IN VITRO RESPONSE BY Terminalia arjuna GENOTYPES DURING MICROPROPAGATION

Terminalia arjuna is an important tree of the medicinal and sericulture industry, commonly known as Arjun. It’s bark is rich in secondary metabolites makes this plant highly valuable in medicine industry to treat cardiovascular disease. Overexploitation due to high demand in medicine, low seed germination, limitations of the conventional method of propagation push this plant towards being endangered. To conserve germplasm of such tree species and meet the requirement in medicinal industry, some non-conventional propagation method like micropropagation has been developed. The present work highlighted the effect of three genotypes (G-1, G-2, and G-3) on tissue culture of T. arjuna situated at Jodhpur, Rajasthan, India. In vitro shoot proliferation was achieved on a modified MS medium enriched with BAP + additives. Among the tested genotypes, Genotype -1 showed maximum bud break response (100%) followed by G-3 (93.33 %) and G-2 (86.66%).  Further multiplication of these shoots on modified MS medium containing BAP + NAA + additives gave 11.38±0.26 (G-1), 9.44±0.21 (G-2) and 10.22±0.32 (G-3) shoots. In vitro rooting was done by pulse treatment with IBA for 10 min prior to transfer on hormone free half strength MS medium containing 0.1% activated charcoal. Maximum in vitro rooting was obtained in G-1 (80%) followed by G-3 (71.11%) and G-2 (68.88%). In the present study, it was observed that optimum growth in all three genotypes required different doses of Plant Growth Regulator. Thus, by identifying and multiplying the best performing genotypes the gap between demand and supply of such medicinal plant can be fulfilled

Vegetative propagation of Azadirachta indica: effect of auxin and rooting media on adventitious root induction in mini-cuttings

Azadirachta indica (Neem), a prodigious multipurpose tree, has immense potential to benefit mankind and to protect the environment. In order to investigate the effects of the auxins hormone and rooting media on root formation in mini-cuttings of A. indica, an experiment was conducted using mist system in poly house in summer 2012. The experiment was established on three different rooting media: sand, vermiculite and soil assembled with six concentrations (100; 250; 500; 750; 1,000 and 1,500 mg L-1) of the auxins IBA (indole-3-butyric acid), IAA (indole-3-acetic acid) and NAA (α-naphthalene acetic acid) with randomized block design, each block containing four replicates. The basal long soak methods were used for treating cuttings. The data revealed significant effect of different auxins and rooting media on the Neem establishment. The rooted mini-cuttings were assessed for rooting percent, number of roots, root length and number of leaves. In present finding among all the auxins and rooting media, , IBA (250 mg L-1) showed better results with sand in terms of rooting percent (80%), number of roots (70.63), root length (11.13) and number of leaves (5.25) per rooted mini-cuttings. The formation of healthy plants after hardening under outdoor conditions showed that A. indica could be successfully propagated by mini-cuttings techniques

Regeneration of Complete Plantlets from Callus Culture of Azadirachta indica A. Juss using Immature Flower Buds

Plant regeneration via callus culture was achieved using immature flower buds of mature tree of Azadirachta indica A. Juss on semi-solid MS medium enriched with 9.05µM 2,4-D + additives. The regenerated callus were amplified on lower concentration of  4.52 µM  2,4-D + additives for micro-shoot regeneration study, the best micro-shoot regenerated from callus was achieved on MS medium supplemented with 8.88  µM  BAP with 3.78±0.50  numbers of shoot and 3.66±0.57 shoot length. The micro-shoots were amplified on 8.88 µM BAP+ 0.27 µM NAA with 5.22±0.53 numbers of shoot and 4.62±0.59 shoot length respectively. Micro shoot were successfully rooted on MS medium supplemented with IBA 4.92 µM with rooting percent (100%), 4.35±0.22 root number and 3.95±0.23 root length. The callus derived plantlets were hardened in vitro and ex vitro and acclimatized in poly house followed by agro-shade house. The efficiency of the protocol will be helpful for propagation of Azadirachta indica plants on a mass scale

VEGETATIVE PROPAGATION OF Azadirachta indica A. JUSS (NEEM) THROUGH CUTTINGS: A REVIEW

PROPAGAÇÃO VEGETATIVA DO NIM INDIANO (Azadirachta indica A. Juss) POR MEIO DE ESTACAS: UMA REVISÃO Azadirachta indica A. Juss (nim indiano) é uma importante espécie florestal que apresenta múltiplos usos. Hoje, o nim está recebendo o reconhecimento mundial por sua grande variedade de compostos bioativos. Considera uma herança biológica significativa, também é reconhecida como um dos presentes da natureza para a humanidade. Este trabalho tem como objetivo realizar uma revisão da literatura sobre os principais métodos científicos utilizados para se obter clones de Azadirachta indica por meio de propagação vegetativa, avaliado perspectivas atuais e as tendências futuras para a aplicação de novas técnicas de clonagem para a produção em grande escala. A literatura descreve métodos de tratamento (mergulhia), tipos de regulador de crescimento (IBA, NAA e IAA), tipos de estacas (madeira dura, madeira semiduro, madeira macia, folhas e mini estacas) e enraizamento (areia, solo, vermiculita e areia + solo + FYM) durante diferentes épocas (estação chuvosa, inverno e verão). A divulgação de tais técnicas podem minimizar os custos, encurtar as etapas de produção e, consequentemente, reduzir o tempo de cultivo em laboratório. Palavras-chave: Azadirachta indica, propagação vegetativa, estacas, auxina. ABSTRACT Azadirachta indica A. Juss (Neem) is one of the important multipurpose tree species. Today, neem is receiving worldwide recognition for its variety of bioactive principle components. Neem, being an important part of our biological heritage and is also recognised as one of nature’s gift to mankind.  This review aims to perform a literature review of the main scientific methods used to obtain clones of Azadirachta indica, such as the processes of vegetative (macro) propagation and present perspectives and future trends for the application of new cloning techniques aiming for large scale for plant production. The literature describes methods (dip treatment), types of plant growth regulator (IBA, NAA & IAA), types of stem cuttings (hard wood, semi hard wood, soft wood, leafy & mini cuttings) and planted in rooting media (sand, soil, vermiculite & sand+soil+FYM) during different season (monsoon, winter & summer). The improvement aiming the disseminating such techniques can minimize costs, shorten production stages and consequently, reduce the cultivation time in the laboratory. Keywords: Azadirachta indica, vegetative propagation, cuttings, auxin. DOI: http://dx.doi.org/10.14583/2318-7670.v02n04a1

VEGETATIVE PROPAGATION OF Azadirachta indica A. JUSS (NEEM) THROUGH CUTTINGS: A REVIEW

PROPAGAÇÃO VEGETATIVA DO NIM INDIANO (Azadirachta indica A. Juss) POR MEIO DE ESTACAS: UMA REVISÃO Azadirachta indica A. Juss (nim indiano) é uma importante espécie florestal que apresenta múltiplos usos. Hoje, o nim está recebendo o reconhecimento mundial por sua grande variedade de compostos bioativos. Considera uma herança biológica significativa, também é reconhecida como um dos presentes da natureza para a humanidade. Este trabalho tem como objetivo realizar uma revisão da literatura sobre os principais métodos científicos utilizados para se obter clones de Azadirachta indica por meio de propagação vegetativa, avaliado perspectivas atuais e as tendências futuras para a aplicação de novas técnicas de clonagem para a produção em grande escala. A literatura descreve métodos de tratamento (mergulhia), tipos de regulador de crescimento (IBA, NAA e IAA), tipos de estacas (madeira dura, madeira semiduro, madeira macia, folhas e mini estacas) e enraizamento (areia, solo, vermiculita e areia + solo + FYM) durante diferentes épocas (estação chuvosa, inverno e verão). A divulgação de tais técnicas podem minimizar os custos, encurtar as etapas de produção e, consequentemente, reduzir o tempo de cultivo em laboratório. Palavras-chave: Azadirachta indica, propagação vegetativa, estacas, auxina. ABSTRACT Azadirachta indica A. Juss (Neem) is one of the important multipurpose tree species. Today, neem is receiving worldwide recognition for its variety of bioactive principle components. Neem, being an important part of our biological heritage and is also recognised as one of nature’s gift to mankind.  This review aims to perform a literature review of the main scientific methods used to obtain clones of Azadirachta indica, such as the processes of vegetative (macro) propagation and present perspectives and future trends for the application of new cloning techniques aiming for large scale for plant production. The literature describes methods (dip treatment), types of plant growth regulator (IBA, NAA & IAA), types of stem cuttings (hard wood, semi hard wood, soft wood, leafy & mini cuttings) and planted in rooting media (sand, soil, vermiculite & sand+soil+FYM) during different season (monsoon, winter & summer). The improvement aiming the disseminating such techniques can minimize costs, shorten production stages and consequently, reduce the cultivation time in the laboratory. Keywords: Azadirachta indica, vegetative propagation, cuttings, auxin. DOI: http://dx.doi.org/10.14583/2318-7670.v02n04a1

Influence of cutting diameter, auxin and rooting substrate on adventitious rooting from hardwood cuttings of Azadirachta indica A. Juss (Neem)

Azadirochta indica A. Juss (Neem) is a native Indian tree, well known for its medicinal features and have important to local economy. For Conservation of the genetic resources of neem, the effect of diameter (> 0.5; 0.5-1.5 and 1.5-2.5 cm), auxin, their different concentration in different rooting media on adventitious root formation (ARF) in hardwood cuttings of Azadirachta indica was studied. Three different rooting substrate (media) (sand, vermiculite and soil) were used and the experiment was established using three types of auxin (IBA, IAA and NAA) and 6 concentrations (100; 250; 500; 750; 1,000 and 1,500 mg L-1). Significant effects of cuttings diameter, auxin, their concentration and rooting substrate on adventitious rooting from neem hardwood cuttings were observed. The rooting percentage, number of roots, root length and number of leaves were evaluated. Among the all diameter of hardwood cuttings used, 0.5-1.5 cm diameter showed maximum rooting. IBA (500 mg L-1) resulted in higher rooting percentage (80%) with 6.82 sprouts, 53.06 roots, 7.13 cm root length and 7.0 leaves per rooted hardwood cuttings, when growing in sand rooting substrate. The determination of proper rooting protocols and the use of hardwood cuttings were proved important for propagation of A. indica

A micropropagation protocol for mass multiplication of Terminalia arjuna - a valuable medicinal tree

A reliable and efficient micropropagation protocol was developed through axillary shoot proliferation from nodal explants of mature Terminalia arjuna. Season of explants collection and maturity of explants showed direct influence on bud-break. Nodal stem segments collected during the months of April and May gave best response. Nodal segments of fresh sprouts originated from lopped tree of T. arjuna were used as explants for establishment of in vitro culture. Surface sterilized explants produced optimum number of shoots through activation of axillary buds - on modified Murashige and Skoog’s (M-MS) medium. Maximum (100%) in vitro shoot proliferation was obtained on M-MS medium supplemented with 8.86 µM BAP + additives (100 mg L-1 of ascorbic acid, 50 mg L-1 of citric acid, 50 mg L-1 of adenine sulphate and 25 mg L-1 PVP). Modified M-MS medium supplemented with 4.44 µM BAP + 0.54 µM NAA + additives was found to be best for 11.38±0.26 shoot multiplication. After four week of culturing the in vitro regenerated shoots were rooted when pulse treated with 984 µM IBA for 10 min and transferred on hormone free half strength MS medium containing 100 mg L-1 activated charcoal. In vitro regenerated plants were transferred to field after gradual hardening and acclimatization procedure. Present method can be used for large scale commercial production of this medicinally important tree

Micropropagation of Barleria prionitis L. var. dicantha: an ethnomedicinal plant

Barleria prionitis var. dicantha Blatt & Hallb is an important endemic, ethno medicinal plant of Rajasthan, commonly known as Vajaradanti. It is especially well known for its antidontalgic properties. It is a rich source of glycosides, steroids, tannins and flavonoids. Natural habitat loss due to urbanization, high habitat specificity and exploitation for medicinal values places considerable pressure on native population of this endemic endangered plant. There is a need for development of non-conventional methods for propagation and conservation of B. prionitis var. dicantha. The present study was undertaken with the aim to set up a protocol for in vitro propagation of this medicinally important plant. For this, nodal segments were obtained from in vitro raised seedling on half strength MS medium supplemented with 2.88 µM GA3. For axillary bud break, nodal segments were inoculated on different concentrations of BA, Kn or TDZ supplemented MS medium. Maximum (75.5%) bud break was obtained on MS medium supplemented with 8.88 µM BA + additives with an average of 3.82±0.13 shoots explant-1 and 1.74±0.08 cm shoot length. The elongated shoots were excised from mother plant and further multiplied on MS medium supplemented with 4.44 µM BA. For rooting in vitro raised shoots were cultured on MS medium supplemented with different concentrations IBA or IAA. The maximum rooting was 75.5% obtained on 2.46 µM IBA with 4±0.13 roots explant-1 and 3.44±0.04 cm root length. The in vitro raised plantlets were successfully hardened and acclimatized in polybags at shade house

Role of tryptophan on in vitro rooting in microshoots of Azadirachta indica A. Juss (Neem)

Development of the root is a major step in the rapid propagation of plants under in vitro conditions. In the present study, an efficient and rapid protocol was developed for the in vitro regeneration of Azadirachta indica through axillary shoot proliferation in nodal segments collected from mature trees. For axillary shoot proliferation nodal explants were inoculated on MS medium supplemented with different concentration of BAP + additives. Maximum 100% axillary shoot proliferation with 3.95 number of shoot regenerated and 3.60 cm length of regenerated shoots was obtained on MS medium supplemented with BAP 8.88 µM. In vitro regenerated shoots were further multiplied on MS medium supplemented with different concentration of BAP + additives. Best shoot multiplication in terms of 4.80 number of shoot and 3.95 cm length of regenerated shoots was observed on MS medium supplemented with 4.44 µM BAP. For in vitro rooting, regenerated microshoots were cultured on MS medium supplemented with different concentration of tryptophan + additives. The highest rooting percent (100%), number of roots (10.30) and mean root length (6.33 cm) was observed on MS medium supplemented with 146.89 µM tryptophan within 7-8 days of inoculation. The in vitro rooted plantlets were successfully hardened and acclimatized in poly house. These plants showed a good survival rate of 95% under field conditions