| Plant latin name | Zingiber officinale Roscoe |
| Literature code | Zingiber_officinale-Ref-7 |
| Reference | Swarnathilaka D.B.R., et al., British Biotechnology Journal 12: 1-7 (2016) |
| Summary | Aims: In-vitro produced microrhizomes in ginger are favored as planting material over other conventional planting materials as they are free from soil born pathogens. This study was conducted to develop an efficient protocol for production of healthy microrhizomes including the
investigation of effect of different growth regulators, sucrose concentration and photoperiod exposure levels.
Place and Duration of Study: The entire study was conducted in plant tissue culture research unit of the Department of Export Agriculture, Walpita, Sri Lanka between November 2013 and August 2015.
Methodology: In-vitro produced shoots established in hormone free medium were cultured in Murashige and Skoog (1962) (MS) medium fortified with eight treatments of 6-benzylaminopurine (BAP) (0.0, 2.0, 0.4 and 6.0 mg L-1 of BAP) and Naphthalene acetic acid (NAA) (0.1 and 0.2 mg L-1
of NAA) structuring in factorial design to study the effect of growth regulators for formation of microrhizomes. In-vitro produced shoots were cultured on MS medium fortified with different concentration of sucrose (30, 60, 90 and 120gL-1) separately. Effect of the solid/liquid nature of the medium and the different photoperiod level on induction of microrhizome was also studied.
Results: Results revealed that medium containing 4.0 mgL-1 BAP with 0.1 mgL-1 NAA showed the best response followed by 6.0 mgL-1 BAP with 0.1 mgL-1 NAA for induction of microrhizomes within 60 days. Increased level of NAA did not enhance microrhizome induction. Results of different
concentration of sucrose revealed that MS medium fortified with 90 g L-1 sucrose recorded the highest fresh and dry weight of microrhizomes followed by the treatment with 60 g L-1 sucrose. However, plantlets supplemented with more than 90 g L-1 of sucrose exhibited lower weight of
microrhizomes, but higher root induction and root fresh weight probably due to accumulation of water. Different photoperiod exposure levels revealed that 16 hrs of light and 4 hrs of dark condition with solid medium produced highest fresh weight (3.72 g) and highest number of microrhizomes
(9.6).
Conclusion: Murashige and Skoog (1962) medium supplemented with 4 mgL-1 BAP, 0.1 mgL-1 NAA and 90 gL-1 sucrose in solid form with 16-h photoperiod for 10 weeks of culture duration were the best conditions for induction microrhizomes in ginger, cultivar Local. |
| Objectives | Effective, reliable and reproducible protocol
for in-vitro production of ginger microrhizomes |
| Materials | Local variety of ginger |
| Explant | Aseptic shoots |
| Initial culture | Aseptic shoots approximately 3-4 cm long, which
were derived from the in-vitro established culture
of Local variety of ginger were used as explants
for induction of microrhizomes. After separating
the fully grown shoots originated from the axillary
rhizome buds, the small shoots with the size of 3-
4 cm long were again cultured on fresh medium
without any growth hormone for another four
weeks to avoid carryover effects of growth
regulators. |
| Shoot multiplication | In-vitro produced shoots with 3-4 cm shoot
height, established in hormone free medium,
were cultured in Murashige and Skoog (1962)
medium fortified with eight treatments
of 6-benzylaminopurine (BAP) (0.0, 2.0, 0.4 and
6.0 mg L-1 of BAP) and Naphthalene acetic acid
(NAA) (0.1 and 0.2 mg L-1 of NAA) structuring in
factorial design to study the effect of growth
regulators for formation of microrhizome. Each
treatment consisted of 10 individual culture
bottles to provide 10 replicates. Data in each
culture bottle were recorded separately on
number of rhizomes, fresh weight of rhizome,
number of buds, shoot height, and number
leaves after 60 days of culturing. Among the 8 different
combinations of growth regulators, 4 mgL-1 BAP
together with 0.1 mgL-1 NAA exhibited a better
response than the other treatments in terms of
mean number of microrhizomes.
In-vitro produced shoots with 3-4 cm shoot height
established in hormone free medium were
cultured on MS medium supplemented with 4
mgL-1BA, 0.25 mgL-1 NAA and different
consecrations of sucrose (30, 60, 90, and 120
gL-1) separately. All the cultures were incubated
at 26±2°C with 16/8 hr light and dark period.Data were collected on number of buds, shoot
height, number of leaves, wet and dry weight of
shoots, roots and rhizome. Ginger plantlets in the MS medium containing 90
gL-1 sucrose showed the highest weight of
microrhizome with mean value of 8.553 g (wet)
and 0.771 g (dry) followed by the medium with
60 gL-1 sucrose.
In-vitro produced shoots with (about 3-4 cm
height established in hormone free medium were
collected and cultured on solid and liquid forms
of MS medium supplemented with 4 mgl-1BA,
0.1 mgL-1 NAA, and 90 gL-1 sucrose. Varying
level of photoperiod 0 (dark) hr, 4 hr, 8 hr and
16 hr were provided to examine their effect on
formation of microrhizome and all the cultures
were incubated at room temperature (26±2°C).
Data were collected on, number of
microrhizomes, fresh weight of microrhizome,
number of shoots, shoot height, shoot and root
fresh weight and percentage of cultures that
induced microrhizomes. Microrhizomes produced under 4, 8 and 16 photo
period in MS solid treatments significantly
increased the fresh weight than the cultures
produced in the liquid medium. The highest performance was obtained in solid
cultures incubated under light condition with 16-
h/day producing 3.72 g of average fresh weight
of rhizomes and 9.6 number of microrhizomes.
MS medium,
supplemented with 4 mgL-1 BAP, 0.1 mgL-1 NAA
and 90 gL-1 sucrose, in solid condition under the
16-h photoperiod with 10 weeks of culture
duration were the best conditions for induction
of microrhizomes in ginger. |
| Rooting | |
| Acclimation | |
| Planting | |
| Cultivation conditions | |
| Traints of regenerants | |
| Ingredients analyzed | |
| Extraction | |
| Analitical methods | |
| Notes | |