Plant Tissue culture and micropropagation

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Some aspects of the tissue culture and micropropagation of Boronia megastigma Nees

Some aspects of the tissue culture and micropropagation of Boronia megastigma Nees

LITERATURE REVIEW 3 The Uses of Plant Tissue Culture and Micropropagation 3 4 Clonal Propagation Difficulties Associated With Micropropagation 6 Plant Health and Germplasm Storage 7 Germ[r]

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A critical appraisal on plant tissue culture studies in medicinally important plants

A critical appraisal on plant tissue culture studies in medicinally important plants

valuable source of different kinds of useful products like wood, medicine, fiber, firewood and timber suitable for furniture, as well as decorative plants, pharmaceutical products and aestic items. In recent years the relevance of plant tissue culture methods has gained thrust to meet the growing demands for pharmaceutical industries since the genetic diversity of medicinal plants is decreasing hap hazardously. Conventional propagation and high demand of planting material are the major constraints for the large scale multiplication of medicinal species which can be met economically and efficiently in short span of time by in vitro propagation. Micropropagation is an alternative means of propagation that can be engaged in the conservation of flora in relatively shorter time. Tissue culture is useful for multiplying and conserving the useful species which are difficult to regenerate and thus can be saved from extinction. There is an only partial advancement explored at the research level to suggest tissue culture studies in Sterculiaceae. Conclusively, this appraisal is the first of its kind which emphasizes the procedures available for in vitro propagation along with some of the remarkable achievements carried in some pharmaceutically and commercially important plants by in vitro culture techniques of plants from this family.
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Standardization of Protocol for in vitro Micropropagation of Phyllanthus niruri:An Important Medicinal Plant

Standardization of Protocol for in vitro Micropropagation of Phyllanthus niruri:An Important Medicinal Plant

Plants are the traditional source for many chemicals used as pharmaceuticals, biochemicals, fragrances, food colours and flavours. Near about 1,000 plant varieties out of 17.000 plants were commonly used for traditional herbal medicines in Ayurveda, Unani, siddha and Amchi. From many years the traditional medicines was made by wildly grown local plants. Phytochemicals extracted from different parts of plant is used for the treatment of various diseases, for supplementing nutrition in foods and cosmetic industries has a great potential in this century. Most valuable phytochemicals are products of secondary metabolism and possess sufficient chemical or structural complexity, so that artificial synthesis is difficult 1 . Plant tissue culture techniques now play an important role in the micropropagation and quantitative improvement of the medicinally important plant. Though the conventional breeding techniques have considerably increased the productivity of modern crops, the application of biotechnology could speed up
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Standardization of micropropagation from nodal segments of costus pictus  an anti diabetic plant

Standardization of micropropagation from nodal segments of costus pictus an anti diabetic plant

The demand for the medicinal plant resources has been on the rise due to their extensive utilization in herbal health care formulations, cosmetic products and nutritional supplements. About 95% of the medicinal plants used are obtained from wild habitats, especially from forests. The continued commercial exploitation of these plants resulted in depletion tion of many important species from their natural habitats. In the recent years, Plant cell and tissue culture has scale multiplication of many plants. Plant tissue culture technology plays a significant shing a reliable protocol for conservation and large scale propagation of many plant species. The C. pictus (Family: Costaceae) is an important medicinal plant widely used in traditional and commercial formulations. It is commonly known as spiral ginger or crepe ginger, used as tonic, stimulant, diuretic, digestive, antiseptic and a major source of diosgenin , 1956, which is antidiabetic in nature and is used
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“A Review on Tissue Culture Studies in Eclipta alba – An Important Medicinal Plant” by Neeti Dhaka, India.

“A Review on Tissue Culture Studies in Eclipta alba – An Important Medicinal Plant” by Neeti Dhaka, India.

There is an increased focus on naturopathy and Ayurveda in recent times, which has lent prime focus upon medicinal plants. Their medicinal importance has gained attraction of firms and commercial exploiters. This has resulted in over-exploitation of these plants or their cultivation to a greater extent that is causing regional agro-ecological imbalances. Therefore, there is a need to propagate such practices that can help meet both increased demand and reduce stress on natural resources, viz., land, water and biotic material. Plant tissue culture technology is the right intervention to optimise the commercial utilisation of Eclipta alba that can maintain the biodiversity balance as well. Tissue culture protocols developed for this purpose are deployed for conserving the endangered species, rare therapeutic species and other identified species for specific purpose. The micropropagation/clonal techniques are used for mass scale multiplication from shoot tips and nodal segments. One can achieve large numbers in relatively short period of time and less space that is attractive to harness their medicinal properties commercially. Both, the yield of crop and its extracts,
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Coscinium fenestratum: Callus and  Suspension Cell Culture of the  Endangered Medicinal Plant Using  Vermicompost Extract and Coelomic  Fluid as Plant Tissue Culture Media

Coscinium fenestratum: Callus and Suspension Cell Culture of the Endangered Medicinal Plant Using Vermicompost Extract and Coelomic Fluid as Plant Tissue Culture Media

Coscinium fenestratum Colebr. (Menispermaceae) is widely spread across the Western Ghats (India) and Sri Lanka. It is commonly known as tree turmeric. The plant has mainly been used for treating diabetes mellitus in the traditional Siddha and Ayurvedic systems of medicine [1]. The stem portions contained many alkaloids like berberine, hentriacontane, ceryl alcohol, sitosterol, oleic acid, palmitic acid and saponin with some resinous ma- terial [2]. From roots, tertiary alkaloids, dihydroberlambine, berlambine and noroxyhydrastinine were isolated [3]. Antidiabetic activity of C. fenestratum has been reported [4]. The stem portion is suggested to have an- ti-inflammatory, thermogenic, antiseptic, tonic effects and used against inflammations, opthalmopathy, ulcers, general debility and jaundice [5]-[7]. Preliminary studies on micropropagation of C. fenestratum were reported [8]. Protocol was developed for obtaining berberine-producing callus and cell suspension cultures established from the petiole segments of C. fenestratum. With 4 mg/L of NAA, highest yield of alkaloid berberine was ac- quired and 2 mg/L of 2,4-D yielded the best cell growth [9]. Intercellular berberine and berberine recovered from the suspension media were studied [10].
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MICROPROPAGATION OF JUSTICIA ADHATODA: MEDICINAL PLANT OF FAMILY ACANTHACEAE

MICROPROPAGATION OF JUSTICIA ADHATODA: MEDICINAL PLANT OF FAMILY ACANTHACEAE

Plant tissue culture is an important method of obtaining valuable single plant or a hybrid produced through protoplast culture, anther culture, or genetic recombination.This can provide easy method of clonal propagation of selected plants. However chance occurring of “ploidy chimeras” as late Professor Neumann has called in his publication (Neumann et al., 2009) may be a problem. This can provide easy method of clonal propagation of selected plants. Recently molecular characterization of tissue culture produced plants has been attempted to check and monitor clonal fidelity (Kumar and Shekhawat 2009; Kumar, 2010). This has been of great importance in case of woody plants and fruit trees with immense economical potential besides horticultural crops and medicinal plants. To confirm the efficacy of the technique and genetic fidelity of the regenerants to make them commercially
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Micropropagation of Kebericho: An  Endandered Ethiopian Medicinal Plant

Micropropagation of Kebericho: An Endandered Ethiopian Medicinal Plant

Roots were not spontaneously induced during culture initiation and shoot multiplication. When microshoots were cultured on growth regulators free half strength MS medium, poor and few (1.0) numbers of roots were elicited with low (6.7%) frequency and 3.25 ± 0.25 cm elongation. NAA was found to be more conducive than IBA for root induction. The effectiveness of NAA in rooting has been reported for medicinal plants like Vitis thunbergii [29], Spilanthes acmella [30] and Artemisia scoparia [31]. In the present study, the highest number (8.30 ± 0.60) of roots per shoot was observed on 1/3 strength MS medium supplemented with 1.5 mg/l NAA. On this medium, 100% shoots developed roots. The present results showed that a reduction in MS salt concentration is the pre-dominant reason for the improved rooting of in vitro shoots. The promotional effect of reducing the salt concentration of MS on in vitro rooting of shoots has been reported in Stevia rebaudiana [32]. Roots formed on medium containing NAA were thick and long whereas those in IBA were thin, short and weak. NAA showed significant difference on all media strengths in terms of root number per shoot after 4 weeks of culture. In this study, rooting was recorded on all strength MS medium containing different concentrations of IBA, although root formation was accompanied with callusing at the base of shoots in all concentrations. Further application of IBA up to 2.5 mg/l decreased the percentage of root induction, root numbers, and root lengths significantly. This observation is in agreement with the report of Thomas [33] in Curculigo orchioides where a higher level of IBA produced a negative effect resulting in lower root number and root length. The reason for low performance of IBA treatments may be due to the higher stability of IBA which induces higher level of degradative metabolites in tissue, and thus, blocking the regeneration process [34]. Studies also showed that, transport velocity of IBA was slower than NAA, which favors callus formation [35]. In this study, 1/3 strength MS medium supplemented with 1.5 mg/l NAA proved to be most effective for inducing a strong and stout root system where roots were high in number (8.30 ± 0.60) with average length of 4.83 ± 0.32 cm. Acclimatization of in vitro rooted plantlets was successful, where 83% plants survived and established as healthy plant. This result was much better than previous report by Balcha [17] where only 25% of in vitro germinated seedlings survived after being transferred to the greenhouse.
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Thinking inside the Box: Tissue Culture for Plant Propagation in a Key Ecological Species, Andropogon gerardii

Thinking inside the Box: Tissue Culture for Plant Propagation in a Key Ecological Species, Andropogon gerardii

Micropropagation was started from greenhouse potted genotypes of A. gerardii selected by M. Avolio from a single population at the Konza Prairie Biological Station in Manhattan, KS [41]. Plants were excised below actively growing me- ristem using tools dipped in 70% isopropyl alcohol. Rhizome was included whe- rever possible. Excised plants were then sterilized using a 0.5% bleach solution with gentle shaking before transporting to a laminar flow hood with HEPA filter. Meristematic regions were identified as rhizomatous nodes and gently scrubbed with 3% hydrogen peroxide to remove any residual soil. Nodes were excised and placed on a rotary shaker in 3% hydrogen peroxide for at least 2 hours. Nodes were then separated into separate tubes and shaken for 10 minutes in sterile wa- ter with 10% bleach and 50 μL per L of Tween-20 solution. Meristematic nodes were then placed on 10 mL solid “3BX” MS-based growth media [50] for shoot initiation (Table 1). Due to fungal or bacterial presence, contaminated tubes were frequently swapped for fresh media during an 8 week period to establish sterile cultures.
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In Vitro Micropropagation of the Ornamental Plant Dieffenbachia A Review

In Vitro Micropropagation of the Ornamental Plant Dieffenbachia A Review

Most of the tissue culture works, for different purposes, in Dieffenbachia were successfully achieved on agar based MS [43] medium (full or half-strength salt formulations). El-Sawy and Bakheet [26], Feng et al. [20], Hui et al. [37], Iqbal et al. [29], Mogollon [27], Jun et al. [25] El-Mahrouk et al. [14], El-Mahrouk et al. [42], Shen and Lee, [23] Elsheikh and Khalfalla [24] and Abass et al., [44] employed full MS for bud proliferation, callus induction and rapid multiplication of Dieffenbachia. However, some modifications on MS medium were stated to improve multiplication such as Shen and Lee [34] which used full MS with 2% sucrose and 1% glucose, El-Mahrouk et al. [3] and Chao and Li-si [22] exploited half-strength MS medium and Shen and Lee [35] as well as added 2% glucose. However, previously, Taylor and Knauss [19] developed DM basic medium for tissue culture of Dieffenbachia which is identical to MS medium except for the addition of adenine sulphate 80 mg/L, and NaH2PO4.H2O 170 mg/L. Also their experimentation showed that doubling the concentration of adenine sulphate and NaH2PO4-H2O did not increase development of the tissue cultures. Moreover the addition of nicotinic acid, 0.5 mg/L, pyridoxine-HCl, 0.5 mg/L, or glycine, 2 mg/L, alone or in combination, to be unnecessary for explant development. More recent, Sierra et al. [21] and Henny et al. [2] used DM for micropropagation of Dieffenbachia. Even though, no study demonstrates the difference between MS and DM media formulas. Beside MS and DM media, LS medium [45] was as well reported for micropropagation of Dieffenbachia spp. [41]. Moreover, other media including B5 [46] and N6 [47] were used for rapid propagation but as modifications to MS medium such as macro elements [40] or B5 vitamins [28].
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MICROPROPAGATION OF AN ENDANGERED AND ENDEMIC MEDICINAL PLANT CAYRATIA PEDATA VAR  GLABRA

MICROPROPAGATION OF AN ENDANGERED AND ENDEMIC MEDICINAL PLANT CAYRATIA PEDATA VAR GLABRA

Tissue culture technique is now oriented toward the commercial concept of large scale production of medicinally active bioactive compounds in important medicinal plants. The potentiality of the plant cell can be enhanced for the production of useful secondary metabolites by applying various in vitro techniques. It is imperative that, viable strategies have to be taken to conserve the surviving population; at least, the critically important medicinal species from further loss [10,11]. Since the species C. pedata var. glabra is rare and endangered due to ecological anthropological pressure, it is need to be conserved. As there is no strong micropropagation studies recorded on this medicinally potent climber, the present work was undertaken to lay down in vitro micropropagation as a conservation strategy to produce large scale plantlets within a short time without any genetic variation.
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Optimization of Culture Media Formulations for Micropropagation of Lepisanthes Fruticosa

Optimization of Culture Media Formulations for Micropropagation of Lepisanthes Fruticosa

In plant micropropagation, a balance between two groups of growth regulators, auxins and cytokinins, controls cell division and elongation (Soledad et al., 2012). The combination of BAP and NAA in tissue culture media is most effective for shoot formation. As reported in previous studies, the used of both BAP (0.5 – 1.0 mg/L) and NAA (0.02 mg/L) promoted shoot proliferation in Sequoia sempervirens (Boulay,1989). According to Hossain et al. (2016), the highest shoot regeneration was obtained using 5.0 mg/L BAP in exotic banana.Root formation was most successful on MS medium supplemented with 4.0 mg/L NAA. Findings by Zuraida et al. (2013) showed that nodal segments of V. planifolia cultured on a medium containing 1 mg/L BAP produced the highest number of shoots and the greatest lengths of shoots.
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Micropropagation of Curcuma Sp , a Threatened Medicinal Plant

Micropropagation of Curcuma Sp , a Threatened Medicinal Plant

In nature, Curcuma sp. has low reserve and is distributed in very narrow area in Kon Bong 2 village, Dakrong commune, Kbang district, Gialai province, Vietnam. It needs to be conserved and cultured. However, natural regeneration is rather limited and needs tissue culture to micropropagation. So, it needs to be propagated by tissue cul- ture. We have used medium MS (Murashige and Skoog, 1962) [2], but if we only use inorganic minerals in supply nutrition, then its tissue could not have division cell to become callus or propagate the root and shoot. We have improved the micropropaga- tion protocol: reducing the level of macro-nutrition and phytohormon, adding humate, and we have had the successful outcome, which is able to create multiple shoot and root induction and plantlet in vitro and move plantlet to ex-vitro . In order to make plantlet in vitro , we need to proceed below steps: 1) select the organs to culture tissue and its sterilized methods; 2) select the suitable culture medium; 3) select the medium and phytohormone for buds forming in vitro ; 4) select medium and phytohormone for root induction in vitro ; and 5) build plantlets in vitro . The duration of research is from 5/2015 to 4/2016 at Faculty of Agriculture and Forestry, Tay Nguyen University, Vietnam.
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A SUITABLE PROTOCOL FOR MICROPROPAGATION OF SPILANTHES CALVA DC  AN ENDEMIC MEDICINAL PLANT

A SUITABLE PROTOCOL FOR MICROPROPAGATION OF SPILANTHES CALVA DC AN ENDEMIC MEDICINAL PLANT

The tissue culture technique has been proved very efficient in rapid mass propagation and conservation of these endangered, threatened and rare species have been grown and conserved by micropropagation because of high coefficient of multiplication and small demands on a number of plant species (Akin-Idowu et al., 2009). Plant cell culture can be an alternative way to produce these compounds continuously under artificially controlled conditions (Ghani, 2003). In particular, the production of pharmaceutically important plant metabolites has been a target for practical application of plant cell culture for the last few decades (Lakshmi and Mythili, 2003).
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PROPAGATION AND CHEMICAL MUTGENIC INDUCTION OF EUSTOMA GRANDIFLORUM PLANT USING TISSUE CULTURE TECHNIQUE

PROPAGATION AND CHEMICAL MUTGENIC INDUCTION OF EUSTOMA GRANDIFLORUM PLANT USING TISSUE CULTURE TECHNIQUE

Generally, propagation of Eustoma grandiflorum is by seed or cutting, but the quality is not uniform due to the variations in flowering time, plant height and the number of flowers. Propagation of Eustoma grandiflorum by tissue culture technique are relatively low. Several factors like genotype, media, plant growth regulators and type of explants should effect the success of the micropropagation method, most of plant growth regulators that have been used were BA, KIN, NAA and IBA [3],[4]

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In vitro shoot micropropagation and plant establishment of an ornamental plant dumb cane (dieffenbachia compacta)

In vitro shoot micropropagation and plant establishment of an ornamental plant dumb cane (dieffenbachia compacta)

several propagation cycles. Also the traditional propagation using cutting is sometimes encountered with various difficulties such as fungal, bacterial and viral diseases (Poole and Chase, 1987). Consequently, the market demand for propagules is hardly met with such cuttings. As the growers of Dieffenbachia are looking for alternate sources, the use of tissue culture is a feasible alternative option for the rapid multiplication and maintenance of germplasm (Smith et al., 1991; Johnson and Emino, 1979). Subsequently, tissue culture was seen as a method whereby Dieffenbachia stock could be free from systemic viral and bacterial pathogens (Knauss, 1976, Taylor and Kanuss, 1978). Tissue culture has already proved to be successful mean for commercial in vitro propagation of several members of Dieffenbachia (Kanuss, 1976; Chase et al., 1981; Voyiatzi and Voyiatzis, 1989; Henny et al., 2000; El-Mahrouk et al., 2006). The present communication describes in vitro multiple shoot regeneration from nodal segment explants, and the rooting and successful greenhouse establishment of Dieffenbachia compacta, a popular indoor foliage plant in Sudan.
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MICROPROPAGATION OF Myrmecodia Tuberosa Jack.:A MEDICINAL PLANT FROM BORNEO

MICROPROPAGATION OF Myrmecodia Tuberosa Jack.:A MEDICINAL PLANT FROM BORNEO

A method of plant propagation by using tissue culture techniques is an alternative way to solve these problems. Tissue culture is a method to isolate parts of the plant such as protoplasm, cells, tissues and organs, and grow them in aseptic conditions, multiplying and regenerating into whole plants. Tissue culture techniques can produce plants in large numbers and uniform [5; 10]. There are several aspects to be considered when using micropropagation tehnique such as explants material culture media (including PGRs). The level concentration and type of PGRs may vary in order to have successful tissue culture [25]. Previous research revealed that the present of PGRs, viz: cytokinins (BA-6 benzylaminopurine/BAP) were proved responsible for cell division, cell elongation and to induce shoots from the explants [24]. Meanwhile, plant propagation with different explants source has been also established in Rubiacordifolia [21], Coffea arabica [8] and Oldenlandia umbellate [24]. [32] reported that M. pendens was successfully in vitro propagated although the multiplication rate was very low. However, the information regarding the effects of Plant growth regulators (PGRs) and source of explant on the Myrmecodia multiplication is limited. Thus, the current study was design to investigate M. tuberosa propagation by using tissue culture methods with different variation of PGRs such as BAP; Gibberrelic acid (GA 3 );
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INDIRECT ORGANOGENESIS OF THE MEDICINAL PLANT SPECIES, CRYPTOLEPIS GRANDIFLORA WIGHT (APOCYNACEAE) BY TISSUE CULTURE TECHNIQUE

INDIRECT ORGANOGENESIS OF THE MEDICINAL PLANT SPECIES, CRYPTOLEPIS GRANDIFLORA WIGHT (APOCYNACEAE) BY TISSUE CULTURE TECHNIQUE

Tissue culture technique is being increasingly exploited for clonal multiplication and in vitro conservation of valuable indigenous germplasm that are threatened with extinction (Boro et al., 1998). Micropropagation method is specifically applicable to species in which clonal propagation is needed (Gamborg and Phillips, 1995). Cryptolepis grandiflora is a climber belongs to the family, Apocynaceae, found in deciduous and moist deciduous forests and endemic to peninsular India (Henry et al., 1987). Its roots, stems and leaves are used for the treatment of bone fracture. It is used for diabetes also. Due to poor availability possibilities of facing threats by this species may be serve in course of time due to its medicinal importance and hence the demand. Therefore increasing the population size and ensuring the greater biomass availability are more essential to meet the demand and conserve the species as well. This paper describes the indirect organogenesis strategies developed by using tissue culture technology for this species under
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IN VITRO MICROPROPAGATION OF LAWSCNIA INERMIS (HENNA) A NATURAL DYE PLANT

IN VITRO MICROPROPAGATION OF LAWSCNIA INERMIS (HENNA) A NATURAL DYE PLANT

Materials required for equipment’s required for plant. Culture the necessary of required for setting up tissues culture laboratory are rested blow (Table-1) for a small scale simpler require can be used table 1.0 equipment require for plant tissues culture. (i) Gas, water, Electricity supply. (ii) Hot plate & magnetic stirrer. (iii) Sensitive balance. (iv) PH meter. (v) Distillation apparatus. (vi) D-ionizer for sterilization process. (vii) Autoclave & pressure cooker. (viii) Flaming instruments. (ix) Culture rake. (x) Tempt. Control unit. (xi) Laminar air flow cabinet. Techniques involve sterilization inoculation and regeneration of plant cell tissue organ under aseptic condition and culture vials constraining Synthetic nutrient medium. Both chemical composition of the medium and e controlled environment condition (temperature, light, humidity, aeration) effectively control the expression of any genotype or phenotypic potential in the explants. A tissue culture laboratory should be equipped to facilitate the following procedures. (i) Sufficient space for washing, sterilization of glass ware and other equipment’s. (ii) Aseptic transfer condition. (iii) Preparation sterilization & storage of nutrient medium. (iv) Bio chemical analysis of the material studies of cells & tissue etc.
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Micropropagation and conservation of Rheum webbianum collected from Zanaskar valley via tissue culture

Micropropagation and conservation of Rheum webbianum collected from Zanaskar valley via tissue culture

Subculturing was done after every 3-5 weeks, depending upon the organogenesis and proliferation potential of the explants of culture and was carried out in the Laminar air flow chamber under aseptic conditions. The plantlets obtained from different explants through repeated sub-cultures were finally left in culture vials with open mouth for three days in the incubation room, transferred to plastic pots containing soil-peat mixture and then taken out of incubation room of the lab. Attempts were made to acclimatize plants under the green house facility. Ten replicates were taken for each treatment and observations were recorded at the end of every week. Data were subjected to analysis of variance using SSPP software version 17.0 (SAS Institute Inc., Cary, NC). The growth response of explants under normal conditions and also the effect of plant growth hormone response (PGPR) treatments were considered significant according to the magnitude of the F value (P ≤ 0.005).
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