Micropropagation of Vanda tricolor Lindl. var. suavis with various concentrations of organic growth supplements

Plant propagation through in vitro culture is increasingly being used to produce hybrid orchids. Plant tissue culture provides a good alternative to produce plants in large numbers in a short time. The provision of organic growth supplements in tissue culture media plays an important role as a substitute for growth regulators in stimulating the growth of explants. In this study, young coconut water, banana extract (cv. ambon), and tomato extract were used to stimulate the growth of the Vanda tricolor Lindl. Var suavis protocorm. Data were analyzed using a completely randomized design (CRD) with five replications using the Kruskal-Wallis test and continued with the Mann-Whitney test if there was a significant difference between each treatment and its concentration. Parameters observed were the percentage of the number of shoots and the number of leaves. The results showed that there was a significant difference between the treatments given to the number of leaves. Mann-Whitney further test results on the number of leaves showed a significant difference in the banana extract treatment and the control treatment.

Structural and Biochemical Features of Human Serum Albumin Essential for Eukaryotic Cell Culture

Serum albumin physically interacts with fatty acids, small molecules, metal ions, and several other proteins. Binding with a plethora of bioactive substances makes it a critical transport molecule. Albumin also scavenges the reactive oxygen species that are harmful to cell survival. These properties make albumin an excellent choice to promote cell growth and maintain a variety of eukaryotic cells under in vitro culture environment. Furthermore, purified recombinant human serum albumin is mostly free from impurities and modifications, providing a perfect choice as an additive in cell and tissue culture media while avoiding any regulatory constraints. This review discusses key features of human serum albumin implicated in cell growth and survival under in vitro conditions.

Nanoscale Strontium-Substituted Hydroxyapatite Pastes and Gels for Bone Tissue Regeneration

Injectable nanoscale hydroxyapatite (nHA) systems are highly promising biomaterials to address clinical needs in bone tissue regeneration, due to their excellent biocompatibility, bioinspired nature, and ability to be delivered in a minimally invasive manner. Bulk strontium-substituted hydroxyapatite (SrHA) is reported to encourage bone tissue growth by stimulating bone deposition and reducing bone resorption, but there are no detailed reports describing the preparation of a systematic substitution up to 100% at the nanoscale. The aim of this work was therefore to fabricate systematic series (0–100 atomic% Sr) of SrHA pastes and gels using two different rapid-mixing methodological approaches, wet precipitation and sol-gel. The full range of nanoscale SrHA materials were successfully prepared using both methods, with a measured substitution very close to the calculated amounts. As anticipated, the SrHA samples showed increased radiopacity, a beneficial property to aid in vivo or clinical monitoring of the material in situ over time. For indirect methods, the greatest cell viabilities were observed for the 100% substituted SrHA paste and gel, while direct viability results were most likely influenced by material disaggregation in the tissue culture media. It was concluded that nanoscale SrHAs were superior biomaterials for applications in bone surgery, due to increased radiopacity and improved biocompatibility.

CONFIRMATION OF ENDOPHYTIC MICROBES CAUSING CONTAMINATION IN WATER SPINACH (Ipomoea aquatica Forssk.) TISSUE CULTURE

Konfirmasi Mikroba Endofit Penyebab Kontaminasi pada Kultur Jaringan Kangkung (Ipomoea aquatica Forssk.) Tanaman kangkung secara alami bersimbiosis dengan mikroba endofit, yang berpotensi menjadi kontaminan pada kultur jaringan kangkung karena berada di dalam jaringan dan sulit dijangkau saat proses sterilisasi eksplan. Tujuan penelitian ini adalah mengonfirmasi mikroba endofit penyebab kontaminasi pada kultur jaringan kangkung ‘Tetraploid’ sehingga dapat menjadi informasi awal untuk metode sterilisasi yang efektif. Sebanyak 14 sampel kontaminan pada media tanam kultur jaringan kangkung diisolasi dan diidentifikasi secara molekuler berdasarkan gen 16S rDNA untuk bakteri, daerah D1/D2 dari gen LSU rRNA untuk khamir, dan daerah ITS dari gen rDNA untuk jamur. Keragaman jenis mikroba yang teridentifikasi dibandingkan dengan keragaman jenis mikroba endofit dari jaringan tanaman kangkung yang ditanam pada media kultur jaringan tidak terkontaminasi mikroba, media kultur jaringan terkontaminasi mikroba, media tanam campuran tanah steril dan tidak steril, serta kangkung yang didapatkan dari pasar. Hasil penelitian ini mengonfirmasi bahwa khamir endofit dari kelompok Ustilaginaceae (basidiomycetous yeast) yang berasal dari jaringan tanaman kangkung sama dengan jenis kontaminan yang mengontaminasi media kultur jaringan kangkung ‘Tetraploid’. Khamir dari kelompok Ustilaginaceae (basidiomycetous yeast) tersebut juga merupakan mikroba paling dominan yang mengontaminasi media tanam kultur jaringan kangkung ‘Tetraploid’. Keywords: Ipomoea aquatica, kultur jaringan, mikroba endofit, morfologi, pohon filogeni Water spinach in nature lives in symbiosis with endophytic microbes, which have the potential to become contaminants in water spinach tissue culture because they are difficult to eliminate during the explant sterilization process. This study aimed to confirm endophytic microbes that cause contamination in the tissue culture of 'Tetraploid' water spinach so that it can provide initial information for an effective sterilization method. Fourteen contaminant samples in water spinach tissue culture media were isolated and identified molecularly based on the 16S rDNA gene for bacteria, the D1/D2 region of the LSU rRNA gene sequences for yeast, and ITS region of the rDNA gene for mold. The diversity of microbial species identified was compared with the diversity of endophytic microbial types from water spinach plant tissue grown on sterile tissue culture media, microbially contaminated tissue culture media, sterile and non-sterile soil mixed planting media, and water spinach obtained from the market. The results confirmed that endophytic yeast from Ustilaginaceae group (basidiomycetous yeast) derived from water spinach plant tissue was the same type of microbe that contaminated the 'Tetraploid' water spinach tissue culture media. The results gave new information that yeast from the Ustilaginaceae (basidiomycetous yeast) group was the most dominant microbe contaminating water spinach ‘Tetraploid’ tissue culture media. This group is endophytic yeast that lives within Ipomoea aquatica tissues.

Application of Date Seeds Powder as Growth Additive for Callus Induction in vitro Using Vigna radiata Hypocotyl Seedling Explant

Date seeds (Phoenix dectylifera) are one of the seeds that is not usable and always end to be disposed. The present study describes date seeds as an additive on callus induction in vitro from hypocotyl explants of Vigna radiata. The objective is to explore the usage of date seed powder as growth additive to promote in plant tissue culture media thereby it can be utilized as fertilizer in vivo for sustainable agriculture. 1% concentrations of date seed powder under controlled conditions highly influenced the callus induction. MS media supplied with different auxins is prepared for callus induction. The highest degree of callus weight was observed in MS media supplemented with (5mg/L) 2, 4-D + (0.5 mg/L) Kn (0.437±0.1). MS media supplied with different concentrations of date seed powder + (5mg/L) 2,4-D + (0.5 mg/L) Kn are prepared for callus induction under controlled conditions (16hrs light and 8hrs dark, 3000 lux light intensity,60% humidity and 25±20c) in a plant growth chamber. MS media supplied with 1% date seed + 3% sucrose + 0.5 mg/l of Kn +5 mg/l of 2,4D gave the highest stimulation of callus growth. Results show that date seed is not replacing sucrose as a carbon source, but it acts as a good additive to promote induction callus. Quantitative nutritional analysis of date seed powder was carried out. The results show date seed powder contains a high amount of elements like: Ca (2994.33), k (1712.33), Si (456.33), Mg (687.33), which plays a major role in callus formation.

High throughput mathematical modeling and multi-objective evolutionary algorithms for plant tissue culture media formulation: Case study of pear rootstocks

Simplified prediction of the interactions of plant tissue culture media components is of critical importance to efficient development and optimization of new media. We applied two algorithms, gene expression programming (GEP) and M5’ model tree, to predict the effects of media components on in vitro proliferation rate (PR), shoot length (SL), shoot tip necrosis (STN), vitrification (Vitri) and quality index (QI) in pear rootstocks (Pyrodwarf and OHF 69). In order to optimize the selected prediction models, as well as achieving a precise multi-optimization method, multi-objective evolutionary optimization algorithms using genetic algorithm (GA) and particle swarm optimization (PSO) techniques were compared to the mono-objective GA optimization technique. A Gamma test (GT) was used to find the most important determinant input for optimizing each output factor. GEP had a higher prediction accuracy than M5’ model tree. GT results showed that BA (Γ = 4.0178), Mesos (Γ = 0.5482), Mesos (Γ = 184.0100), Micros (Γ = 136.6100) and Mesos (Γ = 1.1146), for PR, SL, STN, Vitri and QI respectively, were the most important factors in culturing OHF 69, while for Pyrodwarf culture, BA (Γ = 10.2920), Micros (Γ = 0.7874), NH4NO3 (Γ = 166.410), KNO3 (Γ = 168.4400), and Mesos (Γ = 1.4860) were the most important influences on PR, SL, STN, Vitri and QI respectively. The PSO optimized GEP models produced the best outputs for both rootstocks.

PENGARUH EKSTRAK Sargassum cristaefolium PADA MULTIPLIKASI Dendrobium antennatum Rchb.f SECARA IN VITRO

The purpose of this study was to determine the potential and concentration of Sargassum cristaefolium extract as a natural cytokinin in tissue culture media of Dendrobium antennatum Rchb.f. This study is experimental with a completely randomized design, using several extract concentrations compared with the positive control (BAP 1.5 ppm) and negative control (MS0 media). Extract concentrations used 5 ppm, 10 ppm, 15 ppm, 20 ppm, and 25 ppm. Based on analysis of variance (ANOVA), the effect of Sargassum cristaefolium extract on the growth media significant on all parameters. Sargassum cristaefolium extracts caused different responses at certain levels of concentration. Extract concentration of 10 ppm was able to initiate the highest number of shoots and leaves compared to other extract concentrations, where as the concentration 20 ppm was able to accelerate and increase root growth.

Survival of Lassa Virus in Blood and Tissue Culture Media and in a Small Particle Aerosol

Knowledge of the survival and stability of a pathogen is important for understanding its risk, reducing its transmission, and establishing control measures. Lassa virus is endemic in West Africa, causes severe disease, and is an emerging pathogen of concern. Our study examined the survival of Lassa virus in blood and tissue culture media at two different temperatures. The stability of Lassa virus held within a small particle aerosol was also measured. In liquids, Lassa virus was found to decay more quickly at 30 °C compared to room temperature. Sealed samples protected from environmental desiccation were more stable than samples open to the environment. In a small particle aerosol, the decay rate of Lassa virus was determined at 2.69% per minute. This information can contribute to risk assessments and inform mitigation strategies in the event of an outbreak of Lassa virus.