Tissue culture applied to carnivorous species

The purpose of the review is to comment on available data on the application of plant tissue culture to carnivorous plants. Thus, the review encompassed publications from 1979 to 2017 along in vitro germination studies and micropropagation techniques, such as somatic embryogenesis and organogenesis, which emphasized the responses of plant materials to the stimuli offered during in vitro culture. Tissue culture in carnivorous plants is presented as a tool to promote the increase of the population of these plants either for scientific and commercial purposes or for the conservation and reintroduction in their natural habitat, in order to ensure a sustainable exploitation of this nutritional pattern of plants. In general terms, the studies carried out were limited to the following aspects: cultivation technique, explant source, exogenously applied substances and culture medium. The review also revealed the absence of defined protocols for in vitro multiplication of large-scale carnivorous plants.

Characterization and Potential of Coelogyne rochussenii Orchids from Bukit Rimbang and Bukit Baling Wildlife Sanctuary as Explant Source

Conservation is an effort to return natural resources to their habitat to restore the ecosystem balance, which can be done in-situ and ex-situ. Coelogyne rochussenii orchid conservation efforts are essential to maintain its sustainability. The purpose of this study was to characterize C. rochussenii orchids from Bukit Rimbang and Bukit Baling Wildlife Sanctuary as a source of tissue culture explants to support ex-situ conservation efforts. Orchid plant samples were obtained through exploration in three locations with an altitude of 92 masl, and then the characterization of leaf morphology, pseudobulbs, roots, and fruit were carried out. The characterization results showed that the young pseudobulbs, young leaves, healthy roots, and physiologically ripe fruits of the C. rochussenii orchids obtained could be used as a source of explants to support ex-situ conservation efforts.Keywords: ex-situ conservation, physiologically mature, young pseudobulbs

Genetic Variation Analysis of Hevea brasiliensis Genotype Population of In Vitro Micro-Cutting Culture by RAPD Marker

The rubber seeds are insufficient for producing rootstocks to rubber grafting. It can be overcome by an in vitro micro-cutting culture technique developed in the Indonesian Research Institute for Biotechnology and Bioindustry (IRIBB). However, the origin clone of 57 rubber genotypes used as an explant source in vitro micro-cutting culture is not recognized. The study was to investigate the 57 genotypes that came from mixed GT 1, PB 260, and RRIM 600 as parent clones. We investigated using seven primers of Random Amplified Polymorphic DNA (RAPD), i.e., OPA 02, OPA 07, OPA 15, OPB 04, OPC 05, OPC 11, and OPC 20. The qualitative analyzed by electrophoresis 1% gel agarose. A total of 47 DNA fragments produced with an average of 7 fragments per primer. OPA 02 generated of 13 fragments, whereas OPB 04 only one fragment. The DNA fragment pattern shows the presence of polymorphism. The genetic similarity coefficients obtained in the range of 62-96%. The highest genetic similarity (96%) is genotype 70 and 78. It recognized that 42 genotypes from 57 rubber genotypes had the closest relationship with PB 260 clones. Furthermore, six genotypes had a significant growth response as an explant in vitro micro-cutting culture.

In vitro regeneration and flowering of Portulaca grandiflora Hook

P. grandiflora is a known ornamental plant with abundant flowering. The flowers exhibit varied coloration with distinct forms and simple folded petals and/or multiple. The objective of this work was to induce regeneration via organogenesis and in vitro flowering of P. grandiflora. Nodal segments of seedlings germinated in vitro were used as explant source for regeneration. Kinetin (KIN) and 6-Benzylaminopurine (BA) were used for the induction of organogenesis. The treatments supplemented with 1.0 and 1.5 mg L−1 BA induced the highest number of adventitious shoots with an average number of 7.0 (±1.55) e 5.4 (±0.83), respectively. The microcuttings obtained from regenerated shoots produced floral buds. The floral buds were located in the axillary and terminal regions of the microcuttings and developed in approximately 10 days of cultivation until the anthesis. The highest number of flower buds was observed in the presence of 0.75 mg L−1 of gibberellic acid. This study opens new perspectives for the establishment of biotechnological tools to be applied for this important ornamental species.

Effect of explant source, perlite nanoparticles and TiO2/perlite nanocomposites on phytochemical composition of metabolites in callus cultures of Hypericum perforatum

It appears that the biologically-synthesized nanoparticles (NPs) have potential to perform as effective elicitors for the production of valuable secondary metabolites in plants. Besides, it has been reported that the toxicity of the biologically-synthesized NP is not as much as that of the chemically-synthesized NPs. Therefore, it is necessary to test their advantages aspects. In this study, the physical synthesis of perlite NPs and biologically-synthesis of TiO2/perlite nanocomposites (NCs) were conducted. Subsequently, their effects and explant source influence on the growth characteristics and secondary metabolite profiles of Hypericum perforatum callus cultures were evaluated. According to the obtained results, morphology of the synthesized perlite NPs and TiO2/perlite NCs were mesoporous and spherical with sizes ranging about 14.51–23.34 and 15.50–24.61 nm, respectively. Addition of perlite NPs and TiO2/perlite NCs to the culture medium at the concentration range of 25–200 mg/L showed no adverse impacts on the growth characteristics of H. perforatum calli. According to the GC-MS analysis, the stress caused by perlite NPs and TiO2/perlite NCs led to an increase in the variety, amount and number of volatile compounds. The calli obtained from in vitro grown plants produced more volatile compounds relative to the calli obtained from field grown plants under the nanomaterial stress conditions. The production of hypericin and pseudohypericin were also determined in the callus cultures under desired nanomaterials elicitation. Accordingly, our results suggest that perlite NPs and TiO2/perlite NCs can possibly be considered as effective elicitors for the production of volatile compounds, hypericin, and pseudohypericin in callus cultures of H. perforatum.

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.