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Induction of polyploidy in Dahlia pinnata using various concentrations of Oryzalin under in vitro culture
Inducción de poliploidía en Dahlia pinnata mediante diversas concentraciones de orizalina en cultivo in vitro
DOI:
https://doi.org/10.15446/rfnam.v79.124799Keywords:
Diploid, Explants, Flow cytometry, Micropropagation, Tetraploid (en)Diploide, Explantes, Citometría de flujo, Micropropagación, Tetraploide (es)
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Polyploidy induction is widely used in ornamental breeding to generate novel phenotypes such as larger flowers, thicker stems, and increased vegetative biomass. Therefore, this study aimed to evaluate oryzalin-mediated chromosome doubling in in vitro Culture of Dahlia pinnata using a completely randomized 2×6 factorial design with two explant types (apical shoots and middle nodes) and six oryzalin concentrations (0–75 µM) applied by 24 h immersion. Plant height, node number, and leaf number were recorded weekly for 8 weeks after culture, and ploidy was determined by flow cytometry (BD Accuri C6+, USA). The results showed that oryzalin caused a clear dose-dependent growth suppression. Low doses reduced growth without changing ploidy, while 30 µM produced tetraploid sectors in apical shoot-derived plantlets (16.67% putative tetraploids) but with reduced vigor. Higher concentrations (60–75 µM) nearly arrested growth without further increases in polyploid recovery. Middle-node explants showed greater tolerance to oryzalin than apical shoots. These results identify a narrow concentration window for initial polyploid induction in Dahlia pinnata while indicating the need to balance induction efficiency against phytotoxicity.
La inducción de poliploidía se utiliza ampliamente en el mejoramiento de ornamentales para generar fenotipos novedosos, como flores de mayor tamaño, tallos más gruesos y un incremento de la biomasa vegetativa. En este estudio, evaluamos el duplicado cromosómico mediado por orizalina en cultivos in vitro de Dahlia pinnata mediante un diseño completamente aleatorizado con arreglo factorial 2×6, con dos tipos de explante (brotes apicales y nudos medios) y seis concentraciones de orizalina (0–75 µM) aplicadas por inmersión durante 24 h. Se registraron semanalmente, durante 8 semanas de cultivo, la altura de la planta, el número de nudos y el número de hojas, y la ploidía se determinó mediante citometría de flujo (BD Accuri C6+, USA). La orizalina provocó una supresión del crecimiento claramente dependiente de la dosis; las dosis bajas redujeron el crecimiento sin modificar la ploidía, mientras que 30 µM produjo sectores tetraploides en plántulas derivadas de brotes apicales (16,67% de tetraploides presuntos), aunque con menor vigor. Las concentraciones más altas (60–75 µM) casi detuvieron el crecimiento sin incrementos adicionales en la recuperación de poliploides. Los explantes de nudos medios mostraron mayor tolerancia a la orizalina que los brotes apicales. En conjunto, estos resultados identifican una ventana estrecha de concentración para la inducción inicial de poliploidía en Dahlia pinnata y subrayan la necesidad de equilibrar la eficiencia de inducción con la fitotoxicidad.
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