Percentage Of Shoot Formation Research Articles

TiO2 nanoparticle synthesis, characterization and application to shoot regeneration of water hyssop (Bacopa monnieri L. Pennel) in vitro

ABSTRACT Water hyssop (Bacopa monnieri L. Pennel) is a medicinal aquatic herb used to treat diseases in South Asia. Various regeneration protocols have been developed or modified in vitro to ensure the availability of biomass and secondary metabolites of Bacopa. We applied hydrothermally treated titanium dioxide (TiO2) nanoparticles (NPs) (TiO2-NPs) at different concentrations. Three explants, distal portion of half leaf (DPHL), proximal portion of half leaf (PPHL) and full leaf (FL), were used to evaluate response to TiO2. Regeneration from the three explants in vitro was similar except for shoot length. Application of TiO2-NPs exerted significant, but variable, effects on all parameters except percentage of shoot formation, which was 100%. Interactive effects of explant and TiO2-NPs exhibited significant, but variable, effects on fresh weight and percentage of callus formation. All explants produced more shoots using TiO2-NPs compared to control treatments. DPHL explants with application of 8 mg/l TiO2 produced more shoots than controls. Similarly, FL explant treated with 2 mg/l TiO2-NPs produced more shoots/explant than controls. All concentrations of TiO2-NPs produced significantly longer shoots compared to controls. Increased elongation of shoots justifies use of TiO2-NPs for propagation of plants in vitro during acclimatization. Use of TiO2-NPs for rapid elongation of shoots ultimately fosters survival of plants.

Leaf pigments and concentrations of phenolic compound in Begonia grandis plantlets obtained from the floral explants

The objective of this study was to design a protocol of the successful establishment of plants of Begonia grandis dry. in culture in vitro, and to evaluate the concentrations of chlorophyll a and b, carotenoids, anthocyanins and phenolic compounds, as well as the ratios of chlorophyll a/b, total chlorophyll/carotenoids and total chlorophyll/anthocyanins in the in vitro plantlets in comparison to the same parameters assessed in the leaves of greenhouse plants. A significant increase in the percentage of shoot formation (22.0 ± 1.5%) was recorded on the modified N6 medium supplemented with plant growth regulators in comparison with the standard breeding technology. Begonia grandis plantlets possessed the elevated contents of all photosynthetic pigments per unit mass of plant compared to the greenhouse plants. The increase in chlorophyll b was most substantial. The average anthocyanin concentrations in the plantlets and greenhouse plants were approximately equal, but the essential variation within B. grandis microclones was observed. Phenolic profiles of in vitro plantlets and greenhouse plants was rather similar. Concentrations per unit mass for the majority of components (especially organic and phenolic acids) in the plantlets increased in comparison to those in greenhouse plants (47.23 and 38.22 mg g−1 total phenolics, respectively). Overall data suggest that this protocol can be used for the production of a large number of plants of B. grandis in a good physiological state and with high phenolic content. Graphical abstract

Addition of Benzyl Amino Purine (BAP) to Induction of Drendan Seeds Buds (Lansium domesticum var. aqueum (Jack) Miq.) by In Vitro

Drendan (Lansium domesticum var. aqueum (Jack) Miq. is one of Indonesia's native tropical fruits. Drendan has high antioxidant activity and contains high nutrients, but has started to rarely be found in the field because its management and cultivation is rarely done due to its sour taste in vitro culture is a method or technique of plant propagation which can be an alternative to obtaining drendan seedlings in large quantities and in a short time vegetatively. The purpose of this study was to determine the effect of BAP on shoot induction from drendan seed explants and determine the best concentration of BAP on shoot growth of drendan seed explants in vitro. This study used a completely randomized design with BAP concentrations of 0, 1, 3, 5 and 7 mg / L BAP. The result of this research is the addition of BAP has been able to increase the percentage of live explants and the percentage of shoot formation. Giving BAP did not significantly affect the number of shoots and had a significant effect on shoot emergence time and shoot length. The addition of BAP resulted in slower shoot emergence time. Giving BAP was not able to stimulate shoot induction, but the addition of BAP could increase shoot length growth at a concentration of 5 mg / L BAP of 0.52 cm.

CHIA SEED AS A SOURCE OF IN VITRO ESTABLISHMENT OF Salvia hispanica L. PLANTS

Technique of tissue culture for Chia (Salvia hispanica) micropropagation was achieved, this study investigated the impact of various concentrations of plant growth regulators on shoot multiplication and root induction with the Chia’s mature seed as a source explant. The highest percentage of shoot formation (80%), shoots number per explant(3.20) and shoot length(3.26 cm), were recorded on MS medium enriched with BAP(1.0 mgl-1) after eight weeks of seed culture. The optimal medium for the rhizogenesis was achieved on half strength MS medium fortified with 1.0 mgl-1 IBA after four weeks of culture, which had the highest rooting percentage (100%) with highest mean of roots number (5.6 roots per shoot) with (3.40 cm root length). The rooted plants were successfully adapted ex vitro with a survival rate of 85%.

Efficient plant regeneration and callus induction from nodal and hypocotyl explants of goji berry (Lycium barbarum L.) and comparison of phenolic profiles in calli formed under different combinations of plant growth regulators

Goji berry (Lycium barbarum L.) is a fruit that has many beneficial effects (such as antidiabetic, antioxidant, anticancer, antidepressant, and immunomodulatory) on human health because of several dietary constituents such as phenolics, vitamins, carotenoids, and polysaccharides. In order to develop in vitro culture protocols for callus induction and plant regeneration from different explants of Goji plants and to compare the phenolic composition in calli of different origin, various combinations of plant growth regulators (PGRs) were applied. Various types of explants (leaf, petiole, root, hypocotyl, and node) were cultured on MS medium containing 28 different concentrations and combinations of PGRs [thidiazuron (TDZ) and naphthalene acetic acid (NAA), TDZ and indole-3-acetic acid (IAA), benzyl adenine (BA) and NAA, alone TDZ, and alone BA]. The highest mean number of shoots (23.33 ± 1.86) and percentage of shoot formation (100%) were obtained from nodal explants on medium containing 0.5 mg/l BA alone. The highest mean callus diameter was obtained from hypocotyl explants on medium containing 0.25 mg/l TDZ and 0.1 mg/l IAA (21.40 ± 0.71 mm). The amounts of selected phenolic substances were significantly different in the callus obtained from different PGRs or combinations, individually. The combination of BA/NAA significantly increased the production and accumulation of chlorogenic acid and caffeic acid. The TDZ/IAA combination, TDZ alone, and TDZ/NAA combination significantly increased vanillic acid and rutin, gallic acid, and quercetin synthesis, respectively. These outcomes indicate that different PGRs lead to the production of different kinds of secondary metabolites and affect/accelerate accumulation in the callus of goji berry. The culture protocol described in this paper could be employed for the development of novel methods for the commercial production of goji secondary metabolites.

Regeneration of rice somaclons tolerant to high level of abscisic acid and their characterization via RAPD markers

BackgroundThe main objectives of this study were to compare two in vitro selection procedures for developing drought-tolerant cell lines from rice cultivars, regenerate plants from the selected cells, and use the random amplified polymorphic DNA (RAPD) method for genetic characterization of somaclons. Callus was derived from mature embryo cultures of tow Egyptian rice cultivars, Sakha 102 and Giza 178.ResultsThe direct in vitro selection method indicated that the callus weight of the two cultivars was significantly decreased in response to increasing ABA concentrations. The shoot formation revealed significant differences among ABA concentrations and highly significant for interaction between rice cultivars and ABA concentrations. The percentage of shoot formation either increased or decreased with increasing ABA levels in rice cultivars. However, Giza 178 cultivar was slightly stable for shoot formation than the cultivar Sakha 102. The obtained results for stepwise selection method indicated also that the shoot formation was obtained till 6 mg/L ABA. The results of the RAPD analysis indicated that the rice cultivars and their somaclons were classified into tow clusters. The polymorphic information content (PIC) per primer ranged from 0.715 to 0.906 with an average of 0.860. The results of UPGMA clustering were consistent with those of the PCoA.ConclusionsThe percentage for Sakha 102 was 10.8, 7.7, and 2.2% on 2, 4, and 6 mg/L ABA, respectively, and was 16.4, 10.0, and 3.4% for Giza 178 cultivar, at the ABA concentrations of 2, 4, and 6 mg/L, respectively. The genetic similarity among the fifteen genotypes ranged from 0.30 to 0.82. Rice cultivars and their somaclons were classified into five clusters.

Pertumbuhan Tanaman Pisang Kepok (Musa paradisiaca L.) pada Berbagai Konsentrasi BAP Secara In Vitro

Bananas are one of the horticultural commodities is very popular in the world because it was delicious, the price is relatively cheap and have high nutritional value. Production of banana plants in Hulu Sungai Utara still relatively low because only bananas cultivated on a small and not available the procurement of seeds. To overcome these obstacles do necessary aside propagated using tissue culture techniques. ZPT which are often used to stimulate the formation of shoots in vitro culture banana is cytokinin as BAP. Cytokines in vitro culture has two important roles that stimulate cell division and the formation and propagation of axillary buds and adventitious buds, but the optimum levels of cytokines that can inhibit the growth and root formation. The purpose of this study is to get (i) the effect of various concentrations of BAP on plant growth kepok banana in vitro, (ii) the best BAP concentration for plant growth kepok banana in vitro. This research was conducted in tissue culture laboratory STIPER Amuntai in April 2013 to August 2013 using a completely randomized design (CRD) single factor with 5 treatments and 4 replications, so there are 20 experimental units. Each experimental unit consisted of 2 explants as a sample. Factors that are tested various concentrations of BAP are b1 : 2 mg / l, b2 : 3 mg / l, b3 : 4 mg / l, b4 : 5 mg / l, b5 : 6 mg / l. The results showed that percentage grows, the percentage of shoot formation, root formation percentage, number of shoots and number of leaves had no effect on treatment.

23. Comparison of vitrification-based techniques in the efficacy of cryopreservation of Passiflora suberosa L. and P. foetida L. shoot tips

Passiflora are used as ornamentals and in popular medicine. Passiflora suberosa is commonly used to treat hypertension, diabetes, skin diseases and as a sedative, whereas P. foetida shows analgesic and antibacterial activities. The goal of this study was the development of in vitro conservation strategies for long-term storage for both species through cryopreservation using vitrification and encapsulation-vitrification techniques. The efficacy of vitrification and encapsulation-vitrification protocols was compared, and several parameters were evaluated, including pregrowth on medium with high sucrose concentrations, type of vitrification solution (PVS2 and PVS3), exposure time to vitrification solutions, and recovery conditions. Shoot tips (0.3 cm) excised from in vitro-grown plants were precultured for 24 h on solid 1/2 MSM medium supplemented with different sucrose concentrations (0.3, 0.5 or 0.7 M), at 25 °C. Explants precultured on medium supplemented with 0.3 M sucrose were incubated in the cryoprotectant solutions PVS2 or PVS3 for different periods (0–240 min) at room temperature, and immersed into liquid nitrogen (LN) for 72 h. In the encapsulation-vitrification protocol, precultured shoot tips were encapsulated in calcium alginate beads, followed by incubation in liquid 1/2 MSM medium containing 0.75 M sucrose for 24 h, on a rotary shaker (100 rpm) at 25 ± 2°C. After this period, beads were maintained in cryovials containing PVS2 or PVS3 for 30, 60, 120 or 180 min, before immersion into LN. The materials were rapidly thawed in a warm water bath (38-40 °C) for 2 minutes and transferred to the recovery medium (1/2 MSM or MSM supplemented with 0.44, 2.2 or 4.4 μM BA). Alternatively, explants were kept in the dark for 30 or 60 days before transfer to the presence of light. Plant recovery from the shoot tips before and after cooling in LN was evaluated by assessing the percentage of shoot formation. The shoots were cultured on 1/2 MSM medium for 60–90 days for root induction, and whole plants were transferred to ex vitro conditions. The highest recovery rates for both P. suberosa and P. foetida (28% and 60%) were obtained with the encapsulation-vitrification protocol, after a pre-treatment with 0.3 M sucrose, followed by exposure to PVS2 for 60 or 120 min, respectively, and post-freezing incubation in the absence of light for 30 days on MSM medium supplemented with 0.44 μM BA. These results suggest that alginate beads provided additional protection, reducing the toxicity of the cryoprotectant solution. In addition, they demonstrate that different species from genus Passiflora display distinct responses to cryopreservation treatments.

The effect of liquid organic fertilizer and pearl grass extract on shoot growth of gotu kola (Centella asiatica) in vitro

Hayanti IP, Amiputri RB, Praswanto. 2012. The effect of liquid organic fertilizer and pearl grass extract on shoot growth of gotu kola (Centella asiatica) in vitro. Biofarmasi 10: 54-60. Gotu kola (Centella asiatica L.) is a medicinal crop, needed by local industry of pharmacy. Gotu kola in Indonesia is not enough to supply in market requisites. Tissue culture method was used to multiply gotu kola with a combination of medium from liquid organic fertilizer and pearl grass extract. The purposes of the research were to determine the effect of liquid organic fertilizer and pearl grass extract on the shoot growth of gotu kola in vitro, and to obtain the optimal concentration of liquid organic fertilizer and pearl grass extract combination on the shoot formation of gotu kola in vitro. The research was conducted in November 2010 to June 2011 in Plant Physiology and Biotechnology Laboratory, Faculty of Agriculture, Sebelas Maret University, Surakarta. The experimental design used was a Completely Randomized Design (CRD) with two treatment factors and three replications. The first factor was liquid organic fertilizer concentrations, i.e. 0 mL/L, 2 mL/L, 4 mL/L and 8 mL/L. The second factor was pearl grass extract, i.e. 0 mL/L, 3 mL/L, 6 mL/L and 12 mL/L. Variables observed were the percentage of shoot formation, the time of shoot formation, the number of shoot, the length of shoot, the percentage of leaf formation, the time of leaf formation and the number of leaf. The result of the research showed that not all of treatments provided shoot formation. The combination of liquid organic fertilizer concentrations 0 mL/L and pearl grass extract 12 mL/L showed the best results, in which the average of length of shoot was 2.16 cm, the number of shoot was 8 and the number of leaf was 8.

Shoot regeneration and micrografting of micropropagated hybridtomatoes

SummaryThe use of grafted tomato plants is expanding. An in vitro propagation system was established for three commercial F1 cultivars (‘Garnet 622’, ‘Jumbo’ and ‘Marvel’) to combine micropropagation with micrografting. Nodal tissue explants were excised from the central stem of plantlets and micropropagated on MS medium. A micropropagation rate of 6 shoots per explant was obtained for all three hybrids. Micropropagated shoots were then micrografted onto 3 week-old ‘RT-79’ rootstocks with a union rate of over 80%. After acclimatisation, micropropagated plants were established ex vivo, flowered and matured normally. In addition, regeneration protocols were tested using cotyledon explants from 10 d-old seedlings. Adventitious shoot organogenesis from three parts of the cotyledon explants (proximal, middle, distal) was monitored on MS media supplemented with different concentrations of zeatin (Z) and indole-3-acetic acid (IAA). The highest percentage of shoot formation was observed on MS medium supplemented with 0.5 mg l–1 Z and 0.1 mg l–1 IAA acid with cotyledon explants that originated from the proximal part. Up to 80% of explants formed adventitious shoots, with an average of three shoots per explant. Three week-old regenerated shoots were placed on MS medium without growth regulators for elongation, rooting and further development of plantlets.

Management of chimera through direct shoot regeneration from florets of chrysanthemum (Chrysanthemum morifolium Ramat.)

SummaryThe main bottleneck of mutation induction in vegetatively propagated ornamentals is the development of chimera. Ephemeral small sectors apparent on petals cannot generally be recovered as solid mutant shoots. An efficient technique has been standardized in the present experiment for direct shoot regeneration from individual florets of...chrysanthemum (Chrysanthemum morifolium Ramat.) cv. Colchi Bahar. The efficiency of direct shoot regeneration was tested on 16 media based on MS basal medium containing sucrose, agar and different concentrations and combinations of BAP and NAA. The regenerative potential varied with the developmental stages of the flower head. The highest percentage of shoot formation and most shoots per responding explant were observed on florets taken from stage II flower heads. Direct shoot formation was confirmed by scanning electron microscopy and histological observations. The technique has been tested on 16 other cultivars in the medium containing 0.2 mg l21 NAA and 1.mg.l1 BAP. Gamma ray of 1.5 and 2.0 krad induced yellow and white floret colour sectors in `Maghi' (original colour-mauve) have been established in pure form using this technique. The regenerated plants were rooted in vitro and successfully transferred to soil where they grew vigorously and flowered true to the floret colour type.

An efficient method for adventitious shoot regeneration from stem-segment explants of gypsophila

An efficient adventitious shoot regeneration procedure was developed for Gypsophila paniculata L. Using cultivar Arbel, shoot regeneration from the three upper internodes of the stem was monitored on MS media supplemented with different cytokinins (thidiazuron, benzyladenine, kinetin or zeatin) and an auxin (naphthaleneacetic acid). Thidiazuron was found to be the most efficient cytokinin, with up to 100% of the explants forming shoots, at an average of up to 19 shoots per explant being regenerated. The highest percentage of shoot formation was observed in the stem explants originating from the first internode, with all cytokinins tested. The adventitious origin of shoots regenerated from stem explants was confirmed by scanning electron microscopy. The regeneration procedure was found to be applicable to five other gypsophila cultivars (Perfecta, Golan, Gilboa, Flamingo and Tavor). Regenerating plants were successfully transferred to soil, and did not differ in flower color, size or shape from standard vegetatively propagated plants derived from cuttings.

Plant regeneration via somatic embryogenesis from protoplasts of Uganda cherry orange (Citropsis schweinfurthii)

Protoplasts isolated from embryogenic callus of Citropsis schweinfurthii (Engl.) Swing. & M. Kell were cultured in MT (Murashige and Tucker 1969) basal medium containing 5% sucrose supplemented with 0.0, 0.001, 0.01, 0.1 or 1.0 mg l(-1) BA, 0, 300, 600 or 900 mg l(-1) malt extract and 0.6 M sorbitol. The highest plating efficiency was obtained on MT basal medium containing 5% sucrose supplemented with 0.01 mg l(-1) BA and 600 mg l(-1) malt extract. MT basal medium containing 5% sucrose and supplemented with 0.01 mg l(-1) kinetin was found to be a medium suitable for the development of globular somatic embryos derived from protoplasts into heart-shaped somatic embryos with cotyledon-like structures. The highest percentage of shoot formation was obtained using 0.1 mg l(-1) GA3. A complete protoplast to-plant system was developed for C. schweinfurthii, which could facilitate the transfer of nuclear and cytoplasmic genes from this species into cultivated Citrus through protoplast fusion.

Regeneration of Arabidopsis callus in vitro

This paper reports on an easy and reproducible method of regenerating Arabidopsis plants from callus culture. A combination of 6-benzylaminopurine (BAP) and α-naphthalene acetic acid (NAA) in a Murashige and Skoog's (MS) based medium gives a high percentage of shoot formation in several genotypes.