A protocol for micropropagation using nodal explants from mature Pinus massoniana trees has been developed. Time of explant collection is crucial for the initial success of aseptic culture. Explants collected in early...A protocol for micropropagation using nodal explants from mature Pinus massoniana trees has been developed. Time of explant collection is crucial for the initial success of aseptic culture. Explants collected in early March gave the highest percentage of explant survival (64.5%) and shoot-forming percentage (52.3%). Thidiazuron (TDZ) concentration significantly influenced shoot formation; 4 mu M TDZ was optimum, with 4.8 shoots produced per explant with a mean length of 7.1 cm after 120 days of culture. Regenerated shoots rooted for 60 days in basic medium with 1 mu M NAA were ready for growth in pots. This is the first report on plantlet regeneration in vitro from mature trees of P. massoniana that provides a reliable method for propagating selected elites.展开更多
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 展开更多
Our research work demonstrates the single bead alginate-encapsulation, interim storing and conversion of Tylophora indica(Burm. Fil.) Merrill.Most effective encapsulation of in vitro nodal segments [(4 ± 1) mm lo...Our research work demonstrates the single bead alginate-encapsulation, interim storing and conversion of Tylophora indica(Burm. Fil.) Merrill.Most effective encapsulation of in vitro nodal segments [(4 ± 1) mm long], ensuing in sphere-shaped artificial seeds of similar morphology, was achieved through 75 mmol·L^(-1)calcium chloride(CaCl_2 · 2H_2O) plus 3%(w/v) Na-alginate with 93.3% conversion frequency. The earliest conversion(within 7 days of incubation) of artificial seeds occurred in half-strength liquid Murashige and Skoog medium. Among the three different temperature regimes [(5 ± 1) °C,(15 ± 1) °C, and(25 ± 1) °C], storage of artificial seeds at(15 ± 1) °C executed the highest frequency of conversion(90%) after 15 days of storage. Lengthier storage significantly reduced the conversion frequency of artificial seeds irrespective of storage temperature. Nevertheless, the conversion frequency after 30 days of storage at(15 ± 1) °C was recorded at 70% without further decline even following45 days of storage, which evidently suggests that lower temperature(15 ± 1) °C is apt for storage and subsequent conversion of T. indica artificial seeds. The present protocol could be expedient for short-term storing and swapping of T. indica germplasms between national and international laboratories.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
This study developed an efficient in vitro cultivation and propagation sys- tem for an endangered species Kolkwitzia amabilis using nodal segments as ex- plants. Multiple shoots were induced through axillary bud forma...This study developed an efficient in vitro cultivation and propagation sys- tem for an endangered species Kolkwitzia amabilis using nodal segments as ex- plants. Multiple shoots were induced through axillary bud formation. The highest fre- quency of multiple shoot induction was achieved when the nodal segment explants were incubated in Murashige and Skoog （MS） medium supplemented with 4.44 pM 6-benzyladenine （6-BA） in combination with 0.54 μM a-naphthaleneacetic acid （NAA）, followed by treatment with 4.44 μM 6-BA in combination with 0.27 μM NAA. Shoot multiplication could be induced in MS medium supplemented with stand-alone 6-BA or 6-BA in combination with indole-3-acetic acid （1.71 μM） or NAA （0.27 or 0.54 μM）, with 6-BA and either compound, exhibiting a stronger effect on shoot multiplication. The optimum combination of plant growth regulators for shoot multiplication was 4.44 μM 6-BA with 0.27 μM NAA. The maximum rooting percentage was obtained in a half-strength MS medium supplemented with indole-3-butyric acid alone and in com- bination with NAA and 2,4-dichlorophenoxyacetic acid, but the best combination of plant growth regulators for rooting was 1.48 μM indole-3-butyric acid with 1.08 μM NAA and 0.05 μM 2,4-dichlorophenoxyacetic acid. The rooted shoots were trans- ferred to a greenhouse with a success rate of 100%.展开更多
基金funded by the Science Research and Technology Development from the Department of Science and Technology of Guangxi(14125008-2-17,1598006-5-7)the Natural Science Foundation of China(31360178)+1 种基金the Key Program of Guangxi Forestry Bureau([2015]7)as an independent project by the Key Laboratory of Gaungxi Fine Timber Forest Resources Cultivation(13A-01-01)
文摘A protocol for micropropagation using nodal explants from mature Pinus massoniana trees has been developed. Time of explant collection is crucial for the initial success of aseptic culture. Explants collected in early March gave the highest percentage of explant survival (64.5%) and shoot-forming percentage (52.3%). Thidiazuron (TDZ) concentration significantly influenced shoot formation; 4 mu M TDZ was optimum, with 4.8 shoots produced per explant with a mean length of 7.1 cm after 120 days of culture. Regenerated shoots rooted for 60 days in basic medium with 1 mu M NAA were ready for growth in pots. This is the first report on plantlet regeneration in vitro from mature trees of P. massoniana that provides a reliable method for propagating selected elites.
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
文摘Our research work demonstrates the single bead alginate-encapsulation, interim storing and conversion of Tylophora indica(Burm. Fil.) Merrill.Most effective encapsulation of in vitro nodal segments [(4 ± 1) mm long], ensuing in sphere-shaped artificial seeds of similar morphology, was achieved through 75 mmol·L^(-1)calcium chloride(CaCl_2 · 2H_2O) plus 3%(w/v) Na-alginate with 93.3% conversion frequency. The earliest conversion(within 7 days of incubation) of artificial seeds occurred in half-strength liquid Murashige and Skoog medium. Among the three different temperature regimes [(5 ± 1) °C,(15 ± 1) °C, and(25 ± 1) °C], storage of artificial seeds at(15 ± 1) °C executed the highest frequency of conversion(90%) after 15 days of storage. Lengthier storage significantly reduced the conversion frequency of artificial seeds irrespective of storage temperature. Nevertheless, the conversion frequency after 30 days of storage at(15 ± 1) °C was recorded at 70% without further decline even following45 days of storage, which evidently suggests that lower temperature(15 ± 1) °C is apt for storage and subsequent conversion of T. indica artificial seeds. The present protocol could be expedient for short-term storing and swapping of T. indica germplasms between national and international laboratories.
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.
文摘This study developed an efficient in vitro cultivation and propagation sys- tem for an endangered species Kolkwitzia amabilis using nodal segments as ex- plants. Multiple shoots were induced through axillary bud formation. The highest fre- quency of multiple shoot induction was achieved when the nodal segment explants were incubated in Murashige and Skoog （MS） medium supplemented with 4.44 pM 6-benzyladenine （6-BA） in combination with 0.54 μM a-naphthaleneacetic acid （NAA）, followed by treatment with 4.44 μM 6-BA in combination with 0.27 μM NAA. Shoot multiplication could be induced in MS medium supplemented with stand-alone 6-BA or 6-BA in combination with indole-3-acetic acid （1.71 μM） or NAA （0.27 or 0.54 μM）, with 6-BA and either compound, exhibiting a stronger effect on shoot multiplication. The optimum combination of plant growth regulators for shoot multiplication was 4.44 μM 6-BA with 0.27 μM NAA. The maximum rooting percentage was obtained in a half-strength MS medium supplemented with indole-3-butyric acid alone and in com- bination with NAA and 2,4-dichlorophenoxyacetic acid, but the best combination of plant growth regulators for rooting was 1.48 μM indole-3-butyric acid with 1.08 μM NAA and 0.05 μM 2,4-dichlorophenoxyacetic acid. The rooted shoots were trans- ferred to a greenhouse with a success rate of 100%.
