Different Concentrations Of Indolebutyric Acid Research Articles

MINICUTTING OF CHARDONNAY GRAPEVINE AND SO4 GRAPEVINE ROOTSTOCK

The use of grapevine minicutting is an alternative for the production of quality plants. This technique has been gaining ground as one of the most used forms of vegetative propagation. The objective of this study was to evaluate the rooting of herbaceous minicuttings of Chardonnay grapevine and SO4 grapevine rootstock by using different concentrations of indolebutyric acid (IBA) to form a clonal mini-garden. The experimental design was randomized blocks with four levels of IBA concentrations (0, 500, 1000 and 1500 mg L-1). The minicuttings remained in trays containing vermiculite and kept in a micro sprinkler chamber. After 60 days, the percentage of survival, rooting and callus formation, root number and average length, shoot number and average length were evaluated. For Chardonnay, the means of survival and rooting varied between 93 and 100% in all treatments. In relation to SO4, the IBA concentration of 1500 mg L-1 negatively affected the minicuttings survival and rooting. It is possible to propagate Chardonnay grapevine and SO4 grapevine rootstock through herbaceous minicutting, with no need of apply IBA for the rooting.

Sodium Nitroprusside Stimulates Micropropagation and TDZ Induces Adventitious Shoots Regeneration in Red Flesh Apple Malus niedzwetzkyana Koehne Dieck ex

Abstract The aim of the present investigation was to optimize protocols for micropropagation and adventitious shoot regeneration from leaf explants of two wild ecotypes of red flesh apple Malus niedzwetzkyana for future breeding programs. At the proliferation stage, different concentrations of sodium nitroprusside (SNP) and triacontanol (TRIA) were compared. To optimize shoot regeneration from leaf explants, interactive effects of 1-phenyl-3-(1,2,3-thidiazol-5-yl)-urea – thidiazuron (TDZ), indole-3-butyric acid (IBA) and two explant types were investigated. At rooting stage, the effect of exposure time of microshoots to darkness and exposure time to different concentrations of IBA and α-naphthalene acetic acid (NAA) were compared. The results showed that SNP affected the growth rate significantly and the maximum multiplication rates per explant (9.6 in the first ecotype and 8.8 in the second) were produced in the Quoirin and Lepoivre medium containing 17 SNP µM, in addition to 4 µm 6-benzylaminopurine (BAP) and 3 µm gibberellic acid (GA3). IBA and TDZ affected the adventitious shoot regeneration from leaf explants significantly, the highest number of regenerated shoots (18.3 per explant) was obtained from the basal section of the leaves cultured on the medium containing 2 μM IBA and 15 μM TDZ. At rooting stage, the maximum rooting (88.6%) was obtained in the result of one weak exposure to darkness on medium containing 3 μM IBA.

Rooting of juvenile cuttings of Brazil nut trees using indolebutyric acid in sub-irrigation chamber

The sexual propagation of Brazil nut (Bertholletia excelsa Humb. & Bonpl.) is limited by several factors, including slow and irregular germination, unevenness in root and shoot emergence, and a strategy to overcome these limitations is vegetative propagation. The objective of this study is to evaluate the rooting of Brazil nut cuttings using different concentrations of indolebutyric acid (IBA) in a subirrigation system. The study adopted a completely randomized block design with five concentrations of IBA (0, 1000, 2000, 3000, and 4000 mg L-1). The donor plants of the vegetative material were obtained from seeds of trees selected as highly productive parent plants. Cuttings with approximately 6 cm were obtained from the basal and middle sections of the shoot and contained one pair of nodes and one fully developed leaf. The percentage of survival, callus formation, rooting, and sprouting was determined at 105 days after rooting. IBA improved the analyzed characteristics for the development of Brazil nut plants. The IBA concentration of 1000 mg L-1 provided the highest percentage of survival, rooting, and callus formation. Juvenile cuttings showed a higher rate of survival (69.7%), rooting (49.1%), and callus formation (30.2%) at IBA concentrations between 1.9 and 2.3 g L-1.

Propagation of white mulberry Morus alba L. fruitless cultivar using different cutting times and IBA.

The white mulberry Morus alba L. is an interesting tree that widely planted throughout the world for various uses. The effort was made in this study to propagate the fruitless cultivar of Morus alba L. by hardwood cuttings; comparing different cuttings collection time with using various concentrations of IBA (Indole-3-Butyric Acid) solution. Results showed that taking and planting cuttings on March 1st, gave the best survival percentage of rooted seedlings, which was (20.83%) compared to February 8th (16.33%) and March 22nd (12.5%).Cuttings treated with different concentrations of IBA significantly increased success percentage; 58.33, 66.67, 50.00 and 20.83% for 2000, 1500, 1000 and 0ppm respectively. Furthermore, the combination effect of previous factors had positive effects on studied characters; the heighest values of success rate of rooting (66.67%) from cuttings taken on March 1st and immersed in 1500ppm of IBA solution (T7), while success rate of rooting (58.33%) and all other qualitative parameters of shoots height (70cm), vegetation fresh weight (74.9g), vegetation dry weight (24.06g), root length (41.17cm) roots fresh weight (5.26g), roots dry weights (1.5g), number of leaves per plant (32.33), and leaf area (84.32cm2) were obtained from cuttings taken on the same time but treated with 2000ppm. The results recommend taking cuttings on March 1st, and using 2000ppm of IBA directly before planting to enhance cuttings survival percentage and seedlings quality.

Effect of indolebutyric acid on rooting and budding of cuttings of glyicidium sepium

Studies carried out with the use of gliricidia biomass found that green manure contributes to increasing the productivity of forest crops when compared to the incorporation of other legumes. This study aimed to evaluate the viability of vegetative propagation by cuttings in the development of rooting and budding of Glyicidium sepium in different concentrations of indolebutyric acid (IBA). The experiment was carried out in a greenhouse with completely randomized block design and increasing concentrations of IBA 0, 625, 1250, and 3000 mg.L-1 with six replicates. The cuttings were standardized in length and diameter, then treated with sodium hypochlorite and immersed in different concentrations of IBA. The parameters plant evaluated were the sprouts number, sprouts length, sprouts diameter, sprouts dry weight, and roots dry weight. The concentration of IBA was shown to be effective in increasing all parameters studied at the IBA concentration of 2100 mg.L-1 and the higher concentrations did not offer any cost-benefit advantages for the production of gliricidia by cutting.

Effect of indolebutyric acid on rooting and budding of cuttings of Glyicidium sepium

Studies carried out with the use of gliricidia biomass found that green manure contributes to increasing the productivity of forest crops when compared to the incorporation of other legumes. This study aimed to evaluate the viability of vegetative propagation by cuttings in the development of rooting and budding of Glyicidium sepium in different concentrations of indolebutyric acid (IBA). The experiment was carried out in a greenhouse with an entirely randomized block design and increasing concentrations of IBA 0, 625, 1250, and 3000 mg.L-1 with six replicates. The cuttings were standardized in length and diameter, then treated with sodium hypochlorite and immersed in different concentrations of IBA. The parameters plant evaluated were the sprouts number, sprouts length, sprouts diameter, sprouts dry weight, and roots dry weight. The concentration of IBA was shown to be effective in increasing all parameters studied at the IBA concentration of 2100 mg.L-1, and the higher concentrations did not offer any cost-benefit advantages for the production of gliricidia by cutting.

EFECTO DE DIFERENTES CONCENTRACIONES DE ÁCIDO INDOLBUTÍRICO EN EL ENRAIZAMIENTO DE ESTAQUILLAS DE Annona muricata “GUANÁBANA” EN CÁMARA DE SUBIRRIGACIÓN

La investigación tuvo como objetivo, determinar el efecto de diferentes concentraciones de Ácido Indolbutírico (AIB) en el enraizamiento de estaquillas de Annona muricata «Guanábana» en cámara de subirrigación, el cual se desarrolló en el vivero agroforestal de la Universidad Nacional Intercultural de la Amazonía (UNIA), ubicada en el km 0,63 de la carretera San José de Tushmo, distrito de Yarinacocha, provincia de Coronel Portillo, región Ucayali, Perú, entre los meses de agosto a octubre del 2017. Las estaquillas de guanábana fueron recolectadas, cortando las ramitas con tijera de podar, de 10-12 cm de longitud, y un diámetro de 0,5 cm, para luego cortar la mitad de las hojas con un bisturí. Las estaquillas se desinfectaron con fungicida, por 10 minutos y ventilándolos 10 minutos, luego fueron introducidas cerca de 1 cm de su base, en un envase conteniendo las diferentes soluciones de AIB, por un lapso de 3 segundos, y con la ayuda de un ventilador se secaron durante 1 minuto, con el fin de que el alcohol se evapore y pueda fijarse la hormona, para luego instalarlas dentro de la cámara de subirrigación de acuerdo a los tratamientos estudiados (0 ppm de AIB, 1000 ppm de AIB, 2000 ppm de AIB y 3000 ppm de AIB); concluyendo que, para el número de raíces, el tratamiento testigo (0 ppm de AIB) y el tratamiento con 2000 ppm AIB, presentaron los mejores promedios (3,55 y 3,39 raíces respectivamente), no encontrándose diferencias significativas entre los tratamientos para la longitud de raíces, y para el porcentaje de estaquillas enraizadas, el tratamiento con 1000 ppm de AIB, presento el mejor promedio (41%) y finalmente, para el porcentaje de sobrevivencia, el tratamiento con 2000 ppm de AIB presentó el mejor promedio (99,33 %).

Impact of Indole-3-Butyric Acid (IBA) on the Root Induction of Arbutus pavarii Pamp (Lybian Strawberry Tree) in in vitro Culture

The main objective of this study was to clarify the best concentration of the indole-3-butyric acid (IBA) in order to induce the formation of strong roots of Arbutus pavarii Pamp, an endangered plant in the El-Jabel El-Akhdar region in Libya. A study was carried out to find a protocol for its in vitro propagation. The present paper aimed to investigate the effects of different concentrations of IBA plant growth regulator on the rooting. Three weeks old seedlings obtained with in vitro germination were transferred to Murashige and Skoog (M&S) roots induction medium supplemented with different concentrations of IBA (0, 1, 1.5 and 2 mg L-1). The highest response was obtained with the M&S medium half-strength supplemented with IBA 1 mg L-1 concentration. All the measured growth indicators (rooting percentage, root length and dry weight) significantly enhanced when using this concentration.

Cuttings of Euphorbia phosphorea Mart and Euphorbia enterophora Drake at different concentrations of indole-butyric acid and analysis of economic viability

Abstract Defining seedling production protocols for ornamental species through the use of plant regulators requires economic analysis that demonstrates the real financial returns of such activity to horticulturists. The objective was to evaluate the influence of using indole butyric acid regulator (IBA) on rooting of Euphorbia phosphorea (Mart) and Euphorbia enterophora (Drake) cuttings, as well as to analyze the economic viability of seedling production with the use of IBA. The cuttings were 10 cm long, and treated with IBA at concentrations of 0; 1,000; 3,000; and 5,000 mg L-1. The cuttings were place in vermiculite substrate and were irrigated daily inside a greenhouse. The experimental design was completely randomized, with 4 replications, containing 5 cuttings per experimental unit. Evaluations were performed 150 days after establishing the experiments. The Net Present Value (NPV) and the Internal Return Rate (IRR) were analyzed to determine the economic viability analysis of using IBA. For E. phosphorea there was no significant difference between treatments for cuttings survival, number of roots and length of roots. Callus formation was not observed at the base of the cuttings, and the highest survival percentage was observed in T3 (3,000 mg L-1) and T4 (5,000 mg L-1) treatments. Treatment T1 (control) presented a lower number of roots per cutting (12.8) and higher average length of the three largest roots (12.56 cm). The use of the IBA regulator increased the percentage of rooted cuttings and the number of roots per cutting, and the concentration of 3,000 mg L-1 was recommended for E. phosphorea. Regarding aspects of economic viability, using IBA at the commercial nursery level is recommended. For E. enterophora, IBA is not recommended because it is an easily rooted species (95%), therefore, denotes the economic unfeasibility of its use.

QUALITY OF CLONAL SEEDLINGS OF Tectona grandis Linn F. ROOTED IN DIFFERENT CONCENTRATIONS OF INDOLEBUTYRIC ACID

The general objective of this paper is to evaluate the effect of different IBA concentrations on greenhouse, shade and full sun survival, root vigor and seedling quality in Tectona grandis. To do so, mini-cuttings were prepared containing two pairs of leaves reduced to 25% of their original size (7.0 ± 1.0 cm) in length and bevel cut at the base. They were submitted to five treatments with different concentrations of indolebutyric acid (0, 500, 1000, 1500 e 2000 mg L-1), remaining in greenhouse, house of shade and in full sun for 30, 20 and 15 days, respectively. Survival was evaluated in greenhouse, house of shade and full sun; radicial vigor, height, diameter of the neck, ratio between shoot height and diameter, root dry mass, aerial, and total mass. The experimental design was in randomized blocks, with five replications of 15 plants and five treatments. Data were submitted to the normality test, Bartlett test and polynomial regression of the second degree. As a result, it was verified that the use of IBA did not influence root vigor and survival, however, it provided seedlings with a higher dry mass, being the best the 2000 mg L-1 treatment.

Effect of indolebutyric acid (IBA) and cane position on rooting of rootstock’s cuttings

The aim of this study was to determine the effect of different concentrations of indolebutyric acid (IBA) and to evaluate the influence of the part of the cane from which the cuttings were taken on rhizogenesis of rootstocks (1103 Paulsen, SO4, Dogridge) and grapevine cultivar ‘Vidiano’ (Vitis vinifera L.). Three rootstocks (1103 Paulsen, SO4, Dogridge) with five different auxin treatments (control, 60, 120, 250 and 1000 ppm IBA) were examined under hydroponic conditions. A comparative study of rhizogenesis was performed between grapevine cultivar ‘Vidiano’ and the three rootstocks, as well as the part of the cane. The cuttings were collected from the vineyard of the Laboratory of Viticulture of the Agricultural University of Athens. It was shown that, in general, the middle and, secondarily, the base part of the cane give the best results concerning the percentage of kalogenesis, the percentage of rhizogenesis, the average diameter of roots, the average number of roots per cutting, the total surface of roots and the total length of roots. Among the concentrations, 120 and 250 ppm showed a high percentage of rooting for the three rootstocks studied (Dogridge, 1103P, SO4) and the cultivar ‘Vidiano’. The effect of three factors (cultivar, part of cane from which the cuttings were taken, IBA concentration) was evaluated concerning the percentage of kalogenesis, the percentage of rhizogenesis, the average diameter of roots, the average number of roots per cutting, the total surface of roots and the total length of roots. The results showed that controls had the shortest roots and 60 ppm IBA showed a sharp increase at root length probably due to increased rhizogenesis compared to that of the controls. The bases of the cuttings of the three rootstocks gave the longest roots probably due to better nutrition. The highest percentages of rhizogenesis were observed in the base, middle and top cuttings of 1103 Paulsen at the treatment of 250 ppm (95, 80, 80%, respectively), followed by the treatment of 120 ppm of the middle cuttings of the same rootstock (75%). The experiment showed that the cultivar (‘Vidiano’) and the rootstocks (1103 Paulsen, SO4, Dogridge) which rooted easily need smaller concentrations of IBA. The treatment of 1000 ppm IBA gave the lowest rates (kalogenesis, rhizogenesis, root number, root length, root surface, and average root diameter) at all cuttings of ‘Vidiano’ and the rootstocks SO4 and Dogridge.

English

The traditional way of grapevine (Vitis vinifera L.) propagation is time consuming and allows disease transmission from generation to generation. Moreover, it is difficult to improve this crop through conventional plant breeding methods. Therefore, the objective of this study was to develop efficient in vitro regeneration protocol for ‘Canonannon’ and ‘Chenin Blanc’ varieties of grapevine using leaf explants. MS medium supplemented with different concentrations of thidiazuron (TDZ) alone or in combination with α-naphthalene acetic acid (NAA), and 6-benzyl aminopurine (BAP) alone or in combination with indole-3-butyric acid (IBA) were used for regeneration of shoots from leaves. The regenerated shoots were transferred to shoot multiplication medium and subsequently to rooting medium and the plantlets were acclimatized after rooting. The rooting medium consisted of MS medium containing different concentrations of IBA or indole-3-acetic acid (IAA). The highest number of shoots per leaf explant was obtained from both ‘Chenin Blanc’ (2.3 ± 0.3) and ‘Canonannon’ (2.2 ± 0.2) on medium supplemented with 2.0 mg/L BAP. Among 16 different combinations of TDZ and NAA, the maximum number of shoots per explant (1.5 ± 0.2) was obtained from ‘Canonannon’ on medium containing 1.0 mg/L TDZ and 0.1 mg/L NAA. However, when these shoots were transferred to shoot multiplication medium, 10 ± 0.51 shoots per explant were obtained from ‘Chenin blanc’ on MS medium supplemented with 2.0 mg/L BAP. The highest number of roots per explant (8.3 ± 0.30) was obtained on medium containing 2.0 mg/L IBA. The survival rate of ‘Chenin Blanc’ and ‘Canonannon’ was 83.3 and 75 %, respectively after one month of acclimatization.   Key words:  Callus induction, growth regulators, hyperhydricity, organogenesis.

Indolebutyric acid in “pulse” treatment on the rooting of Luehea divaricata minicuttings

ABSTRACT: The objective of the present research was to evaluate the effect of different concentrations of indolebutyric acid (IBA) on the rooting of Luehea divaricata minicuttings under two different cultivation periods in a greenhouse environment. The minicuttings were immersed in a hydroethanolic solution containing IBA. All minicuttings survived after 30 and 60 days of cultivation. Root formation at 30 days of cultivation with 1000 mg L-1 of IBA was 91.7%, the same rate as that observed with 4000 mg L-1. At the 1000 mg L-1 concentration, the number of roots was the greatest among all treatments, with 8.42 roots per minicutting after 30 days of cultivation. After 30 days of cultivation, the formation and number of roots with the use of 1000 mg L-1 of IBA in the “pulse” treatment produced the best outcome results, allowing the plants to spend less time in a greenhouse at a reduced concentration of auxin when compared with other treatments. However, to obtain higher averages of secondary root formation, root length, and number of leaves, it is necessary to increase the cultivation period to 60 days.

Influence of indole butyric acid (IBA) concentrations on air layerage in guava (Psidium guajava L.) cv. Sufeda

Abstract Air layering is the expedient way to get more number of plants however, root initiation in cuttings is a key success in case of Guava. Indolebutyric acid (IBA) hormone induces rooting but its appropriate concentration and response of guava cv. Sufeda was unknown. This experiment was carried out at Horticultural Research Farm, The University of Agriculture Peshawar, during 2016. One year old healthy branches of guava were selected and wounded by complete removal of 2 inches bark just below the bud for rooting. These wounds were covered with soil media containing different concentrations of IBA hormone, wrapped with plastic and tied at both ends. The air layered plants detached after rooting were shifted to polythene bags and were kept in plastic tunnel to maintain its humidity. Different concentrations of IBA i-e T1 (50 ppm), T2 (100 ppm), T3 (150 ppm), T4 (200 ppm) and control (T0) were applied. According to data analysis layers treated with 150 ppm of IBA (T3) was successful. Data recorded in control was minimum as compared to other treatments. We concluded that, the Guava cv. Sufeda air layered during monsoon season treated with 150 ppm IBA solution gives best results. Keywords: Air layerage; Growth; Guava; IBA concentrations; Rooting http://dx.doi.org/10.19045/bspab.2018.700194

Arbuscular mycorrhizal fungi and indolebutyric acid alter rooting rate and physiological characteristics of Lantana camara L. (Verbenaceae)?

ABSTRACT The objective of this study was to evaluate the rooting and some physiological characteristics of Lantana camara L. (Verbenaceae) stakes in different concentrations of indolebutyric acid (IBA) and its association with the arbuscular mycorrhizal fungi (AMF) Claroideoglomus etunicatum (W.N. Becker & Gerd.) C. Walker & A. Schüssler e Gigaspora albida N.C. Schenck & G.S. Sm. The experimental design was completely randomized in a 3x3 factorial scheme with three replicates, where the first factor was the AMF and the second factor by doses of IBA (0; 1.0 and 1.5 mg .mL-1). The results showed that the presence of AMF in the substrate, as well as the IBA, isolated or in association stimulates the rooting and some physiological characters of L. camara cuttings.

Rooting and acclimatization of grapevine hybridsVitis labrusca×Vitis rotundifolia

Rhizogenesis is an important step in micropropagation and root quality is essential for plant survival during acclimatization. Vitis rotundifolia hybrids show low rooting rate and auxin application is important for cutting propagation. This study was aimed at evaluating different concentrations of naphthalene acetic acid (NAA), 0.0, 0.2, 0.4 and 0.8 µM, in in vitro rooting and different concentrations of indolebutyric acid (IBA), 0, 2.5, 5 and 7.5 mM, in ex vitro rooting of grapevine hybrids and their survival rates after acclimatization. Hybrids between V. labrusca × V. rotundifolia used were: IBCA125 (‘Isabel’ × ‘Carlos’), IBBT481 (‘Isabel’ × ‘Bountiful’), and IBMG628 (‘Isabel’ × ‘Magnolia’). Explants were nodal segments from in vitro plants after the tenth subculture in medium MS/2 without growth regulators. For in vitro rooting after 30 days of cultivation at different NAA concentrations, pre-acclimatization of explants was carried out with gradual opening of bottles. After evaluations on the 34th day plants were transferred to tubes containing vermiculite and left in intermittent mist for 21 days. For ex vitro rooting, shoots were standardized in microcuttings with two leaves and treated with different IBA solutions. Microcuttings were planted in tubes with vermiculite under intermittent mist for 30 days. There was 100% rooting in all treatments for the hybrids tested. For in vitro rooting, survival was high for all hybrids, except for IBBT481, for which the highest survival rate observed was 42.1% in the treatment without NAA. For IBCA125 the survival rate decreased gradually with the increasing NAA concentration and the lowest rate observed was 0.8 µM. For ex vitro rooting, the highest survival rate was observed in the control and in the treatment with 2.5 mM of IBA for IBMG628. Survival of hybrid IBCA125 was not affected by IBA. Hybrids between V. labrusca × V. rotundifolia can easily root both in vitro and ex vitro without exogenous auxins.

Collect Seasons, Indolebutyric Acid and Substrates on Rooting of ‘Bengal’ Lychee Cuttings

Lychee (Litchi chinensis Sonn.) propagation by cuttings is a promising alternative in commercial production of seedlings, since it is an easy procedure, in addition to enabling the standardization in the field and the reduction in production costs. Therefore the objective of this work was to evaluate ‘Bengal’ lychee cuttings rooting, collected in spring and summer, using different concentrations of indolebutyric acid (IBA) and substrates. The experimental design was completely randomized, in a 3 × 3 factorial arrangement, with 5 replications and 10 cuttings per plot, and the following factors were adopted: IBA concentrations (0; 1,000 and 2,000 mg L-1) and types of substrates (carbonized rice husk, vermiculite medium granules and coconut fiber). At 140 days, the following variables were evaluated: cuttings survival; leaf maintenance; cuttings unrooted with calli; cuttings rooted; roots number per cutting; root length and root dry mass. In addition, the pH, electrical conductivity, density and water retention capacity of the substrates were evaluated. There was no difference in IBA concentrations applied in both evaluated periods. However, in the spring, the carbonized rice husk provided highest mean for the measured variables. On the other hand, in the summer, the rooting percentage presented a lower average in all substrates (< 1%), independent of the IBA concentrations. It was concluded that the ‘Bengal’ lychee propagation by cutting can be carried out in the spring, using carbonized rice husk as substrate, without IBA.

EFFICIENCY OF INDOLEBUTYRIC ACID AND DIFFERENT SUBSTRATES IN YERBA MATE CUTTINGS

Ilex paraguariensis A.St.-Hil. is an arboreal species whose leaves are used in the preparation of mate and in the cosmetics and food industries. Due to diffi culty found in the sexual propagation of the species, this paper aimed to test effi ciency of indolebutyric acid (IBA) in saplings production from semi-woody cuttings. On the fi rst experiment, cuttings were taken from young plants, immersed in diff erent concentrations of IBA and established in substrate. On the second experiment, cuttings were established in four diff erent substrates previously submersed in IBA. A concentration of 3.0 mg L-1 of IBA and a substrate composed of biostabilized pine bark, limestone and vermiculite + carbonized rice hull, have shown a greater effi ciency in rooting of young I. paraguariensis saplings, and could be used to facilitate yerba mate propagation.

Comparative analysis of hyoscine in wild-type and <i>in vitro</i> grown <i>Datura innoxia</i> by high performance liquid chromatography

Purpose: To compare the hyoscine contents of Datura innoxia plant grown in the wild and that grown in vitro.Methods: For callus induction from leaf explants, 15 combinations of 2,4-dichlorophenoxy acetic acid, gibberellic acid, benzyl amino purine and kinetin were used. For regeneration of shoots, 8 combinations of BAP, kinetin and indole butyric acid (IBA) were tested. Different concentrations of IBA were used for rooting. The hyoscine contents of the wild and in vitro samples was initially identified by TLC and subsequently quantified by high performance liquid chromatography using hyoscine N-butyl bromide as standard.Results: For callus induction, 2,4-D at 2 mg/L produced maximum calli (96 %). Maximum shoots regeneration was obtained with 2 mg/L BAP and 1 mg/L kinetin. IBA at 0.5 mg/L induced good rooting. Substantial amount of hyoscine was detected in the extracts of in vitro grown plants through TLC and HPLC. Wild root, stem and leaves exhibited higher amounts (approx. 2 μg/mL) of hyoscine than the parts of in vitro grown plants. Green callus grown in vitro contained a maximum concentration of hyoscine (1.01 μg/mL) followed by leaf (0.82 μg/mL) and brown callus (0.432 μg/mL).Conclusion: The callus cultures of D. innoxia are capable of yielding good amounts of hyoscine, and therefore can be exploited to increase hyoscine production using chemical/hormone treatments on a large scale. Thus, this study provides a sustainable and efficient way for the sustainable production of a natural hyoscine product via in vitro-grown D. innoxia cultures.Keywords: Datura innoxia, Hyoscine, Callus, Hyoscine, Explant, Gibberellic acid, Benzyl amino purine, Kinetin

In Vitro Propagation of Agave americana by Indirect Organogenesis

Factors such as slow growth, low rates of sexual and asexual reproduction, and viability of seeds among others limit the massive propagation of Agave americana L. by conventional methods. In this study, callus induction and shoot proliferation was determined in A. americana using Murashige and Skoog (MS) medium supplemented with dicholorophenoxyacetic acid (2,4-D) and 6-benzyl adenine (BA). Meristematic tissue was used as the explants, and were placed on MS medium supplemented with 30.0 g·L−1 sucrose with 0.11, 0.18, or 0.45 μm 2,4-D and 11.0, 22.0, 38.2, 44.0, 58.7, or 73.3 μm BA. Treatments were implemented according to factorial experimental design 3 × 6. After 1 month, the number of explants with callus was determined, whereas the numbers of shoots per explant were monitored after 4, 16, 20, and 36 weeks. The maximum percent of explants with callus was obtained with 0.11 μm 2,4-D and 58.7 and 73.3 μm BA, whereas the maximum numbers of shoots per explant (71) were obtained with 0.11 μm 2,4-D and 73.3 μm BA. The effect of different concentrations of indolebutyric acid (IBA) in the rooting of shoots was evaluated. There were no significant effects of IBA on the number of roots, root length, and axillary roots. Plantlets were acclimatized in the glasshouse and they did not show any phenotypic alteration. This is a highly efficient protocol for the in vitro propagation of A. americana via indirect organogenesis.