Metabolic profiling, in vitro propagation, and genetic assessment of the endangered rare plant Anarrhinum pubescens

Background Anarrhinum pubescens Fresen. (Plantaginaceae) is a rare plant, endemic to the Saint Catherine area, of South Sinai, Egypt. Earlier studies have reported the isolation of cytotoxic and anti-cholinesterase iridoid glucosides from the aerial parts of the plant. The present study aimed to investigate the chemical profiling of the wild plant shoots as well as establish efficient protocols for in vitro plant regeneration and proliferation with further assessment of the genetic stability of the in vitro regenerated plants. Results Twenty-seven metabolites have been identified in wild plant shoots using the Nuclear Magnetic Resonance (NMR) spectroscopy. The metabolites include alkaloids, amino acids, carbohydrates, organic acids, vitamins, and a phenol. In vitro propagation of the plant was carried out through nodal cutting-micropropagation and leaf segment-direct organogenesis. The best results were obtained when nodal cutting explants were cultured on Murashige and Skoog medium with Gamborg B5 vitamins supplemented with 6-benzylaminopurine (BAP) (1.0 mg/L) and naphthaleneacetic acid (NAA) (0.05 mg/L), which gave a shoot formation capacity of 100% and a mean number of shoots of 27.67 ± 1.4/explant. These shoots were successfully rooted and transferred to the greenhouse and the survival rate was 75%. Genetic fidelity evaluation of the micropropagated clones was carried out using random amplified polymorphic DNA (RAPD) and inter simple sequence repeat (ISSR) molecular markers. Jaccard’s similarity coefficient indicated a similarity as high as 98% and 95% from RAPD and ISSR markers, respectively. Conclusions This study provides the chemical profiling of the aerial part of Anarrhinum pubescens. Moreover, in vitro regeneration through different tissue culture techniques has been established for mass propagation of the plant, and the genetic fidelity of the in vitro regenerated plants was confirmed as well. Our work on the in vitro propagation of A. pubescens will be helpful in ex situ conservation and identification of bioactive metabolites.

IN VITRO PROPAGATION OF Eucalyptus citriodora PLANT

This work was carried out in Plant Tissue Culture Laboratory of Prof. Dr. Abd El-Fatah Helmy Belal, Faculty of Environmental Agricultural Sciences, Arish University, North Sinai, Egypt during the period from 2016 to 2018. This study was conducted to study the effect of medium type, explant source and growth regulator type and concentration on micropropagation of E. citriodora plant which is grown in Sinai Peninsula. Results showed that full strength of Murashige and Skoog's medium (MS) was the most suitable medium for seed germination and shoot growth. Meanwhile, addition of BA at 1.00 mg l-1 was effective for improving shoot growth and development. Also, multiplying of shoots was enhanced by using nodal cutting explants cultured on MS medium supplemented with 1.00 mg l-1 BA plus 1.00 mg l-1 NAA. On the other hand, shoot rooting was achieved by using MS medium supplemented with 2.50 mg l-1 IBA. It is worth to mention that obtained plantlets were successfully acclimatized (80% survivability) in a combination of peatmoss, vermiculite and washed sand or peatmoss and vermiculite at equal volumes.

Micropropagation of Stevia Plant from Nodal Segments

The experiment was conducted to develop and establish a reproducible protocol for plantlet regeneration in Stevia. Nodal cutting explants were cultured in Murashige and Skoog (MS) medium supplemented with different concentrations and combinations of ?-Napthaleneacetic acid (NAA) and 6-Benzylamino purine (BAP). The combination of NAA at 1.0 mgl-1 and BAP at 1.0 mgl-1 resulted in the highest percentage (100%) of callus initiation. The maximum shoot regeneration and development of shoot was observed at the same combination. The developed shoots from nodal cuttings, upon transfer to the MS medium containing indole butyric acid (IBA) at 0.1mgl-1 resulted in best rooting within 8 days.DOI: http://dx.doi.org/10.3329/pa.v19i2.16914 Progress. Agric. 19(2): 21 - 26, 2008

Micropropagation of Bauhinia variegata L. from Tissue Culture

Bauhinia variegate L. is a multipurpose tree and its micropropagation holds great promise in agroforestry. The sterilized seeds were first inoculated on Murashige and Skoog (MS) medium. Nodal cuttings from these seedlings grown in vitro were used as explants for micropropagation. Nodal cutting inoculated on the medium with various concentrations of BAP (benzyl-aminopurine) and NAA (£- naphthalene acetic acid) and separately generated varied results. Best propagation was recorded on MS medium supplemented with 1.0 ?M BAP with 0.05 ?M NAA. Propagated plants were successfully acclimatized and rooted in pots (6 cm diameter) containing soil and sand in1:1 ratio and then finally transferred to the field. All the data generated were analysed statistically using SPSS statistical package. Nepal Journal of Science and Technology Vol. 13, No. 1 (2012) 39-41 DOI: http://dx.doi.org/10.3126/njst.v13i1.7397

A Modified Nodal Cutting Bioassay for Assessing Salinity Tolerance in Potato (Solanum sp.)

The potato (Solanum tuberosum L.) is considered moderately sensitive to salinity stress. Yields can be adversely affected by salinity at EC levels of 2 to 3 dS·m–1. In vitro screening and selection for salinity tolerance may be faster and more reliable than traditional field assessment. A modified nodal cutting bioassay was developed based on that of Morpurgo (1991) and used to rank the salinity tolerance of several potato cultivars, wild Solanum species, and their hybrids. Nodal cuttings were cultured in Murashige and Skoog (1962) basal medium which included NaCl at 0, 40, 80, and 120 mM. After 4 weeks at 25C and 16/8 h day/night period, vegetative growth parameters were assessed. Hybrids derived from S. chacoense outperformed hybrids of other wild species, their wild parents, and S. tuberosum cultivars. `Russet Burbank' and `Kennebec' were more salinity-tolerant than the three other cultivars tested.

In Vitro Ranking of Salinity Tolerance in Potato (Solanum sp.)

Globally, salinization of agricultural soils is an increasing problem. At least 25% of the world's cultivated land area is now salt-affected. Although the potato (Solanum tuberosum L.) is among the most important of crops plants, little is known of the range of salt tolerance among cultivars of S. tuberosum, wild Solanum sp., or their hybrids. The objective of this work was to rank the salinity (NaCl) tolerance of 200 North American and European cultivars, 13 hybrids of S. chacoense, S. tuberosum, and primitive cultivated diploids (S. phureja and/or S. stenotomum) and their progenies, and 14 accessions of S. chacoense from different collection sites. This was done in vitro, using a modified nodal cutting bioassay. The parameters evaluated included shoot and root lengths, and fresh and dry weights. Extreme variation for salinity tolerance was noted among the cultivars, the hybrids, and the wild species.