Identification of distinct quantitative trait loci associated with defence against the closely related aphids Acyrthosiphon pisum and A. kondoi in Medicago truncatula

Aphids are a major family of plant insect pests. Medicago truncatula and Acyrthosiphon pisum (pea aphid, PA) are model species with a suite of resources available to help dissect the mechanism underlying plant–aphid interactions. A previous study focused on monogenic and relatively strong resistance in M. truncatula to PA and other aphid species. In this study a moderate resistance to PA was characterized in detail in the M. truncatula line A17 and compared with the highly susceptible line A20 and the more resistant line Jester. The results show that PA resistance in A17 involves both antibiosis and tolerance, and that resistance is phloem based. Quantitative trait locus (QTL) analysis using a recombinant inbred line (RIL) population (n=114) from a cross between A17 and A20 revealed that one locus, which co-segregated with AIN (Acyrthosiphon-induced necrosis) on chromosome 3, is responsible for the reduction of aphid biomass (indicator of antibiosis) for both PA and bluegreen aphid (BGA, A. kondoi), albeit to a lesser degree for PA than BGA. Interestingly, two independent loci on chromosomes 5 and 3 were identified for the plant biomass reduction (indicator of plant tolerance) by PA and BGA, respectively, demonstrating that the plant’s tolerance response to these two closely related aphid species is distinct. Together with previously identified major resistant (R) genes, the QTLs identified in this study are powerful tools to understand fully the spectrum of plant defence against sap-sucking insects and provide opportunities for breeders to generate effective and sustainable strategies for aphid control

Prosudba učinkovitosti celomske tekućine kišne gujavice na cijeljenje rana u punoj debljini kože kunića.

The present study was conducted on 16 New Zealand White rabbits of 10-12 months of age, to evaluate the healing potential of earthworm coelomic fluid in full thickness skin wounds. Under xylazine-ketamine anaesthesia, four rectangular full thickness excisional skin wounds, measuring 2×2 cm2 were created on the dorsum of each animal and designated as groups I, II, III and IV. Wounds were treated by topical coelomic fluid (I), 0.5 % povidone-iodine and coelomic fluid (II), 0.5 % povidone-iodine (III) and normal saline (IV). Healing was evaluated on the basis of gross and histomorphological parameters. The mean wound area was significantly lesser (P<0.05) in the wounds of groups I and II as compared to groups III and IV, up to 21 days. Out of 16 wounds, nine wounds in group I (56.25 %) and 11 wounds in group II (68.75 %) healed completely by day 21, but none in groups III and IV. Histomorphological studies showed more mature and densely placed collagen and better epithelialization in groups I and II as compared to groups III and IV. It was concluded that coelomic fluid of the earthworm Eisenia foetida can accelerate healing of full-thickness skin wounds in rabbits.Istraživanje je provedeno na 16 novozelandskih bijelih kunića u dobi od 10 do 12 mjeseci s ciljem da se prosudi mogući učinak celomske tekućine kišne gujavice na cijeljenje rana u punoj debljini kože u kunića. Kunići su bili podijeljeni u četiri skupine označene I, II, III i IV. Četiri pravokutne ekscizijske kožne rane u punoj debljini veličine 2x2 cm načinjene su na leđima svake životinje pod anestezijom ksilazin-ketaminom. Rane su bile obrađene celomskom tekućinom (skupina I), 0,5 %-tnim povidon-jodom i celomskom tekućinom (skupina II), 0,5 %-tnim povidon-jodom (skupina III) i fiziološkom otopinom (skupina IV). Cijeljenje rana bilo je prosuđivano na osnovi patoanatomskih i patohistoloških nalaza. Prosječna površina rana 21 dan nakon ekscizije bila je značajno manja (P<0,05) u skupinama I i II u usporedbi s površinom skupina III i IV. Devet od 16 rana u skupini I (56,25 %) i 11 u skupini II (68,75 %) u potpunosti su zacijelile 21 dan nakon ekscizije, dok nijedna nije zacijelila u skupini III i IV. Patohistološki nalaz pokazao je više zrelog i gušće raspoređenog kolagena te bolju epitelizaciju u skupinama I i II u usporedbi sa skupinama III i IV. Može se zaključiti da celomska tekućina kišne gujavice Eisenia foetida može ubrzati cijeljenje rana u punoj debljini kože u kunića

Contrasting selective patterns across the segmented genome of bluetongue virus in a global reassortment hotspot

For segmented viruses, rapid genomic and phenotypic changes can occur through the process of reassortment, whereby co-infecting strains exchange entire segments creating novel progeny virus genotypes. However, for many viruses with segmented genomes, this process and its effect on transmission dynamics remain poorly understood. Here, we assessed the consequences of reassortment for selection on viral diversity through time using bluetongue virus (BTV), a segmented arbovirus that is the causative agent of a major disease of ruminants. We analysed ninety-two BTV genomes isolated across four decades from India, where BTV diversity, and thus opportunities for reassortment, are among the highest in the world. Our results point to frequent reassortment and segment turnover, some of which appear to be driven by selective sweeps and serial hitchhiking. Particularly, we found evidence for a recent selective sweep affecting segment 5 and its encoded NS1 protein that has allowed a single variant to essentially invade the full range of BTV genomic backgrounds and serotypes currently circulating in India. In contrast, diversifying selection was found to play an important role in maintaining genetic diversity in genes encoding outer surface proteins involved in virus interactions (VP2 and VP5, encoded by segments 2 and 6, respectively). Our results support the role of reassortment in driving rapid phenotypic change in segmented viruses and generate testable hypotheses for in vitro experiments aiming at understanding the specific mechanisms underlying differences in fitness and selection across viral genomes

INDEL variation in the regulatory region of the major flowering time gene LanFTc1 is associated with vernalization response and flowering time in narrow-leafed lupin (Lupinus angustifolius L.)

© 2018 The Authors Plant, Cell & Environment Published by John Wiley & Sons Ltd. Narrow-leafed lupin (Lupinus angustifolius L.) cultivation was transformed by 2 dominant vernalization-insensitive, early flowering time loci known as Ku and Julius (Jul), which allowed expansion into shorter season environments. However, reliance on these loci has limited genetic and phenotypic diversity for environmental adaptation in cultivated lupin. We recently predicted that a 1,423-bp deletion in the cis-regulatory region of LanFTc1, a FLOWERING LOCUS T (FT) homologue, derepressed expression of LanFTc1 and was the underlying cause of the Ku phenotype. Here, we surveyed diverse germplasm for LanFTc1 cis-regulatory variation and identified 2 further deletions of 1,208 and 5,162 bp in the 5' regulatory region, which overlap the 1,423-bp deletion. Additionally, we confirmed that no other polymorphisms were perfectly associated with vernalization responsiveness. Phenotyping and gene expression analyses revealed that Jul accessions possessed the 5,162-bp deletion and that the Jul and Ku deletions were equally capable of removing vernalization requirement and up-regulating gene expression. The 1,208-bp deletion was associated with intermediate phenology, vernalization responsiveness, and gene expression and therefore may be useful for expanding agronomic adaptation of lupin. This insertion/deletion series may also help resolve how the vernalization response is mediated at the molecular level in legumes

Variability in an effector gene promoter of a necrotrophic fungal pathogen dictates epistasis and effector-triggered susceptibility in wheat

The fungus Parastagonospora nodorum uses proteinaceous necrotrophic effectors (NEs) to induce tissue necrosis on wheat leaves during infection, leading to the symptoms of septoria nodorum blotch (SNB). The NEs Tox1 and Tox3 induce necrosis on wheat possessing the dominant susceptibility genes Snn1 and Snn3B1/Snn3D1, respectively. We previously observed that Tox1 is epistatic to the expression of Tox3 and a quantitative trait locus (QTL) on chromosome 2A that contributes to SNB resistance/susceptibility. The expression of Tox1 is significantly higher in the Australian strain SN15 compared to the American strain SN4. Inspection of the Tox1 promoter region revealed a 401 bp promoter genetic element in SN4 positioned 267 bp upstream of the start codon that is absent in SN15, called PE401. Analysis of the world-wide P. nodorum population revealed that a high proportion of Northern Hemisphere isolates possess PE401 whereas the opposite was observed in representative P. nodorum isolates from Australia and South Africa. The presence of PE401 removed the epistatic effect of Tox1 on the contribution of the SNB 2A QTL but not Tox3. PE401 was introduced into the Tox1 promoter regulatory region in SN15 to test for direct regulatory roles. Tox1 expression was markedly reduced in the presence of PE401. This suggests a repressor molecule(s) binds PE401 and inhibits Tox1 transcription. Infection assays also demonstrated that P. nodorum which lacks PE401 is more pathogenic on Snn1 wheat varieties than P. nodorum carrying PE401. An infection competition assay between P. nodorum isogenic strains with and without PE401 indicated that the higher Tox1-expressing strain rescued the reduced virulence of the lower Tox1-expressing strain on Snn1 wheat. Our study demonstrated that Tox1 exhibits both ‘selfish’ and ‘altruistic’ characteristics. This offers an insight into a complex NE-NE interaction that is occurring within the P. nodorum population. The importance of PE401 in breeding for SNB resistance in wheat is discussed

A specific fungal transcription factor controls effector gene expression and orchestrates the establishment of the necrotrophic pathogen lifestyle on wheat

The fungus Parastagonospora nodorum infects wheat through the use of necrotrophic efector (NE) proteins that cause host-specifc tissue necrosis. The Zn2Cys6 transcription factor PnPf2 positively regulates NE gene expression and is required for virulence on wheat. Little is known about other downstream targets of PnPf2. We compared the transcriptomes of the P. nodorum wildtype and a strain deleted in PnPf2 (pf2-69) during in vitro growth and host infection to further elucidate targets of PnPf2 signalling. Gene ontology enrichment analysis of the diferentially expressed (DE) genes revealed that genes associated with plant cell wall degradation and proteolysis were enriched in down-regulated DE gene sets in pf2-69 compared to SN15. In contrast, genes associated with redox control, nutrient and ion transport were up-regulated in the mutant. Further analysis of the DE gene set revealed that PnPf2 positively regulates twelve genes that encode efector-like proteins. Two of these genes encode proteins with homology to previously characterised efectors in other fungal phytopathogens. In addition to modulating efector gene expression, PnPf2 may play a broader role in the establishment of a necrotrophic lifestyle by orchestrating the expression of genes associated with plant cell wall degradation and nutrient assimilation.Tis study was supported by the Centre for Crop and Disease Management, a joint initiative of Curtin University and the Grains Research and Development Corporation [research grant CUR00023 (Programme 3)]. EJ was supported by the Australian Government Research Training Program Scholarshi

Plasmonic nanoparticle monomers and dimers: From nano-antennas to chiral metamaterials

We review the basic physics behind light interaction with plasmonic nanoparticles. The theoretical foundations of light scattering on one metallic particle (a plasmonic monomer) and two interacting particles (a plasmonic dimer) are systematically investigated. Expressions for effective particle susceptibility (polarizability) are derived, and applications of these results to plasmonic nanoantennas are outlined. In the long-wavelength limit, the effective macroscopic parameters of an array of plasmonic dimers are calculated. These parameters are attributable to an effective medium corresponding to a dilute arrangement of nanoparticles, i.e., a metamaterial where plasmonic monomers or dimers have the function of "meta-atoms". It is shown that planar dimers consisting of rod-like particles generally possess elliptical dichroism and function as atoms for planar chiral metamaterials. The fabricational simplicity of the proposed rod-dimer geometry can be used in the design of more cost-effective chiral metamaterials in the optical domain.Comment: submitted to Appl. Phys.

Evaluation of ground-level and space-borne sensor as tools in monitoring nitrogen nutrition status in immature and mature oil palm

Monitoring nitrogen (N) in oil palm is crucial for the production sustainability. The objective of this study is to examine the capability of visible (Vis), near infrared (NIR) and a combination of Vis and NIR (Vis + NIR) spectral indices acquired from different sensors for predicting foliar N content of different palm age groups. The N treatments varied from 0 to 2 kg per palm, subjected according to immature, young mature and prime mature classes. The Vis + NIR indices from the ground level-sensor that is green + red + NIR (G + R + NIR) was the best index for predicting N for immature palms (R2 = 0.91), while Vis indices blue + red (B + R) and Green Red Index from the space-borne sensor were significantly useful for N assessment of young and prime mature palms (R2 = 0.70 and 0.50), respectively. The application of vegetation indices for monitoring N status of oil palm is beneficial to examine extensive plantation areas

Full-genome sequencing as a basis for molecular epidemiology studies of bluetongue virus in India

Since 1998 there have been significant changes in the global distribution of bluetongue virus (BTV). Ten previously exotic BTV serotypes have been detected in Europe, causing severe disease outbreaks in naïve ruminant populations. Previously exotic BTV serotypes were also identified in the USA, Israel, Australia and India. BTV is transmitted by biting midges (Culicoides spp.) and changes in the distribution of vector species, climate change, increased international travel and trade are thought to have contributed to these events. Thirteen BTV serotypes have been isolated in India since first reports of the disease in the country during 1964. Efficient methods for preparation of viral dsRNA and cDNA synthesis, have facilitated full-genome sequencing of BTV strains from the region. These studies introduce a new approach for BTV characterization, based on full-genome sequencing and phylogenetic analyses, facilitating the identification of BTV serotype, topotype and reassortant strains. Phylogenetic analyses show that most of the equivalent genome-segments of Indian BTV strains are closely related, clustering within a major eastern BTV ‘topotype’. However, genome-segment 5 (Seg-5) encoding NS1, from multiple post 1982 Indian isolates, originated from a western BTV topotype. All ten genome-segments of BTV-2 isolates (IND2003/01, IND2003/02 and IND2003/03) are closely related (&gt;99% identity) to a South African BTV-2 vaccine-strain (western topotype). Similarly BTV-10 isolates (IND2003/06; IND2005/04) show &gt;99% identity in all genome segments, to the prototype BTV-10 (CA-8) strain from the USA. These data suggest repeated introductions of western BTV field and/or vaccine-strains into India, potentially linked to animal or vector-insect movements, or unauthorised use of ‘live’ South African or American BTV-vaccines in the country. The data presented will help improve nucleic acid based diagnostics for Indian serotypes/topotypes, as part of control strategies

Reactive Oxygen Species Play a Role in the Infection of the Necrotrophic Fungi, Rhizoctonia solani in Wheat

Rhizoctonia solani is a nectrotrophic fungal pathogen that causes billions of dollars of damage to agriculture worldwide and infects a broad host range including wheat, rice, potato and legumes. In this study we identify wheat genes that are differentially expressed in response to the R. solani isolate, AG8, using microarray technology. A significant number of wheat genes identified in this screen were involved in reactive oxygen species (ROS) production and redox regulation. Levels of ROS species were increased in wheat root tissue following R. solani infection as determined by Nitro Blue Tetrazolium (NBT), 3,3'-diaminobenzidine (DAB) and titanium sulphate measurements. Pathogen/ROS related genes from R. solani were also tested for expression patterns upon wheat infection. TmpL, a R. solani gene homologous to a gene associated with ROS regulation in Alternaria brassicicola, and OAH, a R. solani gene homologous to oxaloacetate acetylhydrolase which has been shown to produce oxalic acid in Sclerotinia sclerotiorum, were highly induced in R. solani when infecting wheat. We speculate that the interplay between the wheat and R. solani ROS generating proteins may be important for determining the outcome of the wheat/R. solani interaction