Intradermal vaccination with Porcilis® Begonia can clinically protect against fatal PRV challenge with the highly virulent ZJ01 field strain.

Since pseudorabies (PR) re-emerged and rapidly spread in China at the end of 2011, researchers have focused on effective vaccine strategies to prevent and control pseudorabies virus (PRV) infection in pig herds. Due to the extensive application of an attenuated vaccine based on the Bartha-K61 strain isolated in Hungary in 1961 and the variation of the PRV strain, it has been suggested that traditional vaccines based on the Bartha-K61 strain offer only partial protection against variant strains. It was therefore evaluated whether the Porcilis® Begonia vaccine, which is based on the NIA-3 strain with deletions in the gE and TK genes, is efficacious against experimental infection with the virulent, contemporary Chinese PRV strain ZJ01. In this study, piglets were vaccinated with Porcilis® Begonia through either the intradermal (ID) route or the intramuscular (IM) route and subsequently challenged intranasally with strain ZJ01 at 4 weeks post-vaccination. An unvaccinated challenge group and an unvaccinated/nonchallenged group were also included in the study. All animals were monitored for 14 days after challenge. Vaccinated and negative control pigs stayed healthy during the study, while the unvaccinated control animals developed lesions associated with PRV ZJ01 challenge, and 44% of these pigs died before the end of the experiment. This study demonstrated that ID or IM vaccination of pigs with a vaccine based on the NIA-3 strain Porcilis® Begonia clinically protects against fatal PRV challenge with the ZJ01 strain.

Comparative chloroplast genomics reveals the phylogeny and the adaptive evolution of Begonia in China.

BACKGROUND: The Begonia species are common shade plants that are mostly found in southwest China. They have not been well studied despite their medicinal and decorative uses because gene penetration, decreased adaptability, and restricted availability are all caused by frequent interspecific hybridization. RESULT: To understand the patterns of mutation in the chloroplast genomes of different species of Begonia, as well as their evolutionary relationships, we collected seven Begonia species in China and sequenced their chloroplast genomes. Begonia species exhibit a quadripartite structure of chloroplast genomes (157,634 - 169,694 bp), consisting of two pairs of inverted repeats (IR: 26,529 - 37,674 bp), a large single copy (LSC: 75,477 - 86,500 bp), and a small single copy (SSC: 17,861 - 18,367 bp). 128-143 genes (comprising 82-93 protein-coding genes, 8 ribosomal RNAs, and 36-43 transfer RNAs) are found in the chloroplast genomes. Based on comparative analyses, this taxon has a relatively similar genome structure. A total of six substantially divergent DNA regions (trnT-UGU-trnL-UAA, atpF-atpH, ycf4-cemA, psbC-trnS-UGA, rpl32-trnL-UAG, and ccsA-ndhD) are found in the seventeen chloroplast genomes. These regions are suitable for species identification and phylogeographic analysis. Phylogenetic analysis shows that Begonia species that were suited to comparable environments grouped in a small clade and that all Begonia species formed one big clade in the phylogenetic tree, supporting the genus' monophyly. In addition, positive selection sites were discovered in eight genes (rpoC1, rpoB, psbE, psbK, petA, rps12, rpl2, and rpl22), the majority of which are involved in protein production and photosynthesis. CONCLUSION: Using these genome resources, we can resolve deep-level phylogenetic relationships between Begonia species and their families, leading to a better understanding of evolutionary processes. In addition to enhancing species identification and phylogenetic resolution, these results demonstrate the utility of complete chloroplast genomes in phylogenetically and taxonomically challenging plant groupings.

Rutin from Begonia roxburghii modulates iNOS and Sep A activity in treatment of Shigella flexneri induced diarrhoea in rats: An in vitro, in vivo and computational analysis.

In developing countries, diarrhoea is a major issue of concern, where consistent use of antibiotics has resulted in several side effects along with development of resistance among pathogens against these antibiotics. Since natural products are becoming the treatment of choice, therefore present investigation involves mechanistic evaluation of antidiarrhoeal potential of Begonia roxburghii and its marker rutin against Shigella flexneri (SF) induced diarrhoea in rats following in vitro, in vivo and in silico protocols. The roots of the plant are used as vegetable in the North East India and are also used traditionally in treating diarrhoea. Phytochemically standardized ethanolic extract of B. roxburghii (EBR) roots and its marker rutin were first subjected to in vitro antibacterial evaluation against SF. Diarrhoea was induced in rats using suspension of SF and various diarrhoeagenic parameters were examined after first, third and fifth day of treatment at 100, 200 and 300 mg/kg, p.o. with EBR and 50 mg/kg, p.o. with rutin respectively. Additionally, density of SF in stools, stool water content, haematological and biochemical parameters, cytokine profiling, ion concentration, histopathology and Na+/K+-ATPase activity were also performed. Molecular docking and dynamics simulation studies of ligand rutin was studied against secreted extracellular protein A (Sep A, PDB: 5J44) from SF and Inducible nitric oxide synthase (iNOS, PDB: 1DD7) followed by network pharmacology. EBR and rutin demonstrated a potent antibacterial activity against SF and also showed significant recovery from diarrhoea (EBR: 81.29 ± 0.91% and rutin: 75.27 ± 0.89%) in rats after five days of treatment. EBR and rutin also showed significant decline in SF density in stools, decreased cytokine expression, potential antioxidant activity, cellular proliferative nature and recovered ion loss due to enhanced Na+/K+-ATPase activity, which was also supported by histopathology. Rutin showed a very high docking score of -11.61 and -9.98 kcal/mol against iNOS and Sep A respectively and their stable complex was also confirmed through dynamics, while network pharmacology suggested that, rutin is quite capable of modulating the pathways of iNOS and Sep A. Thus, we may presume that rutin played a key role in the observed antidiarrhoeal activity of B. roxburghii against SF induced diarrhoea.

Begonia-Inspired Slow Photon Effect of a Photonic Crystal for Augmenting Algae Photosynthesis.

Plant photosynthesis is considered to be an environmentally friendly and effective measure for reducing carbon dioxide levels to meet the global objective of carbon neutrality. However, the light energy utilization of photosynthetic pigments is insufficient. Begonia pavonine (B. pavonina) with blue leaves exhibits a photosynthetic quantum yield 10% higher than those of other plants by virtue of their photonic crystal (PC) thylakoids. Inspired by this property, we prepared non-angle-dependent PC hydrogels and assembled them with algae Chlorella pyrenoidosa (C. pyre). The band edge of PC hydrogels matched the absorption peaks of C. pyre, and the resulting slow photon effect increased the interaction time between incident light and photosynthetic pigments, which in turn induced the expression of light-harvesting proteins and the synthesis of pigments, thereby improving the light energy utilization. Further, we introduced an artificial antenna into the assembly, which assisted the slow photon effect in increasing the oxygen evolution and carbon sequestration rate by more than 200%. This method avoids the photobleaching problems faced by methods of synthesizing artificial antenna pigments and the biosafety problems faced by genetically engineered methods of editing pigments or proteins.

Polyphenolic Composition and Antioxidant Activity (ORAC, EPR and Cellular) of Different Extracts of Argylia radiata Vitroplants and Natural Roots.

Plant biochemistry studies have increased in recent years due to their potential to improve human health. Argylia radiata is an extremophile plant with an interesting polyphenolic profile. However, its biomass is scarce and occasionally available. Argylia in vitro biomass was obtained from tissue culture and compared with in vivo roots regarding its polyphenolic and flavonoid content. Different solvents were used to prepare extracts from the in vitro tissue of callus and aerial plant organs and in vivo roots. UPLC-MS/MS was used to assess the chemical composition of each extract. ORAC-FL and scavenging of free radicals (DPPH and OH) methods were used to determine the antioxidant capacity of extracts. Furthermore, the biological activity of the extracts was established using the cellular antioxidant activity method. The vitroplants were a good source of polyphenols (25-68 mg GAE/100 g tissue FW), and methanol was the most efficient solvent. Eight polyphenolic compounds were identified, and their antioxidant properties were investigated by different chemical methods with EPR demonstrating its specific scavenging activity against free radicals. All extracts showed cellular dose-dependent antioxidant activity. The methanolic extract of vitroplants showed the highest cellular antioxidant activity (44.6% and 51%) at 1 and 10 µg/mL of extract, respectively. Vitroplants of A. radiata are proposed as a biotechnological product as a source of antioxidant compounds with multiple applications.

Genomes shed light on the evolution of Begonia, a mega-diverse genus.

Clarifying the evolutionary processes underlying species diversification and adaptation is a key focus of evolutionary biology. Begonia (Begoniaceae) is one of the most species-rich angiosperm genera with c. 2000 species, most of which are shade-adapted. Here, we present chromosome-scale genome assemblies for four species of Begonia (B. loranthoides, B. masoniana, B. darthvaderiana and B. peltatifolia), and whole genome shotgun data for an additional 74 Begonia representatives to investigate lineage evolution and shade adaptation of the genus. The four genome assemblies range in size from 331.75 Mb (B. peltatifolia) to 799.83 Mb (B. masoniana), and harbor 22 059-23 444 protein-coding genes. Synteny analysis revealed a lineage-specific whole-genome duplication (WGD) that occurred just before the diversification of Begonia. Functional enrichment of gene families retained after WGD highlights the significance of modified carbohydrate metabolism and photosynthesis possibly linked to shade adaptation in the genus, which is further supported by expansions of gene families involved in light perception and harvesting. Phylogenomic reconstructions and genomics studies indicate that genomic introgression has also played a role in the evolution of Begonia. Overall, this study provides valuable genomic resources for Begonia and suggests potential drivers underlying the diversity and adaptive evolution of this mega-diverse clade.

Alterations in endogenous hormone levels and energy metabolism promoted the induction, differentiation and maturation of Begonia somatic embryos under clinorotation.

The present study provides a visual insight into the effects of simulated microgravity (MG) on somatic embryogenesis (SE) in Begonia through the analysis of phytohormone fluctuations and energy metabolism. To investigate this relationship, thin cell layer culture model was first used. The results showed that MG changed the phytohormone content and stimulated starch biosynthesis to convert into sugar to release energy needed for regeneration and proliferation. Moreover, from the results it is likely that MG accelerated the initiation and subsequently maturation and aging of SE via decrease of AUX and increase of ABA. High content of GA, CKs, starch, sugar and low ABA as well as high CKs/ABA ratio were responsible for the increase in the number of embryos under clinorotation which was 1.57-fold higher than control after 90 days. The increase in fresh and dry weight of somatic embryos and chlorophyll content under MG were confirmed as their adaptive responses to gravitational stress. However, long-term exposure to MG (120 days) stimulated biosynthesis of ABA levels 1.85-fold higher than controls, which resulted in a decrease in chlorophyll content, increase in number of mature embryos and stomata length. These results revealed that MG regulated the induction, differentiation and senescence of somatic embryos via a biochemical interaction pathway.

Multi-tissue transcriptome analysis of two Begonia species reveals dynamic patterns of evolution in the chalcone synthase gene family.

Begonia is an important horticultural plant group, as well as one of the most speciose Angiosperm genera, with over 2000 described species. Genus wide studies of genome size have shown that Begonia has a highly variable genome size, and analysis of paralog pairs has previously suggested that Begonia underwent a whole genome duplication. We address the contribution of gene duplication to the generation of diversity in Begonia using a multi-tissue RNA-seq approach. We chose to focus on chalcone synthase (CHS), a gene family having been shown to be involved in biotic and abiotic stress responses in other plant species, in particular its importance in maximising the use of variable light levels in tropical plants. We used RNA-seq to sample six tissues across two closely related but ecologically and morphologically divergent species, Begonia conchifolia and B. plebeja, yielding 17,012 and 19,969 annotated unigenes respectively. We identified the chalcone synthase gene family members in our Begonia study species, as well as in Hillebrandia sandwicensis, the monotypic sister genus to Begonia, Cucumis sativus, Arabidopsis thaliana, and Zea mays. Phylogenetic analysis suggested the CHS gene family has high duplicate turnover, all members of CHS identified in Begonia arising recently, after the divergence of Begonia and Cucumis. Expression profiles were similar within orthologous pairs, but we saw high inter-ortholog expression variation. Sequence analysis showed relaxed selective constraints on some ortholog pairs, with substitutions at conserved sites. Evidence of pseudogenisation and species specific duplication indicate that lineage specific differences are already beginning to accumulate since the divergence of our study species. We conclude that there is evidence for a role of gene duplication in generating diversity through sequence and expression divergence in Begonia.

Combined effect of silicon and non-thermal plasma treatments on yield, mineral content, and nutraceutical proprieties of edible flowers of Begonia cucullata.

Edible flowers are becoming popular as a nutraceutical and functional food that can contribute to human nutrition with high antioxidant molecules and mineral elements. While comparative studies between different flower species have been performed, less is known about the best agronomical practices to increase yield and nutraceutical proprieties of blooms. Silicon stimulates plant resistance against stress and promotes plant growth while non-thermal plasma (NTP) technology has been applied for the disinfection and decontamination of water, as well as for increasing plant production and quality. The application of silicon and NTP technology through nutrient solution and spraying was investigated in edible flowers given that the combination of these treatments may play a role in promoting their nutritional and nutraceutical proprieties. The treatments were applied on two varieties of Begonia cucullata Willd. (white and red flowers) to explore their effects on different flower pigmentations. Plants with red flowers showed higher nutraceutical proprieties than the white ones but yielded a lower flower number. While the NTP treatment did not improve flower yield and quality, the silicon treatment increased anthocyanins and dry weight percentage in red flowers. NTP treatment increased zinc concentration, while it decreased potassium, magnesium, and manganese, and increased silicon concentration in white flowers. The combination of silicon and NTP showed negative effects on some nutraceutical proprieties of red flowers thus highlighting that the two treatments cannot be combined in edible flower production. In conclusion, the positive effect of silicon use in edible flower production has been demonstrated while the NTP technology showed contrasting results and its use should be explored in greater depth, including a consideration of its role in biotic attack prevention and reduced chemical input.

Antioxidant and antiviral potency of Begonia medicinalis fractions.

OBJECTIVES: This study aims to evaluate the antioxidant and antiviral potency of n-hexane, ethyl acetate and, water fractions of Begonia medicinalis Ardi & D.C.Thomas as well as to identify the chemical constituents. METHODS: Assays for antioxidant and antiviral activity (HIV-1) were carried out on MT-4 cells infected with HIV using the DPPH method and the determination of the cytopathic effect. Meanwhile, GC-MS was used to identify the chemical compounds. RESULTS: The determination of antioxidants showed that all fractions possessed potent activity with the IC50 ranging from 2.61 to 8.26 µg/mL. From the antiviral activity of MT-4 cells infected by HIV, the n-hexane fraction of B. medicinalis showed the most potency with the IC50 of 0.04 ± 0.05 µg/mL. It has less cytotoxicity (11.08 ± 4.60 µg/mL) affording the high selectivity index of 238.80. Furthermore, GC-MS analysis of n-hexane fraction found the major compound of carboxylic acid derivate with the area percentage of 76.4% and the presence of phenolic compounds (8.38%). Meanwhile, in water fraction, terpenoids were found in a higher concentration (10.05%) than others. CONCLUSIONS: Therefore, this study supports the application of B. medicinalis as a herbal medicine for antioxidant and antiviral.

A new steroid glycoside from Begonia sp.: cytotoxic activity and docking studies.

Chemical investigation on the ethyl acetate extract of the aerial parts of Begonia sp. afforded a new steroid glycoside, 9(11)α,16(17)α-dioxirane-20,25-dihydroxy-ß-sitosterol-3-O-ß-glucopyranoside (1) along with a known steroidal glycoside, ß-sitosterol-3-O-ß-D-glucopyranoside (2). The Chemical structures were elucidated by 1D and 2D NMR and mass spectroscopic analysis. Cytotoxicity against four different cancer cell lines (HeLa, T47D, WiDr and Vero) was assessed. Compound 1 was more potent and selective against breast cancer cell line (T47D) than other cell lines with an IC50 value of 0.16 µg/mL. Further docking study of 1 exhibited the preference of molecule to bind in the epidermal growth factor receptor tyrosine kinase (EGFR-TK) binding pockets with docking scores of -97.8800 (PLANTS) and -3.56 kcal/mol (AutoDock 4.2.6).

Complete genome sequence of begonia flower breaking virus, a novel member of the genus Potyvirus.

We determined the complete genomic sequence of begonia flower breaking virus (BFBV), a novel putative member of the genus Potyvirus isolated from Begonia bowerae cv. 'Tiger' plants grown in Kunming. The genomic RNA comprises 9540 nucleotides (nt), excluding the 3'-terminal poly(A) tail, and contains a typical open reading frame (ORF) of potyviruses. The ORF consists of 9219 nucleotides and encodes a 3073-amino-acid polyprotein that is predicted to be proteolytically cleaved into 10 mature peptides. Sequence comparison indicated that BFBV shares 43.9-55.12% amino acid sequence identity with known potyviruses and that BFBV shares the highest amino acid sequence identity (55.12%) with beet mosaic virus. The results from the complete genomic sequence analysis further suggest that BFBV is a member of a novel species in the genus Potyvirus.

Sensory bias and signal detection trade-offs maintain intersexual floral mimicry.

Mimicry is common in interspecies interactions, yet conditions maintaining Batesian mimicry have been primarily tested in predator-prey interactions. In pollination mutualisms, floral mimetic signals thought to dupe animals into pollinating unrewarding flowers are widespread (greater than 32 plant families). Yet whether animals learn to both correctly identify floral models and reject floral mimics and whether these responses are frequency-dependent is not well understood. We tested how learning affected the effectiveness and frequency-dependence of imperfect Batesian mimicry among flowers using the generalist bumblebee, Bombus impatiens, visiting Begonia odorata, a plant species exhibiting intersexual floral mimicry. Unrewarding female flowers are mimics of pollen-rewarding male flowers (models), though mimicry to the human eye is imperfect. Flower-naive bees exhibited a perceptual bias for mimics over models, but rapidly learned to avoid mimics. Surprisingly, altering the frequency of models and mimics only marginally shaped responses by naive bees and by bees experienced with the distribution and frequency of models and mimics. Our results provide evidence both of exploitation by the plant of signal detection trade-offs in bees and of resistance by the bees, via learning, to this exploitation. Critically, we provide experimental evidence that imperfect Batesian mimicry can be adaptive and, in contrast with expectations of signal detection theory, functions largely independently of the model and mimic frequency. This article is part of the theme issue 'Signal detection theory in recognition systems: from evolving models to experimental tests'.

Antifungal compounds with anticancer potential from Trichoderma sp. P8BDA1F1, an endophytic fungus from Begonia venosa.

Fungi in the genus Trichoderma are notorious producers of secondary metabolites with diverse applications, such as antibacterial, antifungal, and plant growth-promoting properties. Peptaibols are linear peptides produced by such fungi, with more than 440 compounds described to date, including tricholongins, longibrachins, trichobrachins, and trichovirins. Peptaibols are synthesized by non-ribosomal peptide synthetases and they have several biological activities. Our research group isolated four peptaibols (6DP2, 6DP3, 6DP4, and 6DP5) with antifungal activity against the plant pathogen Colletotrichum gloeosporioides and the proteasome (a cancer chemotherapy target) from Trichoderma sp. P8BDA1F1, an endophytic fungus from Begonia venosa. The ethyl acetate extract of this endophyte showed activity of 6.01% and 75% against C. gloeosporioides and the proteasome, respectively. The isolated compounds were identified by MS/MS and compared to literature data, suggesting the presence of trilongins BI, BII, BIII, and BIV, which are peptaibols containing 20 amino acid residues. The minimum inhibitory concentration against C. gloeosporioides was 40 µM for trilongin BI, 320 µM for trilongin BII, 160 µM for trilongin BIII, and 310 µM for trilongin BIV. BI-BIV trilongins inhibited proteasome ChTL activity, with IC50 values of 6.5 ± 2.7; 4.7 ± 1.8; 6.3 ± 2.2; and 2.7 ± 0.5 µM, respectively. The compounds were tested ex vivo against the intracellular amastigotes of Leishmania (L.) infantum but showed no selectivity. It is the first report of trilongins BI-BIV with antifungal activity against C. gloeosporioides and the proteasome target.

Introduction of a new method for two-dimensional NMR quantitative analysis in metabolomics studies.

NMR is one of the most important platforms for metabolomic studies. Though 2D NMR has been applied in metabolomics, most applications have mainly focused on metabolite identification whilst limitations causing a bottle-neck for applying high-throughput 2D NMR data for quantity related statistical analysis lies on the data interpretation methods. In this study, instead of using the traditional methods of calculating the 2D NMR data to search for the important features, a new procedure, which applies the high-resolution 1D NMR metabolites chemical shift range to filter the 2D NMR data, was developed. This new method was demonstrated using both a mixture of standard metabolites and a case study on plant extracts using 2D non-uniform sampling (NUS) total correlation spectroscopy (TOCSY) data. As a result, our method successfully filtered out the important features with a high success rate, and the extracted peaks showed high linearity between the calculated intensities and the concentrations of metabolites from a range of 0.05 mM-2 mM. The method was successfully applied to a metabolomics case study which included 18 Begonia samples that showed excellent peak extractions. In summary, our study has provided a practical new 2D NMR data extraction method for use in future metabolomics studies.

Estudio farmacobotánico, etnofarmacológico y micrográfico de drogas vegetales utilizadas para las afecciones de mayor índice de mortalidad, comercializadas en la ciudad de Posadas, Misiones Argentina. I parte / Pharmacobotanical, ethnopharmacological and micrographic study of vegetable drugs used for the affections of greater index of mortality, commercialized in the City of Posadas, Misiones Argentine. Part I

El presente estudio corresponde a la primera entrega donde se realizó el análisis farmacobotánico, etnofarmacológico y micrográfico de drogas vegetales utilizadas en forma empírica para las afecciones con mayor tasa de mortalidad en la provincia de Misiones. En esta primera instancia (Parte I), se exponen seis (6) especies utilizadas para las afecciones cardiovasculares y respiratorias. Las mismas fueron relevadas en los diferentes puntos de comercialización de la Ciudad de Posadas. Se conservaron especímenes de resguardo en el Herbario del Departamento de Farmacia de la Facultad de Ciencias Exactas, Químicas y Naturales de la Universidad Nacional de Misiones y mediante fotografías, se identificaron por medio de procedimientos taxonómicos clásicos, y se analizó la endo- y exomorfología empleando técnicas de rutina para este tipo de trabajo, con la documentación fotográfica. Los objetivos fueron relevar y documentar las especies vegetales utilizadas para las afecciones cardiovasculares y respiratorias, sus usos etnofarmacológicos, las partes utilizadas y formas de administración, además de contribuir a incrementar el conocimiento de la morfo-anatomía y micrografía aplicada de dichas drogas vegetales. Los resultados obtenidos en esta instancia se corresponden con el relevamiento de Syagrus romanzoffiana, Cuphea calophylla subsp. mesostemon, Hypericum connatum, Begonia cucullata, Cecropia pachystachya y Luehea divaricata, especies comercializadas y utilizadas como agentes terapéuticos en el tratamiento de las afecciones cardiovasculares y respiratorias.

The present study corresponds to the first delivery where the pharmacobotanical, ethnopharmacological and micrographic analysis of vegetable drugs used empirically for the conditions with the highest mortality rate in the province of Misiones was carried out. In this first instance (Part I), six (6) species used for cardiovascular and respiratory diseases are exposed. They were surveyed at different points of sale in the City of Posadas. Specimens were kept in the Herbarium of the Department of Pharmacy of the Faculty of Exact, Chemical and Natural Sciences of the National University of Misiones and photographed, were identified by means of classical taxonomic procedures, and the endo- and exomorphology were analyzed using routine techniques for this type of work, with photographic documentation. The objectives were to survey and document the plant species used for cardiovascular and respiratory diseases, their ethnopharmacological uses, parts used and forms of administration, in addition to contributing to increase the knowledge of the morpho-anatomy and applied micrograph of said plant drugs. The results obtained in this instance correspond to Syagrus romanzoffiana, Cuphea calophylla subsp. mesostemon, Hypericum connatum, Begonia cucullata, Cecropia pachystachya and Luehea divaricata survey, commercialized species and used as therapeutic agents in the treatment of cardiovascular and respiratory affections.

Selection of suitable reference genes for qRT-PCR analysis of Begonia semperflorens under stress conditions.

Begonia semperflorens (B. semperflorens), belonging to the family Begoniaceae, has now been widely cultivated worldwide and is famous for its ornamental plants with colourful flowers and distinctive leaves. The selection of appropriate internal reference genes is very important to accurately determine target gene expression via quantitative real-time PCR. However, internal reference gene selection has never been conducted in B. semperflorens. In this study, seven candidate reference genes of B. semperflorens, including 18S ribosomal RNA (Bs18S), pentatricopeptide repeat-containing protein (BsPPR), actin-related protein 5 isoform X2 (BsACT), DNAJ homologue subfamily C member 17 (BsDNAJ), glyceraldehyde-3-phosphate dehydrogenase (BsGAPDH), NAD-dependent malic enzyme 59 kDa isoform, mitochondria (BsNAD-ME), and peptidyl-prolyl cis-trans isomerase CYP26-2, chloroplast (BsCYP), which were obtained from our previous studies, were selected. The stabilities of these genes under stress conditions were analysed using geNorm and NormFinder. Validation of target gene expressions, including phenylalanine ammonia-lyase (BsPAL) and respiratory burst oxidase homologue D (BsRBOHD) under biotic and abiotic conditions, phenylalanine ammonia-lyase (BsPAL), anthocyanidin synthase (BsANS), chalcone synthase (BsCHS), and flavanone-3-hydroxylase (BsF3H) under low temperature, using these seven internal reference genes for normalisation further confirmed the stabilities of the selected genes and indicated the need for reference gene selection for normalising gene expressions in B. semperflorens. Of the seven candidate reference genes, the combination of BsACT, BsDNAJ, and BsNAD-ME was the ideal reference gene set for normalising gene expression in samples under biotic conditions. BsCYP combined with BsACT or BsGAPDH was the best reference gene pair under abiotic conditions. BsACT and BsPPR could be combined to normalise gene expression under low temperature. Our results will benefit future studies on gene expression in plants of Begoniaceae.

Spectral effects of light-emitting diodes on plant growth, visual color quality, and photosynthetic photon efficacy: White versus blue plus red radiation.

Arrays of blue (B, 400-500 nm) and red (R, 600-700 nm) light-emitting diodes (LEDs) used for plant growth applications make visual assessment of plants difficult compared to a broad (white, W) spectrum. Although W LEDs are sometimes used in horticultural lighting fixtures, little research has been published using them for sole-source lighting. We grew seedlings of begonia (Begonia ×semperflorens), geranium (Pelargonium ×horturum), petunia (Petunia ×hybrida), and snapdragon (Antirrhinum majus) at 20°C under six sole-source LED lighting treatments with a photosynthetic photon flux density (PPFD) of 160 µmol∙m-2∙s-1 using B (peak = 447 nm), green (G, peak = 531 nm), R (peak = 660 nm), and/or mint W (MW, peak = 558 nm) LEDs that emitted 15% B, 59% G, and 26% R plus 6 µmol∙m-2∙s-1 of far-red radiation. The lighting treatments (with percentage from each LED in subscript) were MW100, MW75R25, MW45R55, MW25R75, B15R85, and B20G40R40. At the transplant stage, total leaf area, and fresh and dry weight were similar among treatments in all species. Surprisingly, when petunia seedlings were grown longer (beyond the transplant stage) under sole-source lighting treatments, the primary stem elongated and had flower buds earlier under MW100 and MW75R25 compared to under B15R85. The color rendering index of MW75R25 and MW45R55 were 72, and 77, respectively, which was higher than those of other treatments, which were ≤64. While photosynthetic photon efficacy of B15R85 (2.25 µmol∙J-1) was higher than the W light treatments (1.51-2.13 µmol∙J-1), the dry weight gain per unit electric energy consumption (in g∙kWh-1) of B15R85 was similar to those of MW25R75, MW45R55, and MW75R25 in three species. We conclude that compared to B+R radiation, W radiation had generally similar effects on seedling growth at the same PPFD with similar electric energy consumption, and improved the visual color quality of sole-source lighting.

Chloroplast and nuclear DNA exchanges among Begonia sect. Baryandra species (Begoniaceae) from Palawan Island, Philippines, and descriptions of five new species.

The Philippine island of Palawan is highly biodiverse. During fieldwork there in 2011 & 2014 we found five unknown species in the large genus Begonia. The species are similar in their rhizomatous stems, four-tepaled flowers, inferior two- or three-locular ovaries with bilamellate placentas, and are assignable to Begonia sect. Baryandra. Our observations support the recognition of these as five new species endemic to Palawan: B. elnidoensis, B. gironellae, B. quinquealata, B. tabonensis and B. tenuibracteata which are described here. The five new species were added to phylogenies based Bayesian analysis of nrDNA (ITS) and chloroplast DNA (ndhA, ndhF-rpl32, rpl32-trnL, trnC-trnD), along with 45 other allied ingroup species. A majority of the species show incongruent positions in the two phylogenies, with evidence of prevalent chloroplast capture. Models show chloroplast capture is more likely in plant populations with high levels of inbreeding following a reduction in selfing rate after hybridisation; we suggest that this is a possible explanation for the massive amount of chloroplast exchange seen in our phylogeny, as Begonia species often exist as small isolated populations and may be prone to inbreeding depression. Our data also indicate a level of nuclear genetic exchange between species. The high prevalence of hybrid events in Begonia is potentially an important factor in driving genomic change and species evolution in this mega-diverse genus.

Inherent Resistance to 14α-Demethylation Inhibitor Fungicides in Colletotrichum truncatum Is Likely Linked to CYP51A and/or CYP51B Gene Variants.

Anthracnose disease, caused by Colletotrichum truncatum, affects marketable yield during preharvest production and postharvest storage of fruits and vegetables worldwide. Demethylation inhibitor (DMI) fungicides are among the very few chemical classes of single-site mode of action fungicides that are effective in controlling anthracnose disease. However, some species are inherently resistant to DMIs and more information is needed to understand this phenomenon. Isolates of C. truncatum were collected from the United States and China from peach, soybean, citrus, and begonia and sensitivity to six DMIs (difenoconazole, propiconazole, metconazole, tebuconazole, flutriafol, and fenbuconazole) was determined. Compared with DMI sensitive isolates of C. fructicola, C. siamense, and C. fioriniae (EC50 value ranging from 0.03 to 16.2 µg/ml to six DMIs), C. truncatum and C. nymphaeae were resistant to flutriafol and fenbuconazole (with EC50 values of more 50 µg/ml). Moreover, C. truncatum was resistant to tebuconazole and metconazole (with resistance factors of 27.4 and 96.0) and displayed reduced sensitivity to difenoconazole and propiconazole (with resistance factors of 5.1 and 5.2). Analysis of the Colletotrichum spp. genome revealed two potential DMI targets, CYP51A and CYP51B, that putatively encode P450 sterol 14α-demethylases. Both genes were identified and sequenced from C. truncatum and other species and no correlation between CYP51 gene expression levels and fungicide sensitivity was found. Four amino acid variations L208Y, H238R, S302A, and I366L in CYP51A, and three variations H373 N, M376L, and S511T in CYP51B correlated with the DMI resistance phenotype. CYP51A structure model analysis suggested the four alterations may reduce azole affinity. Likewise, CYP51B structure analysis suggested the H373 N and M376L variants may change the conformation of the DMI binding pocket, thereby causing differential sensitivity to DMI fungicides in C. truncatum.