Abstract
The high pigment (hp) tomato mutants increase the fruit pigment content in tomato. The dark green (dg) tomato mutant belonging to this category contains the hp-2dg recessive allele, resulting from a single nucleotide polymorphism (SNP) in the DEETIOLATED1 (DET1) gene. Naturally, identification of this functional SNP is crucial for introgression of this important mutant allele in suitable tomato cultivar(s). Hence, we developed a functional co-dominant marker for detecting the hp-2dg mutant allele using the tetra-primer amplicon refractory mutation system (T-ARMS) strategy. Although the other available techniques for identification of SNPs require specialized costly equipments and reagents, our method allows the reliable identification of this functional SNP in time-, labour- and cost-effective manner, using standard molecular biology equipments and reagents. To the best of our knowledge, this is the first report of T-ARMS strategy for the detection of the dg mutant allele in tomato, which will definitely help in identification and introgression of this allele by the breeders, working even under limited fund and resources.
This is a preview of subscription content, log in via an institution to check access.
Similar content being viewed by others
References
Arens P, Mansilla C, Deinum D, Cavellini L, Moretti A, Rolland S, van der Schoot H, Calvache D, Ponz F, Collonnier C, Mathis R, Smilde D, Caranta C, Vosman B (2010) Development and evaluation of robust molecular markers linked to disease resistance in tomato for distinctness, uniformity and stability testing. Theor Appl Genet 120:655–664. https://doi.org/10.1007/s00122-009-1183-2
Hazra P, Longjam M, Chattopadhyay A (2018) Stacking of mutant genes in the development of “purple tomato” rich in both lycopene and anthocyanin contents. Sci Hortic 239:253–258. https://doi.org/10.1016/j.scienta.2018.05.039
Kumar V, Kumar P, Chattopadhyay T (2017) A rapid and reproducible method for isolating genomic DNA from a few crop plants suitable for polymerase chain reaction-based genotyping. J Appl Nat Sci 9(2):1119–1122. https://doi.org/10.31018/jans.v9i2.1332
Levin I, Frankel P, Gilboa N, Tanny S, Lalazar A (2003) The tomato dark green mutation is a novel allele of the tomato homolog of the DEETIOLATED1 gene. Theor Appl Genet 106:454–460. https://doi.org/10.1007/s00122-002-1080-4
Levin I, de Vos CHR, Tadmor Y, Bovy A, Lieberman M, Oren-Shamir M, Segev O, Kolotilin I, Keller M, Ovadia R, Meir A, Bino RJ (2006) High pigment tomato mutants—more than just lycopene (a review). Isr J Plant Sci 54(3):179–190. https://doi.org/10.1560/IJPS_54_3_179
Mustilli AC, Fenzi F, Ciliento R, Alfano F, Bowler C (1999) Phenotype of the tomato high pigment-2 mutant is caused by a mutation in the tomato homolog of DEETIOLATED1. Plant Cell 11(2):145–157. https://doi.org/10.1105/tpc.11.2.145
Ye S, Dhillon S, Ke X, Collins AR, Day INM (2001) An efficient procedure for genotyping single nucleotide polymorphisms. Nucleic Acids Res 29(17):e88. https://doi.org/10.1093/nar/29.17.e88
Acknowledgements
We thank Prof. Pranab Hazra, BCKV for providing seeds of Aft and dg mutant lines and “purple tomato” genotype. ICAR-IIHR, Bengaluru, ICAR-IIVR Varanasi and NBPGR are acknowledged for providing other seed materials. The authors thank Deepak Maurya, Shubham Kumar and Vikash Kumar for their help. Financial support from BAU in terms of project grant (Code: SNP/CI/Rabi/2018-5) is highly acknowledged. This article bears BAU COMMUNICATION NO. 889/200930.
Author information
Authors and Affiliations
Contributions
TC conceived the idea. AM and TC designed the primers and performed experiments. SA, AM and TC carried out crop production and plant hybridization works. All the authors took part in preparation and correction of the manuscript. All the authors read the final manuscript and approved it.
Corresponding author
Ethics declarations
Conflict of interest
On behalf of all authors, the corresponding author states that there is no conflict of interest.
Research involving human participants and/or animals
No human participants and/or animals were involved in this study.
Informed consent
Not applicable as no human participants and/or animals were involved in this study.
Rights and permissions
About this article
Cite this article
Mukherjee, A., Akhtar, S. & Chattopadhyay, T. Development of tetra-primer amplicon refractory mutation system (T-ARMS) strategy for identification of the dark green mutant allele in tomato. 3 Biotech 11, 182 (2021). https://doi.org/10.1007/s13205-021-02732-9
Received:
Accepted:
Published:
DOI: https://doi.org/10.1007/s13205-021-02732-9