Articles
Single cell induction from embryogenic and non-embryogenic callus of Vigna radiata (L.) Wilczek
Article number
1241_43
Pages
307 – 312
Language
English
Abstract
Legumes comprise a wide range of crop species including herbaceous annuals and perennial trees which are used as food, feed, forage, fiber, industrial and medicinal compounds.
Green gram [Vigna radiata (L.) Wilczek.] is an important food crop which is known to be originated in India or Indo-Burmese region and widespread throughout the tropics.
It is an important source of dietary protein and dietary amino acids.
Because of its nutritional quality it is called queen of pulses. Source seedlings for explants were found to influence the callusing response.
Concentration of phytohormones and growth factors were optimized for developing embryogenic callus.
In the present study, explants from green gram (3-day-old embryonal axis and cotyledonary node) were excised and cultured in MS (Murashige and skoog) medium supplemented with various nutritional concentrations viz., 2,4-D (2-10 µM) and Kn (2-10 µM). Cultures were incubated at 25±2°C for 16/8 h photoperiod.
Periodic observation and evaluation were made up to 28 days.
In this experiment callus was taken with 40 mg mass which was added with 10 mL of MES buffer (2-morpholinoethane – sulphonic acid) for standardization of single cells.
The results showed that embryogenic cells developed from cotyledon explants grown in Kn (4 µM) supplemented medium were passed through stainless wire mesh with sieve size of 60 µm.
Whereas, non-embryogenic cells were passed through stainless wire mesh with sieve size of 30 and 100 µm from cotyledon explants grown in 2,4-D (10 µM) supplemented medium.
Green gram [Vigna radiata (L.) Wilczek.] is an important food crop which is known to be originated in India or Indo-Burmese region and widespread throughout the tropics.
It is an important source of dietary protein and dietary amino acids.
Because of its nutritional quality it is called queen of pulses. Source seedlings for explants were found to influence the callusing response.
Concentration of phytohormones and growth factors were optimized for developing embryogenic callus.
In the present study, explants from green gram (3-day-old embryonal axis and cotyledonary node) were excised and cultured in MS (Murashige and skoog) medium supplemented with various nutritional concentrations viz., 2,4-D (2-10 µM) and Kn (2-10 µM). Cultures were incubated at 25±2°C for 16/8 h photoperiod.
Periodic observation and evaluation were made up to 28 days.
In this experiment callus was taken with 40 mg mass which was added with 10 mL of MES buffer (2-morpholinoethane – sulphonic acid) for standardization of single cells.
The results showed that embryogenic cells developed from cotyledon explants grown in Kn (4 µM) supplemented medium were passed through stainless wire mesh with sieve size of 60 µm.
Whereas, non-embryogenic cells were passed through stainless wire mesh with sieve size of 30 and 100 µm from cotyledon explants grown in 2,4-D (10 µM) supplemented medium.
Authors
S. Maheshwari, K. Manoharan
Keywords
Vigna radiata, embryogenic, non-embryogenic cells, cotyledon, single cell induction
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