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 supplement...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.展开更多
Extraction of quality RNA for molecular biology applications from perennial woody plants like mulberry is complicated due to the presence of high polysaccharides, polyphenols and other secondary metabolites. Since the...Extraction of quality RNA for molecular biology applications from perennial woody plants like mulberry is complicated due to the presence of high polysaccharides, polyphenols and other secondary metabolites. Since the existing methods failed to yield quality RNA in sufficient quantity from leaf and root tissues of mulberry, in this study, we modified the CTAB-based protocol. The standardised protocol yielded high quantity (520.00 μg/g fresh weight of leaf tissue) of quality RNA and the RNA extracted was suitable for all downstream applications such as cDNA synthesis, PCR and whole transcriptome analysis. The method developed was also found to be useful for isolating good quality and quantity total RNA from desiccated and salinity stressed leaf tissues of mulberry. The protocol was also applied successfully to isolate total RNA from leaf tissues of other species such as cardamom, papaya and rice.展开更多
文摘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.
文摘Extraction of quality RNA for molecular biology applications from perennial woody plants like mulberry is complicated due to the presence of high polysaccharides, polyphenols and other secondary metabolites. Since the existing methods failed to yield quality RNA in sufficient quantity from leaf and root tissues of mulberry, in this study, we modified the CTAB-based protocol. The standardised protocol yielded high quantity (520.00 μg/g fresh weight of leaf tissue) of quality RNA and the RNA extracted was suitable for all downstream applications such as cDNA synthesis, PCR and whole transcriptome analysis. The method developed was also found to be useful for isolating good quality and quantity total RNA from desiccated and salinity stressed leaf tissues of mulberry. The protocol was also applied successfully to isolate total RNA from leaf tissues of other species such as cardamom, papaya and rice.
