An in vitro propagation protocol using mature seeds of D. membranaceus was successfully establish... more An in vitro propagation protocol using mature seeds of D. membranaceus was successfully established. Scarcity of seeds in bamboos because of their long flowering periods and irregular seed set resulting in low viability and germination potential, motivated us to undertake the present study. The effects of sterilants, light conditions, exogenous application of plant growth regulators and temperature in overcoming germination barriers in ageing seeds of bamboo were studied. It was found that HgCl2 (0.1%) along with bleach (15%) was more effective in raising aseptic cultures. Dark conditions, high temperatures around 30 degrees C and soaking of seeds in GA3 solution (50 ppm) overnight stimulated high percent of seed germination with corresponding increase in shoot length (2.7 +/- 0.7 mm) and number of sprouts (2.1 +/- 0.7) per explants during culture initiation. 6-benzylaminopurine acted synergistically with kinetin to give optimum germination rate of 70 +/- 13.9% as compared to 63.13%...
BRAR J, SHAFI A, SOOD P, ANAND M & SOOD A. 2014. In-vitro propagation, biochemical studies and as... more BRAR J, SHAFI A, SOOD P, ANAND M & SOOD A. 2014. In-vitro propagation, biochemical studies and assessment of clonal fidelity through molecular markers in Bambusa balcooa. Micropropagation protocol of Bambusa balcooa has been established using nodal explants with 92.5% success after acclimatisation. The extraction of chlorophyll from the epidermis of culms using acetone, dimethylformamide and dimethyl sulfoxide as solvents and the use of auxin in shoot multiplication are also reported for the first time in this species. Due to stringent environmental conditions in the greenhouse, electrolyte leakage percentage (67.4 ± 2.4%) was found to be higher in ex-vitro raised plants whereas relative water content percentage (72.7 ± 2.3%) was higher in in-vitro grown plants. Analysis of total soluble sugars (16.5 ± 0.78 mg g-1) and starch content (6.32 ± 0.55 mg g-1) displayed in-vitro plants to be richer in carbohydrates reserves. However, lignin content (28.74 ± 1.59 mg L-1), stomatal density ...
Somatic embryogenesis is a highly preferred method for in vitro multiplication of plants. It also... more Somatic embryogenesis is a highly preferred method for in vitro multiplication of plants. It also forms an ideal experimental model for investigating the process of differentiation and for understanding the mechanism of totipotency in plants. The phenomenon of recurrent somatic embryogenesis makes these attractive explants for genetic transformation and production of transgenic plants. Somatic embryogenesis in monocots has also been reported to reset the internal calendar at zero; thereby, enabling the germinated plantlets to live their full life span. Moreover, there is an increasingly high depletion of natural stands of bamboos mainly due to its gregarious flowering habit and anthopogenic pressures. However, the plant has emerged as a crop of the future and has a huge potential in paper, wood and textile industry. In fact, bamboos are among the most popular groups of plants owing to their immense utility, phenotypic variability and adaptability. Therefore, there is an urgent need ...
The aim of the present study was to standardize an efficient protocol for mass multiplication of ... more The aim of the present study was to standardize an efficient protocol for mass multiplication of an important bamboo Dendrocalamus membranaceus categorized under “Important” category by United Nations Environment Programme – World Conservation Monitoring Centre, (UNEP – WCMC) and ascertaining the clonal fidelity of in vitro raised plants by molecular markers. Axillary buds from the field grown plants were taken as explants to induce multiple shoots on Murashige and Skoog (MS) medium containing 4.4 µM N 6 -Benzylaminopurine (BAP) alongwith 1.16 µM Kinetin (KN). A clump comprising of a minimum of 3 shoots was inoculated on to half strength MS medium containing 5.4 µM a-Naphthaleneacetic Acid (NAA) in addition to low concentrations of 4.4 µM BAP for root induction. About sixty five percent success was achieved in plant establishment after acclimatization in green house. Total chlorophyll and carotenoids (17.92 a ± 1.33) mg/g FW (fresh weight) and electrolyte leakage percentage (76.01 a...
BRAR J, SHAFI A, SOOD P, ANAND M & SOOD A. 2014. In-vitro propagation, biochemical studies and as... more BRAR J, SHAFI A, SOOD P, ANAND M & SOOD A. 2014. In-vitro propagation, biochemical studies and assessment of clonal fidelity through molecular markers in Bambusa balcooa. Micropropagation protocol of Bambusa balcooa has been established using nodal explants with 92.5% success after acclimatisation. The extraction of chlorophyll from the epidermis of culms using acetone, dimethylformamide and dimethyl sulfoxide as solvents and the use of auxin in shoot multiplication are also reported for the first time in this species. Due to stringent environmental conditions in the greenhouse, electrolyte leakage percentage (67.4 ± 2.4%) was found to be higher in ex-vitro raised plants whereas relative water content percentage (72.7 ± 2.3%) was higher in in-vitro grown plants. Analysis of total soluble sugars (16.5 ± 0.78 mg g-1) and starch content (6.32 ± 0.55 mg g-1) displayed in-vitro plants to be richer in carbohydrates reserves. However, lignin content (28.74 ± 1.59 mg L-1), stomatal density ...
The aim of the present study was to standardize an efficient protocol for mass multiplication of ... more The aim of the present study was to standardize an efficient protocol for mass multiplication of an important bamboo Dendrocalamus membranaceus categorized under "Important" category by United Nations Environment Programme – World Conservation Monitoring Centre, (UNEP – WCMC) and ascertaining the clonal fidelity of in vitro raised plants by molecular markers. Axillary buds from the field grown plants were taken as explants to induce multiple shoots on media containing 4.4 μM N 6 -Benzylaminopurine (BAP) alongwith 1.16 μM Kinetin (KN). A clump comprising of a minimum of 3 shoots was inoculated on to half strength Murashige and Skoog (MS) medium containing 5.4 μM á-Naphthaleneacetic Acid (NAA) in addition to low concentrations of 4.4 μM BAP for root induction. About sixty five percent success was achieved in plant establishment after acclimatization in green house. Total chlorophyll and carotenoids (17.92 a ± 1.33) mg/g FW (fresh weight) and electrolyte leakage percentage (7...
Abstract-An efficient and reproducible protocol has been established through the technique of f... more Abstract-An efficient and reproducible protocol has been established through the technique of forced axillary branching for the propagation of an important edible bamboo species namely Bambusa bambos. High frequency multiple shoot induction was achieved from nodal segments collected from elite genotype on Murashige and Skoog's (MS) medium supplemented with 4.4 µM Benzylaminopurine (BAP) and 1.16 µM Kinetin (Kn). The size of explant and season greatly influenced the frequency of bud break. Rooting posed a major problem to be worked out in this particular species. Best rooting response was observed on 9.80 µM of Indole-3 Butyric acid (IBA) with 60 ± 14.1 % rooting. In vitro raised plants were successfully acclimatized and established in the field conditions where they exhibited normal growth. In a bid to ascertain genetic fidelity, DNA was extracted by CTAB method and samples were analysed in 1.8% agarose gel electrophoresis. In the present study no variation was reported among the in vitro raised progeny and the mother plant in the banding profiles generated by the total of fifteen Random Amplified polymorphic DNA (RAPD) and Inter Simple Sequence Repeats (ISSR) markers. Hence, molecular analysis confirmed that these plants were genetically similar and can be used as elite plants.
An in vitro propagation protocol using mature seeds of D. membranaceus was successfully establish... more An in vitro propagation protocol using mature seeds of D. membranaceus was successfully established. Scarcity of seeds in bamboos because of their long flowering periods and irregular seed set resulting in low viability and germination potential, motivated us to undertake the present study. The effects of sterilants, light conditions, exogenous application of plant growth regulators and temperature in overcoming germination barriers in ageing seeds of bamboo were studied. It was found that HgCl2 (0.1%) along with bleach (15%) was more effective in raising aseptic cultures. Dark conditions, high temperatures around 30 degrees C and soaking of seeds in GA3 solution (50 ppm) overnight stimulated high percent of seed germination with corresponding increase in shoot length (2.7 +/- 0.7 mm) and number of sprouts (2.1 +/- 0.7) per explants during culture initiation. 6-benzylaminopurine acted synergistically with kinetin to give optimum germination rate of 70 +/- 13.9% as compared to 63.13%...
The aim of the present study was to standardize an efficient protocol for mass multiplication of ... more The aim of the present study was to standardize an efficient protocol for mass multiplication of an important bamboo Dendrocalamus membranaceus categorized under "Important" category by United Nations Environment Programme -World Conservation Monitoring Centre, (UNEP -WCMC) and ascertaining the clonal fidelity of in vitro raised plants by molecular markers. Axillary buds from the field grown plants were taken as explants to induce multiple shoots on Murashige and Skoog (MS) medium containing 4.4 µM N 6 -Benzylaminopurine (BAP) alongwith 1.16 µM Kinetin (KN). A clump comprising of a minimum of 3 shoots was inoculated on to half strength MS medium containing 5.4 µM á-Naphthaleneacetic Acid (NAA) in addition to low concentrations of 4.4 µM BAP for root induction. About sixty five percent success was achieved in plant establishment after acclimatization in green house. Total chlorophyll and carotenoids (17.92 a ± 1.33) mg/g FW (fresh weight) and electrolyte leakage percentage (76.01 a ± 3.9) was found to be higher in ex vitro raised plants whereas relative water content percentage (68.8 a ± 4.0) was higher in in vitro grown plants depicting role of physiological factors in overall growth and development of plants. An assessment of clonal fidelity using 35 Random Amplified Polymorphic DNA (RAPD) and 20 Inter Simple Sequence Repeats (ISSR) as molecular markers revealed that the bands were shared by both the parent clump and in vitro raised plants confirming the genetic stability of the Tissue Culture (TC) raised plantlets. Micropropagation protocol reported here along with all the molecular and physiological factors studied is the first report in this very important bamboo species.
An in vitro propagation protocol using mature seeds of D. membranaceus was successfully establish... more An in vitro propagation protocol using mature seeds of D. membranaceus was successfully established. Scarcity of seeds in bamboos because of their long flowering periods and irregular seed set resulting in low viability and germination potential, motivated us to undertake the present study. The effects of sterilants, light conditions, exogenous application of plant growth regulators and temperature in overcoming germination barriers in ageing seeds of bamboo were studied. It was found that HgCl2 (0.1%) along with bleach (15%) was more effective in raising aseptic cultures. Dark conditions, high temperatures around 30 degrees C and soaking of seeds in GA3 solution (50 ppm) overnight stimulated high percent of seed germination with corresponding increase in shoot length (2.7 +/- 0.7 mm) and number of sprouts (2.1 +/- 0.7) per explants during culture initiation. 6-benzylaminopurine acted synergistically with kinetin to give optimum germination rate of 70 +/- 13.9% as compared to 63.13%...
BRAR J, SHAFI A, SOOD P, ANAND M & SOOD A. 2014. In-vitro propagation, biochemical studies and as... more BRAR J, SHAFI A, SOOD P, ANAND M & SOOD A. 2014. In-vitro propagation, biochemical studies and assessment of clonal fidelity through molecular markers in Bambusa balcooa. Micropropagation protocol of Bambusa balcooa has been established using nodal explants with 92.5% success after acclimatisation. The extraction of chlorophyll from the epidermis of culms using acetone, dimethylformamide and dimethyl sulfoxide as solvents and the use of auxin in shoot multiplication are also reported for the first time in this species. Due to stringent environmental conditions in the greenhouse, electrolyte leakage percentage (67.4 ± 2.4%) was found to be higher in ex-vitro raised plants whereas relative water content percentage (72.7 ± 2.3%) was higher in in-vitro grown plants. Analysis of total soluble sugars (16.5 ± 0.78 mg g-1) and starch content (6.32 ± 0.55 mg g-1) displayed in-vitro plants to be richer in carbohydrates reserves. However, lignin content (28.74 ± 1.59 mg L-1), stomatal density ...
Somatic embryogenesis is a highly preferred method for in vitro multiplication of plants. It also... more Somatic embryogenesis is a highly preferred method for in vitro multiplication of plants. It also forms an ideal experimental model for investigating the process of differentiation and for understanding the mechanism of totipotency in plants. The phenomenon of recurrent somatic embryogenesis makes these attractive explants for genetic transformation and production of transgenic plants. Somatic embryogenesis in monocots has also been reported to reset the internal calendar at zero; thereby, enabling the germinated plantlets to live their full life span. Moreover, there is an increasingly high depletion of natural stands of bamboos mainly due to its gregarious flowering habit and anthopogenic pressures. However, the plant has emerged as a crop of the future and has a huge potential in paper, wood and textile industry. In fact, bamboos are among the most popular groups of plants owing to their immense utility, phenotypic variability and adaptability. Therefore, there is an urgent need ...
The aim of the present study was to standardize an efficient protocol for mass multiplication of ... more The aim of the present study was to standardize an efficient protocol for mass multiplication of an important bamboo Dendrocalamus membranaceus categorized under “Important” category by United Nations Environment Programme – World Conservation Monitoring Centre, (UNEP – WCMC) and ascertaining the clonal fidelity of in vitro raised plants by molecular markers. Axillary buds from the field grown plants were taken as explants to induce multiple shoots on Murashige and Skoog (MS) medium containing 4.4 µM N 6 -Benzylaminopurine (BAP) alongwith 1.16 µM Kinetin (KN). A clump comprising of a minimum of 3 shoots was inoculated on to half strength MS medium containing 5.4 µM a-Naphthaleneacetic Acid (NAA) in addition to low concentrations of 4.4 µM BAP for root induction. About sixty five percent success was achieved in plant establishment after acclimatization in green house. Total chlorophyll and carotenoids (17.92 a ± 1.33) mg/g FW (fresh weight) and electrolyte leakage percentage (76.01 a...
BRAR J, SHAFI A, SOOD P, ANAND M & SOOD A. 2014. In-vitro propagation, biochemical studies and as... more BRAR J, SHAFI A, SOOD P, ANAND M & SOOD A. 2014. In-vitro propagation, biochemical studies and assessment of clonal fidelity through molecular markers in Bambusa balcooa. Micropropagation protocol of Bambusa balcooa has been established using nodal explants with 92.5% success after acclimatisation. The extraction of chlorophyll from the epidermis of culms using acetone, dimethylformamide and dimethyl sulfoxide as solvents and the use of auxin in shoot multiplication are also reported for the first time in this species. Due to stringent environmental conditions in the greenhouse, electrolyte leakage percentage (67.4 ± 2.4%) was found to be higher in ex-vitro raised plants whereas relative water content percentage (72.7 ± 2.3%) was higher in in-vitro grown plants. Analysis of total soluble sugars (16.5 ± 0.78 mg g-1) and starch content (6.32 ± 0.55 mg g-1) displayed in-vitro plants to be richer in carbohydrates reserves. However, lignin content (28.74 ± 1.59 mg L-1), stomatal density ...
The aim of the present study was to standardize an efficient protocol for mass multiplication of ... more The aim of the present study was to standardize an efficient protocol for mass multiplication of an important bamboo Dendrocalamus membranaceus categorized under "Important" category by United Nations Environment Programme – World Conservation Monitoring Centre, (UNEP – WCMC) and ascertaining the clonal fidelity of in vitro raised plants by molecular markers. Axillary buds from the field grown plants were taken as explants to induce multiple shoots on media containing 4.4 μM N 6 -Benzylaminopurine (BAP) alongwith 1.16 μM Kinetin (KN). A clump comprising of a minimum of 3 shoots was inoculated on to half strength Murashige and Skoog (MS) medium containing 5.4 μM á-Naphthaleneacetic Acid (NAA) in addition to low concentrations of 4.4 μM BAP for root induction. About sixty five percent success was achieved in plant establishment after acclimatization in green house. Total chlorophyll and carotenoids (17.92 a ± 1.33) mg/g FW (fresh weight) and electrolyte leakage percentage (7...
Abstract-An efficient and reproducible protocol has been established through the technique of f... more Abstract-An efficient and reproducible protocol has been established through the technique of forced axillary branching for the propagation of an important edible bamboo species namely Bambusa bambos. High frequency multiple shoot induction was achieved from nodal segments collected from elite genotype on Murashige and Skoog's (MS) medium supplemented with 4.4 µM Benzylaminopurine (BAP) and 1.16 µM Kinetin (Kn). The size of explant and season greatly influenced the frequency of bud break. Rooting posed a major problem to be worked out in this particular species. Best rooting response was observed on 9.80 µM of Indole-3 Butyric acid (IBA) with 60 ± 14.1 % rooting. In vitro raised plants were successfully acclimatized and established in the field conditions where they exhibited normal growth. In a bid to ascertain genetic fidelity, DNA was extracted by CTAB method and samples were analysed in 1.8% agarose gel electrophoresis. In the present study no variation was reported among the in vitro raised progeny and the mother plant in the banding profiles generated by the total of fifteen Random Amplified polymorphic DNA (RAPD) and Inter Simple Sequence Repeats (ISSR) markers. Hence, molecular analysis confirmed that these plants were genetically similar and can be used as elite plants.
An in vitro propagation protocol using mature seeds of D. membranaceus was successfully establish... more An in vitro propagation protocol using mature seeds of D. membranaceus was successfully established. Scarcity of seeds in bamboos because of their long flowering periods and irregular seed set resulting in low viability and germination potential, motivated us to undertake the present study. The effects of sterilants, light conditions, exogenous application of plant growth regulators and temperature in overcoming germination barriers in ageing seeds of bamboo were studied. It was found that HgCl2 (0.1%) along with bleach (15%) was more effective in raising aseptic cultures. Dark conditions, high temperatures around 30 degrees C and soaking of seeds in GA3 solution (50 ppm) overnight stimulated high percent of seed germination with corresponding increase in shoot length (2.7 +/- 0.7 mm) and number of sprouts (2.1 +/- 0.7) per explants during culture initiation. 6-benzylaminopurine acted synergistically with kinetin to give optimum germination rate of 70 +/- 13.9% as compared to 63.13%...
The aim of the present study was to standardize an efficient protocol for mass multiplication of ... more The aim of the present study was to standardize an efficient protocol for mass multiplication of an important bamboo Dendrocalamus membranaceus categorized under "Important" category by United Nations Environment Programme -World Conservation Monitoring Centre, (UNEP -WCMC) and ascertaining the clonal fidelity of in vitro raised plants by molecular markers. Axillary buds from the field grown plants were taken as explants to induce multiple shoots on Murashige and Skoog (MS) medium containing 4.4 µM N 6 -Benzylaminopurine (BAP) alongwith 1.16 µM Kinetin (KN). A clump comprising of a minimum of 3 shoots was inoculated on to half strength MS medium containing 5.4 µM á-Naphthaleneacetic Acid (NAA) in addition to low concentrations of 4.4 µM BAP for root induction. About sixty five percent success was achieved in plant establishment after acclimatization in green house. Total chlorophyll and carotenoids (17.92 a ± 1.33) mg/g FW (fresh weight) and electrolyte leakage percentage (76.01 a ± 3.9) was found to be higher in ex vitro raised plants whereas relative water content percentage (68.8 a ± 4.0) was higher in in vitro grown plants depicting role of physiological factors in overall growth and development of plants. An assessment of clonal fidelity using 35 Random Amplified Polymorphic DNA (RAPD) and 20 Inter Simple Sequence Repeats (ISSR) as molecular markers revealed that the bands were shared by both the parent clump and in vitro raised plants confirming the genetic stability of the Tissue Culture (TC) raised plantlets. Micropropagation protocol reported here along with all the molecular and physiological factors studied is the first report in this very important bamboo species.
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