| Plant latin name | Zingiber officinale Roscoe |
| Literature code | Zingiber_officinale-Ref-4 |
| Reference | Rout G.R., et al., In Vitro Cell. Dev. Biol.-Plant 37: 814-819 (2001) |
| Summary | Shoot multiplication of Zingiber officinale cv. V3S18 was achieved by meristem culture on a Murashige and Skoog (MS)
basal medium supplemented with 26.6 µM 6-benzylaminopurine (BA), 8.57 µM indole-3-acetic acid (IAA), and
1111.1 µM adenine sulfate and 3% (w/v) sucrose. In vitro rhizome formation from in vitro-raised shoots was achieved on
MS medium supplemented with 4.44 µM BA, 5.71 µM IAA, and 3-8% (w/v) sucrose after 8 wk of culture. Cultural
variations such as photoperiod, carbohydrate, nutrient composition, and growth regulators were tested for the maximum
yield of rhizomes. Among the different photoperiods used, a 24-h photoperiod helped in the formation of more rhizomes as
compared with other photoperiods. Of the different carbohydrates used, sucrose helped to achieve rhizome formation as
compared to other carbohydrates. The microrhizomes sprouted in a soil mixture within 2 wk of planting. The sprouted
plantlets survived under field conditions with normal growth. |
| Objectives | Establichment of efficient protocol of in vitro shoot
multiplication and induction of rhizome formation |
| Materials | Rhizomes of ginger (Zingiber officinale cv. V3S18) collected from the High Altitude Research Station of Orissa University of
Agriculture and Technology, Pattangi, Orissa, India |
| Explant | Rhizomes of ginger incubated in a bed
of sterile, moist sandy soil to induce sprouting |
| Initial culture | The pale white sprouting buds were collected and washed with 2% (v/v) detergent solution `Teepol'
(Qualigen, India) for 15 min, subsequently surface-sterilized using a 0.2% (w/v) aqueous solution of mercuric chloride for 25 min followed by several rinses in sterile distilled water. The shoot buds were dissected under a dissecting microscope. The meristems with the apical dome or with one or two leaf primordia ranging in size from 0.1 to 0.5 mm were removed and
used as explants. MS (Murashige and Skoog) mineral salts plus 555 µM myo-inositol,
4.06 µM nicotinic acid, 2.43 µM pyridoxine-HCl, 0.296 µM thiamine-HCl
supplemented with various concentrations of cytokinins, i.e. 6-benzylaminopurine
(BA: 0.0, 2.22, 4.44, 6.66, 8.88, 13.3, 17.77, 22.2, and 26.6 µM),
kinetin (Kn: 0.0, 2.32, 4.64, 6.96, 9.28, 11.6, 13.9, 18.56, 23.2, and
27.9 µM), adenine sulfate (Ads: 370.4, 740.7, 1111.1, and 1481.5 µM), and
auxins like indole-3-acetic acid (IAA: 0.0, 2.85, 5.71, 8.57, and 11.42 µM),
indole-3-butyric acid (IBA: 0.0, 2.46, 4.92, 7.38, and 9.84 µM) and 1-
naphthaleneacetic acid (NAA: 0.0, 2.68, 5.37, 8.05, and 10.7 µM) for shoot
multiplication. The pH of the medium was adjusted to 5.8 prior to
autoclaving. Each treatment was represented by 25 cultures and the
experiment was repeated three times. The cultures were incubated under a
16-h photoperiod having a light intensity of 55 µE m-2s-1 from cool, white
fluorescent lamps at 25±2℃.
The growth of the meristem and subsequent multiplication could not be achieved in medium without growth regulators. The maximum number of
multiple shoots (32.4) was obtained in the medium containing
26.6 µM BA, 8.57 µM IAA, and 1111.1 µM adenine sulfate 4 weeks
after culture initiation. |
| Shoot multiplication | Three to four centimeter long shoots were
separated from the initial culture and transferred to semisolid MS medium
supplemented with different concentrations of BA (0.0, 2.22, 4.44, 6.7, 8.9,
13.3, and 17.8 µM) or Kn (0.0, 2.32, 4.64, 6.96, 9.28, 11.6, and 18.6 µM)
and IAA (0.0, 2.85, 5.71, and 8.57 µM) or NAA (0.0, 2.68, 5.37, and
8.05 µM) with 3% (w/v) sucrose. The cultures were incubated under a 16-h
photoperiod having a light intensity of 55 µE m-2s-1 from cool, white
fluorescent lamps at 25±2℃. The high
percentage of rhizome formation was noted on MS medium
containing 4.44 µM BA, 5.71 µM IAA, and 3% sucrose.
In a second set of experiments, the MS basal
media was supplemented with 4.44 µM BA, 5.71 µM IAA, and different
types of carbohydrates (sucrose, maltose, glucose, and fructose) for induction
of rhizomes. Among the different carbohydrates used, sucrose helped
in the formation of rhizomes as compared to d-glucose, maltose, and
fructose.
In a third set of experiments, the MS media was supplemented
with 4.44 µM BA, 5.71 µM IAA, and different concentrations of sucrose (1,
2, 3, 4, 5, 6, 7, 8, 9, and 10%) (w/v) for optimization of rhizome formation.
The results
showed that the frequency of rhizome formation was the maximum
in the medium containing 6-8% of sucrose in comparison
with other concentrations used.
In the fourth experiment, the effect of photoperiods [continuous dark (0 h), 8 h,
16 h, continuous light (24 h) photoperiod] having a light intensity of
55 µE m-2s-1 from cool, white fluorescent lamps at 25±2℃ on rhizome
formation were tested. The results indicated that 24-h photoperiod
helped in the production of the maximum number of rhizomes as
compared to 8- or 16-h photoperiod. |
| Rooting | |
| Acclimation | The rhizomes were collected from the in vitro grown
plants, washed in sterile distilled water and planted in earthern pots containing a mixture of soil, sand, and cow dung (1:1:1; v/v/v). The earthern
pots were kept in a greenhouse for sprouting of the rhizome and growth.
Watering was done at 2-d intervals.
The rhizomes derived from different
treatments were planted on a soil mixture for sprouting. Maximum
sprouting was observed in rhizomes cultured in the medium
containing 4.44 µM BA, 5.71 µM IAA, and 8% sucrose incubated
under continuous light for 8 weeks. The sprouted shoots were healthy
and normal in morphology. The rhizomes derived from the
same culture medium and incubated under 8- and 16-h
photoperiods showed delayed sprouting. |
| Planting | |
| Cultivation conditions | |
| Traints of regenerants | |
| Ingredients analyzed | |
| Extraction | |
| Analitical methods | |
| Notes | |