Full Murashige And Skoog Medium Research Articles

Ploidy Status, Nuclear DNA Content and Start Codon Targeted (SCoT) Genetic Homogeneity Assessment in Digitalis purpurea L., Regenerated In Vitro.

Digitalis purpurea L. is a therapeutically important plant that synthesizes important cardiotonics such as digitoxin and digoxin. The present work reports a detailed and efficient propagation protocol for D. purpurea by optimizing various PGR concentrations in Murashige and Skoog (MS) medium. The genetic homogeneity of in vitro regenerants was assessed by the flow cytometric method (FCM) and Start Codon Targeted (SCoT) marker technique. Firstly, the seeds inoculated in full MS medium added with 0.5 mg/L GA3 produced seedlings. Different parts such as hypocotyl, nodes, leaves and apical shoots were used as explants. The compact calli were obtained on BAP alone or in combinations with 2, 4-D/NAA. The hypocotyl-derived callus induced somatic embryos which proliferated and germinated best in 0.75 mg/L BAP-fortified MS medium. Scanning electron microscopic (SEM) images confirmed the presence of various developmental stages of somatic embryos. Shoot regeneration was obtained in which BAP at 1.0 mg/L and 2.0 mg/L BAP + 0.5 mg/L 2,4-D proved to be the best treatments of PGRs in inducing direct and indirect shoot buds. The regenerated shoots showed the highest rooting percentage (87.5%) with 24.7 ± 1.9 numbers of roots/shoot in 1.0 mg/L IBA augmented medium. The rooted plantlets were acclimatized in a greenhouse at a survival rate of 85-90%. The genome size and the 2C nuclear DNA content of field-grown, somatic embryo-regenerated and organogenic-derived plants were estimated and noted to be 3.1, 3.2 and 3.0 picogram (pg), respectively; there is no alteration in ploidy status and the DNA content, validating genetic uniformity. Six SCoT primers unveiled 94.3%-95.13% monomorphic bands across all the plant samples analyzed, further indicating genetic stability among in vitro clones and mother plants. This study describes for the first time successful induction of somatic embryos from hypocotyl callus; and flow cytometry and SCoT marker confirmed the genetic homogeneity of regenerated plants.

Rapid germination and development of Acacia sieberiana DC in vitro

Acacia sieberiana is an ecological and economical important forest tree. The plant has poor germination due to dormancy; therefore, in vitro culture can provide a fast supply of plantlets and can be a basis for further biotechnology research. The effect of sodium hypochlorite, Murashige and Skoog (MS) medium, sucrose and cytokinin were investigated. The results indicated that all the treatments induced significant (P < 0.05) effects on germination and seedling development depending on their concentrations. Germinates free of contamination were established successfully with all the concentrations of sodium hypochlorite, however, the germination was reduced as the concentration increased. A higher germination rate of 98.3% was attained with full MS strength and 3% sucrose concentration. The highest germination percentage obtained was 100% with 2 mg/L thidiazuron. The maximum shoot length (13.5 cm) and seedling dry weight (0.49 g) were recorded on full MS medium while the maximum root length (13.4 cm) was obtained on ½ MS strength. However, when full MS medium augmented with 5% sucrose enhanced shoot length and dry weight to (14.2 cm) and (0.5 g), respectively. While the highest root length (10.2 cm) was obtained by 3% provided in full MS medium. Application of cytokinins improved the number of node and dry weight of seedlings to 6.9 nodes with 4 mg/L thidiazuron and 0.54 g by 2 mg/L benzyladenine. The present study describes the establishment of in vitro seed germination and seedling growth system for A. sieberiana for the first time.

In vitro tissue culture protocol of ancient einkorn (Triticum monococcum ssp. monococum) wheat via indirect shoot regeneration

Einkorn indirect shoot regeneration was obtained from leaf, coleoptile, and root explants cultured on full, ½, and ¼ strength Murashige and Skoog (MS) medium containing 0.5 to 10 mg L−1 2,4-dichlorophenoxyacetic acid (2,4-D). Calluses derived from coleoptile and root explants were cultured on full-strength MS medium supplemented with 0.5 to 5 mg L−1 thidiazuron (TDZ) for 6 wk to induce plant regeneration. Only callus cultures derived from coleoptile explants regenerated shoots. For root induction, 1.0 to 10 mg L−1 indole-3-acetic acid (IAA) was used for 4 wk. The highest rate of callus formation (93%) was from root explants cultured on full-strength MS medium with 4 mg L−1 2,4-D. Coleoptile explants had the highest callus formation (100%) after culture on full-strength MS medium with 3 to 6 mg L−1 of 2,4-D. The highest indirect shoot regeneration (7.0 ± 1.1 shoots produced per callus with a 67% shoot formation frequency) was from calluses derived from coleoptile explant cultured on full MS medium supplemented with 5 mg L−1 of TDZ. Of the different IAA concentrations investigated for rooting, the greatest number of roots per explant (7.7 ± 0.088 roots produced per regenerated shoot with a 100% root formation frequency) was observed on MS medium supplemented with 6 mg L−1 IAA. This indirect shoot regeneration protocol for coleoptile-derived einkorn wheat callus will be useful for future wheat genetic and improvement studies.

A simple method for mass propagation of Spathiphyllum cannifolium using an airlift bioreactor

A method for the micropropagation of Spathiphyllum cannifolium is presented using shoot tip proliferation onto Murashige and Skoog (MS) medium supplemented with different plant growth regulator concentrations and combinations. The proliferation responses were significantly influenced by the cytokinin type and concentrations. Supplementation of the medium with benzyladenine (BA; 4.44–13.32 μM) increased the shoot proliferation rate significantly as compared to other treatments. When cytokinins were used with auxin (indole-3-butyric acid, IBA and naphthalene acetic acid. NAA), the number of shoots per explant increased in comparison with treatments with BA alone. The largest number of shoots, 9.3 per explant, was obtained with 13.32 μM BA and 4.9 μM IBA. Different MS medium strengths and sucrose concentrations were used with the aim to stimulate in vitro shoot proliferation. Full MS medium with 30 gl−1 sucrose was found to be suitable for shoot tip culture of Spathiphyllum. Comparative studies between gelled medium and bioreactor culture [continuous immersion (with or without net) and temporary immersion in liquid media using ebb and flood] revealed that shoot multiplication and growth were more efficient in continuous immersion (with net) bioreactor with low cytokinin-supplemented media. Plantlets from the bioreactor were cultured hydroponically for 30 d and 100% of plants were rooted and acelimatized successfully. Rapid and efficient multiplication rate in bioreactor and successful transfer to greenhouse makes this protocol suitable for large-scale multiplication of this important foliage plant.