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
Plant WRKY transcription factors (TFs) constitute one of the largest families of proteinsinvolved in biotic and abiotic stress responses. These TFs have a conserved 60 amino acid WRKYdomain at the N-terminal and a z...Plant WRKY transcription factors (TFs) constitute one of the largest families of proteinsinvolved in biotic and abiotic stress responses. These TFs have a conserved 60 amino acid WRKYdomain at the N-terminal and a zinc finger motif at the C-terminal. To examine the relevance ofOsWRKY72 in imparting salinity stress tolerance, two indica rice genotypes, Rasi (tolerant genotype)and Tellahamsa (susceptible genotype), were used. In Rasi seedlings at 12 h under 100 mmol/L NaClstress, OsWRKY72 expression was up-regulated, whereas in Tellahamsa, it was highly up-regulated atlethal stress. Full-length OsWRKY72 cDNA was cloned from these two rice genotypes for furtheranalysis. We identified a variant, termed as OsWRKY72b that carries an additional sequence of 111 bpwithin the WRKY domain. Expression of OsWRKY72b was higher under salinity stress in Rasi than inTellahamsa. Disorder prediction of OsWRKY72b showed that the additional sequence in the WRKYdomain is ordered thereby maintaining the tertiary structure that might interact with the major groove ofDNA. Prediction of phosphorylation sites in OsWRKY72b indicated that a few serine residues could bethe potential phosphorylation sites. In this study, we firstly reported a OsWRKY72 variant that couldhave a role in abiotic stress responses.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.
基金Niche Area of Excellence-Indian Council for Agriculture Research[Grant No.10(15)/2012]Department of Science and Technology-Fund for Improvement of Science and Technology Infrastructure Government of India for providing financial support
文摘Plant WRKY transcription factors (TFs) constitute one of the largest families of proteinsinvolved in biotic and abiotic stress responses. These TFs have a conserved 60 amino acid WRKYdomain at the N-terminal and a zinc finger motif at the C-terminal. To examine the relevance ofOsWRKY72 in imparting salinity stress tolerance, two indica rice genotypes, Rasi (tolerant genotype)and Tellahamsa (susceptible genotype), were used. In Rasi seedlings at 12 h under 100 mmol/L NaClstress, OsWRKY72 expression was up-regulated, whereas in Tellahamsa, it was highly up-regulated atlethal stress. Full-length OsWRKY72 cDNA was cloned from these two rice genotypes for furtheranalysis. We identified a variant, termed as OsWRKY72b that carries an additional sequence of 111 bpwithin the WRKY domain. Expression of OsWRKY72b was higher under salinity stress in Rasi than inTellahamsa. Disorder prediction of OsWRKY72b showed that the additional sequence in the WRKYdomain is ordered thereby maintaining the tertiary structure that might interact with the major groove ofDNA. Prediction of phosphorylation sites in OsWRKY72b indicated that a few serine residues could bethe potential phosphorylation sites. In this study, we firstly reported a OsWRKY72 variant that couldhave a role in abiotic stress responses.