In Vitro Production of Microtubers and Artificial Seeds of Solanum tuberosum L.
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Date
2008
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Birsa Agricultural University, Kanke, Ranchi, Jharkhand
Abstract
Potato (Solanum tuberosum L.) is the most important non-cereal food crop of the world. In
monetary terms it ranks fourth in the world after wheat, rice and maize. It produces the
largest quantity of carbohydrates per day per unit area among the food crops. It plays a
remarkable role in human diet as a supplement to wheat and rice. Potato recommended as
food security crop, is composed of 80% water, 2-3% protein and 18% carbohydrate. Basic
problem for potato production is the availability of good quality seed. Tissue culture
techniques are used worldwide to produce pre-basic, virus-free seed potatoes known as
microtubers. The microtubers are sown in a protected environment to produce minitubers
(basic seed). The basic seed enters the seed production chain to produce the certified seed to
be sold to the farmers. Therefore, the main objective of the present study was to optimize the
culture condition and media for plant growth and microtuber induction of two potato cultivars
(Kufri Chipsona 3 and MP-97/644) along with this artificial seeds were also prepared.
In the present work, an efficient in vitro method for plantlet regeneration via shoot tips and
leaves of S. tuberosum (Kufri Chipsona 3 and MP-97/644) were developed. The best survival
percentage during surface sterilization of S. tuberosum (Kufri Chipsona 3 and MP-97/644)
explants was achieved by treating shoot tips with 0.05% HgCl2 for 15 minutes. Shoot
multiplication from mother explants brought from CPRI, Shimla was induced on MS medium
supplemented with 1.0 mg/l BAP and 25.0 mg/l AdSO4. Highest number of shoots per bottle
22.67 (Kufri Chipsona 3) and 18.33 (MP-97/644) was observed after 60 days of inoculation
in the same media. Maximum number of microtubers was induced on the MS medium
supplemented with 1.0 mg/l BAP and 25.0 mg/l AdSO4 and 6.0% sucrose. The maximum
number of microtubers/bottle was 9.67 and 7.33 in the Kufri Chipsona 3 and MP-97/644
respectively. MS media supplemented with 1.0 gm/l activated charcoal either in 1.0 mg/l
BAP or 25.0 mg/l AdSO4 or 1.0 mg/l BAP and 25.0 mg/l AdSO4 was not effective in
inducing microtubers in both the cultivars.
Best callus growth from both the cultivars was observed on MS media containing 3.0 mg/l of
2,4-D and1.0 mg/l of kinetin. Callus was observed from microtubers of Kufri Chipsona 3 and
MP-97/644 on MS media supplemented with 2,4-D (1.0 mg/l), Kinetin (2.0 mg/l) and (1.5
mg/l) IAA. Best shoot regeneration from callus was observed on MS media containing 1.5
mg/l BAP and 25.0 mg/l AdSO4 BAP. The mean number of shoots/callus clump was 21.00
(Kufri Chipsona 3) and 18.67 (MP-97/644) after 60 days of inoculation. The media enriched
with 2,4-D and kinetin also showed good response. The maximum number of shoots
regenerated from callus was 8.33 in Kufri Chipsona 3 and 7.33 in MP-97/644 in the same
media. Adventitious shooting was observed from both cultivars on MS media containing
equal concentration of 2,4-D and Kinetin i.e. 1 mg/l . The shoot regeneration potential of
Kufri Chipsona and MP-97/644 microtubers was best on media supplemented with 1.0 mg/l
BAP and 25.0 mg/l AdSO4. The most important aspect is that the plantlets do not require
rooting media. When the plants were transferred to green house, primary hardening was
achieved within 10 days and about 99% plants survived.
Artificial seeds of Solanum tuberosum (Kufri Chipsona 3 and MP-97/644) was prepared by
encapsulating axillary shoot buds (single node) with 1.0% (w/v) sodium alginate and
different concentrations of CaCl2.2H2O (30, 50 and 70 mM). The microshoots encapsulated
in 70mM CaCl2.2H2O were stored for a longer period in ½ MS basal media. The best
morphogenetic response of the microshoots was obtained when the encapsulated seeds were
placed in Murashige and Skoog's (liquid) medium containing 3.0% sucrose and clonespecific
growth regulators. Maximum germination was about 83.0% (Kufri Chipsona 3) and
78.0% (MP-97/644) of seeds encapsulated in 30 mM CaCl2 after 15 days of storage.
Description
In Vitro Production of Microtubers and Artificial Seeds of Solanum tuberosum L.
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