Overexpression of a cytochrome P450 and a UDP-glycosyltransferase is associated with imidacloprid resistance in the Colorado potato beetle, Leptinotarsa decemlineata.

Current control of insect pests relies on chemical insecticides, however, insecticide resistance development by pests is a growing concern in pest management. The main mechanisms for insecticide resistance typically involve elevated activity of detoxifying enzymes and xenobiotic transporters that break-down and excrete insecticide molecules. In this study, we investigated the molecular mechanisms of imidacloprid resistance in the Colorado potato beetle, Leptinotarsa decemlineata (Say) (Coleoptera: Chrysomelidae), an insect pest notorious for its capacity to develop insecticide resistance rapidly. We compared the transcriptome profiles of imidacloprid-resistant and sensitive beetle strains and identified 102 differentially expressed transcripts encoding detoxifying enzymes and xenobiotic transporters. Of these, 74 were up-regulated and 28 were down-regulated in the resistant strain. We then used RNA interference to knock down the transcript levels of seven up-regulated genes in the resistant beetles. Ingestion of double-stranded RNA successfully knocked down the expression of the genes for three cytochrome P450s (CYP6BQ15, CYP4Q3 and CYP4Q7), one ATP binding cassette (ABC) transporter (ABC-G), one esterase (EST1), and two UDP-glycosyltransferases (UGT1 and UGT2). Further, we demonstrated that silencing of CYP4Q3 and UGT2 significantly increased susceptibility of resistant beetles to imidacloprid, indicating that overexpression of these two genes contributes to imidacloprid resistance in this resistant strain.

Transcriptomic evidence for the control of soybean root isoflavonoid content by regulation of overlapping phenylpropanoid pathways.

BACKGROUND: Isoflavonoids are a class of specialized metabolites found predominantly in legumes. They play a role in signaling for symbiosis with nitrogen-fixing bacteria and inhibiting pathogen infection. RESULTS: A transcriptomic approach using soybean cultivars with high (Conrad and AC Colombe) and low (AC Glengarry and Pagoda) root isoflavonoid content was used to find elements that underlie this variation. Two genes, encoding the flavonoid-metabolizing enzymes, flavonoid 3'-hydroxylase (GmF3'H) and dihydroflavonol 4-reductase (GmDFR), had lower expression levels in high isoflavonoid cultivars. These enzymes compete with isoflavonoid biosynthetic enzymes for the important branch-point substrate naringenin and its derivatives. Differentially expressed genes, between the two sets of cultivars, encode transcription factors, transporters and enzymatic families of interest, such as oxidoreductases, hydrolases and transferases. In addition, genes annotated with stress and disease response were upregulated in high isoflavonoid cultivars. CONCLUSIONS: Coordinated regulation of genes involved in flavonoid metabolism could redirect flux into the isoflavonoid branch of the phenylpropanoid pathway, by reducing competition for the flavanone substrate. These candidate genes could help identify mechanisms to overcome the endogenous bottleneck to isoflavonoid production, facilitate biosynthesis in heterologous systems, and enhance crop resistance against pathogenic infections.

Strain Specific Factors Control Effector Gene Silencing in Phytophthora sojae.

The Phytophthora sojae avirulence gene Avr3a encodes an effector that is capable of triggering immunity on soybean plants carrying the resistance gene Rps3a. P. sojae strains that express Avr3a are avirulent to Rps3a plants, while strains that do not are virulent. To study the inheritance of Avr3a expression and virulence towards Rps3a, genetic crosses and self-fertilizations were performed. A cross between P. sojae strains ACR10 X P7076 causes transgenerational gene silencing of Avr3a allele, and this effect is meiotically stable up to the F5 generation. However, test-crosses of F1 progeny (ACR10 X P7076) with strain P6497 result in the release of silencing of Avr3a. Expression of Avr3a in the progeny is variable and correlates with the phenotypic penetrance of the avirulence trait. The F1 progeny from a direct cross of P6497 X ACR10 segregate for inheritance for Avr3a expression, a result that could not be explained by parental imprinting or heterozygosity. Analysis of small RNA arising from the Avr3a gene sequence in the parental strains and hybrid progeny suggests that the presence of small RNA is necessary but not sufficient for gene silencing. Overall, we conclude that inheritance of the Avr3a gene silenced phenotype relies on factors that are variable among P. sojae strains.

Genome-wide analysis of Cyclophilin gene family in soybean (Glycine max).

BACKGROUND: Cyclophilins (CYPs) belong to the immunophilin superfamily, and have peptidyl-prolyl cis-trans isomerase (PPIase) activity. PPIase catalyzes cis- and trans-rotamer interconversion of the peptidyl-prolyl amide bond of peptides, a rate-limiting step in protein folding. Studies have demonstrated the importance of many PPIases in plant biology, but no genome-wide analysis of the CYP gene family has been conducted for a legume species. RESULTS: Here we performed a comprehensive database survey and identified a total of 62 CYP genes, located on 18 different chromosomes in the soybean genome (GmCYP1 to GmCYP62), of which 10 are multi- and 52 are single-domain proteins. Most of the predicted GmCYPs clustered together in pairs, reflecting the ancient genome duplication event. Analysis of gene structure revealed the presence of introns in protein-coding regions as well as in 5' and 3' untranslated regions, and that their size, abundance and distribution varied within the gene family. Expression analysis of GmCYP genes in soybean tissues displayed their differential tissue specific expression patterns. CONCLUSIONS: Overall, we have identified 62 CYP genes in the soybean genome, the largest CYP gene family known to date. This is the first genome-wide study of the CYP gene family of a legume species. The expansion of GmCYP genes in soybean, and their distribution pattern on the chromosomes strongly suggest genome-wide segmental and tandem duplications.

Crowdsourcing genomic analyses of ash and ash dieback - power to the people.

Ash dieback is a devastating fungal disease of ash trees that has swept across Europe and recently reached the UK. This emergent pathogen has received little study in the past and its effect threatens to overwhelm the ash population. In response to this we have produced some initial genomics datasets and taken the unusual step of releasing them to the scientific community for analysis without first performing our own. In this manner we hope to 'crowdsource' analyses and bring the expertise of the community to bear on this problem as quickly as possible. Our data has been released through our website at oadb.tsl.ac.uk and a public GitHub repository.

Management of isolated orbital floor blow-out fractures: a survey of Australian and New Zealand oral and maxillofacial surgeons.

BACKGROUND: This is the first report of involvement of Australian and New Zealand oral and maxillofacial surgeons in the management of isolated orbital floor blow-out fractures and was conducted to obtain comparisons with the results from a recent similar survey of British oral and maxillofacial surgeons. METHODS: A questionnaire survey was sent to all 113 practising members of the Australian and New Zealand Association of Oral and Maxillofacial Surgeons in April 2002 with a second mailout 1 month later. RESULTS: Sixty-nine per cent of the respondents were referred isolated orbital floor blow-out fractures for management, and just over half of these respondents estimated that 50% or more of the cases went to surgery. The materials most commonly used in orbital floor reconstruction were resorbable membrane for small defects and autologous bone for large defects. CONCLUSION: As in Britain, management of isolated orbital floor blow-out fractures comprises part of the surgical spectrum for many oral and maxillofacial surgeons in Australia and New Zealand. The management protocol was observed to be very similar between the two groups.

Cranial variation in the Marquesas Islands.

The Marquesas Islands have traditionally been divided into a northwestern and a southeastern group, a division which reflects language dialect differences. Additionally, archaeological studies have also suggested that differences in material culture existed between the northwestern and southeastern islands. This study examines Marquesan cranial discrete and metric traits to evaluate the level of intra-archipelago heterogeneity, and to determine if a northwest/southeast division is evident cranially. The data consist of 28 cranial discrete traits and 49 craniofacial measurements of prehistoric Marquesans. Male and female data are pooled for discrete trait and metric data, following a Z-score standardization technique adjustment. The data represent three island samples: Nuku Hiva (northwest), Fatuiva (southeast), and a combined Tahuata/Hiva 'Oa (southeast). Of the 28 discrete traits, 16 are utilized in a mean measure of divergence analysis that provides scores of 0.259 for Fatuiva-Tahuata/Hiva 'Oa, 1.850 for Nuku Hiva-Fatuiva, and 1.491 for Nuku Hiva-Tahuata/Hiva 'Oa. Of the 49 craniofacial measurements, 46 are utilized in RMET/NORM analyses, providing unbiased D(2) values of 0.0433 for Fatuiva-Tahuata/Hiva 'Oa, 0.1328 for Nuku Hiva-Fatuiva, and 0.0813 for Nuku Hiva-Tahuata/Hiva 'Oa. The islands of the southeastern group are closer to each other than either was to the island of the northwestern group. When a sample from 'Ua Huka is included in the craniometric analysis, the unbiased D(2) values of 0.0829, 0.1175, and 0.0431 are calculated for 'Ua Huka and Nuku Hiva, and Fatuiva and Tahuata/Hiva 'Oa pairings, respectively, indicating a close similarity of 'Ua Huka to the southeastern islands. Mean measure of divergence analysis of cranial discrete traits as well as RMET/NORM analyses of craniometric variables reveal that differences exist between the islands of the northwestern and southeastern Marquesas Islands. These results support previous research that documented linguistic and cultural differences between these regions of the archipelago. However, the results indicate that 'Ua Huka, an island traditionally included in the northwestern Marquesas Islands, has an affinity to the southeastern Marquesas Islands, possibly due to its pivotal position as a waypoint in the Marquesas Island interaction sphere.