Rhizobium, a nitrogen-fixing bacteria is the essential feature of leguminous plants which is esse... more Rhizobium, a nitrogen-fixing bacteria is the essential feature of leguminous plants which is essential for the regeneration of nutrient-deficient soil. This study was aimed to isolate, identify, mass culture and immobilize Bradyrhizoium japonicum in encapsulated form and test their viability. Root nodules were sterilized, grinded and cultured aseptically in YEMA media containing Congo red. The obtained colon was sub-cultured to get a pure culture and different biochemical tests were conducted which proved Bradyrhizobium japonicum as the slow-growing species. The test shows a positive result of catalase production and nodulation test whereas the pH tolerance test shows more tolerance to the acidic pH. Similarly, Bradyrhizaobium japonicum can tolerate 1% and 2% NaCl concentration and it doesn’t show resistance to the penicillin disc of 10mg. The mass culture and encapsulation with sodium alginate adding sucrose as nutrient proved the simplicity for handling. Altogether 548 beads were ...
Axenic shoot culture of Mentha spicata L. was established from young shoots of the plant naturall... more Axenic shoot culture of Mentha spicata L. was established from young shoots of the plant naturally growing in the field. Large number of multiple shoots were observed on MS media supplemented with NAA (0.5ppm) and BAP (1ppm) within eight weeks of culture. Extensive root formation was observed on MS medium supplemented with 1 ppm NAA. Essential oils from both in vitro and in vivo samples showed l- carvone as a major constituent, however, the in vitro samples showed higher concentration of menthol than in vivo samples. <i>Nepal Journal of Science and Technology</i> Vol. 7, 2006
Botanica Orientalis: Journal of Plant Science, 2012
In vitro culture of Bergenia ciliata (Haw.) Sternb. was carried out for the examination of bergen... more In vitro culture of Bergenia ciliata (Haw.) Sternb. was carried out for the examination of bergenin content. Leaf explants were cultured in MS (Murashige and Skoog) basal media supplemented with or without phytohormones. The hormonal series maintained were in the range of 0-2 mg l-1 for BAP (6-benzyl amino purine) and 0-1.5 mg l-1 for NAA (α-naphthalene acetic acid). Bergenin content of in vitro grown tissues of B. ciliata was compared with that of wild plants collected from three different localities of Nepal. The best growth of callus and plantlets occurred in the media containing BAP 1.0 mg l-1 + NAA 1.0 mg l-1 and BAP 1.5 mg l-1 + NAA 1.0 mg l-1. Production of bergenin was high in the media supplemented with 1.0 mg l-1 BAP + 1.5 mg l-1 NAA (3.40 μg g-1) and 2.0 mg l-1 BAP + 1.5 mg l-1 NAA (3.05 μg g-1) under experimental condition. The bergenin content in the wild plants collected from Langtang, Jumla and Godawari was found to be 4.28 μg g-1, 4.53 μg g-1 and 3.64 μg g-1 respecti...
Total polyphenol or flavonoid contents in the plant extracts of D. moniliforme (triplicate data).... more Total polyphenol or flavonoid contents in the plant extracts of D. moniliforme (triplicate data). (DOCX 12 kb)
Callus holds great potential for biotechnology applications in plants. In this study, several exp... more Callus holds great potential for biotechnology applications in plants. In this study, several experiments were performed to establish a protocol for in vitro induction of good quality callus by using immature cotyledons and embryos of Juglans regia cv. ‘Xiangling’ as the explants. DKW basal media supplemented with different concentrations of plant growth regulators were applied to determine the induction rate, texture and color of the recovered callus. Results showed that the optimum culture medium for the callus induction from these explants was a DKW basal medium consisting of 1 mg/L BA, 2 mg/L KT 250 mg/L glutamine, 500 mg/L casein hydrolysate, 200 mg/L Vc, 50 g/L sucrose and 2.8 g/L Gelrite (i.e., the T8 medium). The average rate of callus induction on the medium reached 80.7% for the immature cotyledons and 66.7% for the immature embryos, and all the callus was characterized by a friable, creamy or yellow appearance. In vitro proliferation of callus was not very successful ...
A rapid in vitro micropropagation method was successfully established for floriculturally importa... more A rapid in vitro micropropagation method was successfully established for floriculturally important rare orchid Esmeralda clarkei Rchb.f. from shoot tip section derived from in vitro grown seedlings. The immature seeds from green capsule were first germinated on full-strength solidified Murashige and Skoog (MS) basal medium to establish the culture. For multiple shoot induction, the shoot tip section (3-5 mm) excised from 5 months old in vitro grown seedlings were cultured on the MS medium supplemented with various concentrations of 6- benzylaminopurine (BAP) and α-naphthalene acetic acid (NAA) individual and in combinations produced shoot and multiple shoots. Best response for the shoot multiplication was achieved when the medium was supplemented with BAP from 0.5 to 2.0 mg/l produced 9.5 to 11 shoots per culture. Increased concentration of BAP (2.0 mg/l) in combination with NAA (0.5 mg/l) induced less number of shoots as compared to the combination of BAP (0.5-1.5 mg/l) with NAA (...
High frequency direct shoot proliferation was induced from the shoot tip explants derived from th... more High frequency direct shoot proliferation was induced from the shoot tip explants derived from the in vitro grown seedlings of a critically endangered and horticulturally important ground orchid Phaius tancarvilleae (L'Her) Blume. Shoot tip explants cultured on solidified MS with alone or combination of various concentrations of NAA and BAP produced shoots and multiple shoots. The maximum number of healthy shoots was observed on MS with BAP (1.0 mg/l) with an average of 13.3 shoots per culture in 20 weeks; where shoot multiplication was initiated after 4 weeks of culture. Regenerated shoots rooted on MS with various concentrations of NAA, IAA, IBA. MS with NAA (0.5 mg/l) was the most appropriate condition for rooting. The well developed in vitro rooted plantlets were hardened successfully in the potting mixture containing cocopeat and sphagnum moss in the ratio of 2 : 1. Key words: Mass propagation, Phaius tancarvilleae, shoot multiplication D. O. I. 10.3329/ptcb.v21i2.10241...
ABSTRACT Talinum triangulare is a medicinally important herb and various parts of the plant are u... more ABSTRACT Talinum triangulare is a medicinally important herb and various parts of the plant are used pharmaceutically for the treatment of different diseases. In our study, a rapid and efficient protocol for micropropagation has been developed from shoot tip and nodal explants of T. triangulare. High shooting frequency (93.33 %) was achieved with shoot tip explants when cultured on Murashige and Skoog’s (MS) medium supplemented with 1.0 mg/L 6-benzyl amino purine (BAP) producing an average of 12.50 ± 0.23 shoots and 5.07 ± 0.02 cm shoot length per explant. A combination of 0.5 mg/L BAP and 0.5 mg/L kinetin was found to be more effective by producing 15.67 ± 0.25 shoots and 6.22 ± 0.02 cm shoot length per explant. The microshoots were excised and cultured on half-strength MS and full-strength MS medium containing different concentrations of indole-3-acetic acid and indole-3-butyric acid (IBA) for root induction. More number of roots (45.10 ± 0.96) with an average length of 5.46 ± 0.08 cm was obtained on half-strength MS medium supplemented with 0.5 mg/L IBA. The rooted shoots were successfully transplanted from different planting substrates to the field with a 100 % survival rate. Random amplified polymorphic DNA analysis was carried out using four random decamer primers. The amplification products were monomorphic in the micropropagated plants and were similar to the mother plant. Absence of polymorphism revealed that no variation was induced, thus maintaining the genetic integrity of the micropropagated plants of T. triangulare.
Orchids are the source of compounds like phenols, alkaloids, phenanthrenes widely used as a thera... more Orchids are the source of compounds like phenols, alkaloids, phenanthrenes widely used as a therapeutic agent. Inhibition of pancreatic α-amylase could be a better therapeutic approach in decreasing levels of post-prandial hyperglycemia. This study aimed to evaluate α-amylase inhibition of some orchid species to assess their inhibitory potential on PPA (porcine pancreatic α-amylase). Methanol extracts of the whole plant of Gastrochilus distichus (GDW), the pseudobulbs of Otochilus albus (OAP), the whole plant of Papilionanthe uniflora (PUW), pseudobulbs of Eria graminifolia (EGP), the leaves and pseudobulbs of Pholidota articulata (PAL and PAP) and stems of Vanda cristata (VCS) were screened for their phytoconstituents and role in α-amylase inhibition by modified 3,5-Dinitrosalicylic acid method. V. cristata, E. graminifolia and G. distichus extract showed moderate inhibition of α-amylase with IC50 of 582.73 μg/ml, 710.89 μg/ml, 798.78 μg/ml respectively when compared to acarbose (2...
The orchids are well known for their microbial association. Bacillus subtilis has great diversity... more The orchids are well known for their microbial association. Bacillus subtilis has great diversity in Vanda cristata which is regarded as a potential growth enhancer for the development of plants. The isolation of its strain was confirmed as Bacillus subtilis by morphological and molecular sequencing of DNA. The isolate was tested for indole acetic acid (IAA) estimation, ammonia production and phosphate solubilization in qualitative and quantitative manners. Its growth-promoting activities were tested on in vitro-raised plantlets of Cymbidium aloifolium. B. subtilis synthesized the IAA with and without a precursor (tryptophan). A more amount of auxin was found by using more amount of tryptophan. During the qualitative test for ammonia and phosphate solubilization, both compounds were synthesized by B. subtilis, which was further confirmed by the quantification method. Further, B. subtilis was used as a biotic elicitor in the in-vitro mass propagation of C. aloifolium. The development...
Valeriana jatamansi is well known for its medicinal and ethnobotanical values. An efficient and r... more Valeriana jatamansi is well known for its medicinal and ethnobotanical values. An efficient and rapid in vitro propagation system for V. jatamansi is presented. The shoot bud explants from V. jatamansi plants were cultured on Murashige and Skoog (MS) media supplemented with different concentrations of plant growth regulators (PGR's). MS medium supplemented with 2 mg/L of benzyl amino purine (BAP) produced shoot bud regeneration. However, the proliferation rate was slow with fewer shoots. The nodal segments were excised from the in vitro plants raised on BAP (2 mg/L) and multiplied by supplementing MS medium with different PGR's at different concentrations. Among the tested growth regulators, supplementation of 10% coconut water resulted in maximum shoot length (6 cm), shoot number (13.0), root length (7.5 cm) and root numbers (19.6). The genetic fidelity of the in vitro raised plants was confirmed by analysis with RAPD and ISSR markers. GC-MS profiling of the root extract fr...
Percentage of DPPH free-radical scavenging activity by plant extracts of D. moniliforme (triplica... more Percentage of DPPH free-radical scavenging activity by plant extracts of D. moniliforme (triplicate data). (DOCX 12 kb)
Rhizobium, a nitrogen-fixing bacteria is the essential feature of leguminous plants which is esse... more Rhizobium, a nitrogen-fixing bacteria is the essential feature of leguminous plants which is essential for the regeneration of nutrient-deficient soil. This study was aimed to isolate, identify, mass culture and immobilize Bradyrhizoium japonicum in encapsulated form and test their viability. Root nodules were sterilized, grinded and cultured aseptically in YEMA media containing Congo red. The obtained colon was sub-cultured to get a pure culture and different biochemical tests were conducted which proved Bradyrhizobium japonicum as the slow-growing species. The test shows a positive result of catalase production and nodulation test whereas the pH tolerance test shows more tolerance to the acidic pH. Similarly, Bradyrhizaobium japonicum can tolerate 1% and 2% NaCl concentration and it doesn’t show resistance to the penicillin disc of 10mg. The mass culture and encapsulation with sodium alginate adding sucrose as nutrient proved the simplicity for handling. Altogether 548 beads were ...
Axenic shoot culture of Mentha spicata L. was established from young shoots of the plant naturall... more Axenic shoot culture of Mentha spicata L. was established from young shoots of the plant naturally growing in the field. Large number of multiple shoots were observed on MS media supplemented with NAA (0.5ppm) and BAP (1ppm) within eight weeks of culture. Extensive root formation was observed on MS medium supplemented with 1 ppm NAA. Essential oils from both in vitro and in vivo samples showed l- carvone as a major constituent, however, the in vitro samples showed higher concentration of menthol than in vivo samples. <i>Nepal Journal of Science and Technology</i> Vol. 7, 2006
Botanica Orientalis: Journal of Plant Science, 2012
In vitro culture of Bergenia ciliata (Haw.) Sternb. was carried out for the examination of bergen... more In vitro culture of Bergenia ciliata (Haw.) Sternb. was carried out for the examination of bergenin content. Leaf explants were cultured in MS (Murashige and Skoog) basal media supplemented with or without phytohormones. The hormonal series maintained were in the range of 0-2 mg l-1 for BAP (6-benzyl amino purine) and 0-1.5 mg l-1 for NAA (α-naphthalene acetic acid). Bergenin content of in vitro grown tissues of B. ciliata was compared with that of wild plants collected from three different localities of Nepal. The best growth of callus and plantlets occurred in the media containing BAP 1.0 mg l-1 + NAA 1.0 mg l-1 and BAP 1.5 mg l-1 + NAA 1.0 mg l-1. Production of bergenin was high in the media supplemented with 1.0 mg l-1 BAP + 1.5 mg l-1 NAA (3.40 μg g-1) and 2.0 mg l-1 BAP + 1.5 mg l-1 NAA (3.05 μg g-1) under experimental condition. The bergenin content in the wild plants collected from Langtang, Jumla and Godawari was found to be 4.28 μg g-1, 4.53 μg g-1 and 3.64 μg g-1 respecti...
Total polyphenol or flavonoid contents in the plant extracts of D. moniliforme (triplicate data).... more Total polyphenol or flavonoid contents in the plant extracts of D. moniliforme (triplicate data). (DOCX 12 kb)
Callus holds great potential for biotechnology applications in plants. In this study, several exp... more Callus holds great potential for biotechnology applications in plants. In this study, several experiments were performed to establish a protocol for in vitro induction of good quality callus by using immature cotyledons and embryos of Juglans regia cv. ‘Xiangling’ as the explants. DKW basal media supplemented with different concentrations of plant growth regulators were applied to determine the induction rate, texture and color of the recovered callus. Results showed that the optimum culture medium for the callus induction from these explants was a DKW basal medium consisting of 1 mg/L BA, 2 mg/L KT 250 mg/L glutamine, 500 mg/L casein hydrolysate, 200 mg/L Vc, 50 g/L sucrose and 2.8 g/L Gelrite (i.e., the T8 medium). The average rate of callus induction on the medium reached 80.7% for the immature cotyledons and 66.7% for the immature embryos, and all the callus was characterized by a friable, creamy or yellow appearance. In vitro proliferation of callus was not very successful ...
A rapid in vitro micropropagation method was successfully established for floriculturally importa... more A rapid in vitro micropropagation method was successfully established for floriculturally important rare orchid Esmeralda clarkei Rchb.f. from shoot tip section derived from in vitro grown seedlings. The immature seeds from green capsule were first germinated on full-strength solidified Murashige and Skoog (MS) basal medium to establish the culture. For multiple shoot induction, the shoot tip section (3-5 mm) excised from 5 months old in vitro grown seedlings were cultured on the MS medium supplemented with various concentrations of 6- benzylaminopurine (BAP) and α-naphthalene acetic acid (NAA) individual and in combinations produced shoot and multiple shoots. Best response for the shoot multiplication was achieved when the medium was supplemented with BAP from 0.5 to 2.0 mg/l produced 9.5 to 11 shoots per culture. Increased concentration of BAP (2.0 mg/l) in combination with NAA (0.5 mg/l) induced less number of shoots as compared to the combination of BAP (0.5-1.5 mg/l) with NAA (...
High frequency direct shoot proliferation was induced from the shoot tip explants derived from th... more High frequency direct shoot proliferation was induced from the shoot tip explants derived from the in vitro grown seedlings of a critically endangered and horticulturally important ground orchid Phaius tancarvilleae (L'Her) Blume. Shoot tip explants cultured on solidified MS with alone or combination of various concentrations of NAA and BAP produced shoots and multiple shoots. The maximum number of healthy shoots was observed on MS with BAP (1.0 mg/l) with an average of 13.3 shoots per culture in 20 weeks; where shoot multiplication was initiated after 4 weeks of culture. Regenerated shoots rooted on MS with various concentrations of NAA, IAA, IBA. MS with NAA (0.5 mg/l) was the most appropriate condition for rooting. The well developed in vitro rooted plantlets were hardened successfully in the potting mixture containing cocopeat and sphagnum moss in the ratio of 2 : 1. Key words: Mass propagation, Phaius tancarvilleae, shoot multiplication D. O. I. 10.3329/ptcb.v21i2.10241...
ABSTRACT Talinum triangulare is a medicinally important herb and various parts of the plant are u... more ABSTRACT Talinum triangulare is a medicinally important herb and various parts of the plant are used pharmaceutically for the treatment of different diseases. In our study, a rapid and efficient protocol for micropropagation has been developed from shoot tip and nodal explants of T. triangulare. High shooting frequency (93.33 %) was achieved with shoot tip explants when cultured on Murashige and Skoog’s (MS) medium supplemented with 1.0 mg/L 6-benzyl amino purine (BAP) producing an average of 12.50 ± 0.23 shoots and 5.07 ± 0.02 cm shoot length per explant. A combination of 0.5 mg/L BAP and 0.5 mg/L kinetin was found to be more effective by producing 15.67 ± 0.25 shoots and 6.22 ± 0.02 cm shoot length per explant. The microshoots were excised and cultured on half-strength MS and full-strength MS medium containing different concentrations of indole-3-acetic acid and indole-3-butyric acid (IBA) for root induction. More number of roots (45.10 ± 0.96) with an average length of 5.46 ± 0.08 cm was obtained on half-strength MS medium supplemented with 0.5 mg/L IBA. The rooted shoots were successfully transplanted from different planting substrates to the field with a 100 % survival rate. Random amplified polymorphic DNA analysis was carried out using four random decamer primers. The amplification products were monomorphic in the micropropagated plants and were similar to the mother plant. Absence of polymorphism revealed that no variation was induced, thus maintaining the genetic integrity of the micropropagated plants of T. triangulare.
Orchids are the source of compounds like phenols, alkaloids, phenanthrenes widely used as a thera... more Orchids are the source of compounds like phenols, alkaloids, phenanthrenes widely used as a therapeutic agent. Inhibition of pancreatic α-amylase could be a better therapeutic approach in decreasing levels of post-prandial hyperglycemia. This study aimed to evaluate α-amylase inhibition of some orchid species to assess their inhibitory potential on PPA (porcine pancreatic α-amylase). Methanol extracts of the whole plant of Gastrochilus distichus (GDW), the pseudobulbs of Otochilus albus (OAP), the whole plant of Papilionanthe uniflora (PUW), pseudobulbs of Eria graminifolia (EGP), the leaves and pseudobulbs of Pholidota articulata (PAL and PAP) and stems of Vanda cristata (VCS) were screened for their phytoconstituents and role in α-amylase inhibition by modified 3,5-Dinitrosalicylic acid method. V. cristata, E. graminifolia and G. distichus extract showed moderate inhibition of α-amylase with IC50 of 582.73 μg/ml, 710.89 μg/ml, 798.78 μg/ml respectively when compared to acarbose (2...
The orchids are well known for their microbial association. Bacillus subtilis has great diversity... more The orchids are well known for their microbial association. Bacillus subtilis has great diversity in Vanda cristata which is regarded as a potential growth enhancer for the development of plants. The isolation of its strain was confirmed as Bacillus subtilis by morphological and molecular sequencing of DNA. The isolate was tested for indole acetic acid (IAA) estimation, ammonia production and phosphate solubilization in qualitative and quantitative manners. Its growth-promoting activities were tested on in vitro-raised plantlets of Cymbidium aloifolium. B. subtilis synthesized the IAA with and without a precursor (tryptophan). A more amount of auxin was found by using more amount of tryptophan. During the qualitative test for ammonia and phosphate solubilization, both compounds were synthesized by B. subtilis, which was further confirmed by the quantification method. Further, B. subtilis was used as a biotic elicitor in the in-vitro mass propagation of C. aloifolium. The development...
Valeriana jatamansi is well known for its medicinal and ethnobotanical values. An efficient and r... more Valeriana jatamansi is well known for its medicinal and ethnobotanical values. An efficient and rapid in vitro propagation system for V. jatamansi is presented. The shoot bud explants from V. jatamansi plants were cultured on Murashige and Skoog (MS) media supplemented with different concentrations of plant growth regulators (PGR's). MS medium supplemented with 2 mg/L of benzyl amino purine (BAP) produced shoot bud regeneration. However, the proliferation rate was slow with fewer shoots. The nodal segments were excised from the in vitro plants raised on BAP (2 mg/L) and multiplied by supplementing MS medium with different PGR's at different concentrations. Among the tested growth regulators, supplementation of 10% coconut water resulted in maximum shoot length (6 cm), shoot number (13.0), root length (7.5 cm) and root numbers (19.6). The genetic fidelity of the in vitro raised plants was confirmed by analysis with RAPD and ISSR markers. GC-MS profiling of the root extract fr...
Percentage of DPPH free-radical scavenging activity by plant extracts of D. moniliforme (triplica... more Percentage of DPPH free-radical scavenging activity by plant extracts of D. moniliforme (triplicate data). (DOCX 12 kb)
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