Objective:To develop the reproducible in vitro propagation protocols for the medicinally important plants viz.,Achyranthes aspera(A.aspera)L.and Achyranthes bidentata(A.bidentata)Blume using nodal segments as explants...Objective:To develop the reproducible in vitro propagation protocols for the medicinally important plants viz.,Achyranthes aspera(A.aspera)L.and Achyranthes bidentata(A.bidentata)Blume using nodal segments as explants.Methods:Young shoots of A.aspera and A.bidentata were harvested and washed with running tap water and treated with 0.1%bavistin and rinsed twice with distilled water.Then the explants were surface sterilized with 0.1%(w/v)HgCl_2 solutions for I min.After rinsing with sterile distilled water for 3-4 times,nodal segments were cut into smaller segments(1 cm)and used as the explants.The explants were placed horizontally as well as vertically on solid basal Murashige and Skoog(MS)medium supplemented with 3%sucrose,0.6%(w/v)agar(HiMedia,Mumbai)and different concentration and combination of 6-benzyl amino purine(BAP),kinetin(Kin),naphthalene acetic acid(NAA)and indole acetic acid(IAA)for direct regeneration.Results:Adventitious proliferation was obtained from A.aspera and A.bidentata nodal segments inoculated on MS basal medium with 3%sucrose and augmented with BAP and Kin with varied frequency.MS medium augmented with 3.0 mg/L of BAP showed the highest percentage(93.60±0.71)of shootlets formation for A.aspera and(94.70±0.53)percentages for A.bidentata.Maximum number of shoots/explants(10.60±0.36)for A.aspera and(9.50±0.56)for A.bidentata was observed in MS medium fortified with 5.0 mg/L of BAP.For A.aspera,maximum mean length(5.50±0.34)of shootlets was obtained in MS medium augmented with 3.0 mg/L of Kin and for A.bidentata(5.40±0.61)was observed in the very same concentration.The highest percentage,maximum number of rootlets/shootlet and mean length of rootlets were observed in 1/2 MS medium supplemented with 1.0 mg/L of 1BA.Seventy percentages of plants were successfully established in polycups.Sixty eight percentages of plants were well established in the green house condition.Sixty five percentages of plants were established in the field.Conclusions:The results have shown that use of nodal buds is an alternative reproducible and dependable method for clonal propagation of A.aspera and A.bidentata.The high rate of direct shoot-root multiplication and their high rate of post-hardening survival indicate that this protocol can he easily adopted for commercial large scale cultivation.展开更多
In this work, nodal-disk segments (4-6 mm in diameter × 5-6 mm in length) were obtained from established shoot culture, resulted from disinfected tomato seedlings, and they were suitable to induce different organ...In this work, nodal-disk segments (4-6 mm in diameter × 5-6 mm in length) were obtained from established shoot culture, resulted from disinfected tomato seedlings, and they were suitable to induce different organogenic pathway under the influence of specific hormonal treatment. Application of BAP (1-2.5 mg/l) alone or in combination of 0.5 mg/l NAA resulted in induction of shoot formation. Somatic embryogenesis was rarely appeared (6%) when relatively low concentration of BAP (1.5 mg/l) with low concentration of IAA (0.5 mg/l IAA) was applied. Root induction was triggered when nodal explants or shoot cuttings were cultured on MS medium with (1 mg/l IAA, IBA or NAA) or without auxins, but the best result was obtained when 1 mg/l IAA was used. Application of 0.5 mg/l NAA stimulated callus formation but the best result was obtained when the three different phytohormoes were used (0.5 mg/l 2,4-D + 1 mg/l NAA + 0.5 mg/l BAP). These results indicated that nodal segments, as described in this protocol, can be used as alternative to other types of explants such as cotyledon, hypocotyl and leaf explants.展开更多
文摘Objective:To develop the reproducible in vitro propagation protocols for the medicinally important plants viz.,Achyranthes aspera(A.aspera)L.and Achyranthes bidentata(A.bidentata)Blume using nodal segments as explants.Methods:Young shoots of A.aspera and A.bidentata were harvested and washed with running tap water and treated with 0.1%bavistin and rinsed twice with distilled water.Then the explants were surface sterilized with 0.1%(w/v)HgCl_2 solutions for I min.After rinsing with sterile distilled water for 3-4 times,nodal segments were cut into smaller segments(1 cm)and used as the explants.The explants were placed horizontally as well as vertically on solid basal Murashige and Skoog(MS)medium supplemented with 3%sucrose,0.6%(w/v)agar(HiMedia,Mumbai)and different concentration and combination of 6-benzyl amino purine(BAP),kinetin(Kin),naphthalene acetic acid(NAA)and indole acetic acid(IAA)for direct regeneration.Results:Adventitious proliferation was obtained from A.aspera and A.bidentata nodal segments inoculated on MS basal medium with 3%sucrose and augmented with BAP and Kin with varied frequency.MS medium augmented with 3.0 mg/L of BAP showed the highest percentage(93.60±0.71)of shootlets formation for A.aspera and(94.70±0.53)percentages for A.bidentata.Maximum number of shoots/explants(10.60±0.36)for A.aspera and(9.50±0.56)for A.bidentata was observed in MS medium fortified with 5.0 mg/L of BAP.For A.aspera,maximum mean length(5.50±0.34)of shootlets was obtained in MS medium augmented with 3.0 mg/L of Kin and for A.bidentata(5.40±0.61)was observed in the very same concentration.The highest percentage,maximum number of rootlets/shootlet and mean length of rootlets were observed in 1/2 MS medium supplemented with 1.0 mg/L of 1BA.Seventy percentages of plants were successfully established in polycups.Sixty eight percentages of plants were well established in the green house condition.Sixty five percentages of plants were established in the field.Conclusions:The results have shown that use of nodal buds is an alternative reproducible and dependable method for clonal propagation of A.aspera and A.bidentata.The high rate of direct shoot-root multiplication and their high rate of post-hardening survival indicate that this protocol can he easily adopted for commercial large scale cultivation.
文摘In this work, nodal-disk segments (4-6 mm in diameter × 5-6 mm in length) were obtained from established shoot culture, resulted from disinfected tomato seedlings, and they were suitable to induce different organogenic pathway under the influence of specific hormonal treatment. Application of BAP (1-2.5 mg/l) alone or in combination of 0.5 mg/l NAA resulted in induction of shoot formation. Somatic embryogenesis was rarely appeared (6%) when relatively low concentration of BAP (1.5 mg/l) with low concentration of IAA (0.5 mg/l IAA) was applied. Root induction was triggered when nodal explants or shoot cuttings were cultured on MS medium with (1 mg/l IAA, IBA or NAA) or without auxins, but the best result was obtained when 1 mg/l IAA was used. Application of 0.5 mg/l NAA stimulated callus formation but the best result was obtained when the three different phytohormoes were used (0.5 mg/l 2,4-D + 1 mg/l NAA + 0.5 mg/l BAP). These results indicated that nodal segments, as described in this protocol, can be used as alternative to other types of explants such as cotyledon, hypocotyl and leaf explants.