Stomatal and Morphological Characteristics of In-Vitro Grown Tomato (Solanum lycopersicum L.) Treated with Different Concentrations of Colchicine and Pendimethalin

Jessa R. Gines; Catherine C. Arradaza; Marilyn M. Belarmino; Marilou M. Benitez; Rosemarie B. Gonzaga

doi:10.11594/

Authors

Jessa R. Gines Visayas State University
Catherine C. Arradaza Visayas State University
Marilyn M. Belarmino Visayas State University
Marilou M. Benitez Visayas State University
Rosemarie B. Gonzaga Visayas State University

DOI:

https://doi.org/10.11594/

Keywords:

Chemical mutagens, Colchicine, In vitro culture, Pendimethalin, Physiological response, Polyploidy, Stomatal traits, Tomato

Abstract

This research investigated the stomatal and morphological response of in vitro grown tomato (Solanum lycopersicum L.) plantlets to different concentrations of colchicine and pendimethalin. The effects of treatments on stomatal parameters, chlorophyll content and morphological characteristics were studied. The study was conducted using a Completely Randomized Design (CRD) with seven treatments replicated three times with ten samples per replicate at the Plant Tissue Culture Laboratory, Department of Horticulture, Visayas State University. To evaluate the responses of various concentrations of colchicine (1.5, 3.0 and 5.0 mM) and pendimethalin (10, 20 and 30 µM), induced polyploid-associated traits and overall plant growth and development rate on tomato plantlets have also been compared with normal diploid plantlet used as control in subsequent measurement. The results indicated that the mutagen treatments have a substantial effect on explant viability, stomatal traits and chlorophyll content as well as vegetative growth of tomato plants. Moderate concentrations had more profound effect with higher survival percentages of explants found for 20 µM pendimethalin (91%) and 5.0 mM colchicine (84%), which produced regeneration and adaptation, as compared to other concentrations in this study. It was found out with an increased stomatal length, width and aperture on mutation treated plantlets but decreased stomatal density shows induction of polyploid trait. Colchicine resulted in the greatest stomatal size together with favourable impact on chlorophyll content. In contrast, at the highest concentrations of pendimethalin (30 µM), growth was inhibited and chlorophyll was decreased as a result of phytotoxicity.

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References

Aina, O.O., Quensberry, K,H., & Gallo, M. (2012). Colchicine-induced morphologi-cal and cytological changes in lemon grass (Cymbopogon citratus (DC.) (Stapf.). African Journal of Biotechnology, 11(3), 618-625. https://doi.org/10.5897/AJB10.2394.

Arnon, D.I. (1949). Copper enzymes in isolated chloroplasts: Polyphenol oxidase in Beta vulgaris. Plant Physiology, 24(1), 1-15. https://doi.org//10.1104/pp.24.1.1. https://doi.org/10.1104/pp.24.1.1.

Beaulieu, J.M., Leitch, I.J., Patel S, Pendharkar A & Knight, C.A. (2008). Genome size is a strong predictor of cell size and stomatal density in angiosperms. New Phytologist 179(4): 975-986. https://doi.org/10.1111/j.1469-8137.2008.02528.

Calabrese EJ & Baldwin LA. (2003). Hormesis: The dose-response revolution. Annual Review of Pharmacology and Toxicology. https://doi.org/10.1146/annurev.pharmtox.43.100901.140223.

Carputo D, Barone A, Frusciante L & Ricciardi L. (2003). Morphological and cytological characterization of colchicine-induced polyploidy in Solanum species. Euphytica, 130, 259–265. https://10.1002/9780470650202.ch2.

Eng, W. H., Ho, W. S., & Tan, S. G. (2021). Pendi-methalin as an alternative antimitotic agent for polyploid induction in plants. Plants, 10(7), 1431. https://doi.org/10.3390/plants10071431.

Eng, W., & Ho, W. (2019). Polyploidization us-ing colchicine in horticultural plants: A review. Sci. Hortic.-Amst., 246, 604–617. https://doi.org/10.1016/j.scienta.2018.11.010.

Datta, S.K. (2009). Role of induced mutagene-sis for mdvelopment of new varieties in floriculture. Acta Horticulturae, 836, 151-166. https://doi.org/10.17660/ActaHortic.2009.836.18.

del Pozo, J.C., & Ramirez-Parra, E. (2015). Whole genome duplications in plants: An overview from Arabidopsis. Journal of Experimental Botany, 66(22), 6991-7003.

https://doi.org./10.1093/jxberv432.

Dhawan, V., & Lavania, U.C. (1996). Enhance-ment of plant secondary metabolites by mutagenic treatments. Plant Cell, Tissue and Organ Culture, 44, 45–55. https://doi.org./10.1007/BF00021884.

Dhooghe, E., Van Laere, K., Eeckhaut, T., Leus, L., & Van Huylenbroeck, J. (2011). Mitotic chromosome doubling of plant tissues in vitro. Plant Cell, Tissue and Organ Cul-ture (PCTOC), 104(3), 359–373. https://doi.org/10.1007/s11240-010-9786-5.

Doyle, J.J., & Coate, J.E. (2019). Polyploidy, the nucleotype, and novelty: The impact of genome doubling on plant growth and physiology. Current Opinion in PlantBiol-ogy, 49, 1-7. https://doi.org/10.1016/j.pbi.2019.02.001.

Gill, S.S., & Tuteja, N. (2010). Reactive oxy-gen species and antioxidant machinery in abiotic stress tolerance in crop plants. Plant Physiology and Biochemis-try, 48 (12), 909-930. https://doi.org/10.1016/j.plaphy.2010.08.016.

Hassan, J., Miyajima, I., Ozaki, Y., Mizune, Y., Sakai, K., & Wasimullah, Z. (2020). Tetra-ploid Induction by Colchicine Treatment and Crossing with a Diploid Reveals Less-Seeded Fruit Production in Pointed Gourd (Trichosanthes dioica Roxb.). https://doi.org/10.3390/plants9030370.

Kermani, M.J., Sarasan, V., Roberts, A.V., Yoko-ya, K. & Wentworth, J. (2003). Oryzalin-induced chromosome doubling in Rosa and its effect on plant morphology and pollen viability. Theoretical and Applied Genetics, 107(7), 1195–1200. https://doi.org/10.1007/s00122-003-1374-1.

Kumar, V., Singh, G., & Sharma, S. (2022). Sur-face sterilization for isolation of endo-phytes: Ensuring sterility while maintain-ing microbial culturability. Journal of Basic Microbiology, 62(3), 263–287. https://doi.org/10.1002/jobm.202100462.

Lehrer, J.M., Brand, M.H., & Lubell, J.D. (2008). Induction of tetraploidy in meristemati-cally active seeds of Japanese barberry (Berberis unbergii var. Atropurpurea) through exposure to colchicine and oryzalin. Scientia Horticulturae, 119:67–71. https://doi.org/10.1016/j.scienta.2008.07.020.

Maherali, H., Walden, A.E., & Husband, B.C. (2009). Genome duplication and the phys-iology of polyploid plants. New Phytolo-gist, 183(3), 751–760. https://doi.org./10.1111/j.1469-8137.2009.02997.x.

Mangena, P., & Mushadu, N. (2023). Colchi-cine-induced polyploidy in leguminous crops enhances morpho-physiological characteristics for drought stress toler-ance. Life, 13 (10), 1966. https://doi.org./10.3390/life13101966.

Matias, K.M.L., Sotto, R.C., Bautista, N.S., Prota-cio,C.M., & Magdalita P.M. (2024). Influ-ence of Drought Stress and Foliar Appli-cation of Salicylic Acid on Early Vegeta-tive Stage of Cacao. Philippine Journal of Science, 153 (2).

Morejohn, L.C., Bureau, T.E., Mole-Bajer, J., Ba-jer, A.S., & Fosket, D.E. (1987). Oryzalin, a dinitroaniline herbicide, binds to plant tubulin and disrupts microtubules. https://doi.org./10.1126/science.3576224.

Mulyana, A., Purwoko, B.S., Dewi, I.S., & Maha-rijaya, A. (2023). In vitro diploidization of haploid plantlet from anther culture of eggplant (Solanum melongena L.) using pendimethalin. SABRAO Journal of Breed-ing and Genetics, 55 (1) 115-122. https://doi.org/10.54910/sabrao2023.55.1.11.

Murashige, T., & Skoog, F. (1962). A revised medium for rapid growth and bioassays with tobacco tissue cultures. Physiol. Plant. 15(3): 473- 497. https://doi.org/10.1111/j.1399-3054.1962.tb08052.x.

Ramulu, K.S., Verhoeven, H.A., & Dijkhuis, P. (1991). Mitotic blocking, micronuclea-tion, and chromosome doubling by oryza-lin, amiprophos-methyl, and colchicine in potato. Protoplasma 160, 65-71. https://doi.org/10.1007/BF01322611.

Sajid M., & Afab, S. (2013). Low dose pendime-thalin mediated polyploidy induction in plant tissue cultures. Journal of Agricul-tural Science, 5(2), 27-35.

Sakhanokho, H.F., Rajasekaran, K., Kelley, R.Y., & Islam-Faridi, M.N. (2009). Induced polyploidy in diploid ornamental ginger (Hedychium muluense R.M. Smith) using colchicine and oryzalin. HortScience, 44(7), 1809-1814. https://doi.org./10.21273/HORTSCI.44.7.1809.

Sattler, M.C., Carvalho, C.R., & Clarindo, W.R. (2016). The polyploidy and its key role in plant breeding. Planta, 243, 281–296. https://doi.org/10.1007/s00425-015-2450-x.

Singh, B., Yun, S., & Park, M.H. (2025). The role of colchicine in plant breeding. Interna-tional Journal of Molecular Sciences, 26(14), 6743. https://doi.org/10.3390/ijms26146743

Talukdar, D. (2013). Cytogenetic characteriza-tion of induced autopolyploidy in plants. Plant Systematics and Evolution, 299, 631–646. https://doi.org/10.5772/55481.

Thao, N.P., Nakashima, A., & Tanaka, K. (2003). Colchicine-induced polyploidy in ornamental plants: Physiological and morphological outcomes. Plant Cell Re-ports, 21(12), 1151–1156. https://doi.org/10.1023/A:1021280611527.

Tossi, Valeria E.., Nichols, L. A., & other au-thors. (2022). Impact of polyploidy on plant tolerance to abiotic and biotic stresses. Frontiers in Plant Science, 13, 869423. https://doi.org/10.3389/fpls.2022.869423.

Vaughn, K.C., & Lehnen, L.P. Jr. (1991). Mitotic disrupter herbicides. Weed Science, 39(3), 450-457.

Yao, P.Q., Chen, J.H., Ma, P.F., Xie, L.H., & Cheng, S.P. (2023). Stomata variation in the process of polyploidization in Chinese chive (Allium tuberosum). BMC Plant Bi-ology, 23, 595. https://doi.org/101186/s12870-023-04615-y.

Zakariyya, F.M., Setyawan, B., & Wahyo, S. (2017). Stomatal, Proline and Leaf Water Status Characters of Some Cocoa Clones (Theobroma cacao L.) on Prolonged Dry Season. Pelita Perkebunan: a Coffee and Cocoa Research Journal 33(1): 109-117. https://doi.org/10.3390/foods121183519.