Abstract
Chickpea, an important grain legume, suffers from considerable loss of yield due to Fusarium wilt disease. Inaccessibility of resistant gene pool among cultivars and lack of report of resistance, genes from alien sources have been the major constraints for resistance development in this valuable crop. However, along with some other transcription factors, MYB78 was significantly upregulated during chickpea—Fusarium interplay in resistant chickpea genotype. Being a highly recalcitrant species, the transformation of this important crop remained non-reproducible until recently. Following a tissue culture independent plumular meristem transformation protocol, introgression of CaMYB78 TF finally became feasible in chickpea. The overexpressed plants developed resistance against the pathogen but the anthocyanin production in transformed flowers was perturbed. In silico analyses of the anthocyanin biosynthetic key gene promoters reported the occurrence of multiple MYB-binding cis elements. Detailed molecular analyses establish the differential regulatory roles of CaMYB78, resistance response against Foc1 on one hand and suppression of pigmentation during flower development on the other, which is an innovative finding of its kind.
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References
Anwar M, Yu W, Yao H et al (2019) NtMYB3, an R2R3-MYB from narcissus, regulates flavonoid biosynthesis. IJMS 20:5456. https://doi.org/10.3390/ijms20215456
Asharani B, Ganeshaiah K, Kumar A, Udayakumar M (2011) Transformation of chickpea lines with Cry1X using in planta transformation and characterization of putative transformants T1 lines for molecular and biochemical characters. J Plant Breed Crop Sci 3:413–423. https://doi.org/10.5897/JPBCS11.074
Cavallini E, Matus JT, Finezzo L et al (2015) The phenylpropanoid pathway is controlled at different branches by a set of R2R3-MYB C2 repressors in grapevine. Plant Physiol 167:1448–1470. https://doi.org/10.1104/pp.114.256172
Chakraborti D, Sarkar A, Das S (2006) Efficient and rapid in vitro plant regeneration system for Indian cultivars of chickpea (Cicer arietinum L.). Plant Cell, Tissue Organ Cult 86:117–123. https://doi.org/10.1007/s11240-005-9072-0
Chakraborty J, Ghosh P, Sen S et al (2019) CaMPK9 increases the stability of CaWRKY40 transcription factor which triggers defense response in chickpea upon Fusarium oxysporum f. sp. ciceri Race1 infection. Plant Mol Biol 100:411–431. https://doi.org/10.1007/s11103-019-00868-0
Chen C, Chen H, Zhang Y et al (2020) TBtools: an integrative toolkit developed for interactive analyses of big biological data. Mol Plant 13:1194–1202. https://doi.org/10.1016/j.molp.2020.06.009
Choudhury FK, Rivero RM, Blumwald E, Mittler R (2017) Reactive oxygen species, abiotic stress and stress combination. Plant J 90:856–867. https://doi.org/10.1111/tpj.13299
Cui F, Brosché M, Sipari N et al (2013) Regulation of ABA dependent wound induced spreading cell death by MYB 108. New Phytol 200:634–640. https://doi.org/10.1111/nph.12456
Cui J, Jiang N, Zhou X, Hou X, Yang G, Meng J, Luan Y (2018) Tomato MYB49 enhances resistance to Phytophthora infestans and tolerance to water deficit and salt stress. Planta 248:1487–1503. https://doi.org/10.1007/s00425-018-2987-6
De Vos M, Denekamp M, Dicke M et al (2006) The Arabidopsis thaliana transcription factor AtMYB102 functions in defense against the insect herbivore Pieris rapae. Plant Signal Behav 1:305–311. https://doi.org/10.4161/psb.1.6.3512
Dixit G, Srivastava A, Singh N (2019) Marching towards self-sufficiency in chickpea. Curr Sci 116:239–242. https://doi.org/10.18520/cs/v116/i2/239-242
Doyle JJ, Doyle JL A rapid DNA isolation procedure for small quantities of fresh leaf tissue. PHYTOCHEMICAL BULLETIN
Dubos C, Stracke R, Grotewold E et al (2010) MYB transcription factors in Arabidopsis. Trends Plant Sci 15:573–581. https://doi.org/10.1016/j.tplants.2010.06.005
FAOSTAT (2019) www.fao.org/faostat/en/#data/QC
Fujimoto SY, Ohta M, Usui A et al (2000) Arabidopsis ethylene-responsive element binding factors act as transcriptional activators or repressors of GCC box–mediated gene expression. Plant Cell 12:393–404. https://doi.org/10.1105/tpc.12.3.393
Ganguly S, Ghosh G, Ghosh S et al (2020) Plumular meristem transformation system for chickpea: an efficient method to overcome recalcitrant tissue culture responses. Plant Cell Tissue Organ Culture (PCTOC) 142:493–504. https://doi.org/10.1007/s11240-020-01873-8
Garg R, Sahoo A, Tyagi AK, Jain M (2010) Validation of internal control genes for quantitative gene expression studies in chickpea (Cicer arietinum L.). Biochem Biophys Res Commun 396:283–288. https://doi.org/10.1016/j.bbrc.2010.04.079
Gaur PM, Jukanti AK, Varshney RK (2012) Impact of genomic technologies on chickpea breeding strategies. Agronomy 2:199–221. https://doi.org/10.3390/agronomy2030199
Ghosh G, Purohit A, Ganguly S et al (2014) In vitro shoot grafting on rootstock: an effective tool for Agrobacterium-mediated transformation of pigeonpea (Cajanus cajan (L.) Millsp.). Plant Biotechnol 31:301–308. https://doi.org/10.5511/plantbiotechnology.14.0805a
Golldack D, Li C, Mohan H, Probst N (2014) Tolerance to drought and salt stress in plants: unraveling the signaling networks. Front Plant Sci 5:151. https://doi.org/10.3389/fpls.2014.00151
Gupta S, Bhar A, Chatterjee M, Das S (2013) Fusarium oxysporum f. sp. ciceri race 1 induced redox state alterations are coupled to downstream defense signaling in root tissues of chickpea (Cicer arietinum L.). PloS one 8:e73163. https://doi.org/10.1371/journal.pone.0073163
Gupta S, Bhar A, Chatterjee M et al (2017) Transcriptomic dissection reveals wide spread differential expression in chickpea during early time points of Fusarium oxysporum f. sp. ciceri Race 1 attack. PLoS One 12:e0178164. https://doi.org/10.1371/journal.pone.0178164
Jefferson RA, Kavanagh TA, Bevan MW (1987) GUS fusions: beta-glucuronidase as a sensitive and versatile gene fusion marker in higher plants. EMBO J 6:3901–3907. https://doi.org/10.1002/j.1460-2075.1987.tb02730.x
Jiménez-Fernández D, Landa BB, Kang S et al (2013) Quantitative and microscopic assessment of compatible and incompatible interactions between chickpea cultivars and Fusarium oxysporum f. sp. ciceris Races. PLoS ONE 8:e61360. https://doi.org/10.1371/journal.pone.0061360
Jin H, Cominelli E, Bailey P, Parr A, Mehrtens F, Jones J, Tonelli C, Weisshaar B, Martin C (2000) Transcriptional repression by AtMYB4 controls production of UV-protecting sunscreens in Arabidopsis. EMBO J 19:6150–6161. https://doi.org/10.1093/emboj/19.22.6150
Kagale S, Rozwadowski K (2011) EAR motif-mediated transcriptional repression in plants: an underlying mechanism for epigenetic regulation of gene expression. Epigenetics 6:141–146. https://doi.org/10.4161/epi.6.2.13627
Kar S, Basu D, Das S et al (1997) Expression of cryIA(c) gene of Bacillus thuringiensis in transgenic chickpea plants inhibits development of pod-borer (Heliothis armigera) larvae. Transgenic Res 6:177–185. https://doi.org/10.1023/A:1018433922766
Kariola T, Brader G, Li J, Palva ET (2005) Chlorophyllase 1, a damage control enzyme, affects the balance between defense pathways in plants. Plant Cell 17:282–294. https://doi.org/10.1105/tpc.104.025817
Kearse M, Moir R, Wilson A et al (2012) Geneious Basic: an integrated and extendable desktop software platform for the organization and analysis of sequence data. Bioinformatics 28:1647–1649. https://doi.org/10.1093/bioinformatics/bts199
Kondo S, Hiraoka K, Kobayashi S et al (2002) Changes in the expression of anthocyanin biosynthetic genes during apple development. J Am Soc Horticult Sci 127:971–976. https://doi.org/10.21273/JASHS.127.6.971
Kranz HD, Denekamp M, Greco R et al (1998) Towards functional characterisation of the members of the R2R3-MYB gene family from Arabidopsis thaliana. Plant J 16:263–276. https://doi.org/10.1046/j.1365-313x.1998.00278.x
Kumar J, Sethi S, Johansen C et al (1996) Potential of short-duration chickpea varieties. Indian J Dryland Agric Res Dev 11:28–32
Landa BB, Navas-Cortés JA, Hervás A, Jiménez-Díaz RM (2001) Influence of temperature and inoculum density of Fusarium oxysporum f. sp. ciceris on suppression of Fusarium wilt of chickpea by rhizosphere bacteria. Phytopathology 91:807–816. https://doi.org/10.1094/PHYTO.2001.91.8.807
Larkin RP, Fravel DR (1998) Efficacy of various fungal and bacterial biocontrol organisms for control of Fusarium wilt of tomato. Plant Dis 82:1022–1028. https://doi.org/10.1094/PDIS.1998.82.9.1022
Lescot M, Déhais P, Thijs G et al (2002) PlantCARE, a database of plant cis-acting regulatory elements and a portal to tools for in silico analysis of promoter sequences. Nucleic Acids Res 30:325–327. https://doi.org/10.1093/nar/30.1.325
Liu X, Yang L, Zhou X et al (2013) Transgenic wheat expressing Thinopyrum intermedium MYB transcription factor TiMYB2R-1 shows enhanced resistance to the take-all disease. J Exp Bot 64:2243–2253. https://doi.org/10.1093/jxb/ert084
Liu T, Chen T, Kan J, Yao Y, Guo D, Yang Y, Ling X, Wang J, Zhang B (2022) The GhMYB36 transcription factor confers resistance to biotic and abiotic stress by enhancing PR1 gene expression in plants. Plant Biotechnol J 20(4):722–735. https://doi.org/10.1111/pbi.13751
Livak KJ, Schmittgen TD (2001) Analysis of relative gene expression data using real-time quantitative PCR and the 2−ΔΔCT method. Methods 25:402–408. https://doi.org/10.1006/meth.2001.1262
Murthy B, Victor J, Singh RP et al (1996) In vitro regeneration of chickpea (Cicer arietinum L.): stimulation of direct organogenesis and somatic embryogenesis by thidiazuron. Plant Growth Regul 19:233–240. https://doi.org/10.1007/BF00037796
Nayeri FD, Anbuhi MH (2019) Transient expression of etanercept therapeutic protein in tobacco (Nicotiana tabacum L.). Int J Biol Macromol 130:483–490. https://doi.org/10.1016/j.ijbiomac.2019.02.153
Park J-S, Kim J-B, Cho K-J et al (2008) Arabidopsis R2R3-MYB transcription factor AtMYB60 functions as a transcriptional repressor of anthocyanin biosynthesis in lettuce (Lactuca sativa). Plant Cell Rep 27:985–994. https://doi.org/10.1007/s00299-008-0521-1
Pathak MR, Hamzah RY (2008) An effective method of sonication-assisted Agrobacterium-mediated transformation of chickpeas. Plant Cell, Tissue Organ Culture 93:65–71. https://doi.org/10.1007/sI1240-008-9344-6
Polisetty R, Patil P, Deveshwar J et al (1996) Rooting and establishment of in vitro grown shoot tip explants of chickpea (Cicer arietinum L.). Indian J Exp Biol 34:806–809
Polowick P, Baliski D, Mahon J (2004) Agrobacterium tumefaciens-mediated transformation of chickpea (Cicer arietinum L.): gene integration, expression and inheritance. Plant Cell Rep 23:485–491. https://doi.org/10.1007/s00299-004-0857-0
Portieles R, Ayra C, Gonzalez E et al (2010) NmDef02, a novel antimicrobial gene isolated from Nicotiana megalosiphon confers high-level pathogen resistance under greenhouse and field conditions. Plant Biotechnol J 8:678–690. https://doi.org/10.1111/j.1467-7652.2010.00501.x
Rabino I, Mancinelli AL (1986) Light, temperature, and anthocyanin production. Plant Physiol 81:922–924. https://doi.org/10.1104/pp.81.3.922
Rao KS, Sreevathsa R, Sharma PD et al (2008) In planta transformation of pigeon pea: a method to overcome recalcitrancy of the crop to regeneration in vitro. Physiol Mol Biol Plants 14:321–328. https://doi.org/10.1007/s12298-008-0030-2
Rizvi S, Singh RP (2000) In vitro plant regeneration from immature leaflet-derived callus cultures of Cicer arietinum L. via organogenesis. Plant Cell Biotechnol Mol Biol 1:109–114
Roorkiwal M, Nayak SN, Thudi M et al (2014) Allele diversity for abiotic stress responsive candidate genes in chickpea reference set using gene based SNP markers. Front Plant Sci 5:248. https://doi.org/10.3389/fpls.2014.00248
Roy S (2016) Function of MYB domain transcription factors in abiotic stress and epigenetic control of stress response in plant genome. null 11:e1117723. https://doi.org/10.1080/15592324.2015.1117723
Sagare A, Suhasini K, Krishnamurthy K (1999) Comparative study of the development of zygotic and somatic embryos of chickpea (Cicer arietinum L.). In: Kishore PB (Ed) Plant tissue culture and biotechnology–emerging trends. Universities Press, Hyderabad, pp 56–63
Sagare A, Krishnamurthy K (1991) Protoplast regeneration in chickpea, Cicer arietinum L. Indian J Exp Biol 29:930–932
Sen S, Chakraborty J, Ghosh P et al (2017) Chickpea WRKY70 regulates the expression of a homeodomain-leucine zipper (HD-Zip) I transcription factor CaHDZ12, which confers abiotic stress tolerance in transgenic tobacco and chickpea. Plant Cell Physiol 58:1934–1952. https://doi.org/10.1093/pcp/pcx126
Seo PJ, Park C (2010) MYB96-mediated abscisic acid signals induce pathogen resistance response by promoting salicylic acid biosynthesis in Arabidopsis. New Phytol 186:471–483. https://doi.org/10.1111/j.1469-8137.2010.03183.x
Shan T, Rong W, Xu H, Du L, Liu X, Zhang Z (2016) The wheat R2R3-MYB transcription factor TaRIM1 participates in resistance response against the pathogen Rhizoctonia cerealis infection through regulating defense genes. Sci Rep 6:28777. https://doi.org/10.1038/srep28777
Shri P, Davis TM (1992) Zeatin-induced shoot regeneration from immature chickpea (Cicer arietinum L.) cotyledons. Plant Cell, Tissue Organ Cult 28:45–51. https://doi.org/10.1007/BF00039914
Shukla PS, Gupta K, Agarwal P et al (2015) Overexpression of a novel SbMYB15 from Salicornia brachiata confers salinity and dehydration tolerance by reduced oxidative damage and improved photosynthesis in transgenic tobacco. Planta 242:1291–1308. https://doi.org/10.1007/s00425-015-2366-5
Siddique KHM, Brinsmead RB, Knight R, Knights EJ, Paull JG, Rose IA (2000) Adaptation of chickpea (Cicer arietinum L.) and faba bean (Vicia faba L.) to Australia. In: Knight R (eds) Linking Research and Marketing Opportunities for Pulses in the 21st Century. Current Plant Science and Biotechnology in Agriculture, vol 34. Springer, Dordrecht, pp 289–303. https://doi.org/10.1007/978-94-011-4385-1_26
Siefkes-Boer H, Noonan M, Bullock D, Conner A (1995) Hairy root transformation system in large-seeded grain legumes. Israel J Plant Sci 43:1–5. https://doi.org/10.1080/07929978.1995.10676585
Singh NP, Pratap A (2016) Food legumes for nutritional security and health benefits. In: Singh U, Praharaj CS, Singh SS, Singh NP (eds) Biofortification of food crops. Springer, New Delhi, pp 41–50 https://doi.org/10.1007/978-81-322-2716-8_4
Sreevathsa RSR (2008) In planta transformation strategy to generate transgenic plants in chickpea: proof of concept with a cry gene. J Plant Biol 35:201–206
Van der Ent S, Verhagen BW, Van Doorn R et al (2008) MYB72 is required in early signaling steps of rhizobacteria-induced systemic resistance in Arabidopsis. Plant Physiol 146:1293–1304. https://doi.org/10.1104/pp.107.113829
Wan S, Li C, Ma X, Luo K (2017) PtrMYB57 contributes to the negative regulation of anthocyanin and proanthocyanidin biosynthesis in poplar. Plant Cell Rep 36:1263–1276. https://doi.org/10.1007/s00299-017-2151-y
Waterhouse AM, Procter JB, Martin DMA et al (2009) Jalview Version 2–a multiple sequence alignment editor and analysis workbench. Bioinformatics 25:1189–1191. https://doi.org/10.1093/bioinformatics/btp033
Wei Q, Zhang F, Sun F et al (2017) A wheat MYB transcriptional repressor TaMyb1D regulates phenylpropanoid metabolism and enhances tolerance to drought and oxidative stresses in transgenic tobacco plants. Plant Sci 265:112–123. https://doi.org/10.1016/j.plantsci.2017.09.020
Xu H, Zou Q, Yang G et al (2020) MdMYB6 regulates anthocyanin formation in apple both through direct inhibition of the biosynthesis pathway and through substrate removal. Horticulture Research 7:72. https://doi.org/10.1038/s41438-020-0294-4
Zhang H, Zhang D, Chen J et al (2004) Tomato stress-responsive factor TSRF1 interacts with ethylene responsive element GCC box and regulates pathogen resistance to Ralstonia solanacearum. Plant Mol Biol 55:825–834. https://doi.org/10.1007/s11103-004-2140-8
Zhang Z, Liu X, Wang X, Zhou M, Zhou X, Ye X, Wei X (2012a) An R2R3 MYB transcription factor in wheat, Ta PIMP 1, mediates host resistance to Bipolaris sorokiniana and drought stresses through regulation of defense-and stress-related genes. New Phytol 196:1155–1170. https://doi.org/10.1111/j.1469-8137.2012.04353.x
Zhang Z, Liu X, Wang X et al (2012b) An R2R3 MYB transcription factor in wheat, Ta PIMP 1, mediates host resistance to Bipolaris sorokiniana and drought stresses through regulation of defense-and stress-related genes. New Phytol 196:1155–1170. https://doi.org/10.1111/j.1469-8137.2012.04353.x
Zhang M, Wang J, Luo Q et al (2021) CsMYB96 enhances citrus fruit resistance against fungal pathogen by activating salicylic acid biosynthesis and facilitating defense metabolite accumulation. J Plant Physiol 264:153472. https://doi.org/10.1016/j.jplph.2021.153472
Zhu X, Qi L, Liu X et al (2014) The wheat ethylene response factor transcription factor pathogen-induced ERF1 mediates host responses to both the necrotrophic pathogen Rhizoctonia cerealis and freezing stresses. Plant Physiol 164:1499–1514. https://doi.org/10.1104/pp.113.229575
Zhu Y, Hu X, Wang P, Wang H, Ge X, Li F, Hou Y (2022) GhODO1, an R2R3-type MYB transcription factor, positively regulates cotton resistance to Verticillium dahliae via the lignin biosynthesis and jasmonic acid signaling pathway. Int J Biol Macromol 201:580–591. https://doi.org/10.1016/j.ijbiomac.2022.01.120
Acknowledgements
The authors are grateful to Bose Institute for providing the infrastructural facility. SD acknowledges INSA for her Senior Scientist Fellowship. SS acknowledges UGC, Govt of India for Senior Research Fellowship (Serial. No. 2121430422). SP is grateful for the support from CSIR, Government of India (09/015(0526)/20) for her fellowship. The authors are grateful to Dr. Shreeparna Ganguly and Dr. Dipankar Chakraborty for their generous help in providing the in-planta transformation protocol of chickpea.
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Sampa Das and Surbhi Shriti contributed to the study conception, design, and writing. SP helped in writing some parts of the manuscript. SP helped in performing Fungal Bioassay experiments and preparation of the first draft. Material preparation, data collection, and analysis were performed by SS. All authors read and approved the final manuscript.
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Shriti, S., Paul, S. & Das, S. Overexpression of CaMYB78 transcription factor enhances resistance response in chickpea against Fusarium oxysporum and negatively regulates anthocyanin biosynthetic pathway. Protoplasma 260, 589–605 (2023). https://doi.org/10.1007/s00709-022-01797-4
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DOI: https://doi.org/10.1007/s00709-022-01797-4