Preculture sugarcane tissue in sucrose-supplemented culture medium to induce desiccation tolerance

Abstract

Cryopreservation is a promising technique for long term conservation of sugarcane's genetic resources which could be used in breeding programs. Preculture is a key step for success this technique. The purpose of this study was to evaluate the effect of sucrose concentrations and preculture period on dessication tolerance of sugarcane tissue dehydrated to 30, 20 and 10 % moisture. Sugarcane shoot tips were encapsulated in sodium alginate and precultured in a liquid culture medium with 0.3, 0.5, and 0.75 M sucrose. Preculturing in 0.3 M sucrose was ideal to induce desiccation tolerance in tissue. This sucrose concentration reduced the percentage of electrolytic leakage. Shoot tips were sensitive to a sucrose concentration of 0.75 M, resulting in low survival rates after desiccation. Preculturing in 0.5 M sucrose resulted in a higher survival rate after drying for 5.7 and 7.45 h. The percentage of electrolytic leakage was high when shoot tips were desiccated after preculturing in 0.5 and 0.75 M sucrose.

Adding Ascorbic Acid to Reduce Oxidative Stress During Cryopreservation of Somatic Embryos of <i>Paphiopedilum niveum</i> (Rchb.f.) Stein, an Endangered Orchid Species

“…With the V cryoplate method, large sugarcane shoot tips resulted in higher regrowth than smaller ones (Rafique et al, 2015). Melo et al (2011) also found that large explants could accumulate carbohydrate in the form of starch grains that could serve as an energy source for the regeneration of sugarcane shoot tips after cryopreservation. Moreover, Chaireok et al (2016) reported that vigor regrowth could be observed in large postcryopreserved P. niveum calli with high carbohydrate accumulation.…”

Influence of ethylene glycol on Eucalyptus grandis cryopreservation using the V cryo-plate technique

“…Despite the many advantages of cryopreservation, viable protocols that can guarantee good rates of regeneration of the material after cryopreservation are not yet available for most species, thus limiting its application for long-term germplasm conservation (Funnekotter et al 2017). Several factors can influence plant recovery after immersion in liquid nitrogen, including the age of the mother plant that provided the explant, resistance of tissues during dissection, the type of cryoprotectant solution employed, the time of exposure to it, and the specific cryopreservation techniques to be employed (Sakai and Engelmann 2007, Uchendu and Reed 2008, Melo et al 2011, Engelmann 2014.…”

Challenges in the development of a widely applicable method for sugarcane (Saccharum spp.) shoot tip cryopreservation

“…Cryopreservation techniques that make use of shoot tips excised from in vitro-grown plants have been published for many genera, including Saccharum. Paulet et al (1993), González-Arnao et al (1999), González-Arnao and Engelmann (2006) and Melo et al (2011) all published encapsulation-dehydration methods that demonstrated successful cryopreservation of sugarcane cultures. These techniques precultured shoot tips on medium supplemented with 0.5 to 0.75 M sucrose prior encapsulation in calcium-alginate beads.…”

“…Beads were then dehydrated to moisture contents of 20 to 25%, and then plunged into liquid nitrogen (LN). Regrowth levels were highly variable, ranging from 14 to 91% for S. officinarum cultivars (Paulet et al, 1993;González-Arnao et al, 1999;González-Arnao and Engelmann, 2006;Melo et al, 2011).…”