Determination of Genetic Relationships among Phalaenopsis Spp. Using Random Amplified Polymorphic DNA and In Vitro Propagation of Phalaenopsis Gigantea

Phalaenopsis, with long arching sprays of flowers, is among the most beautiful flowers in the world. Phalaenopsis is an important genus and one of the most popular epiphytic monopodial orchids, grown commercially for production of cut flowers and potted plants. Most of them have different and interesting morphological characteristics which have different value to the breeders. Study was carried out using molecular characterization through Random Amplified Polymorphic DNA (RAPD) analyses for 20 species of Phalaenopsis was conducted to determine their genetic variation and relationships. Among the 20 primers used for these analyses, 10 showed polymorphism, with 26 to 54 DNA fragments amplified per primer. A total of 414 polymorphic fragments were generated by these 10 primers and then they were used for correlation grouping analysis. The highest value of Similarity index was 0.28 between Phalaenopsis violacea malaysia and Phalaenopsis violacea witte. The dendrogram resulting from UPGMA hierarchical cluster analysis separated the species into three groups: the first group consisted of five species of Ph. violacea blue, Ph. belina, Ph. violacea malaysia, Ph. violacea witte, and Ph. gigantea; the second group included Ph. lamelligera, Ph. amabilis, Ph. parishii, Ph. labbi nepal, Ph. speciosa, Ph. lobbi yellow, Ph. venosa, Ph. hieroglyphica, and Ph. maculata; the third group consist of Ph. Minho Princess, Ph. Leopard prince, Ph. mannii, Ph. modesta, Ph. cornucervi and Ph. pantherina. RAPD markers can thus be successfully applied to this economically important group of orchids to determine relationship between species of these orchids. Phalaenopsis gigantea one of the most difficult to grow and has the potential of producing beautiful hybrids was selected for further study in developing a rapid and efficient in vitro propagation technique, this technique can also be tested for other species in the future. Ripe capsules of Phalaenopsis gigantea were collected and sterilized. Seed germination was conducted using Vacin and Went (VW) medium supplemented with coconut water (CW) and 6-benzylaminopurine (BAP) and kinetin (KIN). Protocorm-like bodies (PLBs) were successfully induced and plantlets were produced after 60 days. BAP at 1 mgL-1 in combination with 2 mgL-1 KIN produced the highest number of plantlets followed by treatment 1 mgL-1 BAP and 1 mgL-1 KIN. Protocorm-like bodies (PLBs) were successfully induced from leaf segments using New Dogashima medium (NDM) with BAP, Thidiazuron (TDZ), and KIN, each at 0.01, 0.1, 0.5 and 1.0 mg/L alone and in combination with naphthalene acetic acid (NAA), at 0.01, 0.1, 0.5 and 1.0 mgL-1, within 6-8 weeks of culture. The highest percentage of callus formation (100%) was obtained from treatment containing 1 mgL-1 NAA in combination with 0.1 mgL-1 TDZ followed by treatment supplemented with 1 mgL-1 NAA and 0.5 mgL-1 TDZ (76.56%).Plant regeneration from PLBs was achieved in PGR-free NDM basal medium. There was no PLB observed on full and half-strength MS media supplemented with different concentrations of 2, 4-D, BAP and TDZ. Similarly, no PLB was observed on VW and ½ MS media supplemented with different concentrations and combinations with 2, 4.-D, BAP, TDZ and CW or NAA. No PLB was also observed on NDM medium with different concentrations and combinations with BAP and NAA. However multiplication of PLB was observed in liquid media of basal MS, VW, and NDM with or without sucrose. The highest fresh weight of PLBs was obtained from VW medium supplemented with CW

Molecular characterization and phylogenetic relationships among and within species of Phalaenopsis (Epidendroideae: Orchidaceae) based on RAPD analysis

Random amplified polymorphic DNA (RAPD) analysis for 20 species of Phalaenopsis was conducted to determine their genetic distances and relationships. Among 20 different primers used for RAPD analysis, 10 primers showed polymorphism, and according to the primer type, 26 to 54 DNA fragments were amplified. A total of 414 polymorphic fragments were generated by 10 primers and used for correlation group analysis. The highest value of Similarity index was 0.28 between Ph. violacea malaysia and Ph. violacea witte. The dendrogram resulting from UPGMA (Unweighted Pair Group Method using Arithmetic average) hierarchical cluster analysis separated the original species into three groups: The first group had five species of Ph. violacea blue, Ph. belina, Ph. violacea malaysia, Ph. violacea witte, and Ph. gigantea; the second group included Ph. lamelligera, Ph. amabilis, Ph. parishii, Ph. labbi nepal, Ph. speciosa, Ph. lobbi yellow, Ph. venosa, Ph. hieroglyphica, and Ph. maculata; the third group consisted of Ph. minho princess, Ph. leopard prince, Ph. mannii, Ph. modesta, Ph. cornucervi and Ph. pantherina. RAPD markers can thus be successfully applied in this economically important group of orchids for the study of molecular characterization and relationships. The data acquired from this study could be used for identification and classification of other orchid genera and oriental Phalaenopsis

Plant-based expression systems for protein and antimicrobial peptide production

Molecular farming technology offers a unique advantage that almost any protein can be produced economically and safely under very controlled conditions. Besides traditional production systems, such as bacteria, yeasts, insects and mammal cell lines, plants can now be used to produce eukaryotic recombinant proteins, especially therapeutic ones. Their advantages as hosts for protein production include correct post-translational modifications, low-cost maintenance and no risk of contamination by human pathogens. The system is widely applied in agriculture and industry, especially in life science and pharmaceutical industry. The application of transgenic plants in the production of vaccines, antibodies and pharmaceutical proteins has been playing a key role in plant genetic engineering in recent years. The production of recombinant proteins plays a critical role in the production of high amounts of high-quality proteins. In this review, common problems in the production of recombinant proteins and antimicrobial peptides in plant-based expression systems are discussed and strategies for their solution are suggested. Viral vector-mediated transient gene expression in plants enables rapid production of pharmaceutical proteins such as vaccine antigens and antibodies. To conclude, plant-based systems have the potential to bring unique efficacy-enhancing features to increase the utility and effectiveness of vaccines and therapeutics

Transgenic plant

This photograph originally appeared in the 2012 Research student photography and image competition held to celebrate National Science Week (11-19 August)

Optimisation of expression of recombinant puroindolines and analysis of protein-protein interactions in planta

Wheat is the staple food of 35 per cent of the world’s population and the third largest cereal crop after maize and rice. The two puroindoline proteins of wheat, PINA and PINB are largely responsible for grain texture, a property important in food technology and the wheat trade. Furthermore, it has been suggested that PINs may have an in vivo role in seed pathogen defence and antimicrobial properties, which make them very attractive as antimicrobial agents for novel medical, pharmaceutical and food-industry applications. This PhD has led to successful methods for obtaining PINs of appropriate quality essential for potential infection control in diverse areas

In vitro plant regeneration from protocorms-like bodies (PLBs) and callus of Phalaenopsis gigantea (Epidendroideae: Orchidaceae)

Phalaenopsis, with long arching sprays of flowers, are among the most beautiful flowers in the world. Phalaenopsis is an important genus and one of the most popular epiphytic monopodial orchids, grown commercially for the production of cut flowers and potted plants. Most of them have different and interesting morphological characteristics which have different value to the breeders. Phalaenopsis gigantea is one of the most difficult to grow and has the potential of producing beautiful hybrids. An efficient and reproducible method for large-scale propagation of Ph. giganteausing leaf sections has been developed. Leaf sections from in vitro young plants were cultured on New Dogashima Medium (NDM) supplemented with cytokinins (6-Benzylaminopurine (BAP), Thidiazuron (TDZ), and Kinetin (KIN), each at 0.01, 0.1, 0.5 and 1.0 mg/L) alone and in combinations with (auxins a-naphthaleneacetic acid (NAA), at 0.01, 0.1, 0.5 and 1.0 mg/L). The explants developed calli and protocorm-like-bodies (PLBs) within 6 weeks of culture. Treatment TDZ in combination with auxins was found to be the best for the induction of callus and PLBs. In vitroregeneration of Ph. gigantea PLB was achieved by exposure to light and transferring to hormone free NDM solid medium

Plant-based vaccines production and conducted experiments in this case, as an effective strategy to combat infectious diseases, such as Coronavirus disease (COVID-19)

The emergence of new viruses has always been a threat to the health of people around the world, the latest example of it is the new strains of the coronavirus (SARS-CoV-2) and the resulting acute respiratory distress syndrome (ARDS). The current situation underscores the importance of rapidly producing low-cost stable vaccines that do not require refrigeration equipment for storage and transportation. However, most vaccines are not yet available in developing countries due to import costs and storage and transportation needs. Therefore, the vaccine must be affordable for developing countries so that vaccination can be carried out on a large scale. Herbal vaccines are more cost-effective than other types of vaccines and production methods and can be produced in large quantities. In addition, herbal vaccines have other benefits that are discussed in this article. However, given that an herbal medicinal product is to be used as a vaccine in a semi-processed form (such as mashed potatoes or tomato paste), specific regulatory reviews must apply to injectable vaccines. Products should also be applied to evaluate their side effects clinically. The current review article investigates the opportunities and challenges of producing plant-based vaccines to deal with diseases like Coronavirus disease (COVID-19(

Induction of callus and protocorm-like bodies (PLBs) in Phalaenopsis gigantea

An efficient and reproducible method for large-scale propagation of Phalaenopsis gigantea using leaf sections has been developed. Leaf sections from young plants were cultured on New Dogashima medium (NDM) supplemented with cytokinins (6-Benzylaminopurine (BAP), thidiazuron (TDZ), and kinetin (KIN), each at 0.01, 0.1, 0.5 and 1.0 mgL-1) alone and in combinations with (auxins anaphthaleneacetic acid (NAA), at 0.01, 0.1, 0.5 and 1.0 mgL-1). The explants developed calli and protocorm-like-bodies (PLBs) within 8 weeks of culture. Treatment TDZ in combination with auxins was found to be best for the induction of callus and PLBs