Conventional Propagation Methods Research Articles

Micropropagation of banana (Musa spp) using temporary immersion bioreactor system

Banana is an important crop in the tropics which possess the potential for commercial production in Nigeria. Large scale production requires large volume of planting materials which may be difficult to obtain using conventional methods of propagation. Temporary immersion bioreactor system (TIBs) is a cost effective method for micropropagation of plants. The present study was carried out to develop an efficient method for rapid multiplication of banana using temporary immersion bioreactor system (TIBs). Banana microshoots were regenerated from young suckers obtained from field grown plants using conventional plant tissue culture. Microshoots of 2cm length were used as explants for multiplication in temporary immersion bioreactor system. Ten (10) explants were cultured in bioreactor bottles containing Murashinge and Skoog (MS) liquid media supplemented with different concentrations of 6-bezylaminopurine (BAP) with or without 250mg/L Activated Charcoal (AC). Results showed that explants cultured in media supplemented with 2 mg/L or 1mg/L BAP without AC gave the highest shoot multiplication rate of 900% and 800%, respectively compared to hormone free media. Production of competent plants (plants ready for ex vitro establisment) were however, influenced by the presence of AC and the highest percentage of competent plants (80%) were produced when media was fortified with 1mg/L BAP+ 250mg AC. Regenerated plants were successfully established in the field and were morphologically normal and fertile.

Silver birch (Betula pendula Roth.) culture initiation in vitro and genotype determined differences in micropropagation

Micropropagation has several advantages over conventional vegetative propagation methods, but it is limited by genotype responsiveness. We assessed the effect of age of the mother-tree and the time of explant collection on culture initiation, as well as the multiplication ability and effect of different nutrient media and plant growth regulators on silver birch genotypes. Explants collected from 1‐year‐old trees (66%) and explants collected in spring (64–67%) developed a significantly (both p < 0.001) higher proportion of shoots than those from 15‐year‐old trees (39%) and those collected in mid-summer (31%) and autumn (29%), respectively. In a stabilised culture, the length of the main shoot varied from 1.3 to 7.8 cm between genotypes, and the multiplication rate ranged from 1.0 to 6.8 shoots per explant. Hyperhydrated shoots were present in 17 out of 50 clones, and, among the clones, ranged from 14 to 50%. Cultures on the Murashige and Skoog basal medium had a higher multiplication rate than cultures on a Woody Plant Medium, and the application of zeatin provided better results than 6‐benzylaminopurine. The difference between cytokinin types was 11–29% for the multiplication rate and 21–29% for the length of the main stem. The highest multiplication rate was obtained using a zeatin concentration of 0.5 mg L−1. However, better shoot growth and proliferation had a significant positive relation to shoot hyperhydration (all p < 0.001). Therefore, a medium with an optimal balance between the multiplication rate and the number of hyperhydrated shoots should be carefully selected.

English

Conventional propagation methods of sweet potato (Ipomoea batatas L. var Kulfo) through stem cutting require large amount of propagules and large space for preparation. It has high risk of disease transmission to the next generation. In vitro propagation is the best alternative to overcome such limitations. This study was conducted to optimize protocol for in vitro propagation of Kulfo sweet potato variety using 6-benzylaminopurine (BAP) and indole-3-butyric acid (IBA) phytohormones for shoot multiplication and rooting, respectively. The result revealed that the highest shoot initiation (77.78%) and shoot length (4.40 cm) was observed in MS media supplemented with 0.5 mg/l BAP. Best shoot multiplication (5.33 shoots per explants) was obtained in MS medium supplemented with 1 mg/l BAP. MS medium supplemented with 0.5 mg/l IBA showed 100% rooting and average root length of 7.44 cm in vitro. In ex vitro conditions, 93.33% rooting was recorded. During acclimatization, 84 and 93% survival of in vitro and ex vitro rooted plantlets, respectively, were recorded. It could be concluded that MS media without BAP provides optimal condition for shoot initiation. MS supplemented with 1 mg/l BAP provides optimum condition for shoot multiplication. The ex vitro rooting could be better option to reduce in vitro rooting cost and for increased plantlet survival during acclimatization. Based on the result, 0.5 mg/l BAP for shoot initiation, 1 mg/l BAP of shoot multiplication and directly transferring to soil for rooting was recommended for micropropagation of Kulfo sweet potato variety. However, further studies will be needed in ex vitro root induction. Key words: Sweet potato, micropropagation, shoot tip culture.

Synthetic Seed Technology Development and Production Studies for Storage, Transport, and Industrialization of Bracken Spores.

Bracken fern (Pteridium aquilinum var. latiusculum (Desv.) Underw. ex A. Heller) has long been grown industrially in South Korea. Conventional propagation methods, including planting rhizomes and in vitro seedling culture, are labor intensive and expensive, and thus not commercially suitable. We aimed to develop a system to produce synthetic seeds using fern spores (SFS). Synthetic seeds were prepared by mixing bracken spores and alginate matrix. Spore germination and gametophyte and sporophyte growth and development from SFS proceeded normally. Spore density affected gametophyte and sporophyte numbers. SFS prepared using cold (4 °C) long-term storage spores (even 7-year-old spores) could effectively form sporophytes. The highest germination was observed at 25 °C. Soaking-treated SFS successfully formed sporophytes, even after 30 days of storage at 4 °C; indeed, sporophytes formed even after five days of storage at 25 °C during transport conditions. SFS were sown in plug trays for commercial use. Young sporophytes grown from plug seedlings were greenhouse cultivated, and transplanting within eight weeks was effective for root growth and growing-point formation. Developing synthetic seeds is a feasible solution for facilitating efficient transport and the handling of small-sized fern spores; furthermore, this SFS technology provides the basis for fern seedling culture and fern spore industrialization.

Process of Establishment And In Vitro Development of Simaba Cedron Planch Seedlings. (Simaroubaceae)

Simaba cedron, popularly known as "cedron", is largely used for fever and snake bites. Its seeds are used in the treatment of stomach problems and liver infections. The fruits are used for the treatment of pain and malaria while its bark is an antispasmodic. Simaba cedron is generally propagated through seeds, but with limited success, as the low viability of same restricts its propagation. In view of such difficulty, it becomes necessary the study for adequate conditions for the large scale production of these seedlings. Being it known that in several species, the use of micropropagation has made it possible to obtain a large amount of disease-free and more homogeneous seedlings, in reduced time and physical space, in comparison with conventional propagation methods, the objective of this work was to analyze the effect of two culture media on the production of aseptic parent plants as a first step in the development of a micropropagation protocol for Simaba cedron. The seeds were collected from a matrix plant located in the Amazon Biotechnology Center (CBA), in Manaus/AM. The experiment was installed at the Vegetable Tissue Culture Laboratory, where the explants were desinfected and grown in culture medium according to Murashige &amp; Skoog (MS) and in Wood Plant Medium (WPM), during 60 days. The disinfestation rate obtained was 75% and, of the disinfested seeds, 100% germinated. The cultivation medium that was more favorable to the cultivation of simaba was the MS, where the multiplication rate was of 8.0: 1, whose seedlings reached, in average, 4.8 cm and 75% of rooting.

Development of plantlet regeneration pathway using in vitro leaf of Ficus carica L. cv. Panachee supported with histological analysis

Fig ( Ficus carica L.) is a dioecious fruiting tree. However, Fig improvement was found challenging and time-consuming through conventional plant propagation methods due to low survival rate. Development of plant tissue culture technology through optimum regeneration pathway would offer many advantages especially on the mass production of elite and true to type planting materials. In this study, it was found that 24 h of incubation in the absence of light with the abaxial surface of the in vitro leaves contacting the media produced the highest mean of regeneration rate at 6.67 ± 4.63%. Histological sections revealed direct organogenic shoot regeneration was more prominent rather than indirect organogenic and embryogenic pathways. This is the first study on the effect of photoperiod and gravitropism on plantlet regeneration of F. carica. Micropropagation of F. carica involved various factors in photoperiodism and gravitropism on the development of plantlet's regeneration pathway. • First study on the effect of photoperiod and gravitropism on plantlet regeneration of Fig ( Ficus. carica ). • Regeneration pathway confirmed with histological analysis. • Dark condition with positive gravitropism produced the highest regeneration rate. • Histology confirmed organogenic regeneration pathway. • Two layers of palisade mesophyll tissue provided taxonomical information for this chimera plant.

Efficiency of the temporary immersion system versus the conventional in vitro propagation method

The work was carried out with the aim of assessing the efficiency of the methods of mass propagation of vitroplants through results obtained in the Conventional Method and the Temporary Immersion System (SIT) and achieve increases in the multiplication coefficient, in addition to characterizing both methods of in vitro micropropagation by describing them. Considering that conventional propagation methods would be slow to produce and of high production costs. This technique allows for proper planning, from the stage of establishment of aseptic culture in the laboratory, to the establishment and production of the crop in the field. However, in temporary immersion crops, the constitution and physiological proceeding of outbreaks are very similar to those shown in ex vitro environments, due to the evolution of the system environment. Several authors, including Cruzat, (2009), states that the use of THE SIT reduces production costs per plant; this as a result of the mechanization of some stages of micropropagation and Winkelman, et al, (2006), commenting as in bioreactors, obtaining seedlings allows the reduction of production costs by between 50 and 60 %. Results that corroborate the efficiency of the temporary immersion system against the conventional in vitro propagation method.

EMBRYO CULTURE AND IN VITRO CLONAL PROPAGATION OF OAK (Quercus aegilops L.)

An attempt was done to develop a micropropagation protocol for oak using embryo culture. Oak is considered a hard-to-root woody plant by conventional propagation methods, that’s why using tissue culture techniques is a very suitable alternative method. For oak embryo culture, WPM was used and found to be better than MS medium for embryo germination which gave 66.13%. As well as adding of GA3 to the medium improved the germination rate of embryos (43.25% and 82.25 %). At initiation stage, WPM was used and found to be the best medium by giving the highest number of shoots/ explant which was 1.80, the highest number of leaves (15.17 leaves/ explant) and the longest shoots (1.42 cm) followed by MS medium then GD which gave the lowest parameters which gave 0.98 shoots/ explant, 7.20 leaves/ explant and 1.06 cm shoot length. At shoot multiplication stage, BA was better than Kinetin for multiplication of oak explants. The addition of BA at 3 mg.l-l gave the highest number of shoot and leaves which were 3.33 and 26.11 respectively. The longest shoots were achieved when 4.5 mg.l-l of BA was used. Furthermore, kinetin at 3 and 4.5 mg-l gave the lowest parameters which were 1 cm in length and 1.54 leaves/ explant. For rooting stage, NAA was better than IAA in giving better parameters and rooting percentage. The highest number of roots and rooting percentage were achieved when 1 mg.l-l was added by giving 6 roots/ explant and 100% rooting percentage. While the longest roots were achieved when 0.5 mg.l-l of NAA was used (3.67 cm) followed by 1.5 mg.l-l IAA which gave 3.55 roots/ explant with rooting percentage 90%. The produced plantlets were successfully acclimatized and transferred to the open-air conditions with a rate reached 85%.

In Vitro Propagation and Variation of Antioxidant Properties in Micropropagated Vaccinium Berry Plants-A Review.

The berry crops in genus Vacciniun L. are the richest sources of antioxidant metabolites which have high potential to reduce the incidence of several degenerative diseases. In vitro propagation or micropropagation has been attractive to researchers for its incredible potential for mass production of a selected genotype in a short time, all year round. Propagation techniques affect the antioxidant activity in fruits and leaves. Total antioxidant activity was higher in the fruit of in vitro propagated plants compare to the plants grown ex vivo. This review provides critical information for better understanding the micropropagation and conventional propagation methods, and their effects on antioxidant properties and morphological differentiation in Vaccinium species, and fills an existing gap in the literature.

Spatially truncated Gaussian pulsed beam and its application in modeling diffraction of ultrashort pulses from hard apertures.

A new kind of pulsed beam, which we call a spatially truncated Gaussian pulsed beam, is defined to represent a Gaussian pulsed beam that is diffracted from a semi-infinite hard aperture. The analytical equations for the propagation of the spatially truncated Gaussian pulsed beam through a nonrotationally symmetric paraxial system with second-order dispersion is derived starting from the generalized spatiotemporal Huygens integral. The spatially truncated Gaussian pulsed beam is then combined with the conventional Gaussian pulsed beam decomposition method to enable the modeling of diffraction of a general ultrashort pulse from an arbitrarily shaped hard aperture. The accuracy of the analytical propagation equation derived for the propagation of the truncated Gaussian pulsed beam is evaluated by a numerical comparison with diffraction results obtained using the conventional pulse propagation method based on the Fourier transform algorithm. The application of the modified Gaussian pulsed beam decomposition method is demonstrated by propagating an ultrashort pulse after a circular aperture through a dispersive medium and a focusing aspherical lens with large chromatic aberration.

Izvor osvjetljenja i tip citokinina utječu na stopu multiplikacije, sadržaj klorofila i formiranje puči kod izdanaka Amelanchier alnifolia u in vitro uvjetima

Amelanchier alnifolia, a deciduous shrub or small tree with edible berry-like fruit, is gaining importance as a commercial fruit crop. The application of micropropagation could complement conventional propagation methods of A. alnifolia. The present study aimed to investigate how two light treatments—a mix of red and blue LED lights with dominant wavelengths of 457 and 658 nm (RB LED) and conventional fluorescent lamps emitting light at broad wavelengths of 400-700 nm (FL)—in combination with three cytokinins (CK) belonging to either substituted adenines (6-benzylaminopurine, BAP; meta-Topolin, mT) or to phenylurea cytokinins (thidiazurone, TDZ) affected the multiplication rate, chlorophyll content, and stomata formation in in vitro culture of A. alnifolia axillary shoots. The two light sources proved equally effective in producing axillary shoots, but FL favorably influenced the elongation and chlorophyll content. On the other hand, RB LED lights triggered a significantly higher stomata number of A. alnifolia plantlets in vitro compared with those developed under FL. Presented results reveal a negative ratio between the stomata number and chlorophyll content in response to different cytokinins: TDZ induced the highest number of stomata while BAP significantly increased the chlorophyll content. As the largest number of axillary shoots per explant was achieved on the medium with addition of TDZ, it could be beneficial to use TDZ for multiplication, whereas the use of BAP could be advantageous in the last passage of multiplication before rooting.

Desarrollo de plantas micropropagadas de Agave americana var. Oaxacensis durante su aclimatización en invernadero

Agave americana var. Oaxacensis Gentry es una especie longeva que tarda aproximadamente 20 años en llegar a su etapa reproductiva. En Oaxaca, México, esta especie se usa como materia prima para elaborar la bebida destilada mezcal. Para incrementar el área de cultivo con esta especie, se propone propagarla in vitro, en complemento a los métodos de propagación convencionales. Las plantas micropropagadas deben pasar por un periodo de aclimatación, pero se desconocen las condiciones ambientales de adaptación, tales como el sustrato y abastecimiento de nutrientes las cuales afectan la sobrevivencia y magnitud de crecimiento de las plantas. El objetivo de la investigación fue evaluar las características morfológicas de plantas micropropagadas de A. americana var. Oaxacensis que se sometieron a prueba de diferentes mezclas de sustratos: turba-arena: 33.3%+66.6% v/v, 66.6%+33.3%, v/v 75%+25% v/v; así como, el abastecimiento de nutrientes mediante fertirriego con diluciones 0, 50 y 100% de la formulación de Steiner, durante 290 días de su aclimatización para lo que se utilizó un diseño completamente al azar con arreglo factorial, al término de este periodo, de un total de 135 plantas, el 100% de éstas sobrevivió. Las plantas que alcanzaron mayor tamaño tuvieron las siguientes características: 5.5-5.6 hojas, la hoja mayor de 17.7 a 18.2 cm de longitud, 2.4 a 2.5 cm de ancho, el tallo de 2.4 a 2.5 cm de diámetro, y correspondieron a las establecidas en sustrato con 66.6% turba+ 33.3% arena v/v o 75% turba +25% arena v/v, y que además se fertirrigaron. Las plantas fertirrigadas con solución nutritiva al 100% lograron tamaño mayor que las plantas irrigadas con solo agua.

Split crown technique for mass propagation of pineapple (Ananas comosus L.) var. queen

The major problem limiting large scale commercial production and/or expansion of pineapple production is the difficulty in obtaining large quantity of quality planting materials due to slow rate of multiplication by conventional methods of propagation that mostly relies on the use of suckers and slips. The study investigated the effect of the methods of crown preparation on the sucker production of pineapple var. Queen and to evaluate the horticultural qualities of suckers produced from plantlets prepared using different methods of crown preparation. Destroying the growing point without splitting reduced percentage survival of plantlets but destroying the meristem and splitting the crown into two or four sections improved survival comparable to the intact crown. Destruction of the growing point and then splitting the crown promoted emergence and increased the number of suckers relative to the intact crown which did not produce suckers after, 3 months from planting. Increasing the number of sections prepared per crown (from two to four) did not reduce the number of suckers produced per plantlet and thus increased the multiplication rate by 7 and 8 times, respectively. The method of crown preparation did not significantly influence the early growth performance (survival, height, number size of leaves) of the harvested &amp; suckers during the first 6 months potting. Among the three methods of crown after preparation, destroying the meristem and splitting into two or four produced more number of ready-to-plant Queen pineapple suckers (1-ft tall) within a period of 5 months.

Effect of mulch and nutrients on growth and yield in transplanted ginger

The conventional propagation method using ginger rhizome being slow, a suitable method of raising ginger seed material in portrays has been devised by Indian Institute of Spices Research and Kerala Agricultural University. The advantages of this technology are production of healthy uniform planting materials and reduction in seed rhizome quantity which eventually reduced cost on rhizomes. The experiment was carried out in the Instructional Farm, College of Agriculture, Vellayani during April 2016 to January 2017. The ginger variety used was Karthika. Field experiment was laid out in split plot design with four levels of mulches in main plots and fertilizer levels in sub plots with four replications. Two nodded rhizome bits of ginger cultivar was raised in protrays were transplanted at 55 days in beds taken in the interspaces of coconut. Plants that received M1 (30 t ha1) in main plot resulted in maximum plant height, number of tillers, number of leaves/plant shoot weight, fresh yield and dry yield treatment T2 (150:100:100 kg ha1 and their interaction (m1t2) also resulted in highest plant height, number of tillers, number of leaves/plant, shoot weight, fresh yield and dry yield on all periods of observation The results of the study indicated that ginger transplants intercropped in coconut garden, that mulching @ 30 t ha1 ( half at transplanting and half 2 MAT) along with 150:100:100 kg NPK ha-1 and basal application of 30 t ha-1 of farm yard manure could be recommended for higher yield and growth.

Configuration of propagator method for calculation of electron velocity distribution function in gas under crossed electric and magnetic fields

This paper presents a self-contained description on the configuration of propagator method (PM) to calculate the electron velocity distribution function (EVDF) of electron swarms in gases under DC electric and magnetic fields crossed at a right angle. Velocity space is divided into cells with respect to three polar coordinates v, θ and ϕ. The number of electrons in each cell is stored in three-dimensional arrays. The changes of electron velocity due to acceleration by the electric and magnetic fields and scattering by gas molecules are treated as intercellular electron transfers on the basis of the Boltzmann equation and are represented using operators called the propagators or Green’s functions. The collision propagator, assuming isotropic scattering, is basically unchanged from conventional PMs performed under electric fields without magnetic fields. On the other hand, the acceleration propagator is customized for rotational acceleration under the action of the Lorentz force. The acceleration propagator specific to the present cell configuration is analytically derived. The mean electron energy and average electron velocity vector in a model gas and SF6 were derived from the EVDF as a demonstration of the PM under the Hall deflection and they were in a fine agreement with those obtained by Monte Carlo simulations. A strategy for fast relaxation is discussed, and extension of the PM for the EVDF under AC electric and DC/AC magnetic fields is outlined as well.

The Stimulating Effects of Imazalil and Carbendazim Fungicides on in Vitro Propagation and ex Situ conservation of the Medicinal Balkan Range-Restricted Sideritis Raeseri Boiss &amp; Heldr. Subsp. Raeseri

Sideritis raeseri Boiss &amp; Heldr. subsp. raeseri, known in Greece as Mountain tea of Parnassus or Velouchi is a range restricted medicinal plant of the Balkan peninsula. Conventional propagation methods did not allow the mass production of plant material in a short period of time due to both low seed germination rate and rooting of cuttings. Therefore, the aim of this study was to establish a reliable, reproducible and efficient regeneration protocol for mass and large-scale micropropagation, germplasm and ex situ conservation of S. raeseri Boiss &amp; Heldr. using Imazalil and Carbendazim fungicides. After 9 weeks, 2.5-10 mg/l Imazalil stimulated root length by 1 cm but diminished root number and rooting percentage. Optimum shoot number (5.58), shoot length (24.91 mm), shoot multiplication (100%), root number (20.63) and rooting (66.67%) were recorded with 0.5 mg/l kinetin (KN) + 0.05 mg/l ?-napthaleneacetic acid (NAA) (Imazalil-free). After 4 ½ weeks, 1 mg/l Carbendazim + 0.5 mg/l 6-benzyladenine (BA) yielded 5.77 shoots/explant, 16.1 mm shoot length and 100% shoot multiplication. However, Carbendazim did not result in rooting. The ex vitro survival percentage of rooted shoot-tip explants derived from Imazalil experiment was 95%. Carbendazim proved useful in shoot proliferarion and Imazalil in root elongation of S. raeseri Boiss &amp; Heldr. subsp. raeseri micropropagation system.&#x0D;

In vitro cloning via direct somatic embryogenesis and genetic stability assessment of Paphiopedilum niveum (Rchb.f.) Stein: the endangered Venus’s slipper orchid

Wild populations of Paphiopedilum niveum (Rchb.f.) Stein, an endangered orchid species, have been threatened with extinction due to climate change and overcollection. Conventional methods of propagation are unable to meet market demands. The current study reports the production of genetically stable, protocorm-derived protocorm-like bodies (PLBs) of P. niveum that can be used for commercial production. These PLBs are referred to hereafter as somatic embryos (SEs). Factors affecting SE induction and SE proliferation were examined. The highest percentage of SE formation (68.33 ± 11.77%) and the maximum number of SEs per explant (5.19 ± 0.67) were obtained on modified Vacin and Went (MVW) medium containing 0.1 mg L−1 1-naphthaleneacetic acid (NAA). The highest increase in fresh weight (FW) of proliferated SEs (183.33 ± 28.93 mg per 100 mg initial FW) was gained on hormone-free MVW. These SEs eventually developed into vigorous plantlets after culturing on MVW supplemented with 0.2% (w/v) activated charcoal and 50 g L−1 banana homogenate for 12 wk. A histological study revealed that SEs originated from both the epidermal and the sub-epidermal layers of the original protocorm via direct somatic embryogenesis. The genetic homogeneity between the mother generation (V1; the original protocorm) and subsequent generations (V2 and V3; the primary and secondary somatic embryos, respectively) was shown to be identical via random amplification of polymorphic DNA (RAPD) assessment. All regenerated plants grew well under greenhouse conditions.

Establishment of a regeneration system for the production of Calla plants (Zantedeschia spp.) via embryogenic callus culture

Calla lilies (Zantedeschia spp.) are monocotyledonous ornamental plants which belongs to the Araceae family. After the release of elite calla cultivar, an efficient propagation system is needed for commercial use. Despite the use of conventional propagation methods such as splitting of tubers and rhizomes of calla, rapid and efficient propagation system should be developed. In order to achieve this goal, stem segments contained apical meristems derived from calla lily cultivar (cv. Gag-si) were cultured on Murashige and Skoog (MS) medium supplemented with various concentrations of cytokinin and auxin. This was aimed at inducing embryogenic calluses, shoots and multiple shoots. As a result, about 25% of induction rates of yellow embryogenic calluses were observed with MS medium containing both 0.5 mg·L-1 NAA and 1.5 mg·L-1 BA as growth regulators. In the experiments involving the regeneration from embryogenic calluses through shoot formation, MS medium supplemented with 0.5 mg·L-1 IAA and 2.0 mg·L-1 BA showed the highest rates at approximately 85 ~ 90% with regard to the formation of shoots in calla. Moreover, multiple shoots needed for rapid propagation were generated when explants were cultured on MS medium supplemented with 0.5 mg·L-1 IAA and 2.0 mg·L-1 BA with 40% of formation rate. In this study, the combination of auxin and cytokinin showed positive effects on both the induction of embryogenic calluses, the formation of shoots as well as multiple shoots in calla. The regeneration system described here can contribute to the development of breeding programs of calla in the future.

Evaluation of Anti-microbial Activity of Ex vitro and Callus Extracts from Commiphora gileadensis.

Commiphora gileadensis a medicinal plant rare species. A large amount of plant materials were needed to produce secondary metabolite under in vitro culture. Therefore, callus is used in the in vitro culture, since it can proliferate quickly and continuously provide an appropriate amount of plant which used for extracting the antimicrobial compounds from C. gileadensis. Rapid protocol for optimum callus production has been assessed to overcome limitations of the conventional propagation methods. The effect of plant growth regulator (PGR) on the regeneration of C. gileadensis was investigated for callus induction experiment using a standard MS medium with various concentrations of 6-Benzyl adenine (BA), Kinetin (Kn), 2,4-dichlorophenoxy acetic acid (2,4-D) and naphthalene acetic acid (NAA) at 0.0, 0.5, 1.0,1.5, 2.0 and 2.5 mg L-1. The result showed that the maximum regeneration of callus induced the fresh and dry weight were obtained 5675±1321 and 376.7±56.9 mg, respectively on MS media containing 2 mg L-1 2,4D + 0.5 mg L-1 BA after 12 weeks. The anti-bacterial and anti-fungal activities of C. gileadensis were evaluated using the callus and ex vitro extracts, six bacterial species fungal genera were used the agar well diffusion method used of 25, 50, 75 and 100 μL methanolic or ethonlic extracts of ex vitro and callus had considerable inhibition effects on the tested bacteria and fungi. Callus culture technique may be an important tool to get the C. gileadensis quickly as compared to the natural growth phenomenon where it takes many years. Moreover, it's give us an opportunity to get the active constituent without destroying the plant available in nature. The results of the present study can improve our understanding of the economic importance of C. gileadensis as activity ingredient antimicrobial agent and provided methods for its preparation.

Micropropagation of Guadua chacoensis (Rojas) Londoño &amp; P. M. Peterson1

ABSTRACT The bamboo productive chain is still incipient in Brazil, and the low supply of plantlets due to low-efficient conventional propagation methods presents a significant bottleneck to its development. This study aimed to establish a micropropagation protocol for Guadua chacoensis. Explants from donor plants cultivated under controlled environment showed less contamination, if compared to explants from plants grown in the field. The contamination rate was even lower when 2 mL L-1 of Plant Preservative Mixture (PPM™) were added to the culture medium, leading to a higher establishment rate. The obtained cultures were then multiplied using either in vitro-derived nodal segments or clump division in the presence of increasing contents (0 mM, 10 mM, 20 mM, 30 mM or 40 mM) of 6-Benzylaminopurine (BAP). The number of shoots increased with increasing BAP concentrations, but this also resulted in a reduced rooting rate and root length. Plants acclimatized under 0 %, 35 % or 65 % of shading showed a dynamic maximum quantum yield of photosystem II (Fv/Fm), which initially decreased within the first seven days after the transfer to ex vitro conditions, but then increased until reaching stable values of 0.775 after 17 days. Additionally, the shading improved the plant survival rates, if compared to those under non-shaded conditions, which presented photoinhibition and photodamage symptoms.