Development of a temporary immersion system (RITA®) for mass production of sugarcane (Saccharum spp. interspecific hybrids)

Abstract

Commercial micropropagation of sugarcane is largely determined by the clonal fidelity and the cost of plants produced. Rapid production of plants in vitro reduces the frequency of offtypes in many species. By exploiting the concept of transverse thin cell layer culture, we have developed a rapid, high frequency direct plant regeneration system, called SmartSett®, for commercial sugarcane cultivars grown in Australia. Similar to conventional micropropagation, labour remains the major cost of this plant production system. Hence, to reduce the labour component, we have integrated the SmartSett® system with the RITA® temporary immersion bioreactor. Thin transverse leaf sections or fragmented leaves cultured on agar-based SmartSett® shoot induction medium were used as the starting material for RITA®. Shoot initiation on semi-solid medium prior to transferring to RITA®, culture immersion frequency, explant size and genotype determined the productivity (number of plants produced per unit culture) of the system. Results obtained with cultivar Q165 indicate that explants cultured for 45 d on SmartSett® shoot induction medium were the most prolific, producing on average 275 shoots per vessel after 45 d of culture in RITA with 1 min immersion every 12 or 24 h. Using the fragmented tissue, 14-d-old explants and 3-mm leaf tissue fragments were the most productive. Experiments with three cultivars (Q117, Q165 and Q205) showed that RITA® culture conditions need to be optimised for each cultivar for maximum plant production.

Agrobacterium-mediated in planta genetic transformation of sugarcane setts

“…Development of techniques using mature tissue is important for the improvement of commercial cultivars (Pérez-Jiménez et al 2013). In addition, sugarcane is a vegetatively propagated crop and stable transformants could be clonally multiplied for distribution to growers (Mordocco et al 2009). …”

In Vitro Propagation of Sugarcane for Certified Seed Production

“…However, to reduce the labor required and increase efficiency, temporary immersion systems (TISs) have been successfully used to improve in vitro sugarcane multiplication [12][13][14][15][16]. The principle of these systems is the immersion of explants for a determined time and frequency. )…”

Evaluasi varietas, sumber eksplan dan strain Agrobacterium terhadap keberhasilan transformasi tebu dengan gen P5CS Evaluation of varieties, explant sources, and Agrobacterium strains for successful sugarcane transformation using P5CS gene

“…Kalus embriogenik dan embrio somatik varietas PS 881, PS 891 dan KK disiapkan melalui metode Temporary Immersion System (TIS) atau Sistem Perendaman Sesaat (SPS) (Mordocco et al, 2009;Minarsih et al, 2013). Kalus yang berasal dari pucuk tebu ketiga varietas tersebut setelah disubkultur sebanyak 2-4 kali kemudian dimasukkan dalam bejana SPS RITA ® yang dihubungkan dengan pompa dan automatic timer (Autonic) dengan lama perendaman diatur selama tiga menit dengan frekuensi setiap 24 jam hingga terbentuk kalus embriogenik (enam minggu) dan embrio somatik (delapan minggu).…”