Abstract
Genes encoding Rboh (respiratory burst oxidase homolog) have been described in a variety of plant species. Transcriptional regulation of rboh genes has been shown during plant response to stress or phytohormone treatment. In vitro conditions often induce plant stress that leads to slow plant growth, early senescence, or even recalcitrance to in vitro growth. Production of reactive oxygen species (ROS) is an important manifestation of plant environmental stress. Expression of rboh orthologs in plant tissues grown under in vitro conditions and their role in response to abiotic stress is not fully understood. Therefore the aim of this study was to identify rboh homologues in apple (Mdrboh) and to characterize their expression during the senescence of apple shoot culture in vitro. Similarity searches using Arabidopsis Rboh (AtRboh) protein sequences revealed nine homologous rboh genes in the apple genome. Phylogenetic analysis using conservative N-terminal half region sequences of the apple, Arabidopsis, pear, peach and wild strawberry Rboh proteins revealed four groups of related sequences that were linked to AtRboh D, E, F and H. A homologue with unique sequence was annotated as Rboh K in plants of the Rosaceae family. Expression of MdrbohD1–3 and F was detected in apple leaves, in vitro cultures of shoots and cell suspension, and MdrbohE2 and H1–2 varied among the tissues. The MdrbohD1–2 and F genes were differentially expressed during transfer, growth and senescence stages of apple in vitro shoot culture having varying levels of oxidative stress damage, suggesting transcriptional regulation of the Mdrboh genes in the apple shoot culture, and a potential for distinctive functions of the three Rboh D orthologs.
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Abbreviations
- cv.:
-
Cultivar
- FW:
-
Fresh weight
- MDA:
-
Malondialdehyde
- Rboh:
-
Respiratory burst oxidase homolog
- ROS:
-
Reactive oxygen species
- O ·−2 :
-
Superoxide
- H2O2 :
-
Hydrogen peroxide
- SEM:
-
Standard error of the mean
- TBARS:
-
Thiobarbituric acid reactive substances
References
Aldwinckle H, Malnoy M (2009) Plant regeneration and transformation in the Rosaceae. In: Nageswara-Rao M, Soneji JR (eds) Transgenic plant journal 3 (Special Issue 1). Global Science Books, Isleworth, pp 1–39
Altschul SF, Gish W, Miller W, Myers EW, Lipman DJ (1990) Basic local alignment search tool. J Mol Biol 215:403–410. doi:10.1016/S0022-2836(05)80360-2
Amicucci E, Gaschler K, Ward JM (1999) NADPH oxidase genes from tomato (Lycopersicon esculentum) and curly-leaf pondweed (Potamogeton crispus). Plant Biol 1:524–528. doi:10.1111/j.1438-8677.1999.tb00778.x
Andriunas FA, Zhang HM, Xia X, Offler CE, McCurdy DW, Patrick JW (2012) Reactive oxygen species form part of a regulatory pathway initiating trans-differentiation of epidermal transfer cells in Vicia faba cotyledons. J Exp Bot 63:3617–3629. doi:10.1093/jxb/ers029
Apel K, Hirt H (2004) Reactive oxygen species: metabolism, oxidative stress, and signal transduction. Annu Rev Plant Biol 55:373–399. doi:10.1146/annurev.arplant.55.031903.141701
Bairu MW, Kane ME (2011) Physiological and developmental problems encountered by in vitro cultured plants. Plant Growth Regul 62:101–103. doi:10.1007/s10725-011-9565-2
Balen B, Tkalec M, Pavokovic D, Pevalek-Kozlina B, Krsnik-Rasol M (2009) Growth conditions in in vitro culture can induce oxidative stress in Mammillaria gracilis tissues. J Plant Growth Regul 28:36–45. doi:10.1007/s00344-008-9072-5
Benson EE (2000a) Special symposium: in vitro plant recalcitrance do free radicals have a role in plant tissue culture recalcitrance? In Vitro Cell Dev Biol Plant 36:163–170. doi:10.1007/s11627-000-0032-4
Benson EE (2000b) Do free radicals have a role in plant tissue culture recalcitrance? In Vitro Cell Dev Biol Plant 36:163–170. doi:10.1007/s11627-000-0032-4
Blomster T, Salojarvi J, Sipari N, Brosche M, Ahlfors R, Keinanen M, Overmyer K, Kangasjarvi J (2011) Apoplastic reactive oxygen species transiently decrease auxin signaling and cause stress-induced morphogenic response in Arabidopsis. Plant Physiol 157:1866–1883. doi:10.1104/pp.111.181883
Bonasera JM, Kim JF, Beer SV (2006) PR genes of apple: identification and expression in response to elicitors and inoculation with Erwinia amylovora. BMC Plant Biol 6:23. doi:10.1186/1471-2229-6-23
Cassells A, Curry R (2001) Oxidative stress and physiological, epigenetic and genetic variability in plant tissue culture: implications for micropropagators and genetic engineers. Plant Cell Tissue Organ 64:145–157. doi:10.1023/A:1010692104861
Chagne D, Crowhurst RN, Pindo M et al (2014) The draft genome sequence of European pear (Pyrus communis L. ‘Bartlett’). PLoS ONE 9:e92644. doi:10.1371/journal.pone.0092644
Chatzissavvidis C, Veneti G, Papadakis I, Therios I (2008) Effect of NaCl and CaCl2 on the antioxidant mechanism of leaves and stems of the rootstock CAB-6P (Prunus cerasus L.) under in vitro conditions. Plant Cell Tissue Organ 95:37–45. doi:10.1007/s11240-008-9411-z
Cheng C, Xu X, Gao M, Li J, Guo C, Song J, Wang X (2013) Genome-wide analysis of respiratory burst oxidase homologs in grape (Vitis vinifera L.). Int J Mol Sci 14:24169–24186. doi:10.3390/ijms141224169
Czynczyk A, Bielicki P, Bartosiewicz B (2008) Evaluation of the effect of P 14 rootstock propagated in vitro and in stoolbeds on the growth and yielding of three apple cultivars. J Fruit Ornam Plant Res 16:25–30
Debergh PC, Read PE (1991) Micropropagation. In: Debergh PC, Zimmerman RH (eds) Micropropagation: Technology and Application. Kluwer Academic, Dordrecht, pp 1–13
Defilippi BG, Kader AA, Dandekar AM (2005) Apple aroma: alcohol acyltransferase, a rate limiting step for ester biosynthesis, is regulated by ethylene. Plant Sci 168:1199–1210. doi:10.1016/j.plantsci.2004.12.018
Denness L, McKenna JF, Segonzac C, Wormit A, Madhou P, Bennett M, Mansfield J, Zipfel C, Hamann T (2011) Cell wall damage-induced lignin biosynthesis is regulated by a reactive oxygen species- and jasmonic acid-dependent process in Arabidopsis. Plant Physiol 156:1364–1374. doi:10.1104/pp.111.175737
Dobranszki J, da Silva JA (2010) Micropropagation of apple—a review. Biotechnol Adv 28:462–488. doi:10.1016/j.biotechadv.2010.02.008
Doke N (1983) Generation of superoxide anion by potato tuber protoplasts during the hypersensitive response to hyphal cell wall components of Phytophthora infestans and specific inhibition of the reaction by suppressors of hypersensitivity. Physiol Plant Pathol 23:345–357. doi:10.1016/0048-4059(83)90020-6
Doke N (1997) The oxidative burst: roles in signal transduction and plant stress. In: Oxidative stress and the molecular biology of antioxidant defenses, Cold Spring Harbor Laboratory Press, Plainview, pp 785–813
Erturk U, Sivritepe N, Yerlikaya C, Bor M, Ozdemir F, Turkan I (2007) Responses of the cherry rootstock to salinity in vitro. Biol Plant 51:597–600. doi:10.1007/s10535-007-0132-7
Foreman J, Demidchik V, Bothwell JH, Mylona P, Miedema H, Torres MA, Linstead P, Costa S, Brownlee C, Jones JD, Davies JM, Dolan L (2003) Reactive oxygen species produced by NADPH oxidase regulate plant cell growth. Nature 422:442–446. doi:10.1038/nature01485
Franck T, Kevers C, Gaspar T, Dommes J, Deby C, Greimers R, Serteyn D, Deby-Dupont G (2004) Hyperhydricity of Prunus avium shoots cultured on gelrite: a controlled stress response. Plant Physiol Biochem 42:519–527. doi:10.1016/j.plaphy.2004.05.003
Goujon M, McWilliam H, Li W, Valentin F, Squizzato S, Paern J, Lopez R (2010) A new bioinformatics analysis tools framework at EMBL-EBI. Nucleic Acids Res 38:W695–W699. doi:10.1093/nar/gkq313
Groom QJ, Torres MA, Fordham-Skelton AP, Hammond-Kosack KE, Robinson NJ, Jones JD (1996) rbohA, a rice homologue of the mammalian gp91phox respiratory burst oxidase gene. Plant J 10:515–522. doi:10.1046/j.1365-313X.1996.10030515.x
Hu B, Jin J, Guo AY, Zhang H, Luo J, Gao G (2014) GSDS 2.0: an upgraded gene feature visualization server. Bioinformatics. doi:10.1093/bioinformatics/btu817
Ivanchenko MG, den Os D, Monshausen GB, Dubrovsky JG, Bednarova A, Krishnan N (2013) Auxin increases the hydrogen peroxide (H2O2) concentration in tomato (Solanum lycopersicum) root tips while inhibiting root growth. Ann Bot 112:1107–1116. doi:10.1093/aob/mct181
Jagendorf AT, Takabe T (2001) Inducers of glycinebetaine synthesis in barley. Plant Physiol 127:1827–1835. doi:10.1104/pp.010392
Jakubowicz M, Galganska H, Nowak W, Sadowski J (2010) Exogenously induced expression of ethylene biosynthesis, ethylene perception, phospholipase D, and Rboh-oxidase genes in broccoli seedlings. J Exp Bot 61:3475–3491. doi:10.1093/jxb/erq177
Jesaitis AJ, Heners PR, Briggs WR, Hertel R (1977) Characterization of a membrane fraction containing a b-type cytochrome. Plant Physiol 59:941–947. doi:10.1104/pp.59.5.941
Jung S, Ficklin SP, Lee T et al (2014) The genome database for Rosaceae (GDR): year 10 update. Nucleic Acids Res 42:D1237–D1244. doi:10.1093/nar/gkt1012
Kaur G, Sharma A, Guruprasad K, Pati PK (2014) Versatile roles of plant NADPH oxidases and emerging concepts. Biotechnol Adv 32:551–563. doi:10.1016/j.biotechadv.2014.02.002
Keller T, Damude HG, Werner D, Doerner P, Dixon RA, Lamb C (1998) A plant homolog of the neutrophil NADPH oxidase gp91phox subunit gene encodes a plasma membrane protein with Ca2+ binding motifs. Plant Cell 10:255–266. doi:10.1105/tpc.10.2.255
Kumar GN, Iyer S, Knowles NR (2007) Strboh A homologue of NADPH oxidase regulates wound-induced oxidative burst and facilitates wound-healing in potato tubers. Planta 227:25–36. doi:10.1007/s00425-007-0589-9
Kurkcuoglu S, Degenhardt J, Lensing J, Al-Masri AN, Gau AE (2007) Identification of differentially expressed genes in Malus domestica after application of the non-pathogenic bacterium Pseudomonas fluorescens Bk3 to the phyllosphere. J Exp Bot 58:733–741. doi:10.1093/jxb/erl249
Kwak JM, Mori IC, Pei ZM, Leonhardt N, Torres MA, Dangl JL, Bloom RE, Bodde S, Jones JD, Schroeder JI (2003) NADPH oxidase AtrbohD and AtrbohF genes function in ROS-dependent ABA signaling in Arabidopsis. EMBO J 22:2623–2633. doi:10.1093/emboj/cdg277
Lightfoot DJ, Boettcher A, Little A, Shirley N, Able AJ (2008) Identification and characterisation of barley (Hordeum vulgare) respiratory burst oxidase homologue family members. Funct Plant Biol 35:347–359. doi:10.1071/FP08109
Lin F, Ding H, Wang J, Zhang H, Zhang A, Zhang Y, Tan M, Dong W, Jiang M (2009) Positive feedback regulation of maize NADPH oxidase by mitogen-activated protein kinase cascade in abscisic acid signalling. J Exp Bot 60:3221–3238. doi:10.1093/jxb/erp157
Lombardi L, Sebastiani L, Vitagliano C (2003) Physiological, biochemical, and molecular effects of in vitro induced iron deficiency in peach rootstock Mr.S 2/5. J Plant Nutr 26:2149–2163. doi:10.1081/PLN-120024271
Macpherson N, Takeda S, Shang Z, Dark A, Mortimer JC, Brownlee C, Dolan L, Davies JM (2008) NADPH oxidase involvement in cellular integrity. Planta 227:1415–1418. doi:10.1007/s00425-008-0716-2
Marino D, Andrio E, Danchin EG, Oger E, Gucciardo S, Lambert A, Puppo A, Pauly N (2011) A Medicago truncatula NADPH oxidase is involved in symbiotic nodule functioning. New Phytol 189:580–592. doi:10.1111/j.1469-8137.2010.03509.x
Marino D, Dunand C, Puppo A, Pauly N (2012) A burst of plant NADPH oxidases. Trends Plant Sci 17:9–15. doi:10.1016/j.tplants.2011.10.001
Maruta T, Inoue T, Tamoi M, Yabuta Y, Yoshimura K, Ishikawa T, Shigeoka S (2011) Arabidopsis NADPH oxidases, AtrbohD and AtrbohF, are essential for jasmonic acid-induced expression of genes regulated by MYC2 transcription factor. Plant Sci 180:655–660. doi:10.1016/j.plantsci.2011.01.014
Miller G, Schlauch K, Tam R, Cortes D, Torres MA, Shulaev V, Dangl JL, Mittler R (2009) The plant NADPH oxidase RBOHD mediates rapid systemic signaling in response to diverse stimuli. Sci Signal 2:ra45. doi:10.1126/scisignal.2000448
Mittler R, Vanderauwera S, Suzuki N, Miller G, Tognetti VB, Vandepoele K, Gollery M, Shulaev V, Van BF (2011) ROS signaling: the new wave? Trends Plant Sci 16:300–309. doi:10.1016/j.tplants.2011.03.007
Molassiotis AN, Diamantidis GC, Therios IN, Tsirakoglou V, Dimassi KN (2005) Oxidative stress, antioxidant activity and Fe(III)-chelate reductase activity of five Prunus rootstocks explants in response to Fe deficiency. Plant Growth Regul 46:69–78. doi:10.1007/s10725-005-6396-z
Molassiotis AN, Sotiropoulos S, Tanou G, Kofidis G, Diamantidis G, Therios I (2006) Antioxidant and anatomical responses in shoot culture of the apple rootstock MM 106 treated with NaCl, KCl, mannitol or sorbitol. Biol Plant 50:61–68. doi:10.1007/s10535-005-0075-9
Montiel J, Nava N, Cardenas L, Sanchez-Lopez R, Arthikala MK, Santana O, Sanchez F, Quinto C (2012) A Phaseolus vulgaris NADPH oxidase gene is required for root infection by Rhizobia. Plant Cell Physiol 53:1751–1767. doi:10.1093/pcp/pcs120
Morel J, Fromentin J, Blein JP, Simon-Plas F, Elmayan T (2004) Rac regulation of NtrbohD, the oxidase responsible for the oxidative burst in elicited tobacco cell. Plant J 37:282–293. doi:10.1046/j.1365-313X.2003.01957.x
Muller K, Carstens AC, Linkies A, Torres MA, Leubner-Metzger G (2009) The NADPH-oxidase AtrbohB plays a role in Arabidopsis seed after-ripening. New Phytol 184:885–897. doi:10.1111/j.1469-8137.2009.03005.x
Muller K, Linkies A, Leubner-Metzger G, Kermode AR (2012) Role of a respiratory burst oxidase of Lepidium sativum (cress) seedlings in root development and auxin signalling. J Exp Bot 63:6325–6334. doi:10.1093/jxb/ers284
Murashige T, Skoog F (1962) A revised medium for rapid growth and bioassays with tobacco tissue cultures. Physiol Plant 15:473–497. doi:10.1111/j.1399-3054.1962.tb08052.x
Peer WA, Cheng Y, Murphy AS (2013) Evidence of oxidative attenuation of auxin signalling. J Exp Bot 64:2629–2639. doi:10.1093/jxb/ert152
Potocky M, Jones MA, Bezvoda R, Smirnoff N, Zarsky V (2007) Reactive oxygen species produced by NADPH oxidase are involved in pollen tube growth. New Phytol 174:742–751. doi:10.1111/j.1469-8137.2007.02042.x
Proels RK, Oberhollenzer K, Pathuri IP, Hensel G, Kumlehn J, Huckelhoven R (2010) RBOHF2 of barley is required for normal development of penetration resistance to the parasitic fungus Blumeria graminis f. sp. hordei. Mol Plant Microbe Interact 23:1143–1150. doi:10.1094/MPMI-23-9-1143
Remans T, Opdenakker K, Smeets K, Mathijsen D, Vangrosveld J, Cuypers A (2010) Metal-specific and NADPH oxidase dependent changes in lipoxygenase and NADPH oxidase gene expression in Arabidopsis thaliana exposed to cadmium or excess copper. Funct Plant Biol 37:532–544. doi:10.1071/FP09194
Rojas-Martinez L, Visser RGF, De Klerk G-J (2010) The hyperhydricity syndrome: waterlogging of plant tissues as a major cause. Propag Ornam Plants 10:169–175
Rozen S, Skaletsky HJ (2000) Primer3 on the WWW for general users and for biologist programmers. In: Krawetz S, Misener S (eds) Bioinformatics methods and protocols: methods in molecular biology. Humana Press, Totowa, pp 365–386
Sagi M, Fluhr R (2006) Production of reactive oxygen species by plant NADPH oxidases. Plant Physiol 141:336–340. doi:10.1104/pp.106.078089
Sagi M, Davydov O, Orazova S, Yesbergenova Z, Ophir R, Stratmann JW, Fluhr R (2004) Plant respiratory burst oxidase homologs impinge on wound responsiveness and development in Lycopersicon esculentum. Plant Cell 16:616–628. doi:10.1105/tpc.019398
Saher S, Piqueras A, Hellin E, Olmos E (2004) Hyperhydricity in micropropagated carnation shoots: the role of oxidative stress. Physiol Plant 120:152–161. doi:10.1111/j.0031-9317.2004.0219.x
Saitou N, Nei M (1987) The neighbor-joining method: a new method for reconstructing phylogenetic trees. Mol Biol Evol 4:406–425
Schopfer P, Liszkay A (2006) Plasma membrane-generated reactive oxygen intermediates and their role in cell growth of plants. BioFactors 28:73–81
Sen A (2012) Oxidative stress studies in plant tissue culture. In: El-Missiry MA (ed) Antioxidant enzyme. InTech, Rijeka
Shulaev V, Korban SS, Sosinski B et al (2008) Multiple models for Rosaceae genomics. Plant Physiol 147:985–1003. doi:10.1104/pp.107.115618
Shulaev V, Sargent DJ, Crowhurst RN et al (2011) The genome of woodland strawberry (Fragaria vesca). Nat Genet 43:109–116. doi:10.1038/ng.740
Si Y, Dane F, Rashotte A, Kang K, Singh NK (2010) Cloning and expression analysis of the Ccrboh gene encoding respiratory burst oxidase in Citrullus colocynthis and grafting onto Citrullus lanatus (watermelon). J Exp Bot 61:1635–1642. doi:10.1093/jxb/erq031
Sievers F, Wilm A, Dineen D, Gibson TJ, Karplus K, Li W, Lopez R, McWilliam H, Remmert M, Soding J, Thompson JD, Higgins DG (2011) Fast, scalable generation of high-quality protein multiple sequence alignments using Clustal Omega. Mol Syst Biol 7:539. doi:10.1038/msb.2011.75
Simon-Plas F, Elmayan T, Blein JP (2002) The plasma membrane oxidase NtrbohD is responsible for AOS production in elicited tobacco cells. Plant J 31:137–147. doi:10.1046/j.1365-313X.2002.01342.x
Sivritepe N, Erturk U, Yerlikaya C, Turkan I, Bor M, Ozdemir F (2008) Response of the cherry rootstock to water stress induced in vitro. Biol Plant 52:573–576. doi:10.1007/s10535-008-0114-4
Solovyev V, Kosarev P, Seledsov I, Vorobyev D (2006) Automatic annotation of eukaryotic genes, pseudogenes and promoters. Genome Biol 7(Suppl 1):S10–S12. doi:10.1186/gb-2006-7-s1-s10
Sotiropoulos TE (2007) Effect of NaCl and CaCl2 on growth and contents of minerals, chlorophyll, proline and sugars in the apple rootstock M4 cultured in vitro. Biol Plant 51:177–180. doi:10.1007/s10535-007-0035-7
Sotiropoulos TE, Molassiotis AN, Almaliotis D, Mouhtaridou G, Dimassi KN, Therios I, Diamantidis G (2006) Growth, nutritional status, chlorophyll content, and antioxidant responses of the apple rootstock MM111 shoots cultured under high boron concentrations in vitro. J Plant Nutr 29:575–583. doi:10.1080/01904160500526956
Suzuki N, Miller G, Morales J, Shulaev V, Torres MA, Mittler R (2011) Respiratory burst oxidases: the engines of ROS signaling. Curr Opin Plant Biol 14:691–699. doi:10.1016/j.pbi.2011.07.014
Tamura K, Stecher G, Peterson D, Filipski A, Kumar S (2013) MEGA6: molecular evolutionary genetics analysis version 6.0. Mol Biol Evol 30:2725–2729. doi:10.1093/molbev/mst197
Torres MA, Dangl JL (2005) Functions of the respiratory burst oxidase in biotic interactions, abiotic stress and development. Curr Opin Plant Biol 8:397–403. doi:10.1016/j.pbi.2005.05.014
Torres MA, Onouchi H, Hamada S, Machida C, Hammond-Kosack KE, Jones JD (1998) Six Arabidopsis thaliana homologues of the human respiratory burst oxidase (gp91phox). Plant J 14:365–370. doi:10.1046/j.1365-313X.1998.00136.x
Velasco R, Zharkikh A, Affourtit J et al (2010) The genome of the domesticated apple (Malus × domestica Borkh.). Nat Genet 42:833–839. doi:10.1038/ng.654
Verde I, Abbott AG, Scalabrin S et al (2013) The high-quality draft genome of peach (Prunus persica) identifies unique patterns of genetic diversity, domestication and genome evolution. Nat Genet 45:487–494. doi:10.1038/ng.2586
Wang W, Zheng H, Fan C, Li J, Shi J, Cai Z, Zhang G, Liu D, Zhang J, Vang S, Lu Z, Wong GK, Long M, Wang J (2006) High rate of chimeric gene origination by retroposition in plant genomes. Plant Cell 18:1791–1802. doi:10.1105/tpc.106.041905
Wang GF, Li WQ, Li WY, Wu GL, Zhou CY, Chen KM (2013) Characterization of rice NADPH oxidase genes and their expression under various environmental conditions. Int J Mol Sci 14:9440–9458. doi:10.3390/ijms14059440
Waterhouse AM, Procter JB, Martin DMA, Clamp M, Barton GJ (2009) Jalview version 2—a multiple sequence alignment editor and analysis workbench. Bioinformatics. doi:10.1093/bioinformatics/btp033
Wohlgemuth H, Mittelstrass K, Kschieschan S, Bender J, Weigel H-J, Overmyer K, Kangasjarvi J, Sandermann H, Langebartels C (2002) Activation of an oxidative burst is a general feature of sensitive plants exposed to the air pollutant ozone. Plant Cell Environm 25:717–726. doi:10.1046/j.1365-3040.2002.00859.x
Yamamizo C, Kuchimura K, Kobayashi A, Katou S, Kawakita K, Jones JD, Doke N, Yoshioka H (2006) Rewiring mitogen-activated protein kinase cascade by positive feedback confers potato blight resistance. Plant Physiol 140:681–692. doi:10.1104/pp.105.074906
Yoshioka H, Sugie K, Park HJ, Maeda H, Tsuda N, Kawakita K, Doke N (2001) Induction of plant gp91phox homolog by fungal cell wall, arachidonic acid, and salicylic acid in potato. Mol Plant Microbe Interact 14:725–736. doi:10.1094/MPMI.2001.14.6.725
Yoshioka H, Numata N, Nakajima K, Katou S, Kawakita K, Rowland O, Jones JD, Doke N (2003) Nicotiana benthamiana gp91phox homologs NbrbohA and NbrbohB participate in H2O2 accumulation and resistance to Phytophthora infestans. Plant Cell 15:706–718. doi:10.1105/tpc.008680
Zhao S, Fernald RD (2005) Comprehensive algorithm for quantitative real-time polymerase chain reaction. J Comput Biol 12:1047–1064. doi:10.1089/cmb.2005.12.1047
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This research was funded by the Lithuanian Research Council Grant No. MIP-47/2013.
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Conceived and designed the experiments: AJJ, DB, VS. Performed the experiments: DC, DG, GS, IM. Analyzed the data: DB, DC, IM. Wrote the paper: AJJ, DB, DC, IM, VS. All authors read and approved the final manuscript.
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Cepauskas, D., Miliute, I., Staniene, G. et al. Characterization of apple NADPH oxidase genes and their expression associated with oxidative stress in shoot culture in vitro. Plant Cell Tiss Organ Cult 124, 621–633 (2016). https://doi.org/10.1007/s11240-015-0920-2
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DOI: https://doi.org/10.1007/s11240-015-0920-2