Multiple shoot formation in lentil (Lens culinaris) seeds

Abstract

A protocol based on seed culture was developed for efficient in vitro propagation of lentil (Lens culinaris Medik). Benzyladenine (BA), thidiazuron (TDZ), and kinetin all induced multiple shoot formation. In terms of the number of long shoots (>2.0 cm) produced per seed, BA and TDZ at optimum concentrations (0.2-0.4 and 0.1 mg/litre, respectively) had similar efficiency, whereas kinetin produced less shoots. Murashige and Skoog (MS) salt composition was better than that of Gamborge (B5) for shoot induction. Increasing calcium (Ca) concentration was necessary to overcome shoot-tip necrosis. For shoot elongation, fresh medium of the same composition of shoot induction medium could be used for stumps from medium with low BA (<0.8 mg/litre) or TDZ (<0.4 mg/litre). Otherwise, MS medium supplemented with 0.5 mg/ litre gibberellic acid (GA 3 ) or 0.05 mg/litre α-naphthaleneacetic acid (NAA) was appropriate. H00045 Received 30 October 2000; accepted 8 November 2001Multiple shoots were induced from 10 inbred of four species. Best rooting was achieved using MS medium supplemented with 1.5 mg/litre NAA. Using this rooting medium more than 50% shoots from the 10 inbreds rooted. From 80 to 100% plantlets of the 10 inbreds survived after being transferred to potting mixture in glass house.

Effect of Nutrient Media and KNO3 on &lt;i&gt;in Vitro&lt;/i&gt; Plant Regeneration in &lt;i&gt;Saraca asoca&lt;/i&gt; (Roxb.) Willd

“…Earlier reports on micropropagation of different leguminous species have also used B 5 medium for shoot induction [21]- [23]. In contrast to our results, B 5 medium was found to be less effective than MS and WP medium in leguminous species, viz., Swartzia madagascariensis [24] and Lens culinaris [25]. Our results are also in agreement with earlier workers, who have used B 5 medium for in vitro shoot formation in other species.…”

In vitro Regeneration and Over Expression of Pea DNA Helicase 45 (PDH45) Gene into the Local Cultivars of Chickpea (Cicer arietinum L.) through Agrobacteriummediated Genetic Transformation

“…5) and due to these necrotic activities regenerating shoots failed to grow and survive further. To overcome this problem of shoot tip necrosis in chickpea double the concentration of CaCl 2 and KNO 3 were used in MS following the report of Ye et al (2002) as found in case of lentil. Addition of twice the concentration CaCl 2 and KNO 3 in the regeneration medium has been found to prevent the browning of the regenerating shoots during this study thus prevented the necrosis of young shoots.…”

“…In order to overcome shoot tip death and to improve shoot health, increased concentration of macro-salts such as CaCl 2 , NH 4 NO 3 and KNO 3 were used in the medium (Ye et al 2002). All media contained 3.0% sucrose with 0.8% agar with pH 5.8, adjusted before autoclaving.…”

Agrobacterium-mediated Genetic Transformation of Local Cultivars of Chickpea (Cicer arietinum L.)

“…But in most of the cases necrosis was found to occur on the tip of these regenerated shoots and such shoots failed to grow and survive further. To overcome this problem of shoot tip necrosis in chickpea double the concentration of CaCl2 and KNO3 used in MS was added in the regeneration medium following the report of Ye et al (2002) as they could overcome shoot tip death in lentil.…”