Highest Number Of Plantlets Research Articles

Nano-Integrated Plant Tissue Culture to Increase the Rate of Callus Induction, Growth, and Curcuminoid Production in Curcuma longa.

Turmeric has attracted a significant amount of interest in recent years due to its strong antimicrobial properties. The tissue culture of turmeric is preferred to obtain disease-free, highest number of plantlets with good uniform chemistry. However, there is a need to increase the speed of the whole process to meet the growing demand for planting materials and to save time and resources. Iron oxide nanoparticles (Fe3O4 NPs) showed positive effects on callus initiation time, proliferation rate, percent root response, shoot length, percent rooting, and number of roots per explant. Highest callus induction, i.e., 80%, was recorded in cultures that were grown in the presence of 15 mg/L of Fe3O4 NPs. Callus initiated earlier in culture tubes that received green synthesized iron nanoparticles in a concentration between 10-15 mg/L. Biofabricated nanoparticles were characterized for their size, physiochemical, and optical properties through UV-Vis spectroscopy, FTIR, XRD, and SEM. Curcuminoids profiling was performed by implementing LC-Ms that revealed increased quantities in plantlets grown in nano-supplemented media when compared to the control.

Effect of culture media, auxins and genotypes on plantlet regeneration from oil palm (Elaeis guineensis Jacq.) zygotic embryos through somatic embryogenesis

Aim: This study evaluated efficient culture media for the regeneration of elite material through somatic embryogenesis from oil palm zygotic embryos. Methodology: For callus induction, zygotic embryos of four elite genotypes (G1-264T, G2-238DX17P, G3-37DX17P and G4-237T) were cultured on three basal media (Y3, MS and N6) with different auxin 2 mg l-1 (Picloram, 2,4-D and Dicamba) combinations. Subculture was made every month for three passages. It evaluated various callus characters. The embryogenic calli from callus induction media were transferred to the embryo maturation medium and subcultured until the polyembryoids formed. For shoot and root formation, somatic embryo clumps were transferred into regeneration media. In-vitro plantlets with well-grown roots were hardened in pots for six weeks and assessed clonal fidelity using polymorphic SSR primers. Results: Among the treatments, calli from N6+2,4-D, Y3+2,4-D and N6+Picloram showed the highest embryogenic callus potential. G4-237T induced more embryogenic calli (32.982) among genotypes, which was on par with G1-264T (24.196). Embryogenic calli grown on N6 media with Dicamba showed the highest proliferation rate (1.141). After 60 days of culture on regeneration media, the highest number of plantlets per somatic embryogenic clump was obtained from G1-264T on N6 media supplemented with Dicamba. Interpretation: Culture media salt concentration showed a significant difference among media by causing perturbations of auxin flow during somatic embryogenesis affecting callus induction, proliferation and plantlet regeneration. This may be useful for standardizing the genotype-specific regeneration media in oil palm.

OPTIMIZATION PARAMETER FOR PRODUCTION OF PROTOCORM-LIKE BODY AND GROWTH OF GRAMMATOPHYLLUM SPECIOSUM

Micropropagation of orchids are often through the production of Protocorm Like Bodies (PLBs) and this research conducted a study to discover the optimal condition for the growth of Protocorm Like Bodies (PLBs) and also plantlets regeneration of Grammatophyllum speciosum Several aspects such as different carbon sources (sucrose, maltose) and the presence of charcoal alongside with the variety of concentration of agar were taken into account for the optimization. This study discovered that the 30 g/L of sucrose in 1 mg/L of BAP was superior in producing heavy PLBs. However, the trend differs in the number of plantlets where maltose (5 g/L) is seen to produce a higher number of plantlets compared to sucrose. Charcoal plays an important role in the growth of PLBs where the heaviest PLBs were recorded in the medium supplemented with 0.20 % of charcoal along with 4.5 g of agar. Vice versa relationship was recorded for the number of plantlets where without charcoal in 4.5 g of agar produce the highest number of plantlets.

The genotypic response towards haploid induction in Turkish onion (Allium cepa L.) germplasm

Haploid induction efficiency of unpollinated flower buds in twenty-six Turkish onion germplasm were determined on two media, Dunstan and Short medium (BDS) and Murashige and Skoog’s medium (MS). On BDS medium, 5850 flower buds were cultured and the induction rate was 2.10% and a plant survival percentage of 1.62% (2 plants). On MS medium, 5850 flower buds were cultured and 175 plantlets were induced; induction rate was 2.99% and plant survival percentage was 13.71% (24 plants). The highest number of plantlets was obtained from genotype Sanliurfa 2; 27 plantlets on BDS medium while 45 plantlets on MS medium. Flow cytometry confirmed that out of the twenty-six plants, 4 plants were haploid (15.38%), 2 mixoploid (7.69%), 18 (69.23%) diploid and 2 tetraploid (7.69%).

In Vitro Culture of Ovules and Embryos from Seedless Grapes (Vitis vinifera L.)

Ovules of stenospermocarpic seedless grapes were cultured under different conditions. The number of embryos was not significantly increased when ovules of Muscat Seedless were cultured on a medium supplemented with the plant growth regulators indole-3-acetic acid and gibberellic acid compared to the basal medium. No correlation was found between the ovule size of Sultanina and the presence of embryos. Comparable percentages of embryos were found in small, medium and large ovules following weekly culture from four to 10 weeks after bloom. Varying results were obtained when ovules of four open-pollinated seedless cultivars were cultured on two basal media with or without activated charcoal. In all four cultivars the highest number of embryos was found at the later culture date. With three cultivars the best results were obtained when ovules were cultured on the medium of Bouquet & Davis (1989), supplemented with activated charcoal. Isolated embryos of Muscat Seedless and Sunred Seedless were cultured under various conditions and it was found that the absence of growth regulators resulted in the highest number of plantlets with normal leaves.

IN VITRO PROPAGATION OF NATIVE ORCHID DENDROBIUM SPECTABILE (BLUME) MIQ.

The forests of Papua and New Guinea have a rich variety of flowering plants. One orchid species from Papua and New Guinea is the Dendrobium spectabile. The D. spectabile has unique characteristics in which the flowers are slightly different in a florescent light. The flowers of D. spectabile are dissimilar with most of the other Dendrobium sp. Unlike the common Dendrobium, the D. spectabile will not flower without much higher levels of light. They will even take full sunlight for longer periods of time. The D. spectabile will start flowering when it reaches 4-5 years old. Recently, many people exploit this species, causing it to become very rare. Therefore, it is important to study the propagation method in order to conserve this species. In this study, in vitro propagation using different culture media were explored. The aim of this study is to compare two different media, i.e. medium VW with modification and medium W3, which has been known to give highest respond for growth of D. spectabile. The experiment was carried out at Tissue Culture Laboratory, CV Ayu Orchid Bali, from April-August 2012. The experiment employs Completely Randomized Design (CRD) design with 20 replications for each media. The result of the experiment showed that the highest respond for growth of D. spectabile was VW medium. This was indicated by higher number of plantlets (average 306.75), number of leaves (average 3.45) and plant height (average 2.65 cm). However, the leaves of plantlets using medium W3 showed dark green and the explants were stronger than explants in medium VW.

Plant regeneration via somatic embryogenesis in Drimia robusta

A simple efficient in vitro plant regeneration system was developed by direct and indirect somatic embryogenesis of Drimia robusta, a medicinal plant extensively used in South African traditional medicine. Different developmental stages of somatic embryos (SEs: globular embryos, partial pear-shaped embryos and club-shaped embryos), club-shaped cotyledon initiation, plumule initiation and plantlets were directly obtained from leaf explants on Murashige and Skoog (MS) medium containing 3.5 % (w/v) sucrose and different plant growth regulators (PGRs). In MS medium containing 3.5 % (w/v) sucrose and supplemented with 10 μM picloram, 1 μM thidiazuron (TDZ) and 20 μM glutamine, a higher number of SEs and plantlets were achieved. These were established onto half-strength MS medium followed by successful acclimatization (100 %) in the greenhouse. Liquid somatic embryo medium (SEML) containing 500 mg of friable embryogenic callus on MS medium supplemented with different concentrations and combinations of PGRs and organic elicitors produced different stages of SEs. Somatic embryo production was enhanced by 0.5 μM picloram, 1 μM TDZ and mebendazole treatment. The highest number of plantlets (9.0 ± 0.70) was obtained in SEML containing 0.5 μM picloram, 1 μM TDZ and 25 mg l−1 haemoglobin. All the cotyledon and plumule embryos germinated on half-strength MS medium, however 90 % of SEs germinated on half-strength MS medium containing 0.5 μM naphthaleneacetic acid. All plantlets were successfully acclimatized in the greenhouse. This first report of D. robusta somatic embryogenesis provides an opportunity to control extinction threats, ensure germplasm conservation and provides a system for analysis of bioactive compounds and bioactivity.

Enhancement of regeneration efficiency in banana (Musa acuminata cv. Berangan) by using proline and glutamine

Regeneration of Musa acuminata cv. Berangan (AAA) from embryogenic cell suspension was enhanced by using proline and glutamine. Sixteen embryo development media consisting of different concentrations of proline and glutamine were tested. Glutamine was preferred as the source of organic nitrogen supplement compared to proline. The embryos developed in liquid media were shown to produce 10.6-fold higher number of plantlets in lesser duration (1.5 to 2.5 months) as compared to solid media. Optimum somatic embryo development was observed in liquid media supplemented with 400 mg/l glutamine (M4GL) with an average regeneration of 33,844 ± 402 plantlets per ml of settled cell volume (SCV). Although both amino acids have proven to promote the embryo development, concentrations of proline between 200 to 400 mg/l in liquid media caused abnormal embryo differentiation.

Plantlet regeneration through indirect shoot organogenesis and somatic embryogenesis in Justicia gendarussa Burm. f., a medicinal plant

A protocol for the regeneration of a large number of plantlets via indirect shoot organogenesis and somatic embryogenesis has been developed from the stem and leaf explants of Justicia gendarussa Burm. f. The callus was efficiently induced from the explants using Murashige and Skoog (MS) medium supplemented with α-Naphthalene acetic acid (NAA) + Benzyl amino purine (BAP) (1.0 + 0.1 mg/l). The highest number of plantlets through indirect shoot organogenesis was obtained when the callus was subcultured to MS medium with BAP + NAA (0.1 + 1.0 mg/l). The maximum number of plantlets via somatic embryos was obtained in the medium with BAP + NAA (1.0 + 0.1 mg/l) for stem derived calli and Kinetin (Kn) + NAA (2.0 + 0.1 mg/l) for leaf derived calli. The in vitro developed shoots were rooted well in half strength MS medium supplemented with 0.5 mg/l of Indole-3-acetic acid (IAA). The in vitro regenerated plantlets were hardened using a mixture of sterile sand:soil:manure (1:1:1). The present study is the first report on the regeneration of plants through somatic embryogenesis from stem and leaf derived calli of J. gendarussa.

Effect of plant growth regulator combination and culture period on in vitro regeneration of spinach (Spinacia oleracea L.)

The objective of this study was to develop an efficient system for the regeneration of spinach plants (Spinacia oleracea L.) by investigating the factors influencing callus and shoot induction. All plant growth regulator (PGR) combinations tested induced callus with high frequency (73–100 %), and the combination of 5 μM α-naphthaleneacetic acid (NAA), 10 μM 6-benzyladenine (BA) and 0.1 μM gibberellic acid (GA3) had the most significant effect on callus growth in term of weight (120.98 ± 22.56 mg). A high auxin-containing medium induced competent callus for shoot formation, while high cytokinin-containing media enhanced callus growth and made callus incompetent for shoot regeneration. Longer periods of callus induction in a high auxin-containing medium were required to form competent callus and led to a high regeneration capacity. The PGR combination shift from a high auxin to cytokinin ratio (ACR) to a low ACR resulted in highly efficient regeneration. Among the regeneration systems tested, the combination of 10 μM NAA and 0.3 μM GA3 for callus induction for 6 weeks followed by 2 μM NAA and 5 μM BA resulted in the highest plant regeneration frequency (83.33 ± 6.43 %) and the highest number of plantlets per explant (7.93 ± 1.24). Somatic embryos at cotyledonary stage and plantlets were transferred to PGR-free medium to establish whole plants. Regenerated female plants grew well to maturity in the greenhouse (77.17 ± 9.80 %) and produced seeds (175.21 ± 28.01 firm seeds per plant).

Effects of banzyladenine and gibberellin on runner production and some vegetative traits of three strawberry cultivars

Two experiments were conducted to study the effect of Banzyladenin and Gibberellin on increasing runner production and some vegetative traits of three strawberry cultivars in greenhouse. The experiments were carried out in a Completely Randomized Design (CRD), with two factors (hormonal treatments and three strawberry cultivars). In the first year (2006), hormonal treatments had two levels (a combination of BA at 1200 ppm and GA in 300 ppm, and control). Three strawberry cultivars (Pajaro, Queen Eliza and Paros) were sprayed in May and July. In the second experiment (2007), hormonal treatments had five levels (1- Control (0 ppm), 2- BA 1200 ppm + GA 300 ppm sprayed in May, 3- BA 1200 ppm + GA 300 ppm sprayed in July, 4- BA 1200 ppm sprayed in May, 5- GA 300 ppm sprayed in May). Effects of hormonal treatments were not significant on any of the vegetative traits in the first year, but not the second year. Highest number of plantlets with a standard diameter of 6.15 mm was produced by applying the second treatment (mixture of BA 1200 ppm and GA 300 ppm sprayed in May) and it was chosen as the best treatment. However, highest number of runners in each mother plant and highest diameter of plantlets were observed in the control (0 ppm). Cultivars responded differently to hormones. Number of runners in Paros cultivar decreased dramatically with application of hormones, but it did not change in Queen Eliza cultivar.

Optimization of peptidase production conditions in koji making

Two experiments were conducted to study the effect of Banzyladenin and Gibberellin on increasing runner production and some vegetative traits of three strawberry cultivars in greenhouse. The experiments were carried out in a Completely Randomized Design (CRD), with two factors (hormonal treatments and three strawberry cultivars). In the first year (2006), hormonal treatments had two levels (a combination of BA at 1200 ppm and GA in 300 ppm, and control). Three strawberry cultivars (Pajaro, Queen Eliza and Paros) were sprayed in May and July. In the second experiment (2007), hormonal treatments had five levels (1- Control (0 ppm), 2- BA 1200 ppm + GA 300 ppm sprayed in May, 3- BA 1200 ppm + GA 300 ppm sprayed in July, 4- BA 1200 ppm sprayed in May, 5- GA 300 ppm sprayed in May). Effects of hormonal treatments were not significant on any of the vegetative traits in the first year, but not the second year. Highest number of plantlets with a standard diameter of 6.15 mm was produced by applying the second treatment (mixture of BA 1200 ppm and GA 300 ppm sprayed in May) and it was chosen as the best treatment. However, highest number of runners in each mother plant and highest diameter of plantlets were observed in the control (0 ppm). Cultivars responded differently to hormones. Number of runners in Paros cultivar decreased dramatically with application of hormones, but it did not change in Queen Eliza cultivar. Key words: Hormone, daughter plant, plantlet, crown, strawberry.

Morphological, physico-chemical and micropropagation studies in Jatropha curcas L. and RAPD analysis of the regenerants

Four accessions of Jatropha curcas L. were characterized with respect to their yield and physico-chemical properties of seed-oil. Further, an efficient micropropagation protocol was developed followed by RAPD analysis of the regenerants. Although the highest 1000-seed-weight and seed-oil content were recorded in OJC1 and OJC5 accessions respectively, the seed-oil of OJC9 accession was more suited for biodiesel production due to the desirable characteristics like low values of free fatty acids, acid value, moisture content and total unsaturated fatty acids. An accession – independent micropropagation protocol of Jatropha was optimized by using axillary nodal explants with MS medium supplemented with a combination of growth regulators, glutamine and citric acid. All the accessions were amenable to in vitro culture, and the highest number of plantlets could be regenerated from the OJC5 accession. Although no somaclonal variation was recorded in the regenerants of all the accessions, RAPD analysis confirmed their genetic similarity and also the similarity of the clones with their respective mother plants.

EFFECTS OF ORGANIC SUPPLEMENTS IN LOCAL FERTILIZER MEDIUM ON GROWTH AND DEVELOPMENT OF PHALAENOPSIS SILKY MOON PROTOCORMS

Seed-derived protocorms of Phalaenopsis Silky Moon, approximately 5 mg, were used as explants for protocorm-like bodies (PLBs) proliferation and induction of plantlets. Basal culture media contained 2.75 g/L Viking Ship (10N-20P-30K), a local commercial fertilizer, supplemented with 22.2 μM N 6 -Benzyladenine (BA), 20 g/L sucrose, 1.0 g/L activated charcoal and 2.0 g/L Phytagel. Effects of organic supplements on growth and development of explants were investigated by adding 100 g/L boiled potato juice, 2 g/L peptone, 10 g/L polyethylene glycol, 10 mg/L adenine sulphate, 6 g/L malt extract agar or 150 mg/L glutamine in a basal medium with different combinations. All explants were cultured for 18 weeks. The basal medium containing glutamine with peptone and that containing glutamine, peptone with adenine sulfate provided a high number of plantlets (5.1 to 6.2) with significantly higher total fresh weight (FW) (1.06 to 0.94 g), shoot FW (0.60 and 0.52 g) and root FW (0.47 and 0.43 g) than other media. The shoot FW and root FW ratios obtained from both media were 1.2 and 1.3, respectively. Although the highest number of plantlets (6.0 and 9.7) and number of protocorm-like bodies (PLBs) (6.4 and 5.0) were found from the basal medium containing boiled potato juice with peptone and that containing boiled potato juice, malt extract agar, adenine sulphate with polyethylene glycol (PEG), respectively, the shoot FW and root FW ratios were very high at 4.1 and 2.1, respectively. Effects of other organic supplements on PLBs and plantlets formation were discussed.

EFFICIENT PROPAGATION BY BIOREACTOR SYSTEM OF KOREAN NATIVE GINGER IN SEOSAN REGION

To establish an efficient propagation method through airlift bioreactor system of Zingiber officinale Rosc. Korean native Seosanjong, the effect of different factors and bioreactors on cultured plantlets were investigated. The highest number of plantlets, fresh weight per plant was obtained from explants when cultured in MS liquid medium including 0.3 mg/L NAA and 2.0 mg/L kinetin for 40 days. A 10 L bottle type bubble bioreactor, compared with 250 ml Erlenmeyer flask, was more efficient, producing 4.7 plantlets or from 1.5 to 1.6 times more than did the Erlenmeyer flask. The results demonstrate a rapid mass propagation method for scaling up production of normal ginger using the airlift bioreactor.

The relative importance of different regeneration mechanisms in a selectively cut savanna-woodland in Burkina Faso, West Africa

Savanna plant species are capable of regenerating both sexually and asexually. The importance of each regeneration mechanism depends on the species, the type and the intensity of disturbance. This paper describes the relative importance of sexual and asexual regeneration for the restoration of a selectively cut savanna woodland in Burkina Faso. Plantlets regenerated sexually or asexually were determined by excavating the below ground system and assessing basal and aerial sprouts within 144 quadrats along transects in 2, 5, 10 year-cut and control plots. The result shows that seedling sprouts were predominant (83%) compared to root sucker (4%), coppice (5%), water sprout (2%) and layering (less than 1%). True seedlings constituted a minor component (5%) of the plantlet population. A total of 54 woody species was found in the plantlet population with 52 presenting seedling sprouts. Twelve species were found regenerating from both seeds and root buds; of which Dichrostachys cinerea, Pteleopsis suberosa and Detarium microcarpum had the highest number of plantlets. Feretia apodanthera was the only layering species encountered in the sampled area. Coppices, root suckers and water sprouts showed higher height and bigger collar diameter compared to seedling sprouts and true seedlings. It can be concluded that sexual reproduction is the dominant mechanism of seedling recruitment in this disturbed savanna-woodland, and that its success relies on the ability of plantlets of seed origin to resprout abundantly.

EFEITOS DO BAP E DO TDZ NA PRODUÇÃO DE MUDAS DE BANANEIRA - 'MAÇÃ' ATRAVÉS DA PROPAGAÇÃO RÁPIDA "IN VIVO"

O 6-benzilaminapurina (BAP) a 0,0 e 10,0 mg/l e o tidiazuron (TDZ) a 0,0; 10,0 e 20,0 mg/l foram testados quanto aos seus efeitos indutores de brotações visando à produção de mudas de bananeira-'Maçã', através da técnica de propagação rápida "in vivo". O trabalho foi conduzido na Fazenda Experimental da EPAMIG, em Lavras, MG, no período de abril a novembro de 1995. Os rizomas com 16,0 a 20,0 cm de diâmetro, previamente preparados, foram plantados em bancadas contendo substrato de areia e casca de arroz (1:1), sob cobertura plástica transparente, a 1,3 m de altura, em condições de telado. Os resultados obtidos mostraram que nenhum dos reguladores de crescimento testados aumentaram significativamente a quantidade de mudas produzidas em relação à testemunha. O máximo que se conseguiu, foi 2,99 mudas/broto tratado com 10,0 mg/l de BAP mais 20,0 mg/l de TDZ. Entretanto, os tratamentos contendo somente BAP a 10,0 mg/l reduziram o período de brotação (do tratamento dos brotos ao início da brotação) de 32,0 para 24,2 dias (24,4%), em relação à testemunha e de 30,6 para 24,2 dias (17,6%), em relação aos tratamentos contendo somente TDZ. Por outro lado, o período ou ciclo total de produção de mudas foi reduzido de 200,0 para 188,1 dias (6,95%) e de 198,4 para 188,1 dias (5,2%) em relação à testemunha e aos tratamentos contendo somente TDZ, respetivamente.

In vitro propagation of reed grass by somatic embryogenesis

A micropropagation system using regeneration via somatic embryogenesis from immature inflorescences has been optimized. This system is proposed for the production of the macrophyte Phragmites australis (Cav.) Trin. for the construction of wetlands used in wastewater purification. Embryogenic calli were produced in florets from inflorescences in the presence of 2,4-dichlorophenoxyacetic acid in the induction media. Up to 28.4% of the calli were embryogenic. Somatic embryos developed into plantlets when transferred to the regeneration medium lacking growth regulators. The addition of myo-inositol to the induction medium resulted in the highest number of plantlets on the regeneration medium. A decrease in the number of plantlets was observed when the embryogenic calli were maintained longer than three months on the induction medium. Plantlets can be further propagated by node culture. Plantlets were successfully acclimatized and developed normally showing no morphological differences when compared to seed-grown plants.

Production of plantlets for use as artificial seeds from horseradish hairy roots fragmented in a blender

An efficient method for production of plantlets from horseradish hairy roots ( Armoracia rusticana) was developed and the plantlets were encapsulated and used as artificial seeds. Hairy root fragments were obtained by fragmentation of hairy roots in a blender after the root growth. A fragmentation time of 30 s gave the best result on the plantlet formation. Treatment of the hairy roots with naphthaleneacetic acid (NAA, 1.0 mg/ l) prior to fragmentation resulted in a decrease in the time required for hairy root growth. Kinetin supplementation of 0.1 mg/ l after fragmentation (in the plantlet formation stage) resulted in the highest number of plantlets. Furthermore, we found that the kinetin treatment led to a change in the size distribution of plantlets and inhibition of root elongation, which facilitated encapsulation of plantlets. Although the frequency of plantlet development was significantly reduced by encapsulation in 2% calcium alginate beads, dehydration treatment before the encapsulation strongly improved the plantlet development frequency.

Plant Regeneration of Zoysiagrass from Embryo‐Derived Callus

Tissue culture methods could enhance development of improved cultivars of zoysiagrass (Zoysia japonica Steudel). A tissue culture system for callus induction and plant regeneration of zoysiagrass was developed. Embryos were aseptically excised from seeds and over 90% produced callus. Embryogenic callus was produced on Murashige‐Skoog (MS) agar medium supplemented with 2,4‐D (2,4‐dichlorophenoxy) acetic acid, but not on N6 medium. A 2,4‐D concentration of 1.0 mg L−1 resulted in greater callus production than higher concentrations. A 16‐h photoperiod of fluorescent light (65 μmol photon m−2 s−1) increased callus production over dark conditions. Direct elimination of 2,4‐D and dark callus induction resulted in greater regeneration frequency and a higher number of plantlets per callus than gradual elimination and light callus induction. Normal plantlets were transplanted to soil and grown to maturity in the field, where overwinter survival was exhibited.