Combinations Of IAA Research Articles

Application of mannitol as pre-treatment and sucrose supports callus induction through anther culture in Gossypium arboreum

Diploid or tree cotton (Gossypium arboreum) species may provide a breeding source for fiber quality and disease resistance. Developing homozygous lines of Gossypium arboreum through anther culture can be a successful strategy for breeding the parental lines needed for cotton hybrids. This study describes callus induction on four G. arboreum genoytypes under various concentrations of sucrose as well as plant growth regulators. Explant materials were derived from immature anther and grew on modified Murashige and Skoog (MS) media. Prior to main treatment, mannitol application were applied to anthers with concentration of 0.7 M and modified MS media consisted of 3%, 5%, and 7% sucrose, as well as an addition of IAA, NAA, BAP, 2,4-D and Kin. Optimum callus formation was observed in Marvi genotype, in a medium containg IAA as well as sucrose between 5-7%. Furthermore, combination of IAA and NAA with sucrose 7% indicates highest number of embryogenic calli. However, Marvi genotype showed decreased rate of callus formation when the amount of sucrose increased on MS supplemented with NAA medium. The results indicated that the best formula on each genotypes were different. However, MS media with IAA (1 mg/l) + kin (0.2 mg/l) + CM (70 ml/l) + sucrose 5% provided the best performance to produce embryogenic calli on each genotypes. Our report can be utilized as an opportunity and strategy to obtain double haploid lines as a parental source in hybrid production.

Direct Organogenesis of Citrus Cultivars from Shoot Tip Nodal Segments.

Citrus cultivar improvement via conventional breeding strategies is impeded by factors related to its reproductive biology. The orange is a hybrid between pomelo (Citrus maxima) and mandarin (Citrus reticulata). Among various orange cultivars, Valencia oranges have a bit of bitter tang mixed in with their sweetness, as Navel oranges are, the most widely cultivated citrus species, quite sweeter, and also don't contain any seeds. Tangelo mandarin orange cultivar is a hybrid of C. reticulata × C. maxima or × C. paradisi. The present study was undertaken to optimize the hormonal composition of the media with regard to plant growth regulators for in vitro propagation of sweet orange cultivars from nodal segment explants. The nodal segment explants were collected from three citrus cultivars, Washington Navel, Valencia and Tangelo. Murashige and Skoog (MS) medium supplemented with sucrose and different concentrations of growth regulators was used for shoot proliferation and root induction, and the optimum medium composition was assessed. The patent for Citrus Tissue Culture was obtained from the Office of Research Affairs, Haramaya University. The results indicate that the highest shoot response was recorded for Washington's navel with maximum shoot proliferation rate (99.75%), shoot number per explant (1.76), shoot length (10.70 cm), leaf number per explants (3.54) after three weeks of culture. In all experiments, no growth was observed for the basal MS medium. Phytohormone combinations of IAA (1.2 mg/L) and kinetin (2.0 mg/L) were found to be the best for shoot proliferation. Among the cultivars, there were significant differences for the highest rooting rate (81.255), root number (2.22), and root length (2.95 cm) variables for Washington Navel. The lowest rooting rate (48.45%), root number (1.47) and root length (2.26 cm) were observed for Valencia. The highest rooting rate (84.90%), root number per microshoot (2.22) and root length (3.05 cm) was on MS medium supplemented with 1.5 mg/L NAA. A comparison of different concentrations of IAA and NAA on root induction of microshoots from nodal segments of citrus cultivars demonstrated NAA was a more effective hormone than IAA.

Standardization of a protocol for micropropagation of Eupatorium glandulosum L. an important medicinal plant

Eupatorium glandulosum of Asteraceae family is an important medicinal plant used to cure various diseases by tribal populace. In the present study, a protocol for mass propagation of Eupatorium glandulosum has been developed from nodal explants cultured in the MS basal medium supplemented with various concentrations and combinations of IAA and BA. The maximum rate of proliferation of shoots per explants (59.66 ± 1.76) was observed in the medium containing 0.5 mg/l of IAA and 3 mg/l of BA, but medium with 1 mg/l of IAA and 4 mg/l of BA showed the maximum shoot length. When the regenerated shoots were rooted in vitro on MS basal medium supplemented with various concentrations of IAA and IBA, medium with 4 mg/l of IBA induced more number of roots per shoot (32.67 ± 0.89) while maximum root length (6.1 ± 0.12 cm) was recorded in 3 mg/l of IBA. The plantlets, with well developed root systems, were successfully acclimatized and transferred to the pots containing a mixture of Soil, Sand and Coco peat (1:1:1) with 100% survival rate. The research article tends to elucidate the tissue cultural techniques for the mass propagation and sustainable utilization of Eupatorium glandulosum. This technique could be implemented for the continuous extraction of valuable medicinal components in the pharmaceutical industry.

Indirect Organogenesis from Stem Derived Callus of Dregea Volubilis (L.F) Benth

In vitro propagation methods offer a powerful tool for germplasm maintenance and multiplication. This report established the methodology for callus regeneration of Dregea volubilis from stem explants. The effect of various concentrations and combinations of IAA, NAA, 2,4-D, BAP, KIN and IBA were tested for in vitro callus induction, plant regeneration, and rooting of D. volubilis. The maximum number of greenish, organogenic callus was achieved on the MS medium containing 1.5 mg/L 2,4-D+ 0.5 mg/L KIN and then subsequent shoot regeneration was obtained on MS medium supplemented with 3.0 mg/L KIN from stem derived callus. The highest frequency of shoot regeneration (81.93%) with the maximum number of shoots (10.56 + 0.44) was achieved in this medium. In vitro raised micro shoots showed a 90% response with the maximum number of roots (5.31 ± 0.66) per shoot on MS medium supplemented with 2.5 mg/L IBA after 15 days. Then in vitro rooted plantlets were moved to mud pot bearing soil and vermiculite mixture in 1:1 ratio for hardening and hardened plantlets showed 90% survivability in field conditions. Using this protocol, it is possible to clonally produce viable, uniform and healthy plants with a maximal survival rate that can be used for large scale cultivation, genetic transformation and biopharmaceuticals applications.

In vitro Propagation of Mortiño (Vaccinium floribundum Kunth.)

The disinfected axillary shoots of mortino - a wild plant species were cultured in MS with different concentrations (0.002, 0.05 and 0.1%) of methyl N-(1H-benzimidazol-2-yl) carbamate to develop an in vitro protocol for micropropagation through organogenesis. They were then placed in a concentration matrix (0.1, 1.0 and 10.0 μM) whose factorial design involved BAP and IAA in WPM. Results showed a 100% disinfection at 0.05% concentrations of carbendazim and the combination of IAA and BAP at 10 μM yielded a multiplication factor of 3.35, with rooting after seven weeks. This system showed efficient disinfection, multiplication and yielding an acceptable root system.
 Plant Tissue Cult. & Biotech. 30(2): 167-177, 2020 (December)

Somatic embryogenesis and in vitro plant regeneration of Bacopa monnieri (Linn.) Wettst., a potential medicinal water hyssop plant.

Bacopa monnieri (Linn.) Wettst. commonly known as waterhyssop, Brahmi plant, traditionally used for memory enhancement, nerve tonic, epilepsy, central nervous system (CNS), antidepressant, anxiety, blood pressure and antioxidant activities. Due to pharmaceutical demands its lost natural habitat. At this juncture we describe a resourceful protocol for micropropagation of water hyssop plant. Surface sterilized leaf and nodal explants were inoculated on basal MS semi-solid medium added with PGRs; auxins, cytokinins. Highest calli formation from leaf explants was obtained on NAA (2.5 mg−1) and showed (94.22%) accompanied via 2,4-D showed (2.5 mg−1; 82.43%), maximum calli formation in nodal explants was obtained on 2,4-D showed (2.5 mg−1; 71.14%) followed by NAA (2.5 mg−1) showed (62.15%), in internodes explants uppermost calli formation was obtained from 2,4-D showed (2.5 mg−1; 65.21%) followed by NAA (2.5 mg−1) showed (52.14%). The maximum somatic embryogenic callus, calli induction and formation (84%) was observed on 2,4-D + KIN (2.0 + 1.5 mg−1) amended solid medium. Uppermost shoot formation was observed in combination of IAA + BAP (1.0 + 1.0 mg−1) showed (78.54%) shoot formation followed by IBA (2.0 mg−1) alone showed (75.37%). The maximum shoot elongation was noticed from NAA + BAP (3.0 + 3.0 mg−1) with 21.21 cm followed by NAA (2.0 mg−1) showed (15.22 cm) although, chief root formation was obtained from IBA (2.0 mg−1) with 83.75% root formation along higher number of roots (47.43%) per shoot. Followed by IAA (2.0 mg−1) showed root induction (73.43%) and no of roots (38.54%) per shoot. In hardening under pot condition plants survivability (100%) was observed under glass house conditions, the present in vitro PTC techniques is extremely significant to gratifying its natural conservation.

Enhancement of rhamnetin production in Vernonia anthelmintica (L.) Willd. cell suspension cultures by eliciting with methyl jasmonate and salicylic acid.

Vernonia anthelmintica (L.) Willd. belongs to the family Asteraceae and has anthelmintic, anti-diabetic, diuretic and anti-asthmatic properties. Present study describes the production of rhamnetin: an O-methylated flavonol with anti-oxidant and anti-inflammatory activities; in cell suspension cultures of V. anthelmintica. Suspension cultures were established from cream coloured friable calli initiated from leaf explants and cultured in liquid MS medium supplemented with combinations of IAA (1.5mgL-1) and BAP (1.5mgL-1). After 20days of cell suspension culture, the initial inoculum size of 0.5g was increased to 1.05g100mL-1 dry weight at exponential phase. When the culture was treated with 0.8mM methyl jasmonate for 6days; 2.2 fold accumulation of rhamnetin production was obtained on a dry weight basis compared to unelicited control. Likewise, the elicitation with salicylic acid 1mM for 6days brought about 2.7 fold rhamnetin production compared to unelicited control. The rhamnetin content in dried leaf tissue of field grown plants was found to be 0.143mgg-1. Elicitation with methyl jasmonate and salicylic acid brought about 7.7 and 9.4 fold increments in rhamnetin production, compared to leaf tissue of field grown plants on a dry weight basis.

Micropropagation of hop (Humulus lupulus L.) via shoot tip and node culture

Humulus lupulus is a valuable medicinal plant that grows wild in forest areas of northern Iran. In recent decades with extension of village boundaries toward forest, hop habitat is suffering destruction. Optimization of hop micropropagation by shoot tip and nodal segments was conducted towards conservation of Iranian hop germplasm. Nodal segments were provided from the rootstock and after disinfection were established in the MS medium. After six weeks, explants grown in in vitro conditions were isolated, then were incubated in MS medium supplemented with various concentrations of plant growth regulators, including BA (0, 0.1, 0.5, 1 and 2 mg L(-1)), Kin (0, 0.1, 0.5, 1 and 2 mg L(-1)) and TDZ (0, 0.1, 0.5, 1 and 2 mg L(-1)) alone and in combination with IAA. The combination of 1 mg L(-1) BA with 0.1 mg L(-1) IAA was the best treatment applied in all traits evaluated, followed by 5.0 mg L-1 Kin and TDZ with 0.1 mg L(-1) IAA. The best treatments for shoot tip explant proliferation were the best treatments for nodal segments proliferation, except in the case of Kin, where the best results for nodal segments proliferation was observed at a concentration of 1 mg L(-1) Kin in combination with 0.1 mg L(-1) IAA. For shoot rooting, the shoots were grown in vitro; and plantlets were transferred to a rooting medium containing MS medium supplemented with different concentrations IAA (0, 0.5 and 1 mg L(-1)) and BA (0, 0.5 and 1 mg L(-1)). The highest percentage of rooting was observed in half strength MS supplemented with 1 mg L-1 IAA in combination with 1 mg L(-1) BA. Thus we recommend MS medium containing 1 mg L(-1) IAA and 1 mg L(-1) BA as a suitable micropropagation medium for industrial purposes.

Production of biomass and flavonoid of Gynura procumbens (Lour.) Merr shoots culture in temporary immersion system.

Gynura procumbens (Lour.) Merris one of medicinal plant which was carried out used as antioxidant, anticancer, anti-inflammatory, hepatoprotective, and antimicrobial. Many strategies were used to increase the production of biomass and valuable compounds. This study was to investigate the variation effect of growth regulators and immersion frequency on production of biomass and flavonoid contained of G. procumbens shoots culture in temporary immersion bioreactor. Stem nodes were used as an explants and induction of shoots were done in solid MS medium supplemented with many kinds of growth regulator. The best treatments were used to produce biomass and flavonoid compounds in temporary immersion bioreactor; there are combination of IAA 2 mg/L and BA 4, 6, 8 mg/L and immersion frequency (5 min each 3 h; 15 min each 12 h). Results showed that the growths of G. procumbens shoots in solid MS medium were influenced by supplementation of growth regulators. MS medium supplemented with single cytokinine (6 mg/L kinetin) and combination of auxin (IAA) and cytokinine (BA) caused increasing of shoots growth. Production of biomass of G. procumbens in temporary immersion bioreactor was achieved in long immersion interval (12 h) and highest flavonoid production was obtained in combination treatment of immersion frequency 15 min each 12 h and MS medium supplemented with IAA 2 mg/L, BA 8 mg/L.

Micropropagation’s Complete Protocol of Red Araçá (Psidium cattleianum, Myrtaceae) from Germinated Seeds in vitro

Red Araçá’s (Psidium cattleianum) micropropagation processes have shown enormous potential both in terms of research and as a sustainable native resource to be used in the areas of food production, ecology, and pharmacology. Currently, however, despite that potential, research efforts involving this myrtaceae, native to the Brazilian Atlantic Forest, have been scarce. With that in mind, this study set out to establish micropropagation techniques that would allow the development of a feasible protocol to be used with Red Araçá, achieving its mircropropagation from in vitro germinated seeds. Different types of explants were tested for in vitro establishment. For the multiplication of nodal segments, different concentrations of BAP and IAA combinations were tested in an MS medium. Using the same medium, different concentrations of ampicillin were applied in order to determine its influence on the decontamination of the apical segments. The BAP and IAA combinations were also used to test their effects on the in vitro explants’ development and rooting. During pre-acclimatization, survival of in vitro rooted plants was tested in a nebulizer chamber, using a commercial substrate and that same substrate mixed with washed sand (1:1). In essence, it was indeed possible to develop a complete protocol for the micropropagation of the Red Araçá from seedlings obtained by in vitro germination. The in vitro introduction of the Red Araçá was rather efficient, independently of the type of explants used. As the BAP and IAA concentrations increased, so did the in vitro seedlings’ development (7 leaves explant-1) and rooting (67%). Additionally, the in vitro rooted plants exhibited a high rate of survival (80%) in the pre-acclimatization phase, independently of the substrate used.

IAA and flavonoids modulates the association between maize roots and phytostimulant endophytic Aspergillus fumigatus greenish

ABSTRACTThe association between microbes and plant roots involves a complex chemical dialogue featured by IAA and flavonoids in addition to other signals. Current study is focused on the role of IAA and flavonoids as essential elements of the chemical dialogue between an endophytic fungus and maize roots. The strain was isolated from the leaves of Withania somnifera and was identified as Aspergillus fumigatus greenish by 18S rDNA sequence. Culture filtrate of the strain contained phyotstimulant (IAA, ammonia and ACC) and signaling compounds including IAA, flavonoids and flavonols. The strain effectively colonized the maize roots and enhanced its growth. In order, to determine the effect of IAA and flavonoids on the ability of the endophyte to colonize maize roots, we inhibited the release of IAA and flavonoids in maize separately which reduced the colonization of endophyte in maize root to 66% and 55% of the untreated control. Similarly, application of IAA in combination with endophyte enhanced root colonization by 90% in zone of cell division, 66% in the zone of elongation and 30% in the zone of maturation. These findings determined that IAA and flavonoids are among the key players of the complex chemical dialogue between plant root and fungal endophyte.

Pemberian Indole Acetic Acid dan Benzil Amino Purine Terhadap Pembentukan Protocorm dan Tunas Anggrek Vayes Limondok (Phaius tankervilleae (Banks) Bl ) In Vitro

The objective of this research is to identify the best combination of IAA and BAP concentration to support the formation of protocorms and budding of Phaius tankervilleae in vitro. This research were conducted with two treatment factors that are IAA and BAP concentration levels. The results showed that there were no interaction between IAA and BAP concentrations to the budding explant percentage and leaf number while they were affected by BAP concentration only. BAP concentration at 1 ppm produced the highest number of protocorm

Effect of spray combinations of IAA, GA3 and zeatin on berry growth and quality in ‘Superior Seedless’

Effect of spray combinations of IAA, GA₃ and zeatin on berry growth and quality in ‘Superior Seedless’

Influence of zeatin, glutamin and auxins on root and shoot organogenesis of Guava (Psidium guajava L.) cv. safeda seedling explants

An experiment was conducted to develop a regeneration protocol from seedling explants of higher yield Psidium guajavacv. safeda. Initially surface sterilized guava seeds were germinated inside the conical flasks. Explants taken from 49-day old seedlings were cultured on MS-medium modified with different concentration of plant growth regulators. Maximum explants showing shoot response (48%) was observed on MS-medium supplemented with 6-benzyladanin (BA) 1.0 mgl-1combined with 0.5 mgl-1ziatin. On the same media after 38-days of culturing, maximum number of 3.6 shoots per explant, 2.6 cm long was also recorded. On MS containing combination of BA (2.0 mgl-1) and zeatin (1.0 mgl-1), 37% explants responded which produced 2.3 shoot per explant after 42 days of culturing. Whereas on MS containing glutamin 250 mgl-1, 25% explants responded and gave 2.2 shoots per explants after 46-days. However, when the concentration of glutamin alone was increased from 250 to 500 mgl-1 the shoot formation potential of explant decreased to 10%. Addition of BA, glutamin and zeatin alone to MS medium was less effective. However, combination of BA with zeatin was effective in shooting response, number of shoot/explants and shoot length. Maximum number of explants (76.6%) responded to rooting with 1.5 mgl-1 IAA in combination with 1.0 mgl-1 NAA. On average, maximum of (2.5) roots were also recorded per explants in 42 days. In vitro plantlets were acclimatized and shifted to soil successfully. Keywords: Psidium guajava L; Micropropagation; Zeatin; Glutamin and auxins http://dx.doi.org/10.19045/bspab.2017.60014

Seedling Propagation of Kulo Chrysanthemum by Tissue Culture

Aims of this research were to get the right dosage of Plant Growth Regulkator (PGR) and Coconut Water (CW) for shoot multiplication of Kulo chrysanthemums. The study was conducted using a factorial design of three factors namely IAA (0.0; 0.5 and 1.0 ppm), BAP (0, 0.5 and 1.0 ppm) and coconut water (0.0 and 30%). The explants used were clean micro cuttings and were cultured on Murashige and Skoog (MS) media. The media was added IAA, BAP and coconut water. The result showed that the best shoot multiplication of Kulo chrysanthemums was the MS media using combination of 0.5 ppm and 1.0 of IAA, 0.5 ppm of BAP and 30% of coconut water. Good rooting was revealed by the combination of IAA 0.5 and 1 ppm with 30% coconut water and BAP 1 ppm and 30% coconut water.

Direct <i>in vitro</i> Regeneration of <i>Nicotiana plumbaginifolia</i> L. and the Potential for Genetic Transformation

Wild crop relatives are important source of genetic diversity. Plant tissue culture provides proficienct means of conservation of endangered wild plants sustaining biodiversity, improving plant traits by virtue of induced somaclonal variation and trangensis. The objective of study was to establish direct in vitro regeneration and biolistic transformation protocol for wild tobacco N. plumbaginifolia using uidA gene. For direct in vitro regeneration, six combinations of IAA and kinetin were used. Then, biolistic transformation was carried out using BioRad PDS 1 000 (He) gun and 1 100-psi rupture discs. Transformation was confirmed through histochemical gus assay. A proficient and reproducible in vitro regeneration system was standardized for wild tobacco named as Nicotiana plumbaginifolia . In present study 1 mg/L kinetin in combination with 0.25 mg/L IAA proved best for direct in vitro shoot regeneration from mature leaf-based explants. Success in direct in vitro regeneration circumvented the callogensis step, accelerating the process of in vitro regeneration and increasing the chances of obtaining true to type plants. Individual shoots were placed on MS medium with/without IBA for rooting. Individual shoots were successfully rooted on MS medium without IBA. Then, explants were transformed with uidA gene using gene gun. Transient assay of bombarded calli showed gus activity on histochemical assay even after a week. Analysis of gus expression has been found as a principal method of plant transformation optimizations.

Establishment of Efficient Regeneration System from Leaf Discs in Long Pepper an Important Medicinal Plant (Piper longum L.)

Cultivation of Piper longum L. till recently was not very common; still it is extensively collected from the wild, threatening the very existence of the plant. Hence, it is alarming that there is an urgent need not only cultivate this plant but also develop conservation strategies. Conventional propagation is overwhelmed with problems of poor seed viability low percentage of germination, scanty and delayed rooting of vegetative cuttings indicating there is a need for alternative propagation methods. In vitro technique is an alternative approach to solve the problem. Therefore, the current research was aimed at developing a promising in vitro mass propagation protocol for Piper longum L. using leaf as explant. Murashige and Skoog medium was used throughout the experiment. For callus induction, MS medium supplemented with alone or in combination of IAA (1 to 2 mg/l) and BAP (1 to 2 mg/l) were used. Shoot induction was undertaken on MS medium supplemented with different concentrations and combinations of Kinetin and BAP (1 to 3 mg/l). Emerged shoots were transferred onto elongation medium supplemented with 2 mg/l Zeatin and 1 mg/l GA3. In vitro rooting was achieved on ½ MS medium+NAA 1 mg/l. Accordingly, the highest calli were induced on MS+1 mg/L IAA+1 mg/L BAP. Among the various treatments, the maximum percentage (91.50 ± 3.54) in vitro shooting was observed on MS+2 mg/L BAP+1 mg/L Kinetin. In vitro rooted shoots were successfully acclimatized in the greenhouse conditions. Therefore, it is possible to deduce that the current protocol is promising for in vitro mass propagation of Piper longum L. to solve the reproduction and cultivation problem of the plant.

Transcriptome profiling of postharvest strawberry fruit in response to exogenous auxin and abscisic acid.

Auxin and abscisic acid regulate strawberry fruit ripening and senescence through cross-talk of their signal transduction pathways that further modulate the structural genes related to physico-chemical properties of fruit. The physiological and transcriptomic changes in harvested strawberry fruits in responses to IAA, ABA and their combination were analyzed. Exogenous IAA delayed the ripening process of strawberries after harvest while ABA promoted the postharvest ripening. However, treatment with a combination of IAA and ABA did not slow down nor accelerate the postharvest ripening in the strawberry fruits. At the molecular level, exogenous IAA up regulated the expressions of genes related to IAA signaling, including AUX/IAA, ARF, TOPLESS and genes encoding E3 ubiquitin protein ligase and annexin, and down regulated genes related to pectin depolymerization, cell wall degradation, sucrose and anthocyanin biosyntheses. In contrast, exogenous ABA induced genes related to fruit softening, and genes involved in signaling pathways including SKP1, HSPs, CK2, and SRG1. Comparison of transcriptomes in responses to individual treatments with IAA or ABA or the combination revealed that there were cooperative and antagonistic actions between IAA and ABA in fruit. However, 17% of the differentially expressed unigenes in response to the combination of IAA and ABA were unique and were not found in those unigenes responding to either IAA or ABA alone. The analyses also found that receptor-like kinases and ubiquitin ligases responded to both IAA and ABA, which seemed to play a pivotal role in both hormones' signaling pathways and thus might be the cross-talk points of both hormones.

Direct Multiple Shoot Induction and Plantlet Regeneration from Cotyledonary Explants of Sapindus emarginatus Vahl (Soapnut)

Multiple shoot induction in S. emarginatus has been achieved by two methods: (1) Direct germination of S. emarginatus in vitro cotyledon explants in BAP/Kn/TDZ (1.0-3.0 mg/L) supplemented MS medium and (2) in plant treatment with BAP/Kn/TDZ (3.0 mg/ L) in combination of 1AA (0.5 mg/L) of the cotyledon explants of plants and maintained under sterile conditions. While the former method resulted in as many as (7.5±8.6 shoot buds) from the cotyledonary explants within four weeks, the latter yielded on average approximately 8 shoot buds from each treated node in eight weeks. The cytokinin treatment in plant consisted of placing sterile filter paper moistened with sterile distilled water over the node and adding different concentrations of 6benzylaminopurine. The best results for shoot bud regeneration were obtained with cotyledons, when cultured in the presence of (0.5 mg/L) IAA in combination with (3.0 mg/L). The shoots elongated and rooted directly in vermiculite after a pulse treatment with IBA (2.5 mg/L) for 15 min. Original Research Article Srinivas et al.; BBJ, 6(1): 1-9, 2015; Article no.BBJ.2015.021 2 Fungus growth, a serious problem in S. emarginatus tissue culture, was controlled using a fungicide, Bavistin, along with elimination of organic nutrients from the growth medium.

Micropropagation of Hippeastrum hybridum

According to the conventional and traditional propagation of Amaryllis is very low rate and less efficiency, Research in the field of plant micropropagation was necessary. Current research aims to achieve an optimal and efficient micropropagation protocol of this plant. Accordingly, the impact of plant growth regulators of 2,4-D in four levels (0.0, 5.0, 1 and 2 mg/L), IAA in four levels (0.0, 2, 5 and 10 mg/L), BAP with three levels (0.0, 1 and 2 mg/L) and the combination of IAA and BAP in two types of medium including NL and MS was investigated in the regenation of stablishment stage of five different explants . Explants were varied from single scale, twin scale, scape, basal plate and single scale without basal plate. Parameters evaluated were: percentage callusing, percent of bulblet production, produced bulblet number, bulblet diameter and rooting percentage. Current researchwere designed on a factorial experiment based on completely randomized design with four replication.Results of variance analysis showed that highest rate of callus production were seen in the present of 2mg/L of 2,4-D. Moreover highest rate of bulblet production were seen with twin scale and single scale explants and highest number of bulbs were seen with single scale explants that was cultured in NL medium containing 0.5 mg/L of 2,4-D. The greatest diameter of bulblets regardless of growth regulators was observed in both twin scale and single scale explants were cultured on MS medium. Since the main objective of the study was to achieve a higher number of bulblet. Therefore, among the applied treatments, NL medium containing 5.0 mg/L 2,4-D was the most desirable options and the best type of explant to culture the single scale explant was proposed.