Short-Term Storability of Alginate-Encapsulated Persian Violet Microshoots for Germplasm Exchange

Microshoots have been widely used for micropropagation. It may be necessary to store microshoots for a short period of time, for example in germplasm exchange needing transport to other research groups. Here, we investigated the short-term storability of alginate-encapsulated Persian violet (Exacum affine Balf. f. ex Regel) microshoots at 4 °C and 25 °C. After storage, the encapsulated microshoots were sown on basal Murashige and Skoog medium for germination and viability determination using tetrazolium chloride staining. The results showed that one or five microshoots encapsulated with a single alginate layer could be stored at 4 °C for up to 30 days, while the percentages of germination and viability of the microshoots encapsulated with two layers of alginate were greatly reduced upon storage. This is the first report on the storability of alginate-encapsulated multiple microshoots, which could be a more efficient way to encapsulate microshoots used for short-term cold storage.

In vitro propagation from nodal segments of Lippia origanoides (chemotype A)

ABSTRACT: This research described an efficient micropropagation protocol for Lippia origanoides (Verbenaceae). Sterile seeds were used to obtain germinated seedlings in Murashige and Skoog medium (MS) supplemented with sucrose and agar. The nodal segments obtained from seedlings were grown on MS medium supplemented with different concentrations of gibberellic acid (GA), benzylaminopurine (BAP) and 1-naphthalenacetic acid (NAA) with BAP. The callus induction, shoots length, shoots number and root length, were analyzed. The treatments showed high percentage of callus formation at 0.5 to 1.5 mg L-1 of BAP alone or in combination with NAA (0.1 mg L-1). The highest value of shoot number per nodal segments was obtained at 1.5 mg L-1 of BAP (4.3 ± 0.8). The obtained plantlets were better rooted in vitro in the absence of plant growth regulators (PGRs) and they showed acclimatization rate of 90%. We reported a protocol for in vitro propagation and acclimatization of L. origanoides for A chemotypes from Colombia.

The influence of growth regulators and explant position on the growth and development of Mandevilla sanderi (Hemsl.) Woodson in vitro

Mandevilla is a valuable ornamental pot vine. However, due to a low propagation rate, it is difficult to keep up with the demand. Micropropagation would allow to produce lots of plants for the market. The aim of the study was to determine the effect of the growth regulators addition to the media and explants orientation on multiplication of Mandevilla sanderi, an exotic, ornamental pot plant. The shoot tips were placed vertically or horizontally on the Murashige and Skoog medium supplemented with benzyladenine (BA) or isopentenyladenine (2iP), at concentrations of 1, 2.5 or 5 mg·dm–3 singly or in combination with thidiazuron (TDZ) at concentrations of 0.01, 0.025 or 0.05 mg·dm–3. Maximum multiplication rate was noted on the media supplemented with 2.5 mg·dm–3 2iP + 0.025 mg·dm–3 TDZ or 5 mg·dm–3 2iP, when explants were placed horizontally. All the treatments resulted in callus formation. Medium supplemented with the highest concentration of BA combined with TDZ was the most active in callus growth.

First record and verification of Tyrophagus putrescentiae (Acari: Acaridae) causing direct damage on anthurium plants cultivated in vitro

The population size at which Tyrophagus putrescentiae can produce damage on anthurium plants (Anthurium sp.), cultivated in vitro, was determined. Symptoms caused by these mites on the plants are described. Two anthurium varieties coded as ANMIRO and ANWHGE, were cultivated in a Murashige and Skoog medium for four weeks. The treatments for each variety were as follows: a) Absolute Control (AC), b) fungus inoculated in the cultured medium (CF), c) fungus on leaves + mites (FLM), d) fungus in medium + mites (FMM), and e) only mites (OM). Mite density within the vessels increased through time. Fifty-five days after the first evaluation, the highest amount of injury was observed in the treatments FLM, FMM, and OM. The observed symptoms were holes on the foliar lamina produced by the feeding of mites until complete plant deterioration. According to the prediction model, when there is an average population close to 62 mites per vessel, it would be expected to find 20% damaged. The probability of observing damage on the variety ANWHGE was 1.7 times higher than on the variety ANMIRO. Although T. putrescentiae is a known contaminant in tissue culture laboratories as it carries bacteria and fungi on its body, this is the first record of this species causing direct damage to plants cultivated in vitro.

Optimization of biotechnological process clonal micropropagation in vitro of Asparagus officinalis L.

The study presents the results of obtaining regenerated plants of asparagus from seeds. Surface sterilizing the seeds by 0,75% sodium hypochlorite for 30 min is effective, during this obtained 83% viable sterile plants. The Murashige and Skoog medium supplemented with 6‑benzylaminopurine (2 mg/L), inositol (100 mg/L) and thiamine (0,4 mg/L) was found to be the best for seed germination. The expediency of using kinetin (1 mg/L) as a growth regulator to obtain a homogeneous plant material was established. The reproduction coefficient was 6,0. Only 11% of the explants formed callus. For the selection needs and production of somaclonal variants, the use of the culture medium with indole-3-acetic acid (0,2 mg/L) and 6‑benzylaminopurine (1 mg/L) is justified. In this condition reproduction coefficient was 3,7, and the level of different intensity callusogenesis was 59%. The rooting of obtained plants was performed in Murashige and Skoog medium supplemented with a half dose of macro- and micronutrients and growth regulators. Rooting frequency was up to 63%. The knowledge of hormonal requirements helps to promote isolated tissue and cells technologies of asparagus with purpose of rapid propagation and obtaining healthy, high-quality planting material.

Evaluation of the Conditions for the Cultivation of Callus Cultures of Hyssopus officinalis Regarding the Yield of Polyphenolic Compounds

The cultivation of plants in the form of callus cultures constitutes a renewable source of secondary plant metabolites. The conditions for the cultivation of callus cultures affect the yield of target compounds. Callus cultures of Hyssopus officinalis were chosen for study. Nutrient media of various compositions were used for Hyssopus officinalis callus culture. For each culture, data on the quantitative contents of saponins, flavonoids and polyphenolic compounds, as well as antioxidant activity, were obtained. It was found that Murashige and Skoog medium supplemented with 1-naphthylacetic acid and kinetin led to the highest yield of secondary metabolites.

IN VITRO REGENERATION OF ARUNDINA GRAMINIFOLIA (D. DON) HOCHR

We can see Orchids come in a wide variety of shapes, sizes, colours, and textures far beyond the human mind’s imagination. They emerge from seeds in nature, but in the absence of suitable hosts, they do not germinate in sufficient numbers. This problem was solved by using the tissue culture technique for its germination. One of the successful method used for mass propogation of orchid plantlets is in vitro techniques. Therefore, an initial analysis was conducted in order to establish an appropriate procedure for mass multiplication of Arundina graminifolia. MS (Murashige and Skoog) medium was found to be suitable for the asymbiotic seed germination of Arundina graminifolia. Direct protocorm like bodies were induced by using combinations and individual supplement of MS medium with IAA (Indole-3-acetic acid), IBA (Indole-3- butyric acid), BAP (6-Benzylaminopurine) and KIN (Kinetin). Hormone-free MS basal medium was found suitable for the conversion of PLBs (protocorm-like bodies) into complete plantlets

Optimation of Auxin and Cytokinin on Enhanced Quality and Weight of Coffea liberica Somatic Embryos

Coffea liberica is a variety of coffee that tolerant to marginal land, especially peatlands. One of propagation methods in C. liberica is somatic embryogenesis(SE) which producing large number of true-to-type plant seedlings in a short time. This research aimed at studying the effect of application of plant growthregulator (PGR) on quality and weight of somatic embryo of C. liberica. Somatic embryo in development stage was induced by Murashige and Skoog medium containing cytokinin as benzyl amino purin (BAP) and auxin as 2,4-dichlorophe-noxyacetic acid (2,4-D). While cotyledonary embryo in germination stage was induced by Murashige and Skoog medium containing cytokinin (BAP) and auxins as 2,4-D, indole acetic acid (IAA) and naphthaleneacetic acid (NAA). The resultsshowed that the application of auxins and cytokinins on development stage affected the formation of embryos, texture of calli, color of calli and embryos, and weight of somatic embryo. It also influenced the shoot and root formation, color and weight of geminating embryos of C. liberica at the germinating stage. During the development stage, addition of 1 mg/L BAP in the absence of 2,4-D in MS medium produced the highest quality of somatic embryo of C. liberica. This medium also produced heaviest somatic embryos but with lighter callus. While in germination stage, all medium treatments produced a typical germinating embryo. Coffea liberica germinating embryo growth optimally on MS medium containing 0.5 mg/L BAP as a single chemical or 0.5 mg/L BAP in combination with 0.5 mg/L IAA for shooting development. Whereas on rooting development, addition of 0.5 mg/L NAA on MS medium produced an optimal germinating embryo. Moreover, germination embryo of C. liberica recorded the highest in terms of dry weight on MS media with addition of 0.5 mg/L BAP. Application of appropriate concentration of auxin and cytokinin is needed to support the formation of somatic embryo and germinating embryo.

Effect of the Different Concentrations of Protective-stimulating Complex (HFC) on the Growth and Development of Grape Seedlings in the Tissue Culture at the Different Nutrient Levels

Background: Humic substances plays a vital role in the plant tissue culture as a growth hormone for in vitro propagation of many plant seedlings. The aim of this study was to investigate the effect of added humic-fulvate complex (HFC) at the various concentrations on the growth and development of grape seedlings in in vitro at the different nutrient levels. Methods: The cutting of khasansky grape were cultivated on ¼ Murashige and Skoog medium or ½ Murashige and Skoog medium either alone or supplemented with the humic-fulvate complex at the different concentrations at (0.1, 1 and 10 ml/l). Then, they were cultured for 4 weeks under a controlled environment. Result: The data observed that the low concentration of Murashige and Skoog medium (¼ MS) for in vitro rooting of grape cv. ‘Khasansky’ either alone or combined with HFC at the various concentrations significantly increased the rooting percentages and the total length of roots and stimulating the rate of vegetative growth compared with cultivated in ½ MS medium either alone or with supplemented with HFC. ¼ MS+ 10 ml/l HFC was the best treatment for improving the growth of khasansky grape seedlings.

INDUCTION OF ABACA BANANA ROOTS BY IN VITRO USING KINDS OF MEDIA AND THIAMIN

Abaca is a type a banana with high economic value with it stem fiber used in textile and paper industries. As a superior commodity, its number is relatively limited, with the need of a largeplanting area to meet the high market demand. The aim of the research was to observe the abaca banana explants response to various media and Thiamin. The experiment was done at Biotechnology laboratory, UPN “Veteran” Yogyakarta. Treatments were arranged in completely randomized design with 2 factor. The first factor is the growing media: Murashige & Skoog, a half Murashige & Skoog media, Vacint & Went Media and the second factor is the Thiamin concentration: 2 mg/L; 3 mg/L; 4 mg/mL.The results showed there is an interactions on the parameters of planlet height, number of lenghth of root in the combination of Murashige and Skoog and thiamin 3 mg/L medium. Murashige and Skoog medium produced the highest fresh weight and dry weight and Thiamin concentration 3 mg/L produced fresh and dry weight in the highestKey words: abaka, root induction, various media, thiamin