Abstract
Adventitious bud induction and plantlet regeneration were studied in a popular mulberry variety, V1 using leaf as an explant. Fully expanded leaf explants were cultured on Murashige and Skoog’s (MS) medium supplemented with thidiazuron (TDZ) (0.5-4.0 mg/l), 6-benzylaminopurine (BAP) (0.5-2.0 mg/l), indole acetic acid (IAA) (2.0 mg/l), gibberlic acid (GA3) (1.0-2.0 mg/l) silver nitrate (AgNO3) (2.0 mg/l) and different carbon sources such as sucrose, fructose and glucose (10%-30%) either individually or in combination to induce adventitious buds and regeneration. The highest percentage (63%) of adventitious bud formation and regeneration (68%) was achieved in the medium containing MS with TDZ (1.0 mg/l), IAA (2.0 mg/l) and AgNO3 (2.0 mg/l). For subsequent regeneration and shoot elongation the MS medium having BAP (1.0 mg/l), GA3 (2.0 mg/l) and AgNO3 (2.0 mg/l) was found to be suitable. Amongst the carbon sources tested, the most suitable carbon source was found to be sucrose (3%) followed by fructose (2%) for adventitious bud formation. Excised in vitro shoots were rooted (60%-80%) in half strength MS medium supplemented with indole-3-butyric acid (1.0 mg/l). The well rooted plantlets were hardened in soil + sand + farm yard manure (FYM) mixture with a success rate of 70%-90%. Since in vitro regeneration is highly genotype-dependent in mulberry, the standardized protocol can be effectively used for further improvement of this leading genotype using biotechnological approaches.
Highlights
Mulberry (Morus spp.) is a woody perennial tree of importance to the sericulture industry as mulberry leaf is the sole food for the silkworm (Bombyx mori L.) larvae
Expanded leaf explants were cultured on Murashige and Skoog’s (MS) medium supplemented with thidiazuron (TDZ) (0.5 - 4.0 mg/l), 6-benzylaminopurine (BAP) (0.5 - 2.0 mg/l), indole acetic acid (IAA) (2.0 mg/l), gibberlic acid (GA3) (1.0 - 2.0 mg/l) silver nitrate (AgNO3) (2.0 mg/l) and different carbon sources such as sucrose, fructose and glucose (10% - 30%) either individually or in combination to induce adventitious buds and regeneration
Amongst the carbon sources tested, the most suitable carbon source was found to be sucrose (3%) followed by fructose (2%) for adventitious bud formation
Summary
Mulberry (Morus spp.) is a woody perennial tree of importance to the sericulture industry as mulberry leaf is the sole food for the silkworm (Bombyx mori L.) larvae. Targeted manipulation of elite genotypes through incorporation of specific genes encoding desired traits using modern biotechnological methods offers a new opportunity for crop improvement. An efficient in vitro regeneration procedure is pre-requisite for transgenic approach in any crops. Information on development and standardization of in vitro regeneration protocols in promising mulberry genotypes is limited, there are reports. Studies have been made in mulberry to examine the impact of various growth regulators on in vitro organogenesis and plant regeneration by using different explants viz. The shoot differentiation from callus is confined only to a few genotypes and repeatability of protocols developed was not assured due to the recalcitrant nature of the plant. We made an attempt to develop and standardize in vitro regeneration protocol in a widely cultivated mulberry variety, V1 using leaf explants. The major emphasis was given to investigate the effect of Thidiazuron (TDZ), a substituted phenyl urea and different carbon sources in inducing adventitious buds and efficient regeneration in V1
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