Rapid Somatic Embryogenesis of Citrus reticulata Blanco cv. Madu in an Air-Lift Bioreactor Culture

Abstract

Embryogenic cell (EC) growth and proliferation as well as somatic embryo induction were examined in a bioreactor culture using Murashige and Skoog basal medium particularly for the influence of 6-benzyladenine (BA) concentration, initial cell density and aeration rate. Embryogenic callus was induced from seeds of Citrus reticulata Blanco cv. Madu. The cell suspension in a 3-L bioreactor exhibited maximum cell growth following the addition of 1.5 mg/l of BA. The fresh weight (FW) of the cells after 28 days of growth was found to increase from an initial of 5.5 g cell culture to 57.3 g, a 10.4-fold increase and the maximum growth rate (GR) of the cells (0.33 g/day) was attained by the 7 th day of culture. In the cell density experiment, ECs at a concentration of 5.5 g/l constituted the most effective inoculum, reaching the highest GR of ECs (0.52 g/ day), again by the 7 th day of culture. In the aeration experiment, the highest EC GR of 2.6 g/day was obtained at the maximum aeration rate of 1.5 vvm (air volume medium/ volume/min). After 28 days of somatic embryogenesis, 79% of ECs became somatic embryos, of which 29% were at cotyledonary stage.

Effect of Gamma-Ray Irradiation on Vetiver Grass (Vetiveria Zizanioides (L.) Nash.) in Vitro Shoots Growth and Multiplication

“…in vitro shoots occurred at 33 Gy [14], callus of sugarcane (Saccharum officinarum L.) happened with a dosage of 28.8 Gy [26], With an 18 Gy dose, grape variety "Red Globe" in vitro shoots appeared [27], and callus Citrus reticulata cv. Limau Madu occurred at a dose of 30 Gy [13]. This shows that the level of radiosensitivity in each species was different.…”

“…At low doses of irradiation, vetiver in vitro shoot explants was able to survive (high survival rate) and produced new shoots. Low doses of gamma rays were able to stimulate cell division, growth and development in various plant species [13]. Low dose irradiation on banana cv.…”

“…Research on nucellus of Citrus reticulata cv. Limau Madu irradiated at a dose of 100-120 Gy became brown in the second week and died on the 35 days of culture [13]. The browning reaction occurs due to the presence of oxygen, phenol content, and polyphenol oxidase (PPO).…”

Effect of Gamma Rays Irradiation and In Vitro Selection on Citrus nobilis (L.) ‘Siam Banyuwangi’ to Huanglongbing (HLB) Disease

“…The irradiated embryos were then planted on the prepared media, enriched by GA3 at 1 mg/l. Additionally, planting medium plays a pivotal role for the embryos' growth (Agisimanto, Normah, & Ibrahim, 2019). Mutant shoots were sub-cultured every two months.…”

Somatic Embryos Induction of East Kalimantan Local Rice (Oryza sativa L.) Cultivars and In Vitro Selection Against Salinity

“…Somatic embryogenesis is a process whereby embryos develop from somatic cells or tissues (von Arnold, Sabala, Bozhkov, Dyachok, & Filonova, 2002). For mass propagation, somatic embryos have benefits due to the high multiplication rate, easily propagated in liquid media, the handling of large numbers of embryos at once (Agisimanto, Normah, & Ibrahim, 2019;Loyola-Vargas & Ochoa-Alejo, 2016). Therefore, somatic embryos are potential explants for plant developmental purposes, including in vitro selection for developing stress-tolerant plants.…”