Thidiazuron (TDZ) has gained a considerable attention during past decades due to its efficient role in plant cell and tissue culture. Wide array of physiological responses were observed in response to TDZ-application in different plant species. TDZ has shown both auxin and cytokinin like effects, although, chemically, it is totally different from commonly used auxins and cytokinins. A number of biological (physiological and biochemical) events in cells are induced or enhanced by TDZ, but the mode of action of TDZ is yet unknown. However, varieties of underlying mechanisms were revealed by reports showing how morphogenic events were induced by application of TDZ. Other reports showed that TDZ may modify endogenous plant growth regulators, either directly or indirectly and produce reactions in cell/tissue, necessary for its division/regeneration. Other possibilities include modification in cell membrane, energy levels, nutrient absorption, transport and assimilation, etc. In this review, recent advancements in TDZ application in plant sciences are discussed.

	 

	Key words: Thidiazuron, plant growth regulators, somatic embryogenesis, regeneration, cell cultures, metabolism.

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Thidiazuron: A multi-dimensional plant growth regulator

The application of plant growth regulators to improve phytoremediation of contaminated soils: A review.

Chilli peppers--a review on tissue culture and transgenesis.

Synseed technology-a complete synthesis.

Magnolia sirindhorniae Noot. & Chalermglin is an endangered species with high ornamental and commercial value that needs to be urgently protected and judiciously commercialized. In this study, a protocol for efficient regeneration of this species is standardized. The lateral buds of the M. sirindhorniae plant were used as an explant. Half-strength Murashige and Skoog (MS) medium supplemented with 2.0 mg/L 6-benzyladenine (BA), 0.1 mg/L α-naphthaleneacetic acid (NAA), and 2.0 mg/L gibberellic acid (GA3) was found to be the optimal medium for shoot induction. The maximum shoot multiplication rate (310%) was obtained on Douglas-fir cotyledon revised medium (DCR) fortified with 0.2 mg/L BA, 0.01 mg/L NAA, and additives. The half-strength DCR medium supplemented with 0.5 mg/L NAA and 0.5 mg/L indole-3-butyric acid (IBA) supported the maximum rate (85.0%) of in vitro root induction. After a simple acclimatization process, the survival rate of plantlets in a substrate mixture of sterile perlite and peat soil (1:3; v/v) was 90.2%. DNA markers were used for assessment of genetic uniformity, confirming the genetic uniformity and stability of regenerated plants of M. sirindhorniae. Thus, the described protocol can safely be applied for large scale propagation of this imperative plant.

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An efficient micropropagation protocol for an endangered ornamental tree species ( Magnolia sirindhorniae Noot. & Chalermglin) and assessment of genetic uniformity through DNA markers

The effect of thidiazuron (TDZ) was investigated on in vitro shoot proliferation from nodal explants of Rauvolfia tetraphylla. Murashige and Skoog (MS) medium containing TDZ (0.5–10μM) was effective in inducing shoot buds and maintaining high rates of shoot multiplication on hormone free medium. The highest shoot regeneration frequency (90%) and mean number (18.50 ± 1.25) of shoots per explant were achieved from nodal segments cultured on MS medium supplemented with 5μM TDZ for 4 weeks prior to transfer to MS medium without TDZ for 8 weeks. The regenerated shoots rooted best on MS medium containing 0.5μM indole-3-butyric acid (IBA). Micropropagated plantlets were hardened to survive ex vitro conditions and were then established into soil.

Shoot multiplication in Rauvolfia tetraphylla L. using thidiazuron

An efficient protocol is described for the rapid in vitro multiplication of an endangered medicinal plant, Tylophora indica (Burm. f.) Merrill, via enhanced axillary bud proliferation from nodal explants collected from young shoots of a two-year-old plant. The physiological effects of growth regulators [6-benzyladenine (BA), kinetin (Kin) thidiazuron (TDZ), indole-3-acetic acid (IAA), indole-3-butyric acid (IBA) or α-naphthalene acetic acid (NAA)], ascorbic acid (AA), different strengths of Murashige and Skoog (MS) medium and various pH levels on in vitro morphogenesis were investigated. The highest number (8.6 ± 0.71) of shoots and the maximum average shoot length (5.2 ± 0.31 cm) were recorded on MS medium supplemented with 2.5 μM BA, 0.5 μM NAA and 100 mg/l AA at pH 5.8. Rooting was best achieved on half-strength MS medium augmented with 0.5 μM IBA. The plantlets regenerated in vitro with well-developed shoot and roots were successfully established in pots containing garden soil and grown in a greenhouse with a 90% survival rate. The regenerated plants did not show any immediate detectable phenotypic variation. The described method can be successfully employed for large-scale multiplication and long-term in vitro conservation of T. indica.

An efficient micropropagation system for Tylophora indica: an endangered, medicinally important plant

An efficient and improved shoot regeneration technique for the micropropagation of Vitex negundo, an aromatic and medicinal shrub through in vitro culture of nodal segments with axillary buds, is described. Thidiazuron (TDZ) used at 1.0 μM was the most effective in inducing bud break and growth, and also in initiating multiple shoot proliferation at the rate of 25 microshoots per nodal explant with axillary buds, after 4 weeks of culture. By repeated subculturing of nodal explants, a high-frequency multiplication rate was established. Optimum shoot multiplication and elongation was achieved when TDZ exposed explants were subcultured on Murashige and Skoog (MS) media containing a combination of 1.0 μM 6-benzyladenine (BA) and 0.5 μM α-naphthalene acetic acid (NAA). Efficient rooting was achieved directly in soilrite when basal portion of the shoots were treated with 500 μM indole-3-butyric acid for 10 min which was the most effective in inducing roots, as 97% of the microshoots produced roots. Plantlets went through a hardening phase in a controlled plant growth chamber, prior to ex-vitro transfer. Micropropagated plants grew well, attained maturity and flowered. No phenotypical differences for morphogenesis were observed among the regenerants.

Rapid clonal multiplication of a woody tree, Vitex negundo L. through axillary shoots proliferation

In vitro callus induction and plant regeneration from leaf explants of Ruta graveolens L.

Multiple shoots were induced by culturing nodal explants excised from 1-month-old aseptic seedlings of red pepper (Capsicum annuum L. cv. Pusa Jwala) on Murashige and Skoog (MS) medium supplemented with (0.1–10 μM) thidiazuron (TDZ). The rate of multiple shoot induction per explant was maximum (14.4 ± 0.06) on MS medium supplemented with 1.0 μM TDZ. Regenerated shoots were elongated well on growth regulator free MS medium. Adventitious roots were induced two weeks after transfer of elongated shoots to MS medium supplemented with auxins (IAA, IBA or NAA) in different concentrations. Optimum root formation frequency was obtained in medium containing 1.0 μM IBA. Ex-vitro rooting was also achieved by pulse treatment with 300 μM IBA for 10 min. Rooted shoots were transplanted in plastic pots containing garden soil (with 90 % survival rate), where they grew well and attained maturity. Regenerated plants were phenotypically and cytologically normal.

Naseem Ahmad

Papers

Shoot multiplication in Rauvolfia tetraphylla L. using thidiazuron

An efficient micropropagation system for Tylophora indica: an endangered, medicinally important plant

Rapid clonal multiplication of a woody tree, Vitex negundo L. through axillary shoots proliferation

In vitro callus induction and plant regeneration from leaf explants of Ruta graveolens L.

Improved plant regeneration in Capsicum annuum L. from nodal segments