Skip to main content
Log in

Developing resistance against soil-borne Fusarium pathogen causing tomato wilt through in vitro cell line selection

  • Original Article
  • Published:
Plant Cell, Tissue and Organ Culture (PCTOC) Aims and scope Submit manuscript

Abstract

Numerous complex chemicals are being routinely used to prevent pathogens in various commercially important agri-horticultural crops that have some biosafety issues associated with human health and environment. Therefore, the present study was aimed to develop resistance in tomato against soil-borne fungal pathogen Fusarium oxysporum f. sp. lycopersici, through in vitro cell line selection approach. BLASTn analysis of sequenced 16S rRNA region of the fungus isolated from the wilt infected tomato plant was confirmed by ITS gene technology. Highest callus induction (79.63 ± 0.70%) along with shoot regeneration (78.68 ± 0.64%) was recorded on MS medium fortified containing 4.54 µM TDZ + 2.90 µM IAA and 2.27 µM TDZ + 1.0 µM 2,4-D using leaf explants, respectively. To carry out in vitro selection, developed calli using leaf explants were further subjected on selective medium followed by discontinuous selection cycles supplemented with 30% culture filtrate (CF) of targeted pathogen, where 21.32 ± 0.99 per cent average callus survival and 45.98 ± 0.55% average shoot induction (devoid of CF) were recorded. 12 moderately resistant lines were obtained from the total 48 cell lines during in vivo bioassay under glass house conditions. Two somaclones were confirmed at molecular level among the total 12 moderately developed resistant cell lines. The multilocation field trials are being done to check stable inheritance with generation wise advancements. It was inferred from the present study that these findings could be a cutting edge to develop resistance against various stresses in commercially important horticultural crops like tomato without posing any threat to environment.

Key message

Two tolerant tomato somaclones developed against Fusarium oxysporum f. sp. lycopersici causing Fusarium wilt disease using an ecofriendly research methodology of in vitro cell line selection approach.

This is a preview of subscription content, log in via an institution to check access.

Access this article

Price excludes VAT (USA)
Tax calculation will be finalised during checkout.

Instant access to the full article PDF.

Fig. 1
Fig. 2
Fig. 3
Fig. 4
Fig. 5
Fig. 6

Similar content being viewed by others

Data availability

The authors confirm that the data supporting the findings of this study are available within the article.

References

  • Alizadeh M, Singh SK, Patel VB (2010) Comparative performance of in vitro multiplication in four grape (Vitis spp.) rootstock genotypes. Int J Plant Prod 4:41–50

    CAS  Google Scholar 

  • Anonymous (2018) Annual Report. Area and Production of vegetables in Himachal Pradesh. Directorate of Agriculture (H.P.), Shimla-5

  • Baayen RP, DeMatt AL (1987) Passive transport of microconidia of Fusarium oxysporum f. sp. dianthi in carnation after root inoculation. Nether J Plant Pathol 93:3–13

    Article  Google Scholar 

  • Bhardwaj SV, Thakur T, Sharma R, Sharma P (2012) In vitro selection of resistant mutants of ginger (Zingiber officinale Rosc.) against wilt pathogen (Fusarium oxysporum f. sp. zingiberi Trujillo). Plant Dis Res 27:194–199

    Google Scholar 

  • Booth C (1971) Fusarium: laboratory guide to the identification of the major species. The Common Wealth Mycological Institute p 237

  • Borras O, Bermudez RS (2010) The pineapple-Fusarium subglutinans interaction: an early selection system for disease resistance. FAO/IAEA. Mass screening techniques for selecting crops resistant to disease. Inter Atom Ener Agen, Vienna, pp 159–172

    Google Scholar 

  • Bouizgarne B, El-Maarouf-Bouteau H, Frankart C, Reboutier D, Madiona K, Pennarun AM, Bouteau F (2006) Early physiological responses of Arabidopsis thaliana cells to fusaric acid: toxic and signalling effects. New Phytol 169:209–218

    Article  CAS  PubMed  Google Scholar 

  • Curir P, Dolci GL (2000) Fusaric acid production by Fusarium Oxysporum f.sp. lilii and its role in the lily basal rot disease. Euro J Pl Patho 106:849–856

    Article  CAS  Google Scholar 

  • Doyle JJ, Doyle JL (1990) A rapid DNA isolation procedure for small quantities of fresh leaf tissues. Phytochem Bull 19:11–15

    Google Scholar 

  • Flores P, Otoni W, Dhingra O, Sergio P, Santos T, Bruckner CH (2012) In vitro selection of yellow passion fruit genotypes for resistance to Fusarium vascular wilt. Plant Cell Tissue Org Cult 108:37–45

    Article  CAS  Google Scholar 

  • Gengenbach BG, Rines HW (1986) Use of phytotoxins in selection of disease resistant mutants in tissue culture. Iowa State J Res 60:449–476

    Google Scholar 

  • Gomez KA, Gomez AA (1984) Statistical procedure for agricultural research. Wiley, New York, pp 328–332

    Google Scholar 

  • Hafizi R (2013) Morphological and molecular characterization of Fusarium solani and F. oxysporum associated with crown disease of oil palm. Braz J Microbiol 44:959–968

    Article  CAS  PubMed  Google Scholar 

  • Hahn M (2014) The rising threat of fungicide resistance in plant pathogenic fungi: Botrytis as a case study. J Chem Biol 7(4):133–141

    Article  PubMed  PubMed Central  Google Scholar 

  • Kapoor B (2020) Agrobacterium mediated defensin gene transfer studies in tomato (Solanum lycopersicum L.). PhD Thesis. Dr Y S Parmar University of Horticulture and Forestry, Nauni, Solan, India.

  • Kuanar A, Nayak PK, Subudhi E, Nayak S (2014) In vitro selection of turmeric somaclone resistant to Fusarium oxysporum f. sp. zingiberi. Proc Natl Acad Sci 84:1077–1082

    CAS  Google Scholar 

  • Kumari V, Kumar A, Chaudhary HK, Prasad R, Jambhulkar S, Sharma S (2014) In vitro screening method: an efficient method for screening Alternaria blight resistance/tolerance during early generations in Ethiopian mustard (Brassica carinata A. Braun). Afr J Agric Res 9:137–143

    Article  Google Scholar 

  • Kumari C, Sharma M, Kumar V, Sharma R, Kumar V, Sharma P, Kumar P, Irfan M (2022) Genome editing technology for genetic amelioration of fruits and vegetables for alleviating post-harvest loss. Bioengineering 9(4):176

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Lestari EG (2006) In vitro selection and somaclonal variation for biotic and abiotic stress tolerance. Biodiversity 7:297–301

    Google Scholar 

  • Lokesha R, Naik M (2011) Selection of somaclones resistant to alternaria blight in sesame (Sesamum indicum L.) through in vitro cell line method. Indian J Fundam Appl Life Sci 1:338–346

    Google Scholar 

  • Mahlanza T, Rutherford RS, Snyman JS, Watt PW (2013) In vitro generation of somaclonal variant plants of sugarcane for tolerance to Fusarium sacchari. Plant Cell Rep 32:249–262

    Article  CAS  PubMed  Google Scholar 

  • Matsumoto K, Barbosa ML, Souza LA (1995) Race 1 Fusarium wilt tolerance on banana plants selected by fusaric acid. Euphytica 84:67–71

    Article  Google Scholar 

  • Matsumoto Y, Ogawara T, Miyagi M, Watanabe N, Kuboyama T (2010) Response of wild Cucumis species to inoculation with Fusarium oxysporum f. sp. melonis race 1, 2. J Jpn Soc Hortic Sci 80:414–419

    Article  Google Scholar 

  • McCormick S, Niedermeyer J, Fry B, Barnason A, Horch R, Farley R (1986) Leaf disk transformation of cultivated tomato (Lycopersicum esculentum) using Agrobacterium tumefaciens. Plant Cell Rep 5:81–84

    Article  CAS  PubMed  Google Scholar 

  • Mosqueda MA, Iglesias-Andreu LG, Teixeira-da-Silva JA, Luna-Rodríguez M, Noa-Carrazana JC, Bautista-Aguilar JR, Murguia-Gonzalez J (2019) In vitro selection of vanilla plants resistant to Fusarium oxysporum f. sp. vanillae. Acta Physiol Plant 41:1–8

    Google Scholar 

  • Murashige T, Skoog F (1962) A revised medium for rapid growth and bioassays with tobacco tissue cultures. Physiol Plant 15:473–497

    Article  CAS  Google Scholar 

  • Nachimas A, Barash I, Buchner V, Solel Z, Strobel GA (1979) A phytotoxic glycopeptide from lemon leaves infected with Phoma tracheiphila. Physiol Plant Pathol 14:135–140

    Article  Google Scholar 

  • Panno S, Davino S, Caruso AG, Bertacca S, Crnogorac A, Mandic A, Noris E, Matic S (2021) A review of the most common and economically important diseases that undermine the cultivation of tomato crop in the mediterranean basin. Agronomy 11:2188

    Article  Google Scholar 

  • Pennisi AM, Di Pasquale G, Bonforte M, Sesto F (1988) Phytotoxic metabolites of ipo-virulent and virulent Phoma tracheiphila isolates. Proc Sixth Int Citrus Congr 2:817–827

    Google Scholar 

  • Plich M, Rudnicki MR (1979) Studies of the toxins of P. cactorum pathogenic to apple trees. J Phytopathol 94:270–278

    Article  CAS  Google Scholar 

  • Popoola AR, Durosomo AH, Afolabi CG, Idehen EO (2015) Regeneration of somaclonal variants of tomato (Solanum lycopersicum L.) for resistance to Fusarium wilt. J Crop Improv 29:639–649

    Article  Google Scholar 

  • Purwati RD, HarranSudarsono S (2007) In vitro selection of abaca for resistance to Fusarium oxysporum f. sp. cubense. Hayati J Biosci 14:65–70

    Article  Google Scholar 

  • Remotti PC, Loffler HJM, Vloten-Doting L (1997) Selection of cell-lines and regeneration of plants resistant to fusaric acid from Gladiolus grandiflorus cv. ‘Peter Pears.’ Euphytica 96:237–245

    Article  Google Scholar 

  • Scala A, Bettini P, Buiatti M, Bogani P, Pellegrini G, Togoni F (1984) Tomato Fusarium oxysporum f. sp lycopersici interaction in vitro analysis of several possible pathogenic factors. Phytopathol Z 113:90–94

    Article  Google Scholar 

  • Sengar AS, Thind KS, Kumar B, Pallavi M, Gosal SS (2009) In vitro selection at cellular level for red rot resistance in sugarcane (Saccharum sp.). Plant Growth Regul 58:201–209

    Article  CAS  Google Scholar 

  • Sesto F, Grimaldi V, Pennisi AM (1990) Sensitivity of different citrus and non-citrus species protoplasts towards malsecco toxins. Adv Hortic Sci 4:97–102

    Google Scholar 

  • Shahin EA, Spivey R (1986) A single dominant gene for Fusarium wilt resistance in protoplast-derived tomato plants. Theor Appl Genet 73:164–169

    Article  CAS  PubMed  Google Scholar 

  • Sharma A, Rathour R, Plaha P, Katoch V, Khalsa GS, Patial V, Singh Y, Pathania NK (2010) Induction of Fusarium wilt (Fusarium oxysporum f. sp. pisi) resistance in garden pea using induced mutagenesis and in vitro selection techniques. Euphytica 71:345–356

    Article  Google Scholar 

  • Sharma C, Chandel S, Kaur R (2009) In vitro callus multiplication and shoot regeneration of resistant calli of carnation cv. ‘Raggio-de-Sole’ against Rhizoctonia solani Kuhn. Floricult Ornam Biotechnol 3:49–52

    CAS  Google Scholar 

  • Svabova L, Labeda A (2005) In vitro selection for improved plant resistance to toxin producing pathogens. J Phytopathol 153:52–64

    Article  CAS  Google Scholar 

  • Tripathi MK, Tiwari S, Khare UK (2008) In vitro selection for resistance against purple blotch disease of onion (Allium cepa L.) caused by Alternaria porri. Biotechnology 7:80–86

    Article  Google Scholar 

  • Witjaksono (2003) The role of biotechnology in tropical fruit plant breeding. National seminar- The role of biotechnology in fruit development tropika, IPB. Bogor, 9 May 2003

  • Wu YL, Yi GJ, Peng XX (2010) Rapid screening of Musa species for resistance to Fusarium wilt in an in vitro bioassay. Eur J Plant Pathol 128:409–415

    Article  Google Scholar 

  • Wu Y, Huang B, Peng X, Zhang J (2021) Development of an in vitro hydroponic system for studying the interaction between banana plantlet and Fusarium oxysporum f. sp. cubense. Plant Cell Tiss Organ Cult 146:107–114

    Article  Google Scholar 

  • Yoder OC (1980) Toxins in pathogenesis. Annu Rev Phytopathol 18:103–129

    Article  CAS  Google Scholar 

  • Yusnita Y, Widodo W, Sudarsono S (2005) In vivo selection of peanut somatic embryos on medium containing culture filtrates of Sclerotium rolfsii and plantlet regeneration. Hayati 12:50–56

    Article  Google Scholar 

  • Zhang YP, Jiang S, Qu SP, Yang XM, Wang XN, Wu LL, He YQ, Wang JH (2014) In vitro selection for Fusarium resistant oriental lily mutants using culture filtrate of the fungal agent. Acta Hortic 3:205–212

    Article  Google Scholar 

Download references

Acknowledgements

The authors are grateful to the Department of Environment, Science & Technology, Shimla (HP) India for funding the present research work.

Funding

This work was supported by Department of Environment, Science & Technology (DEST), Shimla (HP) India (No. Env.S&T(F)/R&D Project/2017dated 09/02/2018). Author Rajnish Sharma has received research support from DEST, Shimla (HP) India.

Author information

Authors and Affiliations

Authors

Contributions

All authors contributed to the study conception and design. Material preparation, data collection and analysis were performed by IS, RS, PS, MG and RKB. The first draft of the manuscript was written by IS, RS and PS. All authors read and approved the final manuscript.

Corresponding author

Correspondence to Rajnish Sharma.

Ethics declarations

Conflict of interest

The authors declare they have no conflict of interests.

Additional information

Communicated by Ming -Tsair Chan.

Publisher's Note

Springer Nature remains neutral with regard to jurisdictional claims in published maps and institutional affiliations.

Rights and permissions

Springer Nature or its licensor (e.g. a society or other partner) holds exclusive rights to this article under a publishing agreement with the author(s) or other rightsholder(s); author self-archiving of the accepted manuscript version of this article is solely governed by the terms of such publishing agreement and applicable law.

Reprints and permissions

About this article

Check for updates. Verify currency and authenticity via CrossMark

Cite this article

Shaunak, I., Sharma, R., Sharma, P. et al. Developing resistance against soil-borne Fusarium pathogen causing tomato wilt through in vitro cell line selection. Plant Cell Tiss Organ Cult 153, 91–104 (2023). https://doi.org/10.1007/s11240-023-02446-1

Download citation

  • Received:

  • Accepted:

  • Published:

  • Issue Date:

  • DOI: https://doi.org/10.1007/s11240-023-02446-1

Keywords

Navigation