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Banana NAC transcription factor MusaNAC042 is positively associated with drought and salinity tolerance

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Abstract

Banana is an important fruit crop and its yield is hampered by multiple abiotic stress conditions encountered during its growth. The NAC (NAM, ATAF, and CUC) transcription factors are involved in plant response to biotic and abiotic stresses. In the present study, we studied the induction of banana NAC042 transcription factor in drought and high salinity conditions and its overexpression in transgenic banana to improve drought and salinity tolerance. MusaNAC042 expression was positively associated with stress conditions like salinity and drought and it encoded a nuclear localized protein. Transgenic lines of banana cultivar Rasthali overexpressing MusaNAC042 were generated by Agrobacterium-mediated transformation of banana embryogenic cells and T-DNA insertion was confirmed by PCR and Southern blot analysis. Our results using leaf disc assay indicated that transgenic banana lines were able to tolerate drought and high salinity stress better than the control plants and retained higher level of total chlorophyll and lower level of MDA content (malondialdehyde). Transgenic lines analyzed for salinity (250 mM NaCl) and drought (Soil gravimetric water content 0.15) tolerance showed higher proline content, better Fv/Fm ratio, and lower levels of MDA content than control suggesting that MusaNAC042 may be involved in responses to higher salinity and drought stresses in banana. Expression of several abiotic stress-related genes like those coding for CBF/DREB, LEA, and WRKY factors was altered in transgenic lines indicating that MusaNAC042 is an efficient modulator of abiotic stress response in banana.

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Abbreviations

GUS:

Beta-glucuronidase

cDNA:

Complementary DNA

hpt:

Hygromycin phosphotransferase

SSC:

Saline sodium citrate

TAE:

Tris-acetate buffer

ABA:

Abscisic Acid

RT-PCR:

Reverse transcription PCR

NAA:

α-Naphthaleneacetic acid

PCR:

Polymerase chain reaction

MDA:

Malondialdehyde

GFP:

Green fluorescent protein

References

  • Akhtar M, Jaiswal A, Taj G, Jaiswal JP, Qureshi MI, Singh NK (2012) DREB1/CBF transcription factors: their structure, function and role in abiotic stress tolerance in plants. J Genet 91:385–395

    Article  CAS  PubMed  Google Scholar 

  • Al Abdallat AM, Ayad JY, Abu Elenein JM, Al Ajlouni Z, Harwood WA (2014) Overexpression of the transcription factor HvSNAC1 improves drought tolerance in barley (Hordeum vulgare L.). Mol Breed 33:401–414

    Article  CAS  Google Scholar 

  • Arnon DI (1949) Copper enzymes in isolated chloroplasts: poly-phenol oxidase in Beta vulgaris. Plant Physiol 24:1–15

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Balazadeh S, Siddiqui H, Allu AD, Matallana-Ramirez LP, Caldana C, Mehrnia M, Zanor M-I, Kohler B, Mueller-Roeber B (2010) A gene regulatory network controlled by the NAC transcription factor ANAC092/AtNAC2/ORE1 during salt-promoted senescence. Plant J 62:250–264

    Article  CAS  PubMed  Google Scholar 

  • Balazadeh S, Kwasniewski M, Caldana C, Mehrnia M, Zanor MI, Xue GP, Mueller-Roeber B (2011) ORS1, an H2O2-responsive NAC transcription factor, controls senescence in Arabidopsis thaliana. Mol Plant 4:346–360

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Baloglu MC, Oz MT, Oktem HA, Yucel M (2012) Expression analysis of TaNAC69-1 and TtNAMB-2, wheat NAC family transcription factor genes under abiotic stress conditions in durum wheat (Triticum turgidum). Plant Mol Biol Rep 30:1246–1252

    Article  CAS  Google Scholar 

  • Banerjee A, Roychoudhury A (2015) WRKY proteins: signaling and regulation of expression during abiotic stress responses. Sci World J 2015:807560

    Article  Google Scholar 

  • Bowler C, Montagu MV, Inze D (1992) Superoxide dismutase and stress tolerance. Annu Rev Plant Physiol Plant Mol Biol 43:83–116

    Article  CAS  Google Scholar 

  • Cenci A, Guignon V, Roux N, Rouard M (2014) Genomic analysis of NAC transcription factors in banana (Musa acuminata) and definition of NAC orthologous groups for monocots and dicots. Plant Mol Biol 85:63–80

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Choudhury S, Panda P, Sahoo L, Panda SK (2013) Reactive oxygen species signaling in plants under abiotic stress. Plant Signal Behav 8:e23681

    Article  PubMed  Google Scholar 

  • Cote FX, Domergue R, Monmarson S, Schwendiman J, Teisson C, Escalant JV (1996) Embryogenic cell suspensions from the male flower of Musa AAA cv. Grand Nain Physiol Plant 97:285–290

    Article  CAS  Google Scholar 

  • Fang Y, Liao K, Du H, Xu Y, Song H, Li X, Xiong L (2015) A stress-responsive NAC transcription factor SNAC3 confers heat and drought tolerance through modulation of reactive oxygen species in rice. J Exp Bot. doi:10.1093/jxb/erv386

    Google Scholar 

  • Fujita M, Fujita Y, Maruyama K, Seki M, Hiratsu K, Ohme-Takagi M, Tran LS, Yamaguchi-Shinozaki K, Shinozaki K (2004) A dehydration-induced NAC protein, RD26, is involved in a novel ABA-dependent stress-signaling pathway. Plant J 39:863–876

    Article  CAS  PubMed  Google Scholar 

  • Galeano E, Vasconcelos TS, Ramiro DA, De Martin VF, Carrer H (2014) Identification and validation of quantitative real-time reverse transcription PCR reference genes for gene expression analysis in teak (Tectona grandis L.f.). BMC Res Notes 7:464

    Article  PubMed  PubMed Central  Google Scholar 

  • Ganapathi TR, Higgs NS, Balint Kurti PJ, Arntzen CJ, May GD, Van Eck JM (2001) Agrobacterium mediated transformation of embryogenic cell suspensions of the banana cultivar Rasthali (AAB). Plant Cell Rep 20:157–162

    Article  CAS  Google Scholar 

  • Gaxiola RA, Li J, Undurraga S, Dang LM, Allen GJ, Alper SL, Fink GR (2001) Drought- and salt-tolerant plants result from overexpression of the AVP1 H+-pump. Proc Natl Acad Sci U S A 98:11444–11449

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Greenway H, Munns R (1980) Mechanisms of salt tolerance in nonhalophytes. Annu Rev Plant Physiol 31:149–190

    Article  CAS  Google Scholar 

  • Guo Y, Gan S (2006) AtNAP, a NAC family transcription factor, has an important role in leaf senescence. Plant J 46:601–612

    Article  CAS  PubMed  Google Scholar 

  • Hao YJ, Wei W, Song QX, Chen HW, Zhang YQ, Wang F, Zou HF, Lei G, Tian AG, Zhang WK, Ma B, Zhang JS, Chen SY (2011) Soybean NAC transcription factors promote abiotic stress tolerance and lateral root formation in transgenic plants. Plant J 68:302–331

    Article  CAS  PubMed  Google Scholar 

  • Heath RL, Packer L (1968) Photoperoxidation in isolated chloroplasts. I. Kinetics and stoichiometry of fatty acid peroxidation. Arch Biochem Biophys 125:189–198

    Article  CAS  PubMed  Google Scholar 

  • Hoekstra FA, Golovina EA, Buitink J (2001) Mechanisms of plant desiccation tolerance. Trends Plant Sci 6:431–438

    Article  CAS  PubMed  Google Scholar 

  • Hood EE, Gelvin SB, Melchers LS, Hoekama A (1993) New Agrobacterium helper plasmids for gene transfer to plants. Transgenic Res 2:208–218

    Article  CAS  Google Scholar 

  • Hu H, Dai M, Yao J, Xiao B, Li X, Zhang Q, Xiong L (2006) Overexpressing a NAM, ATAF, and CUC (NAC) transcription factor enhances drought resistance and salt tolerance in rice. Proc Natl Acad Sci U S A 103:12987–12992

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Hu H, You J, Fang Y, Zhu X, Qi Z, Xiong L (2008) Characterization of transcription factor gene SNAC2 conferring cold and salt tolerance in rice. Plant Mol Biol 67:169–181

    Article  CAS  PubMed  Google Scholar 

  • Ito Y, Katsura K, Maruyama K, Taji T, Kobayashi M, Seki M, Shinozaki K, Yamaguchi-Shinozaki K (2006) Functional analysis of rice DREB1/CBF-type transcription factors involved in cold-responsive gene expression in transgenic rice. Plant Cell Physiol 47:141–153

    Article  CAS  PubMed  Google Scholar 

  • Kumar S, Nei M, Dudley J, Tamura K (2008) MEGA: biologist-centric software for evolutionary analysis of DNA and protein sequences. Brief Bioinform 9:299–306

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Livak KJ, Schmittgen TD (2001) Analysis of relative gene expression data using real-time quantitative PCR and the 2(−delta delta T)) method. Methods 25:402–408

    Article  CAS  PubMed  Google Scholar 

  • Mahajan S, Tuteja N (2005) Cold, salinity and drought stresses: an overview. Arch Biochem Biophys 444:139–158

    Article  CAS  PubMed  Google Scholar 

  • Nakashima K, Tran LS, Van Nguyen D, Fujita M, Maruyama K, Todaka D, Ito Y, Hayashi N, Shinozaki K, Yamaguchi-Shinozaki K (2007) Functional analysis of a NAC-type transcription factor OsNAC6 involved in abiotic and biotic stress-responsive gene expression in rice. Plant J 51:617–630

    Article  CAS  PubMed  Google Scholar 

  • Negi S, Tak H, Ganapathi TR (2015a) Cloning and functional characterization of MusaVND1 using transgenic banana plants. Transgenic Res 24:571–585

    Article  CAS  PubMed  Google Scholar 

  • Negi S, Tak H, Ganapathi TR (2015b) Functional characterization of secondary wall deposition regulating transcription factors MusaVND2 and MusaVND3 in transgenic banana plants. Protoplasma. doi:10.1007/s00709-015-0822-5

    PubMed  Google Scholar 

  • Negi S, Tak H, Ganapathi TR (2015c) Expression analysis of MusaNAC68 transcription factor and its functional analysis by overexpression in transgenic banana plants. Plant Cell Tiss Organ Cult Doi. doi:10.1007/s11240-015-0929-6

    Google Scholar 

  • Niu X, Bressan RA, Hasegawa PM, Pardo JM (1995) Ion homeostasis in NaCl stress environments. Plant Physiol 109:735–742

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Nuruzzaman M, Manimekalai R, Sharoni AM, Satoh K, Kondoh H, Ooka H, Kikuchi S (2010) Genome-wide analysis of NAC transcription factor family in rice. Gene 465:30–44

    Article  CAS  PubMed  Google Scholar 

  • Olsen AN, Ernst HA, Leggio LL, Skriver K (2005) NAC transcription factors: structurally distinct, functionally diverse. Trends Plant Sci 10:79–87

    Article  CAS  PubMed  Google Scholar 

  • Placide R, Christian CS, Rony S (2012) Development of in vitro technique to screen for drought tolerant banana varieties by sorbitol induced osmotic stress. Afr J Plant Sci 6:416–425

    Article  CAS  Google Scholar 

  • Podevin N, Krauss A, Henry I, Swennen R, Remy S (2012) Selection and validation of reference genes for quantitative RT-PCR expression studies of the non-model crop Musa. Mol Breed 30:1237–1252

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Ravi I, Uma S, Vaganan MM, Mustaffa MM (2013) Phenotyping bananas for drought resistance. Front Physiol. 4:9

  • Ray DL, Johnson JC (2014) Validation of reference genes for gene expression analysis in olive (Olea europaea) mesocarp tissue by quantitative real-time RT-PCR. BMC Res Notes 7:304

    Article  PubMed  PubMed Central  Google Scholar 

  • Reid KE, Olsson N, Schlosser J, Peng F, Lund ST (2006) An optimized grapevine RNA isolation procedure and statistical determination of reference genes for real-time RT-PCR during berry development. BMC Plant Biol 6:27

    Article  PubMed  PubMed Central  Google Scholar 

  • Rhodes D, Nadolska-Orczyk A (2001) Plant stress physiology. In Encyclopaedia of Life Sciences (eLS). doi:10.1038/npg.els.0001297

    Google Scholar 

  • Robinson JC, Sauco VG (2010) Bananas and plantains, 2nd edn. CABI, Wallingford

    Book  Google Scholar 

  • Rorat T (2006) Plant dehydrins—tissue location, structure and function. Cell Mol Biol Lett 11:536–556

    Article  CAS  PubMed  Google Scholar 

  • Saga H, Ogawa T, Kai K, Suzuki H, Ogata Y, Sakurai N, Shibata D, Ohta D (2012) Identification and characterization of ANAC042, a transcription factor family gene involved in the regulation of camalexin biosynthesis in Arabidopsis. Mol Plant Microbe Interact 25:684–696

    Article  CAS  PubMed  Google Scholar 

  • Schmittgen TD, Livak KJ (2008) Analyzing real-time PCR data by the comparative C(T) method. Nat Protoc 3:1101–1108

    Article  CAS  PubMed  Google Scholar 

  • Shahnejat-Bushehri S, Mueller-Roeber B, Balazadeh S (2012) Arabidopsis NAC transcription factor JUNGBRUNNEN1 affects thermomemory-associated genes and enhances heat stress tolerance in primed and unprimed conditions. Plant Signal Behav 7:1518–1521

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Shan W, Kuang JF, Chen L, Xie H, Peng HH, Xiao YY, Li XP, Chen WX, He QG, Chen JY, Lu WJ (2012) Molecular characterization of banana NAC transcription factors and their interactions with ethylene signalling component EIL during fruit ripening. J Exp Bot 63:5171–5187

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Shan W, Kuang JF, Lu WJ, Chen JY (2014) Banana fruit NAC transcription factor MaNAC1 is a direct target of MaICE1 and involved in cold stress through interacting with MaCBF1. Plant Cell Environ 37:2116–2127

    Article  CAS  PubMed  Google Scholar 

  • Shan W, Chen JY, Kuang JF, Lu WJ (2015) Banana fruit NAC transcription factor MaNAC5 cooperates with MaWRKYs to enhance the expression of pathogenesis-related genes against Colletotrichum musae. Mol Plant Pathol. doi:10.1111/mpp.12281

    PubMed  Google Scholar 

  • Silveira ED, Alves-Ferreira M, Guimarães LA, da Silva FR, Carneiro VT (2009) Selection of reference genes for quantitative real-time PCR expression studies in the apomictic and sexual grass Brachiaria brizantha. BMC Plant Biol 9:84

    Article  PubMed  PubMed Central  Google Scholar 

  • Smart RE, Bingham GE (1974) Rapid estimates of relative water content. Plant Physiol 53:258–260

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Surendar KK, Devi DD, Ravi I, Krishnakumar S, Kumar SR, Velayudham K (2013) Water stress in banana—a review. Bull Env Pharmacol Life Sci 2:01–18

    Google Scholar 

  • Surender Reddy P, Jogeswar G, Rasineni GK, Maheswari M, Reddy AR, Varshney RK, Kavi Kishor PB (2015) Proline over-accumulation alleviates salt stress and protects photosynthetic and antioxidant enzyme activities in transgenic sorghum [Sorghum bicolor (L.) Moench]. Plant Physiol Biochem 94:104–113

    Article  CAS  PubMed  Google Scholar 

  • Székely G, Abrahám E, Cséplo A, Rigó G, Zsigmond L, Csiszár J, Ayaydin F, Strizhov N, Jásik J, Schmelzer E, Koncz C, Szabados L (2007) Duplicated P5CS genes of Arabidopsis play distinct roles in stress regulation and developmental control of proline biosynthesis. Plant J 53:11–28

    Article  PubMed  Google Scholar 

  • Tran LS, Nakashima K, Sakuma Y, Simpson SD, Fujita Y, Maruyama K, Fujita M, Seki M, Shinozaki K, Yamaguchi-Shinozaki K (2004) Isolation and functional analysis of Arabidopsis stress inducible NAC transcription factors that bind to a drought responsive cis-element in the early responsive to dehydration stress. Plant Cell 16:2481–2498

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • van Asten PJA, Fermont AM, Taulya G (2011) Drought is a major yield loss factor for rainfed East African highland banana. Agric Water Manage 98:541–552

    Article  Google Scholar 

  • Vanhove AC, Vermaelen W, Panis B, Swennen R, Carpentier SC (2012) Screening the banana biodiversity for drought tolerance: can an in vitro growth model and proteomics be used as a tool to discover tolerant varieties and understand homeostasis. Front Plant Sci 3:176

    Article  PubMed  PubMed Central  Google Scholar 

  • Wu A, Allu AD, Garapati P, Siddiqui H, Dortay H, Zanor MI, Asensi-Fabado MA, Munné-Bosch S, Antonio C, Tohge T, Fernie AR, Kaufmann K, Xue GP, Mueller-Roeber B, Balazadeh S (2012) JUNGBRUNNEN1, a reactive oxygen species-responsive NAC transcription factor, regulates longevity in Arabidopsis. Plant Cell 24:482–506

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Xia N, Zhang G, Liu X, Deng L, Cai G et al (2010) Characterization of a novel wheat NAC transcription factor gene involved in defense response against stripe rust pathogen infection and abiotic stresses. Mol Biol Rep 37:3703–3712

    Article  CAS  PubMed  Google Scholar 

  • Xie Q, Sanz-Burgos AP, Guo H, García JA, Gutiérrez C (1999) GRAB proteins, novel members of the NAC domain family, isolated by their interaction with a geminivirus protein. Plant Mol Biol 39:647–656

    Article  CAS  PubMed  Google Scholar 

  • Xu Y, Hu W, Liu J, Zhang J, Jia C, Miao H, Xu B, Jin Z (2014) A banana aquaporin gene, MaPIP1;1, is involved in tolerance to drought and salt stresses. BMC Plant Biol 14:59

    Article  PubMed  PubMed Central  Google Scholar 

  • Xue G, Way H, Richardson T, Drenth J, Joyce P et al (2011) Overexpression of TaNAC69 leads to enhanced transcript levels of stress up-regulated genes and dehydration tolerance in bread wheat. Mol Plant 4:697–712

    Article  CAS  PubMed  Google Scholar 

  • Yeo AR (1998) Molecular biology of salt tolerance in the context of whole plant physiology. J Exp Bot 49:915–929

    CAS  Google Scholar 

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Acknowledgment

Authors thank Head, Nuclear Agriculture and Biotechnology Division, BARC, for his motivation and support. The study was funded by Department of Atomic Energy, India. Authors are thankful to Dr. P. Suprasanna for critically going through the manuscript.

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  1. Plant Cell Culture Technology Section, Nuclear Agriculture and Biotechnology Division, Bhabha Atomic Research Centre, Trombay, Mumbai, India

    Himanshu Tak, Sanjana Negi & T. R Ganapathi

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  1. Himanshu Tak
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  2. Sanjana Negi
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  3. T. R Ganapathi
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Correspondence to T. R Ganapathi.

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Tak, H., Negi, S. & Ganapathi, T.R. Banana NAC transcription factor MusaNAC042 is positively associated with drought and salinity tolerance. Protoplasma 254, 803–816 (2017). https://doi.org/10.1007/s00709-016-0991-x

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