Tomato Growth in Soil Amended with Sugar Mill By-Products Compost

Sugar mill by-products compost may be a good soil amendment to promote tomato (Lycopersicon esculentum L.) growth. In addition, the compost may further promote plant growth by inoculation with N₂-fixing bacteria. Compost from sugar-mill waste was prepared with and without the N₂-fixing bacteria, Azotobacter vinelandii, Beijerinckia derxii and Azospirillum sp. and incubated for 50 days. Each compost type was added to 10 kg of soil in pots at rates of 0, 15, and 45 g with and without fertilizer N at rates of 0, 0.75, and 1.54 g. A blanket application of P and K was applied to all pots. Shoot and root dry weights and N content of the whole plant was measured at 55 days. Dry weight of tomato shoots was increased by 40% by addition of fertilizer N and root weight was increased by 66%. Without fertilizer N the high rate of inoculated compost increased shoot growth 180% and uninoculated compost increased shoot growth 112%. For most treatments with and without fertilizer N, inoculated compost enhanced shoot growth and nitrogen content more than uninoculated compost. Root weights were nearly doubled by addition of either compost in comparison to the 0 N treatment. At the low rate of compost addition without fertilizer N, root weight was the same for uninoculated and inoculated compost but at the high rate of compost addition root weight was 32% higher for inoculated compost. The N₂-fixing bacteria colonized roots when inoculated compost was used. Sugar mill by-products compost proved to be an effective soil amendment for promoting the growth of tomato plants.     

<Emphasis Type="Italic">In vitro</Emphasis> regeneration and plant establishment of <Emphasis Type="Italic">Tylophora indica</Emphasis> (Burm. F.) Merrill: Petiole callus culture

Summary A protocol has been developed for high-frequency shoot regeneration and plant establishment of Tylophora indica from petiole-derived callus. Optimal callus was developed from petiole explants on Murashige and Skoog basal medium supplemented with 10μM2,4-dichlorophenoxyacetic acid +2,5μM thidiazuron (TDZ). Adventitious shoot induction was achieved from the surface of the callus after transferring onto shoot induction medium. The highest rate (90%) of shoot multiplication was achieved on MS medium containing 2.5μM TDZ. Individual elongated shoots were rooted best on halfstrength MS medium containing 0.5μM indole-3-butyric acid (IBA). When the basal cut ends of the in vitro-regenerated shoots were dipped in 150μM IBA for 30 min followed by transplantation in plastic pots containing sterile vermiculite, a mean of 4.1 roots per shoot developed. The in vitro-raised plantlets with well-developed shoot and roots were successfully established in earthen pots containing garden soil and grown in a greenhouse with 100% survival. Four months after transfer to pots, the performance of in vitro-propagated plants of T. indica was evaluated on the basis of selected physiological parameters and compared with ex vitro plants of the same age.     

In vitro regeneration of Solanum nigrum with enhanced solasodine production

High frequency of shoot formation was achieved from Solanum nigrum L. leaves on Murashige and Skoog (MS) medium without any callusing stage. Shoot forming ability was more pronounced on leaves positioned dorsally. For shoot induction, 2.0 mg dm⁻³ benzylaminopurine and 1.5 mg dm⁻³ kinetin were observed to be the most effective plant growth regulators (PGRs). The present paper also describes first successful induction of in vitro flowering in S. nigrum. The leaf derived shoots were excised and treated with various root promoting PGRs and 0.25 mg dm⁻³ indole-3-butyric acid produced maximum number of roots (15.2 per plant). Plants were later transplanted in field with 100 % survival. Solasodine content was higher in in vitro raised shoots and leaf derived callus, compared to ex vitro grown shoots.     

Colonization of root tissues and protection against Fusarium wilt of rye (Secale cereale) by nonpathogenic rhizosphere strains of Fusarium culmorum

Colonization of rye (Secale cereale) tissues by nonpathogenic rhizosphere Fusarium culmorum isolates DEMFc2 and DEMFc5 and a pathogenic strain DEMFc37, and their effect on plant fresh weight were studied in pot experiments. Both rhizosphere isolates colonized the epidermis and the cortex but were not found in vessels, while the pathogen colonized all three layers of root cells. The numbers of pathogen CFU isolated from plant tissues were much higher than those of the rhizosphere isolates in spite of the same number of macroconidia used as inoculum (1 × 10⁵ g⁻¹ of soil). Inoculation of seedlings with DEMFc2 resulted in a 20% increase, with DEMFc5 in more than a 20% reduction, and with DEMFc37 in a 38% reduction of shoot fresh weight of 14-day-old plants. Pre-colonization of plants with (either of) the rhizosphere isolates and subsequent inoculation with the pathogen resulted in plant weights the same as those observed in plants inoculated with the rhizosphere strain alone. The disease severity index for shoots of plants pre-colonized with DEMFc2 was reduced from class 4 (86% diseased plants) observed for plants inoculated with the pathogen alone to class 2 (average of 8% diseased plants) when pre-treated with the rhizosphere strain. The CFU number of the pathogen isolated from the interior of roots of plants pre-colonized with the rhizosphere isolates was as low as 10% of the number isolated from plants inoculated with the pathogen alone. A study of in vitro interactions between the rhizosphere isolates and the pathogen suggests that changes in plant colonization by the pathogen and its effect on fresh weight of plants pre-colonized with the rhizosphere isolates were not connected with inhibition of its growth by a direct action of the rhizosphere isolates. The results suggest that strain DEMFc2 can be considered as a potential biocontrol agent.     

Effects of cadmium and nickel on in vivo carbon dioxide exchange rate of pigeon pea (Cajanus cajan L.)

Effects of acidic soil factors (Al, H-ion, Mo, and Mn) upon the soybean (Glycine max (L.) Merr. cv. Essex)/Bradyrhizobium japonicum symbiosis were examined in acidified soil. Plants were grown under full sunlight in pots containing N-deficient soil (pH 6.7) or similar soil amended with sufficient Al₂(SO₄)₃ or elemental S to give soil pH values of 4.8 and 4.6, respectively, and water-extractable Al levels of 30 and 14 M, respectively. Other treatments consisted of the addition of inorganic N or inoculation with commercial or locally-isolated B. japonicum. Acidification did not reduce shoot or root weights of plants receiving inorganic N but reduced (P0.05) shoot and root dry weights, nodule dry weights and numbers, shoot N concentrations, and chlorophyll levels of inoculated plants. Shoot dry weights and nodulation of inoculated plants were greater (P0.05) in Al₂(SO₄)₃-amended soil than in S-amended soil. Addition of Mo was not beneficial. It was concluded that reduced plant growth was caused by the effects of acidified soil on nodulation and that H-ion toxicity was probably the most limiting factor. Effects of Al, Mn, or Mo appeared less likely.     

Contribution of the saprobic fungi Trametes versicolor and Trichoderma harzianum and the arbuscular mycorrhizal fungi Glomus deserticola and G. claroideum to arsenic tolerance of Eucalyptus globulus

The presence of high concentrations of arsenic (As) decreased the shoot and root dry weight, chlorophyll and P and Mg content of Eucalyptus globulus colonized with the arbuscular mycorrhizal (AM) fungi Glomus deserticola or G. claroideum, but these parameters were higher than in non-AM plants. As increased the percentage of AM length colonization and succinate dehydrogenase (SDH) activity in the root of E. globulus. Trichoderma harzianum, but not Trametes versicolor, increased the shoot and root dry weight, chlorophyll content, the percentage of AM root length colonization and SDH activity of E. globulus in presence of all As concentrations applied to soil when was inoculated together with G. claroideum. AM fungi increased shoot As and P concentration of E. globulus to higher level than the non-AM inoculated controls. The contribution of the AM and saprobe fungi to the translocation of As from root to shoot of E. globulus is discussed.     

Micropropagation of photinia employing rhizobacteria to promote root development

An alternative protocol was developed for in vitro propagation of photinia (Photinia × fraseri Dress), an ornamental shrub, using the plant growth-promoting rhizobacteria (PGPR) Azospirillum brasilense and Azotobacter chroococcum during rhizogenesis. Shoot tips from four-year-old mature plants, cut in spring and summer, were used as initial explants. They were cultured on Murashige–Skoog (MS) medium with Gamborg’s vitamins, N⁶-benzyladenine (BA: 11.1 μM) and gibberellic acid (GA₃: 1.3 μM), obtaining 63% of established explants. The highest shoot length (22.9 mm) and multiplication rate (4.3) was achieved by cultivating for four weeks in the same basal medium supplemented with 4.4 μM BA. Both auxin induction and bacterial inoculation were used for rooting. Elongated shoots were treated with two concentrations of indole-3-butyric acid (IBA: 4.9 or 49.2 μM) during 6 days for auxin induction. Then, the shoots were transferred to an auxin-free medium and inoculated with A. brasilense Cd, Sp7 or A. chroococcum (local strain). Bacterial inoculation induced earlier rooting of photinia shoots. A. brasilense Cd with 49.2 μM IBA pulse showed a significant increase (P ≤ 0.05) in root fresh and dry weight (105%, 137%), root surface area (65%) and shoot fresh and dry weight (32%, 62%). A. brasilense Sp7 enhanced the root fresh weight (34%) and root surface area (41%) while no significant differences with A. chroococcum inoculation were detected. The PGPR inoculated micro-cuttings in combination with auxin induction pulses may play a useful role in root organogenesis of micropropagated plants.     

Endophytic fungus Piriformospora indica induced systemic resistance against rice sheath blight via affecting hydrogen peroxide and antioxidants

In this study, the effect of endophytic fungus Piriformospora indica on Rhizoctonia solani AG1-IA, causal agent of sheath blight disease, was investigated. In addition, plant defence responses activated in P. indica-inoculated rice plants were analysed. Two-week-old seedlings were inoculated by dipping their roots in P. indica chlamydospore suspension and transferred to pots containing sterilized soil. After two weeks, the seedlings pre-inoculated with P. indica were inoculated with R. solani. Statistical analysis of biological indicators showed that application of P. indica increased both fresh and dry weight of rice shoots and roots, compared to those of uninoculated healthy controls and the samples only inoculated with R. solani. Accumulation of hydrogen peroxide (H₂O₂) and activity of antioxidants such as superoxide dismutase (SOD) and guaiacol peroxidase (GPX) in plants inoculated with P. indica, R. solani, and P. indica-R. solani were investigated. The obtained results revealed that P. indica not only increased the plant biomass, but also delayed the infection process of R. solani and decreased sheath blight severity. Decreased severity of the disease was associated with decreased levels of H₂O₂ and increased SOD activity. Considering the necessity of reducing fungicide application, using P. indica in seedling bed before transplantation to the field could be a novel and effective method to increase rice production and decrease sheath blight progress.     

印度梨形孢对铁皮石斛种子萌发和原球茎生长的影响

为探究印度梨形孢(Piriformospora indica)对铁皮石斛(Dendrobium officinale)种子萌发和原球茎生长的影响,在铁皮石斛种子离体培养和原球茎生长阶段分别接种印度梨形孢,对其形态发育特征和生理特性进行研究。结果表明,接种印度梨形孢的铁皮石斛种子的起始萌发时间提前,接种印度梨形孢的铁皮石斛原球茎的株高、鲜质量、干质量、叶片数、叶长、叶宽、根数、根长等都高于对照。在接种60 d的种子萌发率为68.8%,高于对照(28.6%);在接种80 d的铁皮石斛原球茎的叶绿素a、叶绿素b、总叶绿素含量显著高于对照,叶绿素a/b小于对照,净光合速率、气孔导度、蒸腾速率均高于对照,药用多糖含量显著高于对照。因此,接种印度梨形孢能促进铁皮石斛种子萌发和原球茎的生长,提高铁皮石斛的品质和产量。     

Plant growth‐promoting effects of native Pseudomonas strains on Mentha piperita (peppermint): an in vitro study

Plant growth‐promoting rhizobacteria (PGPR) affect growth of host plants through various direct and indirect mechanisms. Three native PGPR (Pseudomonas putida) strains isolated from rhizospheric soil of a Mentha piperita (peppermint) crop field near Córdoba, Argentina, were characterised and screened in vitro for plant growth‐promoting characteristics, such as indole‐3‐acetic acid (IAA) production, phosphate solubilisation and siderophore production, effects of direct inoculation on plant growth parameters (shoot fresh weight, root dry weight, leaf number, node number) and accumulation and composition of essential oils. Each of the three native strains was capable of phosphate solubilisation and IAA production. Only strain SJ04 produced siderophores. Plants directly inoculated with the native PGPR strains showed increased shoot fresh weight, glandular trichome number, ramification number and root dry weight in comparison with controls. The inoculated plants had increased essential oil yield (without alteration of essential oil composition) and biosynthesis of major essential oil components. Native strains of P. putida and other PGPR have clear potential as bio‐inoculants for improving productivity of aromatic crop plants. There have been no comparative studies on the role of inoculation with native strains on plant growth and secondary metabolite production (specially monoterpenes). Native bacterial isolates are generally preferable for inoculation of crop plants because they are already adapted to the environment and have a competitive advantage over non‐native strains.     

环链虫草Cordyceps cateniannulata对番茄生长和抗氧化酶活性的影响

【目的】评估环链虫草Cordyceps cateniannulata对植物促生和植物抗氧化酶活性的影响。【方法】本研究利用浸种法将环链虫草接种于番茄植物体,在接种后的第30天和60天,通过番茄株高、根长、地上和地下部分的干鲜重指标评价其对番茄生长的影响;在接种后第10、20、30、60和90天,通过选择性培养基分析其在番茄不同组织中的生存情况,使用形态学及DNA序列比对的方法检验所分离菌株与原有菌株的一致性。在处理后的第30天,检测番茄叶片中的过氧化物酶(POD)、过氧化氢酶(CAT)、超氧化物歧化酶(SOD)及丙二醛(MDA)含量,观察环链虫草对番茄的抗氧化酶活性影响。【结果】环链虫草可定殖于番茄幼苗且对番茄生长有显著促进作用,菌株对植株的定殖偏好性分别为根部>茎部>叶部。酶活检测结果表明,处理组番茄叶片防御酶活性均呈显著升高的趋势,其中POD、CAT、SOD活性分别比对照增加了52.21%、75.31%和158.59%,MDA含量下降了35.15%。【结论】环链虫草可以通过浸种的方法感染并定殖番茄幼苗的根、茎、叶,促进番茄幼苗的生长并提高番茄抗氧化酶活性,具有较好的田间...     

Effect of Arbuscular Mycorrhizal Fungi On Yield and Phytoremediation Performance of Pot Marigold (Calendula officinalis L.) Under Heavy Metals Stress

In order to study the effect of mycorrhizal fungi (inoculated and non-inoculated) and heavy metals stress [0, Pb (150 and 300 mg/kg) and Cd (40 and 80 mg/kg)] on pot marigold (Calendula officinalis L.), a factorial experiment was conducted based on a randomized complete block design with 4 replications in Research Greenhouse of Department of Horticultural Sciences, University of Tehran, Iran, during 2012–2013. Plant height, herbal and flower fresh and dry weight, root fresh and dry weight and root volume, colonization percentage, total petal extract, total petal flavonoids, root and shoot P and K uptakes, and Pb and Cd accumulations in root and shoot were measured. Results indicated that with increasing soil Pb and Cd concentration, growth and yield of pot marigold was reduced significantly; Cd had greater negative impacts than Pb. However, mycorrhizal fungi alleviated these impacts by improving plant growth and yield. Pot marigold concentrated high amounts of Pb and especially Cd in its roots and shoots; mycorrhizal plants had a greater accumulation of these metals, so that those under 80 mg/kg Cd soil^?1 accumulated 833.3 and 1585.8 mg Cd in their shoots and roots, respectively. In conclusion, mycorrhizal fungi can improve not only growth and yield of pot marigold in heavy metal stressed condition, but also phytoremediation performance by increasing heavy metals accumulation in the plant organs.     

Impact of Piriformospora indica on tomato growth and on interaction with fungal and viral pathogens

Piriformospora indica is a root endophytic fungus with plant-promoting properties in numerous plant species and induces resistance against root and shoot pathogens in barley, wheat, and Arabidopsis. A study over several years showed that the endophyte P. indica colonised the roots of the most consumed vegetable crop tomato. P. indica improved the growth of tomato resulting in increased biomass of leaves by up to 20%. Limitation of disease severity caused by Verticillium dahliae by more than 30% was observed on tomato plants colonised by the endophyte. Further experiments were carried out in hydroponic cultures which are commonly used for the indoor production of tomatoes in central Europe. After adaptation of inoculation techniques (inoculum density, plant stage), it was shown that P. indica influences the concentration of Pepino mosaic virus in tomato shoots. The outcome of the interaction seems to be affected by light intensity. Most importantly, the endophyte increases tomato fruit biomass in hydroponic culture concerning fresh weight (up to 100%) and dry matter content (up to 20%). Hence, P. indica represents a suitable growth promoting endophyte for tomato which can be applied in production systems of this important vegetable plant not only in soil, but also in hydroponic cultures.     

Influence of in vitro growth conditions on in vitro and ex vitro photosynthetic rates of easy- and difficult-to-acclimatize sea oats (Uniola paniculata L.) genotypes

Net photosynthetic rates (P _n) of easy (EK 16-3) and difficult-to-acclimatize (EK 11-1) sea oats genotypes were examined under the following culture conditions: (1) photoautotrophic [sugar-free medium, high photosynthetic photon flux (PPF), high vessel ventilation rates and CO₂ enrichment, (PA)]; (2) modified photomixotrophic [sugar-containing medium diluted with sugar-free medium over time, high PPF, and high vessel ventilation rates (PM)]; (3) modified photomixotrophic enriched [same as PM with CO₂ enrichment, (PME)]; or (4) conventional photomixotrophic [sugar-containing medium, low PPF, and low vessel ventilation rates (control)]. Regardless of genotype, plantlets cultured under PA conditions died within 2 wk, whereas under PM and PME conditions, plantlets increased their P _n. After 6 wk, P _n per gram dry weight was 1.7 times greater in EK 16-3 than EK 11-1 plantlets cultured under PME conditions. In vitro-produced leaves of EK 16-3 plantlets were elongated with expanded blades, whereas EK 11-1 produced short leaves without expanded blades, especially under control conditions. After in vitro culture, EK 16-3 PME plantlets exhibited the highest dry weights among treatments. EK 16-3 PME and EK 16-3 PM had similarly high survivability, shoot and root dry weights and leaf lengths ex vitro compared to EK 16-3 control and EK 11-1 PM and PME plantlets. Ex vitro growth, survivability and P _n per leaf area of either genotype were not affected by CO₂ enrichment under modified photomixotrophic conditions. These results suggest that growth and survivability of sea oats genotypes with different acclimatization capacities can be enhanced by optimizing culture conditions.     

Efficient shoot formation on internodal segments and alkaloid formation in the regenerates of Cephaelis ipecacuanha A. Richard

Rapid shoot proliferation was established by adventitious shoot formation on internodal segments. Cross sections of the shoot initiation area were observed microscopically and adventitious shoots were studied under the scanning electron microscope. Shoots were directly formed on the epidermis of internodal segments in vitro without callusing, but not on that of nodal segments with axillary buds. The use of media containing 0.01 – 0.1 mg/l 6-benzyladenine or 0.1 mg /l kinetin and culture under 16 h light increased the number of shoots per segment. The shoots thus obtained were rooted on phytohormone-free Woody Plant or Gamborg B5 solid medium, and were then transferred to soil. When potted, these grew well in a greenhouse. The emetic alkaloid content of adventitious shoots and regenerated plants was determined by HPLC. In vitro shoots cultured in Woody Plant liquid medium supplemented with 0.01 – 0.1 mg/l 6-benzyladenine contained 0.04 – 0.07 % dry wt. emetine and 0.4 – 0.5 % dry wt. cephaeline. One-year old regenerated plants cultivated in a greenhouse demonstrated the same alkaloid content (roots contained 0.82 % dry wt. emetine and 2.16 % dry wt. cephaeline) as the parental plant.Abbreviations MS Murashige — Skoog (Murashige and Skoog 1962) - 1/2 MS half strength MS - B5 Gamborg B5 (Gamborg et al. 1968)] - WP woody plant (Lloyd and McCown 1980) - RC root culture (Thomas and Davey 1982) - HF phytohormone free - BA 6-benzyladenine - Kin kinetin - SEM scanning electron microscopy - RDF rotating drum fermenter     

Smoke-water improves shoot growth and indigo accumulation in shoots of Isatis indigotica seedlings

Although smoke treatments have successfully been used for promoting the germination of many seeds, there are no reports on the effect of smoke on secondary metabolite production in plants. This study highlights the effects of smoke-water on shoot growth and accumulation of indigo in shoots of Isatis indigotica Fort., a Chinese medicinal plant. Results showed that seedlings treated with smoke-water at a dilution of 1:1000 achieved the highest fresh and dry shoot mass, which was significantly (P < 0.05) different from the control and other smoke-water dilutions tested. A higher concentration of smoke-water (1:500) significantly increased (P < 0.05) the indigo concentration in shoots compared with untreated seedlings. The effect of 1:1000 and 1:2000 dilutions gave non-significant increases in the concentration of indigo. These findings suggest the possible use of smoke-water for promoting the growth and accumulation of secondary metabolites in plants.     

Reducing the allelopathic effect of Partheniumhysterophorus L. on wheat (Triticumaestivum L.) by Pseudomonasputida

The present study evaluates the role of Pseudomonas putida NBRIC19 in alleviating biotic stress of Parthenium hysterophorus (Parthenium) in Triticum aestivum. Due to presence of Parthenium there was 43.76, 53.08 and 78.65% inhibition in root length, shoot length and dry weight of wheat respectively. This inhibition was recovered when P. putida NBRIC19 treatment resulted in 52.29, 28.73 and 76.31% increase in root length, shoot length and dry weight respectively as compared to control. P. putida NBRIC19 was able to form more biofilm under toxic environment of allelochemicals and enhanced expression of stress responsive genes in wheat. Inoculated wheat plants showed lower activity of catalase and ascorbate peroxidase under biotic stress of Parthenium indicating that inoculated plants felt less stress as compared to uninoculated plants. Microbial community structure in bacterized and nonbacterized wheat rhizosphere in presence and absence of Parthenium, was investigated using Biolog. There was significant increase in microbial diversity in P. putida NBRIC19 bacterized wheat rhizosphere. Functional microbial diversity revealed that P. putida NBRIC19 had shifted the microflora in such a manner that utilization of phytotoxic allelochemicals increased to lessen its toxic effect and finally it resulted in better growth of wheat in presence of Parthenium. Principal component analysis showed that microbial community function in nonbacterized wheat rhizosphere in presence (WPC) and absence (WC) of Parthenium is totally different from each other but due to P. putida NBRIC19 treatment there was close clustering of WPT and WT indicating a total shift in microbial community structure.     

内生真菌KLBMP0506对拟南芥的促生作用

【目的】研究植物内生菌Wickerhamomyces sp.KLBMP0506对拟南芥（Arabidopsis thaliana）的促生作用及潜在的促生机制。【方法】本研究以野生型拟南芥为试验材料,将其与菌株KLBMP0506进行平板共培养及盆栽接种试验,并测定拟南芥鲜重、干重、主根长、侧根数、叶绿素含量和可溶性糖含量等生长、生理指标,同时对筛选的与拟南芥侧根、主根形成及生长素合成和运输相关的11个基因进行实时荧光定量聚合酶链式反应（quantitative real-time polymerase chain reaction,qRT-PCR）分析。【结果】接种目的菌株KLBMP0506后,平板试验的拟南芥鲜重及侧根、盆栽试验的拟南芥鲜重、干重、茎长、叶绿素含量和可溶性糖含量均有一定程度的增加;分隔平板试验及菌株KLBMP0506发酵液中促生活性物质分析显示,该菌株产生的挥发性有机物质及其发酵液中的正丁醇和乙酸乙酯提取物均对拟南芥有明显的促生作用;此外,qRT-PCR分析显示KLBMP0506处理后,拟南芥中与侧根形成相关基因ABI4、FLA1的表达出现不同程度的下调,与生长素合成、运输相关基因AUX1、EIR1、YUC4的表达整体呈上调趋势,表明菌株KLBMP0506可能通过调控拟南芥中与侧根形成以及与生长素合成和运输相关基因的表达,而实现对拟南芥的促生作用。【结论】本研究明确了菌株KLBMP0506对模式植物拟南芥的促生作用,为其开发成为微生物菌肥提供理论依据。     

In vitro rapid regeneration of plantlets from nodal explants of Mucuna pruriens– a valuable medicinal plant

A rapid and efficient protocol for the large‐scale propagation of a potential medicinal plant, Mucuna pruriens, through in vitro culture of nodal segment explants obtained from 15‐day‐old aseptic seedlings is described. Of the three different cytokinins, 6‐benzyladenine (BA), kinetin (Kin) and 2‐isopentenyl adenine (2‐iP) evaluated as supplements to Murashige and Skoog (MS) medium, BA at an optimal concentration of 5.0 μM was effective in inducing multiple shoots. Strength of the basal media also influenced the efficiency of shoot regeneration. The frequency of shoot regeneration tended to increase when the salt concentration in the basal media was reduced. Highest number of multiple shoots (23.3) and maximum average length (5.6 cm) were standardised on half‐strength MS medium supplemented with 5.0 μM BA along with 0.5 μM α‐naphthalene acetic acid (NAA) at pH 5.8. Rooting was best induced in shoots excised from proliferated shoot cultures on MS medium augmented with an optimal concentration of 1.0 μM indole‐3‐butyric acid (IBA). The in vitro‐raised plantlets with well‐developed shoots and roots were successfully established in earthen pots containing garden soil and were grown in greenhouse with 90% survival rate. The results of this study provide the first report on in vitro plant regeneration of M. pruriens.