Rapid micropropagation of <Emphasis Type="Italic">Ocimum basilicum</Emphasis> using shoot tip explants pre-cultured in thidiazuron supplemented liquid medium

An efficient protocol has been developed for rapid micropropagation of Ocimum basilicum. Multiple shoots were induced by culturing shoot tip explants excised from mature plants on a liquid Murashige and Skoog (MS) medium supplemented with 5–100 μM of thidiazuron (TDZ) for different treatment duration (4, 8, 12 and 16 d). The optimal level of TDZ supplementation to the culture medium was 50 μM for 8 d induction period followed by subculturing in MS medium devoid of TDZ as it produced maximum regeneration frequency (78 %), mean number of shoots (11.6 ± 1.16) and shoot length (4.8 ± 0.43 cm) per explant. A culture period longer than 8 d with TDZ resulted in the formation of fasciated or distorted shoots. The regenerated shoots rooted best on MS medium containing 1.0 μM indole-3-butyric acid (IBA). The micropropagated shoots with well developed roots were successfully established in pots containing garden soil and grown in greenhouse with 95 % survival rate. The regenerated plants were morphologically uniform and exhibited similar growth characteristics and vegetative morphology to the donor plants.     

RETRACTED ARTICLE: Influencing micropropagation in Clitoria ternatea L. through the manipulation of TDZ levels and use of different explant types

A comparative performance of two explants types (CN and Nodal) for their efficiency to induce multiple shoot regeneration in Clitoria ternatea has been carried out. Thidiazuron (TDZ) in different concentrations (0.05–2.5 μM) was used as a supplement to the Murashige and Skoog’s (MS) basal media. Explant type apart, two factors viz. concentration and exposure duration to TDZ played an important role in affecting multiple shoot regeneration. Cotyledonary node explants produced the best results at 0.1 μM TDZ, while in nodal explants the highest rate of shoot formation was achieved on MS medium supplemented with 1.0 μM TDZ. In both the explants, shoot multiplication increased when the regenerated shoots were subcultured on hormone free MS medium after 4 weeks of exposure to TDZ. Among the two, cotyledonary node explants produced considerably higher number of shoots at a comparatively lower concentration of TDZ than nodal explants. The regenerated shoots rooted best on MS medium containing 1.0 μM indole-3-butyric acid (IBA) and were successfully established in pots containing garden soil with 88 % survival rate. All the regenerated plants showed normal morphology and growth characteristics.     

Thidiazuron-induced Shoot Multiplication and Plant Regeneration in Bamboo (Dendrocalamus strictus Nees)

Thidiazuron (TDZ) stimulated shoot proliferation from different seedling explants (i.e., shoot, basal node, node and apical segment) of bamboo (Dendrocalamus strictus) when incorporated in half-strength Murashige and Skoog (MS) medium having 2% (w/v) sucrose. All the concentrations of TDZ (0.01 to 1.0 mg l^?1) tried were effective in shoot proliferation. Maximum shoots (14.8 ± 1.0) were obtained from the shoot explants cultured in 0.5 mg l^?1 TDZ supplemented halfstrength MS liquid medium for 21 days and subsequently transferred to the same medium devoid of TDZ. The longer culture period (i.e. 28 and 35 days) in TDZ medium caused reduction in shoot proliferation. The shoots regenerated with lower concentrations of TDZ treatment (i.e. 0.01 to 0.1 mg l^?1) rooted in half-strength MS liquid medium. The shoots formed with 0.5 mg l^?1 TDZ treatment did not root in basal medium and required auxin supplementation in the medium for rooting and about 55% shoots produced roots in 1.0 mg l^?1 IBA supplemented medium. The shoots formed with 1.0 mg l^?1 TDZ did not root even after auxin treatment. The well rooted shoots transplanted to plastic pots filled with sand and garden soil (1:1) mixture showed 98% establishment.     

In vitro regeneration of Minthostachys andina (brett) Epling - a Bolivian native species with aromatic and medicinal properties

Various explants of Minthostachys andina (Brett.) were evaluated for their morphogenic potential under in vitro culture conditions. Axillary buds derived from 2 year-old plants grown in MS-medium supplemented with 4.4 or 8.8 μM BA and 0.054 μM NAA, initiated shoot growth and new shoot formation. Under subculture in NN medium, shoots were rooted in the presence of NAA (1.6, 2.7 or 5.3 μM) alone or in combination with IBA (9.8 μM), and the regenerated plantlets were later acclimatised in the greenhouse. Also, polynodal segments from seedlings initiated multiple shoots and plantlets when initially cultured in presence of NN-liquid salt medium supplemented with 2.2-17.7 μM BA or 4.5-13.6 M TDZ in combination with different auxin-like growth regulators and after a final transfer for root initiation. The same types of responses were found in hypocotyl and leaf explants, which produced adventitious shoots in the presence of TDZ. The use of antioxidants helped to prevent browning and favoured organogenesis. This revised version was published online in June 2006 with corrections to the Cover Date.     

Thidiazuron-induced high frequency of shoot induction and plant regeneration in protoplast derived pea callus

Protoplasts isolated from lateral shoot buds of cotyledon-free pea embryo axes were regenerated to callus. Protoplast derived calluses with a diameter of about 1cm were transferred to shoot induction media, containing different concentrations (1–50µM) of thidiazuron. Shoot formation was observed after 16 weeks up to 12% efficiency. Thidiazuron (10µM) was the most effective concentration in all experiments. Shoot buds elongated in medium supplemented with N-isopentenyl adenine and indole-3-butyric acid. Since rooting was almost impossible in these thidiazuron-induced shoots, shoots were grafted onto young pea seedlings and regenerated to fertile plants.Abbreviations 2ip N-isopentenyl adenine - IAA indole-3-acetic acid - IBA indole-3-butyric acid - MES 2-(Nmorpholino)ethane acid - NAA naphthaleneacetic acid - TDZ thidiazuron - BAP 6-benzylaminopurine - PEG polyethylene glycol     

Plant regeneration from in vitro leaf tissues of <Emphasis Type="Italic">Viburnum dentatum</Emphasis> L

Shoots were regenerated from in vitro leaf tissues of two genotypes of Viburnum dentatum, a popular shrub species for landscape use. Adventitious shoots were induced when leaf tissues were cultured on woody plant medium (WPM) supplemented with either benzyladenine (BA) or thidiazuron (TDZ). Effects of cytokinin concentration, indole-3-butyric acid (IBA), and dark treatment on shoot regeneration were investigated. Dark treatment for the first 4 weeks of leaf explants cultured in the regeneration medium significantly increased the frequency of regeneration. The highest frequency of shoot regeneration (70%) for ‘Synnesvedt’ was obtained when leaf tissues were cultured in the medium with 40 μM BA or 8 μM TDZ with 4 weeks dark treatment. The highest frequency of shoot regeneration (90%) for ‘MN34’ was found in the 4 μM TDZ medium with 4 weeks dark treatment. Addition of IBA significantly enhanced shoot regeneration. Ethyl methanesulfonate (EMS) treatment inhibited callus proliferation, particularly in the early stage of callus recovery; however, no significant difference in shoot regeneration among different treatments was observed, indicating that the inhibitory effect of EMS was minimal after calluses re-acquired their capacity to grow and regenerate in the regular medium. Regenerated shoots (>1.5 cm) were rooted in the half-strength MS medium containing 5-10 μM IBA or naphthalene acetic acid (NAA). Rooted plants were transferred to the potting medium and grown in the greenhouse.     

TDZ诱导甘露子茎段高频再生

以茎段为外植体,经TDZ诱导后,建立了甘露子的高频再生体系,其中以附加1．0mg/L TDZ的效果最好,经过8周的培养平均每个外植体可以获得37．5个不定芽,在MS培养基上生根以后,经过炼苗的再生植株可有效地定植于土壤中,组织学观察表明再生植株多数是通过器官发生途径获得的,也有少数是通过体胚途径发生的。     

Artemisinin production by shoot regeneration of Artemisia annua L. using thidiazuron

An efficient in vitro method for multiple shoot bud induction and regeneration has been developed in Artemisia annua L. using leaf and stem explants in various concentrations and combinations of plant growth regulators to evaluate the frequency of regeneration. The sources of explants as well as plant growth regulators in the medium were found to influence the multiple shoot induction. The result shows that the stem segment cultured on Murashige and Skoog (MS) medium supplemented with 0.1 mg/l thidiazuron (TDZ) gave a perfect shoot formation (100%) and good shoot multiplication (57 shoots/explant) after 2 weeks of culture. Healthy regenerated shoots were elongated and rooted in MS medium without hormones. The artemisinin content in plants regenerated from stem explants using 0.1 mg/l TDZ was (3.36 +/- 0.36) microg/mg dry weight and two-fold higher than that of in vitro grown plants of the same age [(1.73 -/+ 0.23) microg/mg DW]. This system exhibited a potential for a rapid propagation of shoots from the stem explant and makes it possible to develop a clonal propagation of A. annua.     

白及假鳞茎薄片诱导不定芽及快繁体系的构建

以白及(Bletilla striata)假鳞茎为外植体,根据上、中、下部位切取薄片,探索假鳞茎不同部位BA、NAA和TDZ对假鳞茎薄片诱导不定芽的影响,比较假鳞茎薄片不同厚度对褐化率和出芽数的影响,采用正交试验,研究了BA和NAA对不定芽增殖的效果,并对组培苗进行壮苗、生根和移栽。结果表明:假鳞茎的部位对诱导不定芽作用极显著,下部的出芽率显著高于上部和中部,BA和TDZ对诱导不定芽作用显著,NAA对诱导不定芽作用不显著。最佳诱导不定芽的方式为假鳞茎下部薄片在基本培养基+2.0 mg·L^-1BA+1.0 mg·L^-1TDZ的培养基上培养4周,出芽率为93.3%,出芽数为15个,厚度为1.6~2.0 mm的假鳞茎薄片其褐化率最低。最佳的增殖培养基为基本培养基+1.5 mg·L^-1BA+0.3 mg·L^-1NAA,增殖系数达4.3,平均苗高为7.8 cm。本研究成功建立了白及假鳞茎薄片诱导芽为关键技术的快繁技术体系,为白及种质资源创新奠定了基础。     

Micropropagation of <Emphasis Type="Italic">Cotoneaster wilsonii</Emphasis> Nakai—a rare endemic ornamental plant

A simple and efficient micropropagation system was developed for Cotoneaster wilsonii through node and shoot tip explants obtained from mature field-grown plants. Of the two explants, node explants were found to be the most effective for axillary shoot proliferation. The highest frequency of shoot induction was achieved when nodal explants were incubated on Murashige and Skoog (MS) medium supplemented with 0.5 mg L⁻¹ thidiazuron (TDZ) and 0.1 mg L⁻¹ α- naphthaleneacetic acid (NAA) with an average of 34 shoots per explant. The microshoots were separated from the multiple shoots and subcultured on MS medium supplemented with 3% (w/v) sucrose and 0.8% (w/v) agar for further shoot growth. Maximum rooting was obtained on half-strength MS medium supplemented with 0.5 mg L⁻¹ indole-3-butyric acid (IBA). The in vitro-grown plantlets were successfully acclimatized in a glasshouse with 98% of survival. High concentrations of TDZ (1.5–2.0 mg L⁻¹) and repeated subcultures resulted hyperhydric shoots. Supplementation of the culture medium with silicon significantly reduced the induction of hyperhydric shoots. Increasing silicon concentration significantly decreased malondialdehyde content of the regenerated shoots. Data indicate that addition of silicon to the culture medium can effectively control hyperhydricity.     

Thidiazuron induced high frequency axillary shoot multiplication in Psoralea corylifolia

The effect of thidiazuron (TDZ) was studied on in vitro axillary shoot proliferation from nodal explant of Psoralea corylifolia - an endangered medicinal plant. Proliferation of shoots was achieved on Murashige and Skoog (MS) medium supplemented with 0.5, 1, 2, 3, 4 and 5 μM TDZ. The maximum number (13.6 ± 1.4) of shoots per explant were obtained from nodal segment cultured on 2 μM TDZ for 4 weeks and this increased to 29.7 ± 2.1 on hormone free MS medium after 8 weeks. The in vitro proliferated and elongated shoots were transferred individually on a root induction medium containing 0.5 μM indole-3-butyric acid (IBA) and within 4 weeks 4.5 ± 0.5 roots per shoot were produced. The regenerated plantlets were transferred to 1:1 soil and vermiculite mixture and acclimatized with 80 % survival rate. Fully acclimatized plants were grown in garden soil in greenhouse and their morphological and physiological parameters were comparable with seedlings.     

In vitro shoot multiplication and plantlet regeneration from nodal explants of <Emphasis Type="Italic">Cassia angustifolia </Emphasis>(Vahl.): a medicinal plant

An efficient, rapid and reproducible plant regeneration protocol was successfully developed for Cassia angustifolia using nodal explants excised from 14-day-old aseptic seedlings. Of the two cytokinins, 6-benzyladenine (BA) and thidiazuron (TDZ) evaluated as supplements to Murashige and Skoog (MS) medium, TDZ at an optimal concentration of 5.0 μM was effective in inducing multiple shoots. The highest rate of shoot multiplication was achieved on MS medium supplemented with 5.0 μM TDZ and 1.0 μM indole-3-acetic acid (IAA) at pH 5.8. The regenerated shoots when subcultured on hormone free MS medium considerably increased the rate of shoot multiplication and shoot length by end of fourth subculture passage. Rooting was achieved on the isolated shoots using MS medium with 60 μM indole -3- butyric acid (IBA) and 1% activated charcoal for 1 week and subsequently transferring the shootlets to half strength MS liquid media without IBA and activated charcoal. The in vitro raised plantlets with well-developed shoot and roots were successfully established in earthen pots containing garden soil and grown in greenhouse.     

In vitro organogenesis from internode derived callus cultures of Capsicum annuum L.

An efficient protocol of shoot organogenesis and plant regeneration from internode derived callus has been developed for Capsicum annuum. Optimal callus was developed from internodal segments on Murashige and Skoog (MS) medium supplemented with 10 μM 2,4-dichlorophenoxy acetic acid (2,4-D) and 2.0 μM 6-benzyladenine (BA). Shoot differentiation was achieved from the surface of callus when transferred on shoot induction medium containing BA and thidiazuron (TDZ) alone or in combination. The highest number of de novo adventitious shoots (25.4?±?1.42) and shoot length (4.6?±?0.37 cm) was recorded on MS medium supplemented with 5.0 μM BA and 2.5 μM TDZ. The individual elongated shoots were rooted well on MS medium supplemented with 1.0 μM Indole-3-butyric acid (IBA). The in vitro raised plantlets with properly developed shoot and roots were acclimatized successfully and grew well in the greenhouse. All the regenerated plants appeared normal with respect to morphology and growth characteristics with 85% survival rate.     

Efficient plant regeneration through callus in Zanthoxylum armatum DC: an endangered medicinal plant of the Indian Himalayan region

Abstract

An efficient in vitro propagation protocol for Zanthoxylum armatum DC has been developed via indirect organogenesis using aseptic leaf explants. The explants were soaked for different time duration (12, 24 or 36?h) in liquid woody plant medium (WPM) supplemented with various concentrations (15.0, 25.0 or 50.0?μM) of thidiazuron (TDZ). The pre-exposed explants transferred for callus induction onto WPM supplemented with different concentrations of TDZ (2.0, 4.0, 6.0, 8.0 and 10.0?μM) either alone or in combination with varied concentrations (0.5, 1 and 1.5?μM) of naphthaleneacetic acid (NAA). Of the tested concentrations and combinations, best response for pretreated (15?μM TDZ for 24?h) explants was achieved on WPM augmented with 6.0?μM TDZ and 0.5?μM NAA after 8?weeks of incubation. For shoot induction, the callus clumps were excised into small pieces (～0.5?g) and were transferred onto WPM fortified with different concentrations (2.0–9.0?μM) of benzylaminopurine (BA), indole-3-acetic acid (IAA, 1.0?μM) and gibberellic acid (GA_3, 0.5–3.0?μM). Maximum shoot number (10.4?±?0.74) and average shoot length (4.75?±?0.71?cm) were observed in WPM enriched with 2.0?μM BAP, 1.0?μM IAA and 1.5?μM GA₃ after 8?weeks of incubation. The developed shoots (4?cm) were excised, pulse-treated for 24?h in half-strength WPM containing indole-3-butyric acid (IBA, 50.0?μM) prior to their transfer on hormone-free MS medium, where 100% rooting was achieved. The regenerated plants were implanted in soil-filled poly bags, acclimatized properly and subsequently placed under sunlight with 80% survival rate after 60?days recorded. This is the first report for propagation of Z. armatum via callus phase with high rate of shoot proliferation and can be effectively utilized for generating sufficient planting material in promoting its re-cultivation and conservation programme.     

甜瓜属种间杂交中3种不同倍性资源的微繁

以3种不同倍性的甜瓜属种间杂交资源(双二倍体Cucumis hytivus、异源三倍体黄瓜和种间杂种F1)无菌苗的顶芽或腋芽为试材,比较它们再生能力的差异,寻求各自增殖的最适培养基,从而建立其离体繁殖体系.结果表明:倍性越高,增殖能力越小,增殖速度也最小(双二倍体种转接的间隔时间比其它两种材料长1～2周).双二倍体、异源三倍体黄瓜和种间杂种F1的最大繁殖系数分别为4.6、7.1和8.4;相对应的培养基分别为MS 0.05 mg·L-1TDZ、MS 2.2 mg·L-1 6-BA和MS 0.5 mg·L-1 6-BA).3种材料的生根结果基本一致,在1/2MS 0.2 mg·L-1IBA上的生根率均能达84.5%,驯化成活率达90%以上.同一材料的田间性状表现整齐一致,没有发现变异.     

High frequency multiple shoot induction from nodal segments and rhinacanthin production in the medicinal shrub Rhinacanthus nasutus (L.) Kurz

High frequency multiple shoots have been induced from nodal segments of Rhinacanthus nasutus (L.) Kurz., a potent anticancerous ethnomedicinal plant. For initiation of cultures, nodal segments were cultured on MS medium supplemented with various concentrations (1.0–5.0 μM) of 6-benzyladenine or thidiazuron (TDZ) alone or in combination with α-naphthalene acetic acid (NAA 0.5–1.0 μM). The optimum frequency of response (85 %) and shoot number (3.3) was observed on MS medium supplemented with 4.0 μM TDZ and 0.8 μM NAA. The shoots developed on initiation media were excised and nodal segments were subcultured on MS medium supplemented with TDZ (4.0 μM) and NAA (0.5–1.0 μM). This subculturing process was repeated thrice, each with 45 days of duration and the multiple shoot formation was recorded at the end of every subculture stage. The highest frequency of response (100 %) and number of multiple shoots (24.1) per explant were recorded at the end of the third subculture passage on MS medium supplemented with 4.0 μM TDZ and 0.8 μM NAA. The optimum rooting of shoots was observed on ½ MS medium fortified with 3.0 μM indole-3-butyric acid. The rooted plants were successfully transplanted to soil. The estimation of rhinacanthin (RC) content in shoots and roots was carried out in 6-month-old ex vitro plants (i.e., plants regenerated via in vitro culture) and field grown natural plants by high performance liquid chromatography. Both shoots and roots of naturally grown plants showed slightly higher RC content than ex vitro grown plants. The highest RC content (4.6 mg/g DW RC-C, 0.14 mg/g DW RC-D and 0.10 mg/g DW RC-N) was recorded in roots of naturally grown plants.     

Influence of thidiazuron (TDZ) pretreatment of shoot tips on shoot multiplication and ex vitro acclimatization of Harpagophytum procumbens

The effects of thidiazuron (TDZ) pretreatment of shoot tips on Harpagophytum procumbens shoot proliferation and successive stages of micropropagation, i.e. rooting of regenerated shoots and acclimatization of plantlets to ex vitro conditions, were described in the present study. The best response in terms of shoot proliferation (about seven shoots/explant) and shoot length (3.2 ± 0.4 cm) was obtained when explants pretreated with 25 µmol L^?1 TDZ for 6 h were cultured on Schenk and Hildebrandt medium containing indole-3-acetic acid (IAA) (0.57 µmol L^?1) and 6-benzylaminopurine (BAP) (8 µmol L^?1). Under these conditions, a 330 % increase in shoot multiplication over TDZ non-pretreatment culture was achieved and TDZ pretreatment shoots were longer compared to those in control culture (2.6 ± 0.3 cm). The TDZ pretreatment did not affect the percentage of rooted shoots, length of roots and number of roots formed per shoot. The rooted plantlets were transplanted from in vitro to pots with soil and grown during 1 year in the greenhouse. The hardening process was difficult and time-consuming. We found that the plants developed from the TDZ pretreated culture were superior to plants from non-pretreated culture in terms of survival rate and morphological features, such as shoot length, leaf size, flowering and earlier root tuberisation. Random amplified polymorphic DNA and inter-simple sequence repeat analyses of pretreatment with TDZ plants showed genetic similarity to non-pretreatment plants. We conclude that applying the strategy of initial explant pretreatment with TDZ may be valuable for the improvement in H. procumbens in vitro propagation.     

Peroxidase and catalase activities are involved in direct adventitious shoot formation induced by thidiazuron in eastern white pine (Pinus strobus L.) zygotic embryos.

We reported establishment of an efficient plant regeneration procedure through direct adventitious shoot (DAS) formation from cotyledons and hypocotyls of eastern white pine (Pinus strobus L.) mature embryos in this investigation. Multiple DASs were initiated from cotyledons of embryos on PS medium containing N6-benzyladenine (BA), thidiazuron (TDZ), or kinetin (KIN). Among different concentrations of casein enzymatic hydrosylate (CH) and glutamine used in this study, 500 mg l(-1) CH or 600 mg l(-1) glutamine induced the highest frequency of DAS formation. Rooting of regenerated shoots was obtained on PS medium supplemented with 0.01-0.1 microM indole-3-acetic acid (IAA) with the highest frequency on medium containing 0.01 muM IAA. No DASs were obtained on medium without TDZ. Measurement of peroxidase (POD) and catalase (CAT) activity during direct shoot induction and differentiation demonstrated that the lowest POD activity appeared in the 5-6th week of culture and lowest CAT activity occurred in the 7-8th week of culture on medium with TDZ. No such a change in POD and CAT activities was observed on medium without TDZ. These results demonstrated that POD and CAT activities were involved in DAS formation induced by TDZ in eastern white pine.     

A highly efficient plant regeneration system through multiple shoot differentiation from commercial cultivars of barley (Hordeum vulgare L.) using meristematic shoot segments excised from germinated mature embryos

A fast and highly efficient short-term in vitro regeneration system was developed for barley (Hordeum vulgare L.) based on readily available explants. Clumps of multiple shoots and buds suitable for transformation were obtained 9–10 weeks after culture initiation from model and current commercial cultivars. Meristematic shoot segments (MSSs) excised from mature embryo-derived seedlings and subsequently cultured on MS-based medium containing 2 mg/l Picloram and 3 mg/l thidiazuron (TDZ) differentiated up to ten multiple shoots after 3–4 weeks with no or very little callus formation. Sectors of the already multiplied shoot clumps were further multiplied on proliferation-maintenance medium containing 2 mg/l Picloram and 2.5 mg/l TDZ. Biweekly subcultures resulted in a continuous process of multiplication of these highly differentiating green sectors without any loss of morphogenic potential. The differentiated small shoots and shoot buds gave rise to normal shoots on medium with 0.1 mg/l Picloram and 1 mg/l TDZ. After rooting on basal medium with 0.5 mg/l or 1 mg/l IBA the plants were transferred to soil and showed normal growth and fertility compared to the seed-grown plants. All of the genotypes tested formed multiple shoots. The percentage of relative MSS multiplication was 63–83%, and the average number of multiplied shoots per MSS ranged from 16 to 34 among the genotypes after 9–11 weeks.Abbreviations BAP 6-Benzylaminopurine - 2,4-D 2,4-Dichlorophenoxyacetic acid - Dicamba 3,6-Dichloro-2-methoxybenzoic acid - IBA Indole-3-butyric acid - MSS Meristematic shoot segment - NAA -Naphthaleneacetic acid - Picloram 4-Amino-3,5,6-trichloropicolinic acid - TDZ Thidiazuron     

Improvement of shoot morphogenesis in vitro and assessment of changes of the activity of antioxidant enzymes during acclimation of micropropagated plants of Desert Teak

The aim of the study was to develop an improved and rapid regeneration system via axillary shoot proliferation for Tecomella undulata, an important endangered medicinal tree in India. The Murashige and Skoog (MS) medium augmented with different concentrations (0.1, 0.3, 0.5, 0.7, 1.0, 2.5, 5.0, 7.5 and 10.0 μM) of Thidiazuron (TDZ) was tested for the ability to induce axillary shoot development from cotyledonary node explants excised from 7-day-old sterile seedlings. Among the tried concentrations of TDZ, 0.7 μM showed optimum response in inducing maximum number (25.00 ± 2.30) of shoots and shoot length (4.06 ± 0.58 cm) after 3 weeks of incubation. The regenerated shoots when subcultured on MS medium lacking TDZ gave twofold shoot multiplication rate with maximum number (43.00 ± 2.86) of shoots per explant and longer shoot length (7.40 ± 0.34 cm) during the fourth subculture passage. Attempts were also made to study the morphogenetic effect of medium pH and sucrose concentration on axillary shoot induction and proliferation. The highest efficiency of shoot regeneration was recorded in MS medium supplemented with 0.7 μM TDZ and 3% sucrose at pH 5.8 after 3 weeks of culture. Different concentrations of Indole-3-butyric acid (IBA) were tested to determine the optimum conditions for ex vitro rooting of microshoots. The best result was accomplished with IBA (200 μM) pulse treatment given to the basal end of the microshoots for 30 min followed by their transfer in plastic cups containing soilrite and eventually established in natural soil with 80% survival rate. During acclimatization of plants, catalase (CAT) activity increased reaching maximum at 28th day after transplantation, while superoxide dismutase (SOD) activity increased reaching a maximum in the 21 days. Likewise, changes in the glutathione reductase (GR) and Ascorbate peroxidase (APX) were also detected. The observed changes reflect the plant’s capacity to develop antioxidant mechanisms during acclimatization which determine the ability to survive in oxidative stress. The standardized protocol for mass propagation of Tecomella undulata should eliminate the dependence on natural stands of plants for pharmaceutical purposes, and will also serve as a means of conservation as the species is highly overexploited.