Development of High Frequency Multiple Shoots in The Yellow Cactus, Selenicereus Megalanthus

Journal of Applied Horticulture, 13(1): 42-43, 2011 Journal
Appl
Development of high frequency multiple shoots in the
yellow cactus, Selenicereus megalanthus
S.W. Goveas*, S.K. Nivas and L.D. Souza

Laboratory of Applied Biology, St. Aloysius College, Mangalore-575003, Karnataka, India.
*E-mail: [email protected]
Abstract
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An efficient micropropagation system via multiple shoot induction directly from seedlings of S. megalanthus was developed. Seeds
were germinated on Murashigae and Skoog (MS) basal medium supplemented with 6-Benzyl amino purine (BAP). The shoot tips of
the germinating seedlings started callusing when left on the same medium. After two weeks, the entire shoot apex portion of epicotyl
gave rise to a friable callus, portions of which started becoming green and morphogenetic. The callus gave rise to multiple shoots.
The highest number of shoots i.e.18.6 was produced on MS medium supplemented with 1.5 mgL-1 BAP. The shoots, 3-4 cm in length,
when transferred to MS basal medium supplemented with 1.0 mgL-1 IBA, rooted within one week.
Key words: Selenicereus megalanthus, in vitro, yellow pitaya, multiple shoots, MS-Murashige and Skoog
Introduction chloride for 3 min. The sterilized seeds were washed and rinsed
thoroughly with sterile double-distilled water to remove any
Selenicereus megalanthus is a cactus species native to northern residue of the sterilents. The sterilized seeds were germinated on
South America. The species is grown commercially for its fruit, MS medium (Murashige and Skoog, 1962) supplemented with
but is also an impressive ornamental vine with large yellow 100 mgL-1 m-inositol, 3% w/v sucrose as the carbon source, pH
flowers. According to Anderson (2001) these are the largest was adjusted to 5.6, and solidified with 0.8% (w/v) agar. Seeds
flowers within the cactus family. Because of its yellow flowers were inoculated into test tubes containing 10 mL of MS medium
it is known as yellow pitaya. It is a climbing or procumbent supplemented with different concentrations of BAP. Each test tube
cactus, the fruit is a medium sized oblong berry with a yellow was inoculated with single seed of S. megalanthus. 30 seeds were
peel bearing tubercles and thorns that are shed during ripening. taken for each treatment which was replicated three times. For
The pulp is white, lacking distinct aroma and contains numerous rooting, the regenerated shoots were harvested and transferred
small edible black seeds. In recent years yellow pitaya has been to MS medium supplemented with various concentrations of
seen as a marketable fruit crop with an increasing demand. 3-indole butyric acid (IBA). The cultures were maintained at a
temperature of 25 ± 2°C and relative humidity of 75 ± 5%. A 16
The earliest reported work on shoot proliferation and somatic
hour photo period with photon flux density of 10-15 μEm-2s-1 from
embryogenesis in yellow pitaya is that of Infante (1992), who
cool daylight fluorescent tubes (40W) was provided.
used epicotyls as explants in a culture medium containing
various combinations of napthaleneacetic acid (NAA) and
Results
6-benzyldadenine (BAP). Pelah et al. (2002) reported the
organogenesis in the yellow pitaya using thidiazuron (TDZ). The seeds germinated on all media (Fig. 1A). When left on the
In both cases, the numbers of regenerated plants were limited same medium containing various levels of BAP, the shoot tip of
in number. The present study reports a novel method for the the germinating seedling started callusing whereas the cotyledons
production of large number of plantlets from a single seedling. and the hypocotyls remained unaffected (Fig.1B). After two
weeks, the entire shoot apex gave rise to a friable callus, portions
Materials and methods of which started becoming green and morphogenetic (Fig.1C).
The morphogenetic callus regenerated shoots, their number
Ripe fruits of S. megalanthus were procured from the local
depending on the amount of BAP present in the medium (Fig.1D).
market. The seeds were manually separated from the flesh by
The highest number of shoots i.e.18.6 were produced on medium
rubbing slices of the fruit on a fine sieve and centrifuging the
having 1.5 mgL-1 BAP after three weeks of culture (Fig. 1E). The
resulting pulp at 2000 rpm for 10 min. The seed layer, settled at
shoots elongated on the same medium except on the medium
the bottom of the centrifuge tube, was collected after decanting
containing 2.5 mgL-1 BAP where they remained stunted. The best
the supernatant and the layer of the pulp. The seeds were surface
elongation was obtained with 1.5 mg/L BAP (Table 1). Shoots
sterilized by immersing them in 0.1% Bavistin (w/v) (A broad
which had reached a length of about 3 to 4 cms were harvested.
spectrum fungiside, Carbendizine) for 15 min and washed with
distilled water. The seeds were then treated with 0.1% sodium Rooting: The harvested regenerated shoots were transferred to
hypochlorite solution (4%w/v available chlorine) for 3 min. the rooting medium containing MS basal medium supplemented
They were washed in distilled water and kept in 0.1% mercuric with 0.5, 1.0, 1.5 and 2.0 mg/L IBA. All cultures were incubated
Development of high frequency multiple shoots in the yellow cactus S. megalanthus 43

Fig. 1. Plant regeneration of S. megalanthus, A. Germination of seeds, B. Initiation of shoot tip callusing, C. Entire shoot tip forming friable callus,
D. Shoot initiation from callus, E. Formation of multiple shoots, F. Rooting of the harvested shoot, G. Rooted plant transferred to soil, H. Plants after
two months.
Table 1. Effect of different concentrations of BAP on in vitro shoot Table 2. Effect of different concentrations of IBA on in vitro rooting
multiplication of germinating seedlings of S. megalanthus of S. megalanthus
IBA(mg/L) Rooting (%) No of roots/shoot* Root length (cm)
BAP(mg/L) Number of shoots/ seed Shoot length (cm)
0 mg/L 0 0 0
0 mg/L 4.4±2.4 2.3
0.5 mg/L 26 2.2 1.9
0.25 mg/L 3.8±1.9 3.1
1.0 mg/L 80 3.5 2.6
0.5 mg/L 3.0±1.5 2.8
1.5 mg/L 62 1.8 2.74
1.0 mg/L 2.8±0.8 4.3
2.0 mg/L 54 1.4 1.7
1.5 mg/L 18.6±6.0 6.0 *Total of 50 micro shoots involved in each treatment
2.0 mg/L 8.4±4.1 5.4
to induce bud formation in the cactus Cercis candensis, but the
2.5 mg/L 1.4±0.5 5.0 buds showed only slight or no elongation (Yusnita et al., 1990).
5.0 mg/L 1.0±0.3 2.0 In our study, we have used the entire germinating seedling as
The data are represented as the mean ±SD from 30 samples in three explant. We could induce callus and obtain regeneration of a large
independent experiments. number of shoots as well as their elongation using only BAP.
at 25 ± 2°C with 16-h photoperiod (10-15 μEm-2s-1). After one The use of a single medium for all the 3 steps is an advantage in
week of culture rooting was observed in IBA 1.0 mg/L (Fig.1F). commercial micropropagation. About 19 shoots could be obtained
Two weeks after transfer, 100% of the shoots had rooted. from a single seed with 1.5 mgL-1 BAP.
Hardening and acclimatization of rooted plants: After 4 Acknowledgements

weeks the rooted plants were removed from the culture medium
and thoroughly washed with autoclaved double distilled water The authors are thankful to Mangalore Jesuit Education Society
and transferred to sterile plastic cups containing a mixture of and St. Aloysius College, Mangalore, Karnataka, for the Lab
sterilized soil: sand (1:1) (Fig. 1G). The cups were covered with facilities and financial assistance.
polythene bags for 10-12 days and later transferred to the green
house for acclimatization. Two months old established plants References
were transferred to pots. Survival of the transferred plants was Anderson, E.F. 2001. The Cactus family. Timber Press, Portland, Oregon,
95% after 2 months (Fig.1H). USA.
Infante, R. 1992. In vitro axillary shoot proliferation and somatic
Discussion embryogenesis of yellow pitaya Mediocactus coccineus (Salm-
Dyck). Plant Cell Tiss.Org.Cult., 31: 155-159.
S. megalanthus is not only grown commercially for its fruit, but Murashige, T. and F. Skoog, 1962. A revised medium for rapid growth
is also an impressive ornamental vine producing large yellow and bioassays with tobacco tissue cultures. Physiol. Plant., 15:
flowers. Few studies have been done on in vitro multiplication of 473-497.
this useful horticultural plant (Infante,1992; Pelah et al., 2002). Pelah, D., A.R. Kaushik, Y. Mizrahi and Y. Sitrit, 2002. Organogenesis
in the vine cactus S. megalanthus using thidiazuron. Plant Cell Tiss.
Excised epicotyls were used as explants in both these studies. Org.Cult., 71: 81-84.
Infante (1992) used NAA and BAP whereas Pelah et al. (2002) Yusnita, S., R.L. Geneve and S.T. Kester, 1990. Micropropagation of
used TDZ to induce multiple shoots. The number of shoots white flowering eastern redbud (Cercis canadensis var. alba L.). J.
obtained in both cases were few. TDZ has been shown previously Environ. Hort., 8: 177-179.
Received: July, 2010; Revised: December, 2010; Accepted: December, 2010

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Journal of Applied Horticulture, 13(1): 42-43, 2011 Journal

S.W. Goveas*, S.K. Nivas and L.D. Souza

Complementary Copy-Not for Sale

Complementary Copy-Not for Sale

Hardening and acclimatization of rooted plants: After 4 Acknowledgements

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