[show abstract][hide abstract] ABSTRACT: A reliable regeneration system for faba bean (Vicia faba L.) has been difficult to establish, delaying its genetic improvement. In the present work a rapid, reproducible and efficient regeneration method was developed for faba bean using single cotyledon explants with half embryonal axis. MS medium supplemented with 6 mu M TDZ (thidiazuron), 10 mu M 2-iP and 4 mu M kinetin induced 30 to 50 adventitious buds /shoots after two weeks of culture, which were elongated on MS medium supplemented with 6 mu M 2-iP and 2 mu M kinetin. With healthy and strong roots established, shoots were transferred to MS medium supplemented with 5 mu M IBA within 10 to 14 days. Shoots of 5 cm long were most suitable for rooting. Potting-mixture with good aeration and less capacity to retain water was most suitable for successful establishment of faba bean plantlets. Garden soil mixed with sand (gravel) and bio-manure (1:1:1) was most suitable for transplantation. TDZ promoted adventitious bud formation while 5 mu M IBA was most suitable for rooting, higher concentrations were toxic to plantlets. Aeration of the potting mixture was important for rapid micropropagation and successful establishment. The efficient regeneration protocol reported here allows for successful micropropagation of faba bean, which is essential for future genetic improvement of plants via transformation protocols.
Journal of Medicinal Plants Research. 01/2011; 5(28):6460-6467.
[show abstract][hide abstract] ABSTRACT: Chloroplasts from plants of transgenic lines expressing prokaryotic choline oxidase gene (the codA(ps) gene; GenBank accession number-AY589052) and wild-type of chickpea and Indian mustard were evaluated for their efficacy to withstand photoinhibitory damage, by exposing them to high light intensity ( approximately 1200micromolm(-2)s(-1) photon flux density) at 10 and 25 degrees C. Western analysis confirmed presence of choline oxidase in chloroplasts of only transgenic lines. The loss in PS II activity in chloroplasts of wild-type exposed to high light intensity was significantly higher than that in chloroplasts of transgenic chickpea as well as Indian mustard. Although, chloroplasts of both wild-type and transgenic chickpea as well as Indian mustard were more sensitive to photoinhibitory damage at 10 than at 25 degrees C, the damage recorded in chloroplasts harboring choline oxidase was significantly lower than those of wild-type. High light promotes H(2)O(2) production in chloroplasts more significantly at low temperature (10 degrees C) than at 25 degrees C. We compared low temperature accelerated photoinhibition of chloroplasts with that caused due to exogenously applied H(2)O(2). Although exogenous H(2)O(2) accelerated high light intensity induced loss in PS II activity of chloroplasts of wild-type, it caused only a little alteration in PS II activity of chloroplasts from transgenic lines of both chickpea and Indian mustard, demonstrating that the chloroplasts harboring choline oxidase are better equipped to resist photoinhibition. We hypothesize that H(2)O(2) produced by choline oxidase as a byproduct during synthesis of glycinebetaine is responsible for building stronger antioxidant system in chloroplasts of transgenic lines compared to that of wild-type.
Plant Physiology and Biochemistry 02/2009; 47(5):391-6. · 2.78 Impact Factor
[show abstract][hide abstract] ABSTRACT: A rapid, reproducible and efficient regeneration method was developed for chickpea (Cicer arietinum L.) using single cotyledon with half embryonal axis as explants. MS medium supplemented with 4 ìM TDZ, 10 ìM 2-iP and 2 ìM kinetin induced 50-100 adventitious buds/shoots after 14 days of culture and elongated on MS medium supplemented with 5 ìM 2-iP and 2 ìM kinetin. Healthy, strong and 100 % rooting was achieved by exposing cut ends of the shoots to 10 sec pulse treatment with 100 ìmoles/ml IBA followed by their transfer to liquid MS basal medium within 10-14 d. 2-3 cm long shoots were most suitable for rooting. Potting-mixture with good aeration and lesser capacity to retain water was most suitable for achieving successful establishment of chickpea plantlets. Garden soil mixed with sand (gravel) and bio-manure in the ratio of 1:1:1 is most suitable for achieving cent percent transplantation success. Cent percent of plantlets got acclimatized, survived in the pots and showed normal growth, development, flowering followed by podding and seeds setting. Harvesting of seeds was done after the pods were fully matured and dry. In this communication, we have demonstrated for the first time that shoot length, pulse treatment of cut ends of shoots with 100 ìmoles/ml IBA and aeration of potting mixture are key factors for rapid micro-propagation and successful establishment of chickpea.
Physiology and Molecular Biology of Plants 10/2008; 14(4):329-35.