In Vitro Growth of Curcuma longa L. in Response to Five Mineral Elements and Plant Density in Fed-Batch Culture Systems

Abstract

Plant density was varied with P, Ca, Mg, and KNO3 in a multifactor experiment to improve Curcuma longa L. micropropagation, biomass and microrhizome development in fed-batch liquid culture. The experiment had two paired D-optimal designs, testing sucrose fed-batch and nutrient sucrose fed-batch techniques. When sucrose became depleted, volume was restored to 5% m/v sucrose in 200 ml of modified liquid MS medium by adding sucrose solutions. Similarly, nutrient sucrose fed-batch was restored to set points with double concentration of treatments’ macronutrient and MS micronutrient solutions, along with sucrose solutions. Changes in the amounts of water and sucrose supplementations were driven by the interaction of P and KNO3 concentrations. Increasing P from 1.25 to 6.25 mM increased both multiplication and biomass. The multiplication ratio was greatest in the nutrient sucrose fed-batch technique with the highest level of P, 6 buds/vessel, and the lowest level of Ca and KNO3. The highest density (18 buds/vessel) produced the highest fresh biomass at the highest concentrations of KNO3 and P with nutrient sucrose fed-batch, and moderate Ca and Mg concentrations. However, maximal rhizome dry biomass required highest P, sucrose fed-batch, and a moderate plant density. Different media formulations and fed-batch techniques were identified to maximize the propagation and storage organ responses. A single experimental design was used to optimize these dual purposes.

In vitro arrow cane (Gynerium sagitatum Aubl.) multiplication in double phase medium

“…Considering the medium quantity needed for each system, gelling agent amount, labor related to subculture and washing glassware; it is estimated that double phase medium system can lower the total cost of plants by 20% compared to semisolid medium cultures system (Table 1) López y Suárez -In vitro multiplication of cane arrow Culture media for in vitro tissue culture can be formulated as semisolid, liquid or double phase (liquid on top of semisolid) (Akhtar, 2013;Mehta et al, 2014;Litwińczuk, 2013). Semisolid media favor explant stability and allow to expand the time for subculture because slow nutrient and water release to the tissues; in contrast, they are usually more expensive because of gelling agents costs and nutrient absorption by tissues is slower; in contrast, liquid media are less expensive for absence of gelling agents, nutrient and plant regulators absorption by explants is faster, especially in cell suspension systems; (El-Hawaz et al, 2015;Perera et al, 2015). For liquid cultured tissues subcultures are more frequent because of nutrient depletion, tissue vitrification and phenol leakage in the medium that negatively affects growth and survival (Dong et al, 2016;Jones and Saxena, 2013).…”

In vitro mineral nutrition of Curcuma longa L. affects production of volatile compounds in rhizomes after transfer to the greenhouse

“…Turmeric rhizomes were grown on 40 mL modified MS media [ 21 ] in cylindrical glass jar (180 mL) with plant density (3, 6, 9 buds/vessel) for 3 cycles of 35 days per cycle then rhizomes were transferred to liquid modified MS media in bioreactor (2.5 L Liquid Lab Vessels, Southern Sun Inc., Hodges, SC, USA) with plant density (6, 12, 18 buds/vessel) [ 17 ]. The bioreactors were set on an intermittent immersion rocker system [ 15 , 17 , 20 ] with one rotation per min to allow rhizomes dry and wet in the thin film liquid media (Fig. 1 ).…”

“…1 ). Liquid MS media [ 21 ] was modified with (NH 4 ) 2 SO 4 , (5 mM), sucrose (5% m/v), benzyladenine (3 μM) [ 22 ] different combinations of in vitro factors (Additional file 1 : Table S1), plant density and mineral concentrations, PO 4 3− (1.25, 3.75, 6.25 mM), Ca 2+ (3, 6, 9 mM), Mg 2+ (1.5, 3, 4.5 mM), and KNO 3 (20, 40, 60 mM), were selected by d-optimal criteria [ 17 ]. The set of experimental treatments was duplicated with each subset receiving two different fed-batch techniques, sucrose fed-batch (SF) and nutrient-sucrose fed-batch (NSF) applied twice during the 5 months culture (Fig.…”

“…In vitro cultures were recognized as potential methods to produce secondary metabolites from plant cells, tissues, and organs for industry [ 12 , 13 ]. A large mass of plant material is a primary requirement to produce chemical compounds [ 14 ] and increasing the plant mass could be achieved by either using large vessels in “bioreactors” with liquid media [ 15 , 16 ] or adding more initial plants mass with fed-batch supplementation during the culture cycle [ 17 ]. Commercial scale fed-batch bioreactors (10,000 to 20,000 L) used to produce ginseng saponins from Panax ginseng root culture have achieved high yield [ 18 ].…”

“…In vitro treatments including minerals, plant density, and fed-batch techniques (in vitro treatments) applied during 5 months of micropropagation in fed-batch bioreactors [ 17 ] effects the plant quality during the subsequent 6 months of greenhouse growth where rhizomes continue to attain mass. These effects include both the relative fresh mass gain of nursery plants [ 20 ] and the concentration of curcuminoids in the rhizome following a season of growth [ 19 ].…”