Concentrations Of 6-benzylaminopurine Research Articles

English

The objective of this study was to optimize a protocol for in vitro micropropagation of selected grape vine varieties. Preliminary studies were conducted to optimize the duration of sterilization of the explants. For shoot initiation experiment nodes of the three varieties of grape vines were cultured on MS medium (Murashige-Skoog’s, 1962) supplemented with five different concentrations of 6-Benzylaminopurine (BAP) and the control. Various experiments were carried out to optimize shoot multiplication using MS medium supplemented with different concentrations of BAP alone or in combination with Indole-3-butyric acid (IBA). To optimize root induction, different concentrations of Indole-3-acetic acid (IAA) were used. Sterilization of explants using 1% of NaOCl for 7 min duration was optimum. Chenin blanc showed high percentage of survival rate (96%) followed by Ugni blanc and Canonannon (88%) at 0.5 mg/L BAP. Among the different concentrations and combination of Plant Growth Regulators (PGRs) used for multiplication, maximum mean number of shoots 7.2, 6.7, and 6.1 was achieved at 1 mg/L BAP combined with 0.1 mg/L IBA for Chenin blanc, Canonnanon and Ugni blanc, respectively. All varieties induced root for all the treatments used including the control but good roots were found on MS medium supplemented with 2 and 4 mg/L of IAA. The plantlets were acclimatized in the glasshouse and survival percentage was 92% for Chenin blanc followed by 78.6 and 73.9% for Ugni blanc and Canonannon, respectively. Thus, the achievements of this study will play a big role in the grape vine culture program. Key words: Vitis vinifera L., acclimatization, explant, cytokinin, auxin.

In Vitro Adventitious Shoot Proliferation of Three Basil Species (Ocimum sp. L.) by Addition of Naphthalene Acetic Acid (NAA) and Benzyl Amino Purine (BAP)

Basils are mainly used as food flavoring and source of aromatic oil, but the leaves have also been used for herbals and cosmetics. Basil is propagated by seeds but the germination is often low in the field. This study aims to investigate the effect of different concentrations of NAA (Naphthalene Acetic Acid) and BAP (Benzyl Amino Purine) to induce the adventitious shoot proliferation of three species of basil (Ocimumsp. L.) in vitro. The research was conducted at the Tissue Culture Laboratory II of the Department of Agronomy and Horticulture, Bogor Agricultural University (IPB) from November 2016 till October 2017. This research consisted of three separate experiments with one basil species for each experiment, Lemon Basil, “Greek Minette”, and “Thai Siam Queen”. The experiments were organized in a completely randomized block design with two factors: NAA concentrations (0, 0.5, and 1 mg.L-1) and BAP (0, 1, 2, and 3 mg.L-1) using Murashige and Skoog (MS) medium supplied with 25 ml.L-1 coconut water. The three basil species responded differently to NAA and BAP in the media. The optimum concentration of NAA and BAP to induce callus growth in Lemon Basil was 0.5 mg.L-1 and 3 mg.L-1, respectively, with BAP alone showing the best effect on proliferation. In “Greek Minette”, however, the addition of NAA and BAP had no significant effect on the callus growth, proliferation, height, and number of leaf, node and root per explant. Media without growth regulators provide the best results in increasing height, explant proliferation rate, number of nodes and shoots, and percentage of proliferating shoots of “Thai Siam Queen”.Keywords: cytokinin, “Greek Minette”, Lemon Basil, “Thai Siam Queen”, synthetic auxin

Benzylaminopurine and indol-3-acetic acid concentrations in in vitro proliferation of Agave angustifolia adventitious shoots

Today, a micropropagation method for Agave angustifolia exists, but for the multiplication of propagules, more information is needed on the diverse components of the culture medium, cytokinins and auxins and their effect on organogenetic response. The objective of this study was to evaluate dosages of benzylaminopurine and indol-3-acetic acid (IAA) in a culture medium and their effect on the formation of new adventitious shoots (organogenesis). The explants used were clusters of shoots on a common base of A. angustifolia stem tissue. Clusters of three to four shoots were established in different variants of a Murashige and Skoog culture medium that had different concentrations of benzylaminopurine (BAP; 0.5, 1, 2, and 4 mg L-1) and IAA) (0.1, 0.3, and 1 mg L-1). After 16 wk, it was found that a higher proliferation of shoots was positively related to the concentration of growth regulators. Explants formed a greater quantity of shoots in a culture medium with an increase in BAP of up to 4 mg L-1 and in IAA of up to 1 mg L-1, and there was a synergistic effect with BAP to induce the formation of the greatest number of shoots.

The induction of shoots, multiplication, and rooting of Gyrinops versteegii (Gilg.) Domke By in vitro

Gyrinops versteegii (Gilg.) Domke is including one of superior agarwood-producing plants and naturally growing in Eastern Indonesia as Nusa Tenggara and Papua. Indonesia has been trading agarwood products both domestically and overseas which one of them is agarwood produced by G.versteegii. This study purpose was to develop an in vitro culture method for mass propagation of G. versteegii. Shoot induction conducted on MS medium supplemented with Benzyl Amino Purine (BAP) 0.7; 1.0; 1.5; and 2.0 mg/l. The shoots multiplication conducted on MS medium supplemented with the best concentration of BAP from shoot induction phase. The rooting of shoots conducted on half strength MS medium supplemented with interaction of Napthalene Acetic Acid (NAA) 0.01 mg/l with concentration of Indole-3-Butyric Acid (IBA) 1.0; 2.0; 3.0; and 4.0 mg/l. Epicotyl explant with a given concentration level of BAP 0.7 mg/l produce the highest rates number of shoots and shoot length compared to other explant respectively 4.8 shoots and 0.41 cm within 6 weeks. The best explant developments in the best medium able to promote the growth of the length and number of shoots are 0.28 shoots and 0.3 cm within 4 weeks. Explants easiest, quickest and most high sprouting ability as a factor of success in terms of multiplication is epicotyl. The combination treatment of material explant with concentration of BAP only affect to growth of shoots length. The combination treatment of NAA with concentration of IBA has no effect against root formation and growth rootlength.

THE INDUCTION OF SHOOTS, MULTIPLICATION, AND ROOTING OF Gyrinops versteegii (Gilg.) DOMKE BY IN VITRO

Gyrinops versteegii (Gilg.) Domke is including one of superior agarwood-producing plants and naturally growing in Eastern Indonesia as Nusa Tenggara and Papua. Indonesia has been trading agarwood products both domestically and overseas which one of them is agarwood produced by G.versteegii. This study purpose was to develop an in vitro culture method for mass propagation of G. versteegii. Shoot induction conducted on MS medium supplemented with Benzyl Amino Purine (BAP) 0.7; 1.0; 1.5; and 2.0 mg/l. The shoots multiplication conducted on MS medium supplemented with the best concentration of BAP from shoot induction phase. The rooting of shoots conducted on half strength MS medium supplemented with interaction of Napthalene Acetic Acid (NAA) 0.01 mg/l with concentration of Indole-3-Butyric Acid (IBA) 1.0; 2.0; 3.0; and 4.0 mg/l. Epicotyl explant with a given concentration level of BAP 0.7 mg/l produce the highest rates number of shoots and shoot length compared to other explant respectively 4.8 shoots and 0.41 cm within 6 weeks. The best explant developments in the best medium able to promote the growth of the length and number of shoots are 0.28 shoots and 0.3 cm within 4 weeks. Explants easiest, quickest and most high sprouting ability as a factor of success in terms of multiplication is epicotyl. The combination treatment of material explant with concentration of BAP only affect to growth of shoots length. The combination treatment of NAA with concentration of IBA has no effect against root formation and growth rootlength.

Establishing sterilization protocol and shoot induction medium for in vitro regeneration of sweetbriar (Rosa rubiginosa L.)

Rosa rubiginosa L. is popular in Lesotho where rural communities harvest the rosehips for sale to the local companies which process them into various products. There is, however, a challenge in establishing plantations of the species due to deep seed dormancy and so difficulty in germination. In this study, nodal explants of R. rubiginosa L. were sterilized with 20 g·L-1 Ridomil Gold MZ (RGM) and cultured on Murashige and Skoog (MS) medium supplemented with various concentrations of 6-benzylaminopurine (BAP). The explants treated with 20 g·L-1 RGM showed resistant against fungal contamination giving 73% survival explants. Moreover, addition of BAP in shoot induction medium increased shoot formation as compared to 29% response recorded on MS without BAP. Response of 100% shoot formation was obtained on MS medium with 2 mg·L-1 BAP. Key words: 6-Benzylaminopurine, nodal explants, ridomil gold mz.

PERTUMBUHAN KALUS DAUN MELON (Cucumis melo) VARIETAS MAI 119 DENGAN PEMBERIAN BAP (BENZYL AMINO PURINE) DAN 2,4-D (2,4 DICHLOROPHENOXYACETIC ACID)

This study aims to determine the concentration of growth regulators BAP and 2,4-D which have the highest effect in stimulating the formation of callus melon plants (Cucumis melo) Mai 119 variety. Completely randomized design (CRD) was used in this research. Media used on callus induction was MS medium with addition of several concentration of BAP (0 mg/L, 1 mg/L, 2 mg/L, 3 mg/L) and 2,4-D (0 mg/L, 1 mg/L, 2 mg/L, 3 mg/L) either alone or in a combination of both. Parameters measured were the time appearing of callus, callus diameter, callus texture, and callus color. Anova followed by Tukey's test was used to the analyse of time appearing of callus. Data of callus diameter was analyzed using Kruskal Wallis test followed by Mann-Whitney test. In the analysis of parameter related to the callus texture and callus color, descriptive test were used. The results showed that there were differences in the effect of growth regulators on the callus formation. The fastest callus induction and the largest diameter of callus were obtained on media with concentration of 2 mg/L BAP and 3 mg/L BAP.Keywords: BAP (benzyl amino purine), 2,4-D (2,4-dichlorophenoxyacetic acid), callus induction, melon (Cucumis melo) varieties Mai 119

Rooting of Stem Cuttings with Different Indole 3 Butyric Acid (IBA) Treatments and Development of Micropropagation Protocol for <i>Piper betle</i> L. Node Culture

The present study, conducted during 2016 and 2017 seasons, aimed to investigate the effect of IBA on rooting of Piper betle L. stem cuttings (softwood and semi-hardwood). The experiment was undertaken in misting house field 2 UPM using the sand media to determine the adventitious roots initiation and development using the histological method. The cuttings were treated with different IBA concentrations (0, 500, 1000, 1500 and 2000 mg/L). The nodes explants were used in the development of a protocol for in vitro propagation of P. betle L., with different concentrations of Clorox with different times of immersion (20% Clorox 10 minutes, 30% Clorox 10 minutes, 20% Clorox 20 minutes, and 30% 20 minutes). In multiplication of the plantlets, Murashige and Skoog (MS) medium with different concentrations of BAP (0, 0.5, 1.0, 2.0 mg/L) were used to investigate the rooting of the explants. The results indicated that the types of the cuttings were different in the rooting capacity and the length of the roots. Moreover, it was found that in comparison with the control treatment, by a rise in the concentrations of the IBA, there was a significant upsurge in the rooting percentage, the root diameter, and the number of the roots. The results indicated that the types of cutting with 1000, 1500 and 2000 mg/L IBA perform better in the root percentage (100%) in the semi hardwood cuttings. The best results, however, were 2000 mg/L IBA in the semi hardwood cuttings, with the number of the roots to be 35.05, and the fresh weight of the roots to be 3.94 g, the dry weight of the roots to be 0.33 g, the length of the roots to be 391.88 cm, the roots diameter to be 1.21 mm, the surface area of the roots to be 121.83 cm2, and the root volume to be 2.99 cm3. Nonetheless, the optimal concentration of Clorox with the time immersion was 20% with the 20-minute immersion time, which produced a shoot induction percentage of 30% dead explants and a mean number of 70.00 shoots per explant and the optimal concentration of benzylaminopurine (BAP) at 1.0 mg/L. It is of note that a shoot induction percentage of 22.29% and a mean number of 4.1% number of auxiliary bud per treatment. P. betle shoots in MS medium without PGR MS (0.0) yielded a good rooting.

English

Our previous study indicated that the tiger lily (Lilium lancifolium var. Flore Pleno) has a great ability to produce inflorescence bulbils in nature as a form of natural phenomenon of floral reversion in plants. This present research was carried out to investigate the artificial floral reversion in in vitro culture of two lilies (Asiatic hybrid cv.”Black out"), and (Lilium longiflorum cv "White heaven") based on the type and developmental stage of explants plus the different concentrations of naphthalene acetic acid (NAA) and benzyl aminopurine (BA). Developmental changes were observed in both lilies in response to floral reversion which was enhanced by growth regulators under in vitro condition. The regeneration of vegetative organs was associated with certain degeneration of floral organs. Large bulblets and multiple shoots were formed only in specific regions in floral organs, precisely in two attached points: the boundary region between the receptacle with other floral organs and the branching point where the peduncle joins the pedicel. This direct organogenesis was highly dependent on type of lily, type and developmental stage of explants in addition to the concentration of BA and NAA in in vitro culture. However, 1 mg/L BA combined with 0.1 mg/L NAA was the optimum for regenerating shoots and bulblets in in vitro culture of both lilies after six weeks. Key words: Floral reversion, organogenesis, lilies, active points, growth regulators, in vitro, bulblets.

Improved Micropropagation of Plantlets from Nodal Explants of Anchote (Coccinia abyssinica)—A Calcium- and Protein-rich Tuber

The effect of phytohormones on breaking of dormancy of axillary buds in anchote and their subsequent shoot proliferation were examined. Anchote is an annual trailing endemic plant with high calcium content grown for its edible tuberous roots in Ethiopia. Nodal explants harvested from the greenhouse were sterilized using various concentrations of a commercial bleach (JIK) which contains 3.85% sodium hypochlorite (NaOCl) and time duration. The highest (85%) clean explants were obtained when 5% JIK was used for 15 minutes. The explants were cultured on Murashige and Skoog (MS) medium supplemented with various concentrations of benzylaminopurine (BAP), kinetin, and thidiazuron (TDZ). The highest frequency of microshoot induction (84%) and mean number of microshoots (3.4) were recorded from explants cultured on medium supplemented with TDZ 0.025 µm. Hyperhydrated shoots were observed on media supplemented with high concentrations of BAP and kinetin but interestingly not on TDZ media. Induction of roots was highest (86%; 4.6 roots per shoot) when shoots were transferred to half strength MS medium containing 0.5 μm α-naphthalene acetic acid (NAA) after 12 days. A survival rate of 83% was recorded in the greenhouse and the plantlets appeared to be morphologically normal. This is the first report on use of TDZ for in vitro propagation of anchote.

English

Melia dubia Cav. (Meliaceae) is a multipurpose tree of tropical and subtropical regions mainly cultivated for its medicinal and industrial importance. Due to its versatile properties, it has been depleted in its natural environment. Moreover due to sluggish and poor seed germination, there is a threat of its gene pool exclusion from the natural habitat. The alternative method for conservation and efficient mass propagation is thus need of the hour. As per the extensive literature survey there is no report on efficient protocol for mass propagation of M. dubia through callus organogenesis. Therefore, the present work was aimed to develop in vitro organogenesis protocol for rapid and large scale production of planting material. From our results, maximum callus percentage, callus weight and fragile callus was observed on 1.0 mg/l benzylaminopurine (BAP) in combination with 0.5 mg/l naphthalene acetic acid (NAA). The callus differentiation was achieved at different concentrations of BAP and indole acetic acid (IAA). Multiple Shoot number per callus propagule 5.30 was observed on 0.5 mg/l BAP and 1 mg/l IAA concentration. The maximum rooting percentage (78.5%), root number per explant (4.33) and root length per explant (4.41 cm) was observed at 0.5 mg/l indol butyric acid (IBA) after 30 days of inoculation. Further the total flavonoid content, phenolic content and antioxidant properties of leaves of in-vitro regenerated plants where studied. Total flavonoids and phenolic content in leaves of in vitro Melia dubia was 0.56 ± 0.8 mg quercitin equivalent (QE) and 2.97 ± 0.17 mg gallic acid equivalent (GAE) respectively. The antioxidant property was further assed through measurement of DPPH radical scavenging activity. The in-vitro regeneration protocol can be exploited for commercial cultivation and fulfilling the growing demand for fresh explant material through mass propagation of M. dubia an economically important plant species. Key words:  Melia dubia, antioxidant, indole-3-butyric acid, flavonoids and phenolics.

In vitro multiplication of pear tree cultivar Cascatense

One main obstacle to the development and expansion of pear tree cultivation in Brazil has been the lack of cultivars adapted to the edaphoclimatic conditions of regions with the potential for pear production. Intending to overcome this hindrance, Embrapa Clima Temperado (Embrapa Temperate Climate) developed and released the cultivar Cascatense, with a low cold requirement (~400 h) better suited for Rio Grande do Sul`s and Santa Catarina`s climate. Still, seedlings of high genetic and sanitary quality are necessary for the establishment of a high-yield orchard. Plants with both sanitary quality and uniformity can be obtained through in vitro micropropagation. Thus, the present study aimed to assess the influence of different culture media (MS, MS¾, WPM, QL, and Himédia), concentrations of sucrose (0, 15, 30, 45, and 60 g L-1), and concentrations of benzylaminopurine (BAP) (0, 0.4, 0.8, 1.2, and 1.6 mg L-1) upon the in vitro multiplication of cultivar Cascatense, to establish an efficient protocol to enhance its propagation and reproducibility. The study included three sequential experiments in which the number of shoots per explant, the length of shoots, the number of axillary buds per shoot, fresh mass, and dry mass of shoots were evaluated. Cultivar Cascatense can be best propagated in vitro with the use of Himédia medium, supplemented with 0.8 mg L-1 of BAP and 30 g L-1 of sucrose.

In Vitro micropropagation of Acacia auriculiformis from selected juvenile sources

The effects of 6- Benzylaminopurine (BA), different basal medium, sucrose concentration and gelling agent were investigated for shoot induction and multiplication of Acacia auriculiformis. Nodal ex¬plants derived from 5-month-old seedlings yielded the highest shoot multiplication rate in Murashige and Skoog medium (MS) with 0.44 μM BA, 30 g/L sucrose and 2 g/L Gelrite. The highest mean number of shoots (10) and mean length of shoots (5.07mm) were also obtained in this medium. Qualitative obser¬vation of the shoots cultured in 0.44 μM BA were greener and vigorous in growth as compared to shoots cultured on higher concentrations of BA (22.2 μM). MS medium produced a significantly higher number of shoots (18) compared to Woody Plant Medium (WPM) (11) and B5 medium (10). Media solidified with different gelling agents also produced a significantly different number of shoots with 2 g/L Gelrite produced the highest number of shoots (23). The highest percentage of shoots rooted was found in the MS medium without any growth regulators (40.0%) followed by medium supplemented with Indole-3-bu¬tyric acid (IBA) at 9.84 μM and the combination of 9.84 μM IBA with 5.37 μM α-naphthalene acetic acid (NAA) (33.3%). MS medium without any plant growth regulators produced the highest mean root length (84.33mm), whereas medium supplemented with 9.84 μM IBA produced the highest mean number of roots per shoot (4.33). Out planting of in vitro rooted shoots in shredded coconut husk as the substrate gave the highest percentage of survival (90%) during acclimatization in the greenhouse.

Effect of Antioxidants, Amino Acids and Plant Growth Regulators on in vitro Propagation of Rosa centifolia.

Rosa centifoliais commercially propagated by asexual means but in vitro propagation ensure the production of disease free and healthy plants and browning of explants creates hurdle in their multiplication. The aim was to reduce oxidative browning of shoots of R. centifolia in MS medium during in vitro propagation. Axillary buds of R. centifolia were sterilized with 70% ethyl alcohol for 4 min and 5% sodium hypochlorite for 2 min followed by three washing with sterilized double distilled water. In order to control oxidative browning, Ascorbic acid (100 mg.L-1), citric acid (100 mg.L-1) and activated charcoal (3 g.L-1) were used while to control withering of shoots, different concentrations (3.0 mg.L-1, 6.0 mg.L-1, 9.0 mg.L-1) of either glutamine, asparagine and proline were put into trial. Different concentrations of Benzyl aminopurine (BAP) and naphthalene acetic acid (NAA) were used for in vitro shoot and root formation. Minimum browning percentage (20%) was achieved in the presence of activated charcoal (3.0 g.L-1) and pretreatment of explants with running tap water. Asparagin (9.0 mg.L-1) produced maximum shooting (93%), minimum withering (6.67%), and it took longer period (27 days) for shoots to wither. BAP (3.0 mg.L-1) + NAA (0.5 mg.L-1) was produced the highest number of shoots (1.63), in a shortest periods (9 days). For root production, NAA (1.5 mg.L-1) + BAP (0.5 mg.L-1) reduced the time to 11 days with maximum number of roots (4.33) and root length (4.20 cm). The supplement of activated charcoal (3.0 g.L-1), a sparagin (9.0 mg.L-1) and combination of BAP and NAA in the MS medium is effective for in vitro propagation of R. centifolia.

Somatic embryogenesis of Sandalwood (Santalum album L.)

Sandalwood (Santalum album L.) is native species of Indonesia, especially in East Nusa Tenggara, is oneof the twenty two species of the genus Santalum in the world. Sandalwood is an important tree because it hashigh economic value can produce sandal oil these can be used for perfumes, cosmetics, pharmaceuticals, andare often used in religious ceremonies. In vitro particularly somatic embryogenesis has been widely appliedin the propagation of sandalwood. The Objective of this research is to obtain regeneration of sandalwoodthrough somatic embryogenesis using leaves explant from various clones. Medium for embryo induction is MS(Murashige and Skoog, 1962) solid medium containing treatment of 2,4-D (2,4-Dichlorophenoxyacetic acid)at various concentrations. To the media 0,15 mg /l kinetin, 40 g/l sucrose, and 2,5 g/l gelrite were added.Culture were incubated in the dark. Medium for Embryo development (maturation) is MS solid mediumcontaining treatment of BAP (Benzyl-amino-purine) at various concentrations. To the media 0,01 mg /l NAA(Napthalene-acetic-acid), 40 g/l sucrose, and 2,5 g/l gelrite were added. Culture were incubated in the light. Tostudy the specifi c structure of sandalwood somatic embryo early detection was conducted using histologicalanalysis. Results of anova showed that the clones, media, and interaction between clones with media did notsignifi cantly affect the development of sandalwood callus percentage. Results of anova showed that the clonesand BAP concentration signifi cantly effect to the embryo development of sandalwood.

Root Cell Proliferation from Broccoli Root Tips in vitro Culture Using Indole Acetic Acid (IAA), Indole Butyric Acid (IBA) and Benzylaminopurine (BAP)

Significance of the Study: Cell or tissue culture from root tips, shoot tips and leaf cuttings plays a significant role in micro-propagation in the plant production industry. Nowadays, it has been successfully done from many fruits, vegetables, ornamental and forest plants. Millions of explants can be produced by tissue or cell culture per year in any plant production industry. Aim: The study was conducted to investigate the root, shoot and leaf formation from the root tip cultures using different IAA, IBA and BAP concentrations. Methodology: IAA, IBA and BAP concentrations of 0.25, 0.50, 1.0, 1.5, 2.0, 2.5 mg/l combined with MS media were used in broccoli root tip culture. Results: The results showed that root and callus formation were done successfully. But there was Original Research Article Hossain et al.; ARRB, 9(1): 1-6, 2016; Article no.ARRB.22417 2 no shoot and leaf formation. The highest number (3.5) of callus proliferation was found in the concentration of IAA 1.5 and IBA 1.5 mg/l combination having BAP 1.0 mg/l. Moreover, root formation was found initially in the concentration of IAA 1-2.0 and IBA 1-2.0 mg/l combined with 1.5 mg/l BAP. In addition, later on it was found in the concentration of IAA 0.25-2.5 and IBA 0.25-2.5 mg/l combined with 1.5-2.5 mg/l BAP compared to other combination of concentrations. Conclusion: Therefore present results showed that it was better to use the combination of the IAA, BAP and IBA concentration to produce root proliferation and callus formation of broccoli using root tip cultures.

<i>In Vitro</i> Micropropagation of Himalayan Weeping Bamboo, <i>Drepanostachyum falcatum</i>

Plant growth hormone BAP (benzyl amino purine), KIN (kinetin), NAA (1-naphthalene acetic acid) and IBA (indole-3 butyric acid) effect was studied on in vitro multiplication of shoots and rooting of Drepanostachyum falcatum. In vitro micropropagation of himalayan weeping bamboo is explained by in vitro shoot induction and proliferation. Excised explant with axillary bud is surface sterilized with 0.1% HgCl2 for 10 - 12 minutes, cleaned with 90% ethanol and inoculated on liquid Murashige and Skoog (MS) culture medium supplemented with different concentrations of BAP/ KIN. Effect of BAP/KIN on shoot induction is with different rate and number of shoots produced by explants with axillary bud cultured on MS media supplemented with 0.0 mg/L BAP/KIN - 5.5 mg/L BAP/KIN. Shoot multiplication with highest rate is achieved on MS medium supplemented with 3.5 mg/L BAP after 4th sub-culturing. The most effective with highest rate and number of root induction combination is 6.5 mg/L IBA after 5 weeks. The roots produced by 6.5 mg/L IBA is best compared with other combination of auxin NAA (1-naphthalene acetic acid).

Effect of Different Carbon Sources and Elicitors on Shoot Multiplication in Accessions of Centella asiatica

Centella asiatica is a medicinal herb which has been extensively used in the treatment of nervous disorders and skin diseases. It is distributed throughout the tropical and sub-tropical region of the world. This plant contains various bioactive constituents such as asiaticoside, madecassic acid, flavonoids etc. Centella is in huge demand in International market for the preparation of pharmaceuticals and cosmetics which has resulted in its overexploitation. This situation demands the use of biotechnological methods to conserve as well as increase the production of biomass and bioactive compounds. The present study focused the use of different carbon sources (sucrose and fructose) and elicitors (Malt extract, Salicylic acid and Jasmonic acid) for the enhancement of biomass in five accessions of Centella asiatica. It was concluded that Murashige and Skoog (MS) medium supplemented with 1 mg/L concentration of Malt extract and 1.5 mg/L concentration of 6-Benzyl amino purine (BAP) showed highest biomass production among all elicitors. Among two carbon sources, MS media supplemented with Sucrose (3 mg/L) along with BAP (1.5 mg/L) showed better result than fructose.

Influence of light-emitting diodes and benzylaminopurin on adventitious shoot regeneration of water hyssop (Bacopa monnieri (L.) pennell) in vitro

Water hyssop (Bacopa monnieri (L.) Pennell) is a medicinal plants. Its upper and lower halves of leaf explants were incubated in Murashige and Skoog (MS) medium supplemented with 0.25, 0.50 and 1.0 mg/L benzylaminopurine (BA) for 8 weeks; the explants were exposed to white (W) and red and blue (R and B, respectively) light-emitting diodes (LEDs), at 4:1, 3:1, 2:1 and 1:1 R and B light ratios, respectively. Shoot regeneration (100%) was achieved from all explants at all applied concentrations of BA and LED types. All explants showed different BA concentration requirements for regeneration of the maximum number of shoots. Longer shoots were obtained on medium with 0.25 mg/L BA. The W LED lighting system was found to be more effective for regenerating the maximum number of shoots (26.11) per explant (on the upper half of the leaf). Conversely, longer and shorter shoots were generated under 1:1 R:B and W LEDs, respectively. The number of shoots per explant ranged from 9.67-24.0 (full leaf), 6.33-25.92 (lower half of the leaf) and 7.33-27.33 (upper half of the leaf), respectively, in response to BA and LED light. Shoot length ranged from 0.94-1.90 cm (full lamina), 0.70-2.11 cm (lower half of the leaf) and 0.93-1.83 cm (upper half of the lamina) in response to BA and LED lifght. Regenerated shoots were successfully rooted using indole-3-butyric acid (IBA) and acclimatized in the aquarium provided with tap water.

Naphthalene Acetic Acid and Benzylaminopurine Enhance Growth and Improve Quality of Organic Spinach in Kitchen Gardens

The present study was conducted at the Kitchen Gardening Training Centre, Faisalabad, Pakistan to evaluate the effect of the plant growth regulators Naphthalene Acetic Acid (NAA) and Benzylaminopurine (BAP), on the growth and yield of organic spinach (Spinacia oleracea L.) in kitchen gardening. Different combinations and concentrations of NAA and BAP were tested to evaluate different vegetative parameters. The data were recorded at 40 and 60 days after sowing (DAS). Maximum plant height at 40 DAS was recorded for the combined effect of NAA and BAP at a concentration of 1000 μM (each), while BAP alone at 100 μM concentration showed maximum plant height at 60 DAS. Maximum number of leaves was shown by NAA (10 μM) both at 40 and 60 DAS. Increase in leaf length was observed for NAA (10 μM) and BAP (100 μM) both at 40 and 60 DAS. Significant increases in root length were recorded where maximum root length was in plots treated with BAP (1000 μM) applied 40 DAS while the mixture of BAP (10 μM) and NAA (10000 μM) induced significant increases in root length when applied 60 DAS. Maximum fresh weight of shoot was observed for NAA (1000 μM) and BAP (1000 μM) while maximum shoot dry weight was observed in plots treated with NAA and BAP at a concentration of (1000 μM) each. Similarly, a significant effect of plant growth regulators was observed on root dry weight where highest dry weight was noted in the plants treated with NAA at the rate of 1000 μM.