RESUMEN
Bunium persicum is a valuable medicinal plant with limited production but high market demand. It thrives predominantly in high-altitude regions. The main challenges hindering its widespread cultivation are seed dormancy and a lengthy seed-to-seed cycle, making its large-scale cultivation difficult. Six genotypes of Bunium persicum were collected from different altitudes to evaluate its germination behavior and seed dormancy. The study was conducted during 2020-23 and comprised three experiments (viz., seed germination under an open field, controlled conditions, and micro-tuberization). Under open field conditions, germination percent was genotype dependent, and the highest germination percentage, root length, and shoot length were recorded in Shalimar Kalazeera-1. Germination behavior assessment of the Bunium persicum revealed that treatment T9 (GA3 (25 ppm) + TDZ (9 µM/L)) is effective in breaking the dormancy of Bunium persicum as well as in obtaining a higher germination percent for early development of the tubers. Similarly, with regard to the effect of temperature and moisture conditions, stratification under moist chilling conditions showed effectiveness in breaking seed dormancy as the germination percentage in stratified seeds was at par with the most efficient growth hormone. With regard to the in vitro micro-propagation, direct regeneration showed multiple shoot primordia at the base of the tubers without intervening callus phase from the MS medium supplemented with BA (22.2 µM) and NAA (13.95 µM) 4 weeks after sub-culturing. Similarly, medium supplemented with JA (8.0 mg/L) and BA (22.2 µM) produced well-organized somatic embryos with shiny surfaces, which appeared at the swelled basal portion of apical stems. Further, the combination of JA (6.0 mg/L) and BA (22.2 M) was effective in developing the micro-tubers and also enhanced the weight and length of Bunium persicum micro-tubers.
RESUMEN
Potato microtuber productions through in vitro techniques are ideal propagules for producing high quality seed potatoes. Microtuber development is influenced by several factors, i.e., high content sucrose and cytokinins are among them. To understand a molecular mechanism of microtuberization using osmotic stress and cytokinin signaling will help us to elucidate this process. We demonstrate in this work a rapid and efficient protocol for microtuber development and gene expression analysis. Medium with high content of sucrose and gelrite supplemented with 2iP as cytokinin under darkness condition produced the higher quantity and quality of microtubers. Gene expression analysis of genes involved in the two-component signaling system (StHK1), cytokinin signaling, (StHK3, StHP4, StRR1) homeodomains (WUSCHEL, POTH1, BEL5), auxin signaling, ARF5, carbon metabolism (TPI, TIM), protein synthesis, NAC5 and a morphogenetic regulator of tuberization (POTH15) was performed by qPCR real time. Differential gene expression was observed during microtuber development. Gene regulation of two component and cytokinin signaling is taking place during this developmental process, yielding more microtubers. Further analysis of each component is required to elucidate it.
RESUMEN
RESUMEN Existen numerosos factores que afectan la micropropagación y la microtuberización en papa; entre ellos, las hormonas vegetales y el fotoperiodo. Para estudiar el efecto de estos dos factores en las variedades 'Arbolona negra' (AN) y 'Granola' (G), se cultivaron microesquejes de cada variedad en medio MS líquido con o sin giberelinas (GA) y con 25 g/L de sacarosa e incubados bajo condiciones de luz blanca continua. Para inducir la microtuberización, las vitroplántulas obtenidas fueron sub-cultivadas en medio MS suplementado con 50 g/L sacarosa, tres concentraciones de BA (0, 1 y 5 mg/L) e incubadas bajo diferentes regímenes lumínicos. El pre-tratamiento con GA favoreció el alargamiento del vástago en AN pero no en G. Ambas variedades produjeron el mayor número de microtubérculos en medio MS suplementado con 5 mg/L de BA, bajo condiciones fotoperiódicas, sin la adición previa de GA. El cultivo in vitro de microesquejes de papa en medios de cultivo suplementados con BA y sacarosa, y la incubación bajo condiciones de días cortos permite obtener microtubérculos de papa en condiciones in vitro, en un tiempo más corto que el que podría esperarse en condiciones tradicionales de cultivo.
ABSTRACT Micropropagation and microtuberization in potato plants are both affected by plant hormones and photoperiod. To study the effect of these factors on potato cultivars 'Arbolona negra' (AN) and 'Granola' (G), microcuttings of each cultivar were cultured on MS liquid medium supplemented with sucrose 25 g/L, with or without gibberellins (GA). These microcuttings were incubated under continuous light. In order to induce microtuberization, the obtained vitroplantlets were subcultured on MS medium supplemented with sucrose 50 g/L, three concentrations of BA (0, 1 and 5 mg/L) and incubated under different light patterns. GA pre-treatment induced shoots elongation on AN but not on G cultivar. The highest microtubers production for both cultivars was achieved on MS medium supplemented with BA 5 mg/L under short day light condition without GA pretreatment. The in vitro culture of microcuttings on culture media supplemented with BA and sucrose, incubated under short day conditions, allowed microtubers production in a shorter time lapse.
RESUMEN
Twenty-two genotypes of potato (Solanum tuberosum L.) were induced to form microtubers under six in vitro culture conditions. Cultures maintained under a short photoperiod (10 h of 6-12 µmol m-2 s-1) and low temperatures (day 20°±2°C and night 18°±2°C) had both a higher yield (255 mg/plantlet) and a greater number (2/plantlet) of microtubers than those maintained under long days (16 h of 38-50 µmol m-2 s-1) combined with high temperatures (day 28°±2°C and night 25°±2°C) (yield 207 mg/plantlet; microtuber number, 0.9/plantlet), over a wide range of genotypes. After the plantlets had been cultured under long days for an initial period of 60 days, continuous darkness advanced microtuberization by 2-3 months in various genotypes. Under short-day and low-temperature conditions the addition of 6-benzylaminopurine increased microtuber yield from 255 mg/plantlet to 645 mg/plantlet and average microtuber weight from 115 mg to 364 mg. A similar pattern was observed under conditions of long days and high temperature, and continuous darkness and low-temperature. Microtubers produced under light had a greater number of eyes (maximum average: 5.96/microtuber) than those produced in the dark (maximum average: 3.50/plantlet). The genotype × cultural conditions interactions were significant indicating the importance of developing genotype-specific protocols to maximize microtuberization.
