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1

Embryological observations on unpollinated ovary culture of mulberry [Morus alba L.]

100%
Thomas T. D.
Acta Biologica Cracoviensia. Series Botanica
|
2004
|
tom 46
87-94
A three-step procedure was adopted for the induction of gynogenesis in two cultivars of mulberry (Morus alba L.). This includes in vitro flowering, inflorescence segment culture and isolated ovary culture. In the third step, that is, isolated ovary culture, the cultured ovaries burst and an embryo emerged from the ovary. The present paper investigates the ontogeny of the developing gynogenic embryo. The study confirmed that the gynogenic embryo emerged from the egg cell. Before the onset of division of the egg, all other cells of the embryo sac degenerated in the majority of ovules, but in exceptional cases the polar nuclei will be retained along with the dividing egg cell. The presence of the gynogenic embryo along with free-nuclear autonomous endosperm is the most important feature of the present investigation. Autonomous endosperm is formed from either the polar nuclei or secondary nucleus. In both cultivars used for the experiment, the percentage of ovaries showing proembryo induction during inflorescence segment culture is much higher than that of ovaries producing gynogenic plants during isolated ovary culture. This suggests the degeneration of some gynogenic embryos during the initial stages of induction.
2

Pretreatment in thidiazuron improves the in vitro shoot induction from leaves in Curculigo orchioides Gaertn., an endangered medicinal plant

100%
Thomas T. D.
Acta Physiologiae Plantarum
|
2007
|
tom 29
|
nr 5
Leaf regeneration via direct induction of adventitious shoots obtained from an endangered medicinal plant, Curculigo orchioides Gaertn. by pretreating with thidiazuron. C. orchioides is an endangered medicinal herb belonging to the family Hypoxidaceae. Direct inoculation of leaf pieces on MS medium supplemented with various concentrations of BAP (2–8 µM) or TDZ (2–8 µM) alone or in combination with NAA (0.5 and 1.0 µM) produced low shoot induction both in terms of % response and number of shoots per explant. Hence, leaf explants were pretreated with 15, 25 or 50 µM thidiazuron (TDZ), for 6, 24 or 48 h with the aim of improving shoot regeneration from cultured explants. After pretreatment, explants were transferred to an agar solidified MS medium that was supplemented with BAP (4 µM), TDZ (6 µM), BAP (4 µM) + NAA (1.0 µM), TDZ (6 µM) + NAA (0.5 µM). Control explants were incubated directly on the medium without any pretreatment. The pretreatment of explants with 15 µM TDZ for 24 h significantly promoted the formation of adventitious shoots and the maximum response was observed on MS medium supplemented with 6 µM TDZ. In this medium, 96 % cultures responded with an average number of 16.2 adventitious shoots per explant. The percentage of leaf explants producing shoots and the average number of shoots per explant were significantly improved when TDZ pretreated leaves were cultured onto MS medium supplemented with BAP or TDZ alone or in combination with NAA. The rooted plantlets were successfully transplanted to soil with 90% success. The present investigation indicated the stimulatory role of TDZ pretreatment in regulating shoot regeneration from leaf explants of C. orchioides.
3

In vitro propagation for the conservation of a rare medicinal plant Justicia gendarussa Burm. f. by nodal explants and shoot regeneration from callus

63%
Thomas T. D., Yoichiro H.
Acta Physiologiae Plantarum
|
2010
|
tom 32
|
nr 5
Justicia gendarussa is a valuable medicinal plant and various parts of this plant are pharmaceutically used for the treatment of different diseases. In vitro regeneration of shoot buds was obtained from culture of nodal cuttings as well as shoot regeneration from callus. The nodal cuttings differed in shoot proliferation in terms of percentage of explants that responded and average shoot length with various concentrations (4.4, 8.9, 13.3, 17.7, 22.2 µM) of 6-benzyladenine (BA), kinetin (Kn) and thidiazuron. In all treatments, one shoot was invariably present. Optimum 87% of cultures responded with an average shoot length of 4.4 cm on Murashige and Skoog (MS) medium supplemented with 17.7 µM BA. Callus was induced from the mature leaf segments on MS medium supplemented with Kn (4.7, 13.9, 23.2 µM) alone or in combination with 2, 4-dichlorophenoxyacetic acid (2, 4-D; 2.3 µM, 4.5 µM). Optimum callus induction (78%) was obtained on MS medium supplemented with 14 µM Kn and 4.5 µM 2, 4-D. When the callus was subcultured on MS medium fortified with BA (8.9, 17.7, 26.6 µM) or Kn (9.3, 18.6, 27.9 µM) alone or in combination with α naphthalene acetic acid (NAA; 2.7, 5.4 µM), shoot regeneration was obtained. The highest response (92%) was observed on MS medium containing 17.7 µM BA and 5.4 µM NAA. On this medium, an average number of 12.2 shoots were obtained per responding callus. The shoots obtained from callus and nodal cuttings were rooted with α frequency of 73% on MS medium augmented with 9.8 µM indole-3-butyric acid. The rooted shoots were successfully transplanted to soil and sand mixture (1:1) with 90% survival rate. The protocol standardized for shoot proliferation and regeneration in J. gendarussa from nodal cuttings and leaf-derived callus is suitable for micropropagation and conservation of this essential medicinal plant.
4

In vitro propagation and conservation of Indian sarsaparilla, Hemidesmus indicus L. R. Br. through somatic embryogenesis and synthetic seed production

51%
Cheruvathur M. K., Najeeb N., Thomas T. D.
Acta Physiologiae Plantarum
|
2013
|
tom 35
|
nr 03
A protocol has been developed for achieving somatic embryogenesis from callus derived from nodal cuttings and production of synthetic seeds in Hemidesmus indicus L. R. Br. a highly traded ethnomedicinal plant. Proembryogenic, friable, light yellowish callus was induced from the basal cut end of the nodal cuttings on Murashige and Skoog (MS) medium supplemented with 3 μM indole-3- butyric acid (IBA). The highest rate of somaticembryogenesis (92 %) was observed when the callus was subcultured on half strength MS medium supplemented with 2 μM IBA. On induction medium somatic embryos were developed up to the torpedo stage. Further elongation and germination of somatic embryos were obtained in MS medium supplemented with 4 μM 6-benzylaminopurine (BA) in combination with 1.5 lM gibberellic acid (GA3). Somatic embryos were collected and suspended in a matrix of MS medium containing sodium alginate (3 % W/V) dropped into 75 mM calcium chloride (CaCl2·2H2O) solution for the production of synthetic seeds and later transferred to MS medium for germination. The synthetic seeds were successfully germinated on medium even after 120 days of storage at 4ºC. The plantlets were eventually transferred to soil with 92 % success.
5

Pulvinus: an ideal explant for plant regeneration in Caesalpinia bonduc (L.) Roxb., an important ethnomedicinal woody climber

51%
Cheruvathur M. K., Britto J., Thomas T. D.
Acta Physiologiae Plantarum
|
2012
|
tom 34
|
nr 2
This study demonstrates the morphogenic potential of pulvinus, an important organ situated at the base of the petiole or rachis of leguminous plants. Plant regeneration via pulvinus-derived calli of Caesalpinia bonduc has been achieved. Organogenic calli have been derived from the explant 45 days after culture on Murashige and Skoog (MS) medium supplemented with 2,4-dichlorophenoxyacetic acid (2,4-D) alone or in combination with 6-benzylaminopurine (BA). Optimum callus induction (100%) occurred when the pulvini were cultured on MS medium fortified with 6 mg l⁻¹ 2,4-D and 1 mg l⁻¹ BA. The highest shoot induction was obtained when the calli were transferred to MS medium supplemented with 5 mg l⁻¹ BA and 1 mg l⁻¹ indole-3-acetic acid (IAA). On this medium, 87% cultures responded with an average number of 4.2 shoots per culture. The maximum root induction from the regenerated shoots was observed on half strength MS medium containing 6 mg l⁻¹ indole-3-butyric acid (IBA). Here 100% shoots rooted with a mean number of 6.3 roots per shoot. The regenerated plantlets were acclimatized and subsequently showed normal growth. This efficient protocol will be helpful for propagating elite clones on a mass scale and could be utilized for genetic transformation study.
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