderauwera S, Gollery M, Van Breusegem F (2004) Reactive oxygen gene network of plants. Trends Plant Sci 9:490–498
Mittova V, Volokita M, Guy M, Tal M (2000) Activities of SOD and the ascorbate-glutathione cycle enzymes in subcellular compartments in leaves and roots of the cultivated tomato and its wild salt-tolerant relative Lycopersicon pennellii. Physiol Plant 110:42–51
Niki T, Mitsuhara I, Seo S, Ohtsubo N, Ohashi Y (1998) Antagonistic effect of salicylic acid and jasmonic acid on the expression of pathogenesis-related (PR) protein genes in wounded mature tobacco leaves. Plant Cell Physiol 39:500–507
Pan Y, Seymour GB, Lu C, Hu Z, Chen X, Chen G (2012) An ethylene response factor (ERF5) promoting adaptation to drought and salt tolerance in tomato. Plant Cell Rep 31:349–360
Popova OV, Dihn HQ, Aufsatz W, Jonak C (2013) The RdDM pathway is required for basal heat tolerance in Arabidopsis. Mol Plant 6:396–410
Qian Y, Cheng Y, Cheng X, Jiang H, Zhu S, Cheng B (2011) Identification and characterization of Dicer-like, Argonaute and RNA-dependent RNA polymerase gene families in maize. Plant Cell Rep 30:1347–1363
Sahu PP, Rai NK, Chakraborty S, Singh M, Chandrappa PH, Ramesh B, Chattopadhyay D, Prasad M (2010) Tomato cultivar tolerant to tomato leaf curl New Delhi virus infection induces virus-specific short interfering RNA accumulation and defence-associated host gene expression. Mol Plant Pathol 11:531–544
Seo JK, Wu J, Li Y, Li Y, Jin H (2013) Contribution of small RNA pathway components in plant immunity. Mol Plant Microbe Interact 26:617–625
Singh M, Singh S, Randhawa H, Singh J (2013) Polymorphic homoeolog of key gene of RdDM pathway, ARGONAUTE4_9 class is associated with pre-harvest sprouting in wheat (Triticum aestivum L.). PLoS One 8:e77009
Song Y, Ji D, Li S, Wang P, Li Q, Xiang F (2012) The dynamic changes of DNA methylation and histone modifications of salt responsive transcription factor genes in soybean. PLoS One 7:e41274
Szekely G, Abraham E, Cseplo A, Rigo G, Zsigmond L, Csiszar J, Ayaydin F, Strizhov N, Jasik J, Schmelzer E, Koncz C, Szabados L (2008) Duplicated P5CS genes of Arabidopsis play distinct roles in stress regulation and developmental control of proline biosynthesis. Plant J 53:11–28
Tricker PJ, Gibbings JG, Rodriguez-Lopez CM, Hadley P, Wilkinson MJ (2012) Low relative humidity triggers RNA-directed de novo DNA methylation and suppression of genes controlling stomatal development. J Exp Bot 63:3799–3813
Vaucheret H (2008) Plant ARGONAUTES. Trends Plant Sci 13:350–358
Wang M, Qin L, Xie C, Li W, Yuan J, Kong L, Yu WL, Xia GM, Liu SW (2014) Induced and constitutive DNA methylation in a salinity-tolerant wheat introgression line. Plant Cell Physiol 55:1354–1365
Wheeler BS (2013) Small RNAs, big impact: small RNA pathways in transposon control and their effect on the host stress response. Chromosom Res 21:587–600
Xian ZQ, Yang YW, Huang W, Tang N, Wang XY, Li ZG (2013) Molecular cloning and charactersition of SlAGO family in tomato. BMC Plant Biol 13:126
Xian ZQ, Huang W, Yang YW, Tang N, Zhang C, Ren MZ, Li ZG (2014) miR168 influences phase transition, leaf epinasty, and fruit development via SlAGO1s in tomato. J Exp Bot 65:6655–6666
Xiong L, Schumaker KS, Zhu JK (2002) Cell signaling during cold, drought, and salt stress. Plant Cell 14:165–183
Zhai LH, Sun W, Zhang K, Jia HT, Liu L, Liu ZJ, Teng F, Zhang ZX (2014) Identification and characterization of Argonaute gene family and meiosis-enriched Argonaute during sporogenesis in maize. J Integr Plant Biol 56:1042–1052
Zhang XM, Zhao HW, Cao S, Wang WC, Katiyar-Agarwal S, Huang HD, Raikhel N, Jin HL (2011) Arabidopsis Argonaute 2 regulates innate immunity via miRNA393*-mediated silencing of a golgi-localized SNARE gene, MEMB12. Mol Cell 42:356–366
Zilberman D, Cao XF, Jacobsen SE (2003) ARGONAUTE4 control of locus-specific siRNA accumulation and DNA and histone methylation. Science 299:716–719
Acknowledgments
This work was supported by grants from the National Basic Research Program of China (2013CB127101), the National High Technology Research and Development Program of China (2012AA101702), the National Natural Science Foundation of China (31272166), the Fundamental Research Funds for the Central Universities (CDJXS10231118).
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Wei Huang and Zhiqiang Xian have contributed equally to this work.
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Huang, W., Xian, Z., Hu, G. et al. SlAGO4A, a core factor of RNA-directed DNA methylation (RdDM) pathway, plays an important role under salt and drought stress in tomato. Mol Breeding 36, 28 (2016). https://doi.org/10.1007/s11032-016-0439-1
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DOI: https://doi.org/10.1007/s11032-016-0439-1


