Exploring fungal RiPPs from the perspective of chemical ecology.

Since the initial detection, in 2007, of fungal ribosomally synthesised and post-translationally modified peptides (RiPPs), this group of natural products has undergone rapid expansion, with four separate classes now recognised: amatoxins/phallotoxins, borosins, dikaritins, and epichloëcyclins. Largely due to their historically anthropocentric employment in medicine and agriculture, novel fungal proteins and peptides are seldom investigated in relation to the fungus itself. Therefore, although the benefits these compounds confer to humans are often realised, their evolutionary advantage to the fungus, the reason for their continued production, is often obscure or ignored. This review sets out to summarise current knowledge on how these small peptide-derived products influence their producing species and surrounding biotic environment.

Identification of native endophytic Trichoderma spp. for investigation of in vitro antagonism towards Armillaria mellea using synthetic- and plant-based substrates.

AIMS: To isolate endophytic Trichoderma species and investigate the potential for biological control of the root rot pathogen Armillaria mellea. METHODS AND RESULTS: In all, 40 Trichoderma isolates were obtained from a range of host plants and identities were confirmed by ITS, rpb2 and tef1 sequence. When tested in dual culture assays for antagonism against A. mellea, Trichoderma isolates overgrew the A. mellea colonies within four days and by eight days 38 Trichoderma isolates significantly reduced A. mellea colony size. Armillaria mellea was unable to be recovered from five of eight co-cultivations tested, suggesting Trichoderma had killed the A. mellea in these cases. Pre-colonized hazel disks were used to determine what happens in a more heterogeneous situation with A. mellea and a refined set of eight Trichoderma isolates. Similar to plate-based assays, Trichoderma quickly covered A. mellea stopping any further growth and two Trichoderma isolates were able to eradicate A. mellea. CONCLUSIONS: Of the Trichoderma spp. tested, endophytic isolates of Trichoderma virens and T. hamatum offered the greatest antagonism towards A. mellea. Using pre-colonized hazel disks was of great importance for this work to demonstrate the fungal interactions in plant material. SIGNIFICANCE AND IMPACT OF THE STUDY: Controlling Armillaria root rot is difficult with chemical treatments, thus an environmentally benign and cost-effective alternative is required. This study highlights the prospect of biological control as an effective, environmentally friendly alternative to chemicals.

Weight bias internalization in a commercial weight management sample: prevalence and correlates.

OBJECTIVE: Weight bias internalization (WBI) is associated with poor weight-related health. The purpose of this study was to identify the prevalence and correlates of WBI in a large sample of adults in a commercial weight management programme. METHODS: WW (the new Weight Watchers) members participated in an online survey. Participants (N = 18,769) completed the 10-item Weight Bias Internalization Scale - Modified (WBIS-M) and the Weight Self-Stigma Questionnaire (WSSQ). Participants reported details about weight-stigmatizing experiences, including the onset, frequency and distress, and interpersonal sources of weight stigma. Participants self-reported their demographics, weight history, and height and weight (to compute body mass index [BMI]). RESULTS: Weight bias internalization was relatively high compared with the general population (mean WBIS-M score = 4.3 ± 1.4; mean WSSQ total score = 35.2 ± 9.7). WBI was higher among participants who were female, younger and had higher BMIs (p < 0.001) and lower among those who were Black and were widowed or had a romantic partner (p < 0.001). Onset of weight stigma in childhood and young adulthood, and recent distress due to weight stigma, predicted higher WBI. Extended family and school sources of weight stigma had weaker associations with WBI than did other interpersonal sources. CONCLUSIONS: Weight bias was internalized by a significant proportion of adults enrolled in a commercial weight management programme. A phenotype of WBI includes demographic characteristics and the timing and sources of weight stigma.

Strategies for the Construction of Cassava Brown Streak Disease Viral Infectious Clones.

Cassava brown streak disease (CBSD) has major impacts on yield and quality of the tuberous roots of cassava in Eastern and Central Arica. At least two Potyviridae species cause the disease: Cassava brown streak virus (CBSV) and Ugandan cassava brown streak virus (UCBSV). Cloned viral genome sequences known as infectious clones (ICs) have been important in the study of other viruses, both as a means of standardising infectious material and characterising viral gene function. IC construction is often technically challenging for Potyviridae due to sequence instability in E. coli. Here, we evaluate three methods for the construction of infectious clones for CBSD. Whilst a simple IC for in vitro transcription was made for UCBSV isolate 'Kikombe', such an approach failed to deliver full-length clones for CBSV isolates 'Nampula' or 'Tanza', necessitating more complex approaches for their construction. The ICs successfully generated symptomatic infection in the model host N. benthamiana and in the natural host cassava. This shows that whilst generating ICs for CBSV is still a technical challenge, a structured approach, evaluating both in vitro and in planta transcription systems should successfully deliver ICs, allowing further study into the symptomology and virulence factors in this important disease complex.

Weight loss and frequency of body-weight self-monitoring in an online commercial weight management program with and without a cellular-connected 'smart' scale: a randomized pilot study.

Objective: Evaluate the effects of an online commercial weight management program, with and without provision of a 'smart' scale with instructions to weigh daily and weekly tailored feedback, on weight loss and the frequency of body-weight self-monitoring. Methods: Participants (N = 92; body mass index 27-40 kg/m2) were randomized to 6 months of no-cost access to the Weight Watchers Online (WWO) platform alone, or enhanced with a cellular-connected 'smart' scale, instructions to weigh daily and weekly pre-scripted email feedback (Weight Watchers Online Enhanced [WWO-E]). The number of days that weight was self-monitored (via 'smart' scale in WWO-E and manually in WWO) was recorded automatically across the 6-month trial. Objective weight was measured at baseline, 3 and 6 months. Results: While both groups achieved statistically significant weight loss, mean ± standard error weight loss did not differ between WWO-E and WWO at 3 months (5.1 ± 0.6 kg vs. 4.0 ± 0.7 kg, respectively; p = 0.257) or 6 months (5.3 ± 0.6 kg vs. 3.9 ± 0.7 kg, respectively; p = 0.116). However, a greater proportion of WWO-E lost ≥5% of initial body weight at 3 months (52.2% vs. 28.3%; p = 0.033), but not 6 months (43.5% vs. 30.4%; p = 0.280), compared with WWO. Mean ± standard deviation days with self-monitored weight was higher in WWO-E (80.5 ± 5.6; 44.7% of days) than WWO (12.0 ± 1.0; 6.7% of days; p < 0.001) across the 6-month study period. Conclusions: This is the first study to show that provision of a 'smart' scale with weekly tailored feedback substantially increased the frequency of self-weighing and the proportion of participants achieving an initial clinically significant ≥5% weight loss (52% vs. 28%) in an online commercial weight management program. Both WWO and WWO-E produced significant weight loss over 6 months. While mean weight losses were slightly greater in the enhanced group, the difference was not statistically significant in this small sample. This study provides support for the clinical utility of online commercial weight management programs and the potential for supporting technology such as 'smart' scales to improve adherence to body-weight self-monitoring and clinical outcomes.

The effects of surface structure mutations in Arabidopsis thaliana on the polarization of reflections from virus-infected leaves.

The way in which light is polarized when reflected from leaves can be affected by infection with plant viruses. This has the potential to influence viral transmission by insect vectors due to altered visual attractiveness of infected plants. The optical and topological properties of cuticular waxes and trichomes are important determinants of how light is polarized upon reflection. Changes in expression of genes involved in the formation of surface structures have also been reported following viral infection. This paper investigates the role of altered surface structures in virus-induced changes to polarization reflection from leaves. The percentage polarization of reflections from Arabidopsis thaliana cer5, cer6 and cer8 wax synthesis mutants, and the gl1 leaf hair mutant, was compared to those from wild-type (WT) leaves. The cer5 mutant leaves were less polarizing than WT on the adaxial and abaxial surfaces; gl1 leaves were more polarizing than WT on the adaxial surfaces. The cer6 and cer8 mutations did not significantly affect polarization reflection. The impacts of Turnip vein clearing virus (TVCV) infection on the polarization of reflected light were significantly affected by cer5 mutation, with the reflections from cer5 mutants being higher than those from WT leaves, suggesting that changes in CER5 expression following infection could influence the polarization of the reflections. There was, however, no significant effect of the gl1 mutation on polarization following TVCV infection. The cer5 and gl1 mutations did not affect the changes in polarization following Cucumber mosaic virus (CMV) infection. The accumulation of TVCV and CMV did not differ significantly between mutant and WT leaves, suggesting that altered expression of surface structure genes does not significantly affect viral titres, raising the possibility that if such regulatory changes have any adaptive value it may possibly be through impacts on viral transmission.

Initial weight loss goals: have they changed and do they matter?

OBJECTIVE: Nearly 20 years ago, participants in behavioural weight loss programmes reported goals that greatly exceeded the amount of weight typically produced by these programmes. Whether having unrealistic weight loss goals impacts weight loss or attrition is unclear. The intent of the current study was to revisit current weight loss goals and examine whether goals impact outcomes. METHODS: Adults (N = 308, BMI = 33.7 ± 4.2 kg/m2) participated in a 12-month behavioural weight management programme and completed questionnaires about their goals. RESULTS: Participants' weight loss goal was 19.8 ± 7.9% of their body weight, and 90.4% selected a goal ≥10%. Weight goals were not associated with weight loss at 3 (p = 0.75) or 12 months (p = 0.47), or from 3 to 12 months (p = 0.55). Weight loss goals were not related to attrition at 3 (p = 0.91) or 12 months (p = 0.86). Participants believed that weight reduction would positively impact their health and psychosocial functioning. CONCLUSION: Weight loss goals have decreased, but still greatly exceed what can be expected by most. Unrealistic goals, however, had no impact on weight loss or attrition. These results question the utility of counseling people with obesity to set more realistic weight loss goals, which is typically practiced in behavioural weight management.

Effect of liraglutide 3.0 mg in individuals with obesity and moderate or severe obstructive sleep apnea: the SCALE Sleep Apnea randomized clinical trial.

BACKGROUND: Obesity is strongly associated with prevalence of obstructive sleep apnea (OSA), and weight loss has been shown to reduce disease severity. OBJECTIVE: To investigate whether liraglutide 3.0 mg reduces OSA severity compared with placebo using the primary end point of change in apnea-hypopnea index (AHI) after 32 weeks. Liraglutide's weight loss efficacy was also examined. SUBJECTS/METHODS: In this randomized, double-blind trial, non-diabetic participants with obesity who had moderate (AHI 15-29.9 events h(-1)) or severe (AHI ⩾30 events h(-1)) OSA and were unwilling/unable to use continuous positive airway pressure therapy were randomized for 32 weeks to liraglutide 3.0 mg (n=180) or placebo (n=179), both as adjunct to diet (500 kcal day(-1) deficit) and exercise. Baseline characteristics were similar between groups (mean age 48.5 years, males 71.9%, AHI 49.2 events h(-1), severe OSA 67.1%, body weight 117.6 kg, body mass index 39.1 kg m(-2), prediabetes 63.2%, HbA1c 5.7%). RESULTS: After 32 weeks, the mean reduction in AHI was greater with liraglutide than with placebo (-12.2 vs -6.1 events h(-1), estimated treatment difference: -6.1 events h(-1) (95% confidence interval (CI), -11.0 to -1.2), P=0.0150). Liraglutide produced greater mean percentage weight loss compared with placebo (-5.7% vs -1.6%, estimated treatment difference: -4.2% (95% CI, -5.2 to -3.1%), P<0.0001). A statistically significant association between the degree of weight loss and improvement in OSA end points (P<0.01, all) was demonstrated post hoc. Greater reductions in glycated hemoglobin (HbA1c) and systolic blood pressure (SBP) were seen with liraglutide versus placebo (both P<0.001). The safety profile of liraglutide 3.0 mg was similar to that seen with doses ⩽1.8 mg. CONCLUSIONS: As an adjunct to diet and exercise, liraglutide 3.0 mg was generally well tolerated and produced significantly greater reductions than placebo in AHI, body weight, SBP and HbA1c in participants with obesity and moderate/severe OSA. The results confirm that weight loss improves OSA-related parameters.

The good, the bad and the tasty: The many roles of mushrooms.

Fungi are often inconspicuous in nature and this means it is all too easy to overlook their importance. Often referred to as the "Forgotten Kingdom", fungi are key components of life on this planet. The phylum Basidiomycota, considered to contain the most complex and evolutionarily advanced members of this Kingdom, includes some of the most iconic fungal species such as the gilled mushrooms, puffballs and bracket fungi. Basidiomycetes inhabit a wide range of ecological niches, carrying out vital ecosystem roles, particularly in carbon cycling and as symbiotic partners with a range of other organisms. Specifically in the context of human use, the basidiomycetes are a highly valuable food source and are increasingly medicinally important. In this review, seven main categories, or 'roles', for basidiomycetes have been suggested by the authors: as model species, edible species, toxic species, medicinal basidiomycetes, symbionts, decomposers and pathogens, and two species have been chosen as representatives of each category. Although this is in no way an exhaustive discussion of the importance of basidiomycetes, this review aims to give a broad overview of the importance of these organisms, exploring the various ways they can be exploited to the benefit of human society.

"Snacks are not food". Low-income, urban mothers' perceptions of feeding snacks to their preschool-aged children.

Snacking has become more frequent among US preschool-aged children in recent decades and represents a significant proportion of daily energy intake. Social influences on snacking among children, however, are not well understood. This qualitative research described low-income, urban mothers' perceptions of feeding snacks to their preschool-aged children using data from 7 focus groups with 32 participants. Focus group transcripts were analyzed using a constant comparative method to identify themes. Mothers described snacks as involving less preparation, balance, and sustenance than meals (Theme 1). Mothers also made reference to some snacks as not being "real food" (Theme 2). At the same time, snacks had significant hedonic value as reflected in mothers' enjoyment of those foods (Theme 3), the effectiveness of snacks to manage children's behavior (Theme 4), and the variety of restrictions that mothers placed on children's access to snacks, such as locking cabinets, offering small servings, and reducing the number of snacks in sight (Theme 5). Two overarching themes highlighted distinctions mothers made in feeding children snacks vs. meals as well as the powerful hedonic appeal of snacks for both mother and child. These observations suggest that low-income, urban mothers of preschool-aged children may perceive snacks as serving a more important role in managing children's behavior than in providing nutrition. Child feeding interventions should address non-food related ways of managing children's behavior as well as encouraging caregivers to see snacks as structured opportunities for nutrition and connecting with their children.

Cardio-metabolic risk screening among adolescents: understanding the utility of body mass index, waist circumference and waist to height ratio.

BACKGROUND: Few studies have assessed how well body mass index (BMI), waist circumference (WC), or waist to height ratio (WtHR) perform in identifying cardio-metabolic risk among youth. OBJECTIVE: The objective of this study was to evaluate the utility of BMI and WC percentiles and WtHR to distinguish adolescents with and without cardio-metabolic risk. METHODS: A cross-sectional analysis of data from 6097 adolescents aged 10-13 years who participated in the HEALTHY study was conducted. Receiver operating characteristic curves determined the discriminatory ability of BMI and WC percentiles and WtHR. RESULTS: The discriminatory ability of BMI percentile was good (area under the curve [AUC] ≥ 0.80) for elevated insulin and clustering of ≥3 risk factors, with optimal cut-points of 96 and 95, respectively. BMI percentile performed poor to fair (AUC = 0.57-0.75) in identifying youth with the majority of individual risk factors examined (elevated glucose, total cholesterol, low-density lipoprotein, blood pressure, triglycerides and high-density lipoprotein). WC percentile and WtHR performed similarly to BMI percentile. CONCLUSIONS: The current definition of obesity among US children performs well at identifying adolescents with elevated insulin and a clustering of ≥3 cardio-metabolic risk factors. Evidence does not support WC percentile or WtHR as superior screening tools compared with BMI percentile for identifying cardio-metabolic risk.

18-month outcomes of a community-based treatment for childhood obesity.

BACKGROUND: Few studies have evaluated community-based interventions for childhood obesity and even fewer provide efficacy data 1 year later. OBJECTIVES: This study assessed changes in relative weight 1 year after a 6-month treatment for childhood obesity. METHODS: Participants were 155 overweight/obese children/adolescents and their caregivers. The primary outcome was change in child percent over body mass index (BMI) from 6 to 18 months. The primary outcome was also assessed from 0 to 18 months and changes in secondary outcomes (BMI z-score, guardian weight, health-related quality of life [HRQoL]) were examined from 6-18 to 0-18 months. RESULTS: From 6 to 18 months, there were no significant changes in any outcome. From 0 to 18 months, there were improvements in BMI z-score (P < 0.001), HRQoL (P < 0.001) and guardian weight (P = 0.02). CONCLUSIONS: Changes in relative weight and HRQoL observed after 6 months persisted 1 year later. The programme produced reductions in BMI z-score and obesity prevalence but not in percent over BMI from 0 to 18 months.

Similar effects on cognitive performance during high- and low-carbohydrate obesity treatment.

OBJECTIVE: Low-carbohydrate (L-CHO) diets are often used for weight loss but their effects on cognitive function are not well understood. The present study compared the effects of a L-CHO and high-carbohydrate (H-CHO) weight-loss diet on cognitive function adults. DESIGN: PARTICIPANTS were randomized to either a L-CHO (n=22) or H-CHO (n=25) weight-loss diet. Cognitive function was evaluated by four computerized cognitive tasks (Stroop Task, Continuous Performance Task, Word Recall and Wisconsin Card Sorting Task) presented in random order before and at 1, 4, 12 and 24 weeks after the initiation of the L-CHO or H-CHO diet. PARTICIPANTS: Forty-seven adults (25 males) with a mean±s.d. age of 47.4±8.7 years and body mass index of 35.3±3.4 kg m(-2). RESULTS: There were no significant differences in weight loss between groups at any time point. There were significant improvements on color Stroop task accuracy over time in both diet groups (P<0.05), but there were no differences in performance between groups on this or any other cognitive task at any time period. CONCLUSION: These findings suggest that weight loss has neither a positive nor a negative effect on cognitive function and that L-CHO and H-CHO weight-loss diets have similar effects on cognitive performance.

A randomized comparison of a commercially available portion-controlled weight-loss intervention with a diabetes self-management education program.

OBJECTIVE: This study examined the efficacy of a commercially available, portion-controlled diet (PCD) on body weight and HbA1c over 6 months in obese patients with type 2 diabetes. RESEARCH DESIGN AND METHODS: One-hundred participants with a mean±s.d. age of 55.6±10.6 year, body weight of 102.9±18.4 kg and HbA1c of 7.7±1.3% were randomly assigned to a 9-session group lifestyle intervention that included a PCD or to a 9-session group program of diabetes self-management education (DSME). Participants in the two groups were prescribed the same goals for energy intake (1250-1550 kcal per day) and physical activity (200 min per week). RESULTS: While both groups produced significant improvements in weight and HbA1c after 6 months of treatment, PCD participants lost 7.3 kg [95% confidence interval (CI): -5.8 to -8.8 kg], compared with 2.2 kg (95% CI: -0.7 to -3.7 kg) in the DSME group (P<0.0001). Significantly more PCD than DSME participants lost 5% of initial weight (54.0% vs 14.0%, P<0.0001) and 10% (26.0% vs 6.0%, P<0.0001). HbA1c declined by 0.7% (95% CI: -0.4 to -1.0%) in the PCD group, compared with 0.4% (95% CI: -0.1 to -0.7%) in DSME (P<0.026). Across both groups, larger weight losses were associated with greater reductions in HbA1c (r=0.52, P<0.0001). CONCLUSIONS: These findings demonstrate that a commercially available portion-controlled meal plan can induce clinically meaningful improvements in weight and glycemic control in obese individuals with type 2 diabetes. These data have implications for the management of obesity in primary care, as now provided by the Centers for Medicare and Medicaid Services.

Factor structure of the Emotional Eating Scale in overweight and obese adults seeking treatment.

The purpose of this study was to examine the factor structure and anthropometric correlates of the Emotional Eating Scale in overweight and obese adults presenting for weight loss. Participants were 217 men and women with a mean body-mass index of 33.1 (±3.4) kg/m². Results indicated a four factor structure: depression, anger, anxiety, and somatic arousal. These factors demonstrated strong internal consistency, and together accounted for approximately 60% of the total variance. Women had significantly higher depression and total scores than did men. There were no significant correlations between the Emotional Eating Scale scores and anthropometric measures. This work begins to add to the literature base regarding the applicability of the original design of the Emotional Eating Scale for samples consisting of men and African Americans.

RNA-mediated gene silencing of superoxide dismutase (bcsod1) in Botrytis cinerea.

Gene silencing is a powerful tool utilized for identification of gene function and analysis in plants, animals, and fungi. Here, we report the silencing of superoxide dismutase (bcsod1) in Botrytis cinerea through sense and antisense-mediated silencing mechanisms. Because superoxide dismutase (SOD) is a virulence factor, transformants were tested for phenotypic silencing in vitro and reduction in pathogenicity in planta. Plate-based assays with and without paraquat were performed to screen initial silencing efficiency, and a subset of transformants was used for in planta studies of virulence. Transformants exhibiting strongly decreased transcripts levels were recovered with both constructs but none of those exhibited a reduction in virulence in planta. Our investigations may help optimize a high-throughput gene silencing system useful for identifying potential gene targets for future fungal control.

Single-step RT real-time PCR for sensitive detection and discrimination of Potato virus Y isolates.

Potato virus Y (PVY) has a worldwide distribution and infects several economically important crops from the Solanaceae family. The emergence and spread of the PVYNTN strain, which is the causative agent of potato tuber necrotic ringspot disease (PTNRD), has lead to large economic losses and highlighted the need for accurate discrimination of the different PVY strains. Detection and differentiation of PVY isolates is mainly based on a combination of ELISA, RT-PCR and bioassays; however, PVYNTN isolates are particularly difficult to differentiate from standard PVYN without the use of time-consuming bioassays. A strong correlation has been identified previously between the ability to induce PTNRD and the presence of a recombination point in the virus coat protein. An RT real-time PCR assay has been developed to enable detection of isolates with the recombination point, therefore, enabling rapid differentiation between potentially tuber necrotic PVYNTN isolates and standard PVYN isolates. The assay is also able to detect the presence of PVYO isolates. To aid with routine testing, immuno-capture and post-ELISA virus release were introduced; when coupled with RT real-time PCR the sensitivity of the assays were up to seven orders of magnitude higher than ELISA. The assay was shown to be a suitable method for rapid large-scale diagnostic testing of PVY in different types of plant material including tubers, and specific screening for potentially tuber necrotic recombinant isolates.

The role of patients' expectations and goals in the behavioral and pharmacological treatment of obesity.

OBJECTIVE: To investigate weight loss expectations and goals among obese treatment seekers and to examine the relationships of these expectations and goals to treatment outcomes. METHOD: Participants were 180 obese men and women (age 43.8+/-10.1 years; body mass index 37.6+/-4.2 kg/m(2)) who received one of four-year-long treatments that combined behavioral and pharmacological methods. Before treatment, they reported the amount of weight they realistically expected to lose after 4, 12, 26 and 52 weeks of treatment, as well as their ultimate weight loss goals. Expectations and goals were compared across treatment groups and examined in relation to previous weight loss efforts, weight loss and regain in treatment, attrition, satisfaction with treatment and mood. RESULTS: Participants in all treatment groups expected reductions at week 52 that were significantly greater than the 5-15% of initial weight they were told was realistic and significantly more than they had ever lost before. Weight loss expectations were unrelated to achieved weight loss in all groups but one, in which greater expectations were associated with greater losses. Failure to meet weight loss expectations for the first 26 weeks of treatment was related to lower satisfaction ratings, but was not related to weight regain or attrition over the next 26 weeks. Symptoms of depression were reduced from baseline, regardless of whether participants achieved or failed to achieve their expected weight losses. CONCLUSION: Across groups, we observed no negative consequences of having (and failing to meet) unrealistic expectations for weight loss.

Investigations into the taxonomy of the mushroom pathogen Verticillium fungicola and its relatives based on sequence analysis of nitrate reductase and ITS regions.

The full sequence of the nitrate reductase gene was obtained from a type isolate of Verticilliumfungicola var. fungicola and used for phylogenetic analysis against other ascomycete fungi. Sequencing obtained 2749 bp of coding region, 668 bp of 5' flanking sequence and 731 bp of 3' flanking sequence. In silico analysis indicated that the coding region contains a single intron and translates into an 893 amino acid protein, with BLAST analysis identifying five conserved nitrate reductase domains within the protein. The 5' flanking sequence contains numerous conserved sites putatively involved in binding nitrogen regulatory proteins, indicating that the regulation of the gene is likely to be subject to the same regulation as that of model fungi such as Aspergillus nidulans. The central portion of this gene was amplified and sequenced from a number of V.fungicola isolates and related fungi and the resulting phylogenies compared to those obtained from analysis of the rDNA internal transcribed spacer regions for these fungi. Both nitrate reductase and ITS analyses provide additional evidence that reinforces previous findings that suggest the mushroom pathogenic Verticillium species are more related to other chitinolytic fungi such as the insect pathogens Verticillium lecanii and Beauveria bassiana than to the plant pathogenic Verticillia.

Evaluation of agrobacterium-mediated transformation of Agaricus bisporus using a range of promoters linked to hygromycin resistance.

There is interest in establishing genetic modification technologies for the cultivated mushroom Agaricus bisporus, both for improved crop characteristics and for molecular pharming. For these methods to be successful, it is necessary to establish a set of transformation systems that include robust and reliable vectors for gene manipulation. In this article, we report the evaluation of a series of promoters for driving expression of the Escherichia coli hph gene encoding hygromycin phosphotransferase. This was achieved using the Aspergillus nidulans gpdA and the A. bisporus gpdII and trp2 promoters. The Coprinus cinereus beta-tubulin promoter gave contrasting results depending on the size of promoter used, with a 393-bp region being effective, whereas the longer 453-bp fragment failed to yield any hygromycin-resistant transformants. The C. cinereus trp1 and the A. bisporus lcc1 promoters both failed to yield transformants. We also show that transformation efficiency may be improved by careful selection of both appropriate Agrobacterium strains, with AGL-1 yielding more than LBA1126 and by the choice of the binary vectors used to mobilize the DNA, with pCAMBIA vectors appearing to be more efficient than either pBIN19- or pGREEN-based systems.