Quality assessment of Urochloa (syn. Brachiaria) seeds produced in Cameroon.

Urochloa (syn. Brachiaria) is the most popular fodder of livestock farmers in Cameroon for hay and seed productions. Farmers in Cameroon have been producing Brachiaria seeds for decades for own uses and surplus are sold to neighbours, and to traders from Cameroon and neighbouring countries. However, there is no information available about qualities of these seeds. Fifteen Urochloa seeds samples were collected from farmers and/or government stations in five regions (Adamaoua, East, North, North West, and West) and analysed for major seed quality parameters along with seeds of improved Urochloa cultivar Basilisk imported from Brazil as a check. Study showed significant differences among treatments for various seed quality parameters tested (P < 0.0001). The highest thousand grains weight was recorded in Basilisk (5.685 g), followed by W12 (3.555 g), A05 (3.153 g) and N01 (2.655 g). Caryopsis number and caryopsis weight were highest in Basilisk followed by E09, A06, and W12. Of three conditions tested for seed germination, mean germination was the highest in greenhouse (7.39%) where Basilisk had the highest germination (25.5%) followed by N01 (18.50%), A05 (14.50%) and W12 (12.75%). The seed viability ranged from 18% (E09) to 81% (N01), and there were a positive and highly significant relationships between seed germination and viability traits (r = 0.883; P < 0.0001). This study showed a marked difference in seed quality parameters of Urochloa grass seeds produced in Cameroon, and the potential of developing Urochloa grass seed business in the Northern, Adamaoua and Western regions of Cameroon.

Comparative TMT Proteomic Analysis Unveils Unique Insights into Helicoverpa armigera (Hübner) Resistance in Cajanus scarabaeoides (L.) Thouars.

Pigeonpea [Cajanus cajan (L.) Millspaugh] is an economically important legume playing a crucial role in the semi-arid tropics. Pigeonpea is susceptible to Helicoverpa armigera (Hübner), which causes devastating yield losses. This pest is developing resistance to many commercially available insecticides. Therefore, crop wild relatives of pigeonpea, are being considered as potential sources of genes to expand the genetic base of cultivated pigeonpea to improve traits such as host plant resistance to pests and pathogens. Quantitative proteomic analysis was conducted using the tandem mass tag platform to identify differentially abundant proteins between IBS 3471 and ICPL 87 tolerant accession and susceptible variety to H. armigera, respectively. Leaf proteome were analysed at the vegetative and flowering/podding growth stages. H. armigera tolerance in IBS 3471 appeared to be related to enhanced defence responses, such as changes in secondary metabolite precursors, antioxidants, and the phenylpropanoid pathway. The development of larvae fed on an artificial diet with IBS 3471 lyophilised leaves showed similar inhibition with those fed on an artificial diet with quercetin concentrations with 32 mg/25 g of artificial diet. DAB staining (3,3'-diaminobenzidine) revealed a rapid accumulation of reactive oxygen species in IBS 3471. We conclude that IBS 3471 is an ideal candidate for improving the genetic base of cultivated pigeonpea, including traits for host plant resistance.

Processing methods affect phytochemical contents in products prepared from orange-fleshed sweetpotato leaves and roots.

Phytochemicals enhance human health by acting antagonistically on incidences of cancer and other chronic diseases. They are considered indispensable in a variety of nutraceutical, pharmaceuticals, and medicinal and cosmetic applications. This study evaluated the effects of common processing methods on inherent phytochemical content in the roots and leaves of orange-fleshed sweetpotato (OFSP) varieties called Kabode and SPK031. Yellosp and Whitesp, which are local sweetpotato varieties, were also included as check for roots and leaves, respectively. The sweetpotato products prepared for phytochemical analysis were boiling roots and leaves, frying chips and crisps, baking bread (for roots only), and fermenting and dehydrating leaves. Phytochemicals that were assessed included vitamin C, total phenolics and flavonoids, tannins, phytates, and soluble oxalates. Results indicated that retention of vitamin C was highest in boiled roots (85%-95%), followed by fries (71%-94%) and crisps (44%-76%), whereas the least retention was in bread (4%-11%) and leaves (0%-27%). Total phenolics, flavonoids, and antioxidant activity in leaves significantly (p < .05) varied with the type of processing. Higher retention of these phytochemicals was observed in processed roots but was lowest in bread. Boiling retained more than 100% of all carotenoids, while fermenting and drying the leaves retained 58-62 and 22%-48%, respectively. Frying retained more than 100% of the ß-carotene in the roots, while boiling retained 96%-100%. All processing methods significantly (p < .05) reduced antinutrients in leaves and roots. Fermentation of leaves had higher reduction of oxalates, tannins, and phytates, while boiling had the least effect. It is concluded that traditional boiling enhances phytochemical retention in roots but degrades most of them in leaves.

Study of Ectopic Pregnancy at Tertiary Care Hospital in Province 1 of Nepal.

BACKGROUND: Ectopic pregnancy can be managed expectantly, medically or surgically. Laparoscopic surgery is considered gold standard. The aim of this study was to find incidence, risk factors, clinical presentation and mode of management of ectopic pregnancy. METHODS: Prospective descriptive study was conducted at Nobel Medical College and Teaching Hospital, Biratnagar. All the cases of diagnosed ectopic pregnancy were enrolled in the study. All the relevant data of the patients were recorded in pre-designed Performa. The collected data were entered in MS Excel and exported into SPSS 21 version for statistical analysis. RESULTS: The incidence of ectopic pregnancy was 0.8%. Most of the patients (35 ;( 58.3%) belonged to 20-30 age group. The classic triad of pain abdomen, amenorrhea and per vaginal bleeding was present in 41 (68.3%) cases. The most common risk factor associated was PID (39 ;-( 65%) followed by abortion 98 ;-( 13.3%). USG findings were suggestive of ruptured ectopic pregnancy, which was seen in 52(86.7%) cases and. 93.3% of the cases were managed surgically. The most common site of ectopic pregnancy was found to be ampulla 35(58.3%) followed by isthmus 11(18.3%). There was no mortality. CONCLUSIONS: Pelvic inflammatory disease was the commonest risk factor, laparotomy was the commonest modality of surgical management and ampulla was the commonest site of ectopic pregnancy. Early diagnosis, referral and treatment in the tertiary care center prevents morbidity and mortality related to ectopic pregnancy.

Complete Recovery of Renal Function among Obstetric Patients with Acute Kidney Injury at a Tertiary Care Hospital: A Descriptive Cross-sectional Study.

INTRODUCTION: Acute kidney injury  is a rare complication of pregnancy and is associated with high maternal morbidity and mortality. Obstetric factors associated with it are preeclampsia/eclampsia, sepsis, hemorrhage and dehydration. Here, we aim to find out the prevalence of complete recovery of renal function among obstetric patients with acute kidney injury. METHODS: This is a descriptive cross-sectional study conducted in a tertiary care hospital from 1st July 2020 to 30th June 2021 where obstetric patients who had developed acute kidney injury were included and followed till 6 weeks of diagnosis. Ethical approval was obtained from Institutional Review Committee of Nobel Medical College and Teaching Hospital (IRC- NMCTH 437/2020). The convenience sampling method was used. Data entry and analysis were done using Statistical Package for Social Sciences version 21. Point estimate at 95% Confidence Interval was calculated along with frequency and proportion for binary data. RESULTS: Out of total 66 obstetric patients with acute kidney injury, 45 (68.2%) (57-79.3 at 95% Confidence Interval) had complete recovery of renal function. Rate of renal function recovery in Stage 1, Stage 2 and Stage 3 acute kidney injury were 19 (90%), 19 (86%) and 7 (58%) respectively. The most common causes of acute kidney injury were Preeclampsia/eclampsia 18 (40%), sepsis 23 (28.8%) and hemorrhage 10 (22.2%). CONCLUSIONS:  The prevalence of complete recovery in obstetric patients with acute kidney injury was similar to findings from other studies done in similar settings.

Assessment of Fungal Contamination in Fish Feed from the Lake Victoria Basin, Uganda.

The emergence of commercial fish farming has stimulated the establishment of fish feed factories in Uganda. However, no information is available on the safety of the feed, mainly due to lack of mycotoxin testing facilities and weak regulatory systems. A study was carried out to examine fungal colonization and mycotoxin contamination in fish feed samples (n = 147) of different types collected from nine fish farms (n = 81) and seven fish feed factories (n = 66) in the Lake Victoria Basin (LVB). Fungi were isolated in potato dextrose agar, grouped into morphotypes and representative isolates from each morphotype were identified based on the internal transcribed spacer (ITS) region of ribosomal DNA sequences. Aflatoxin B1 (AFB1) and total fumonisin (combinations of B1, B2 and B3; hereinafter named fumonisin) levels in feed samples were determined by enzyme-linked immunosorbent assay (ELISA). A wide range of fungi, including toxigenic Aspergillus flavus and Fusarium verticillioides, were isolated from the fish feed samples. AFB1 was detected in 48% of the factory samples and in 63% of the farm samples, with toxin levels <40 and >400 µg/kg, respectively. Similarly, 31% of the factory samples and 29% of the farm samples had fumonisin contamination ranging between 0.1 and 4.06 mg/kg. Pellets and powder had higher mycotoxin contamination compared to other commercially available fish feed types. This study shows AFB1 as a potential fish feed safety issue in the LVB and suggests a need for more research on mycotoxin residues in fish fillets.

Phytochemicals in Leaves and Roots of Selected Kenyan Orange Fleshed Sweet Potato (OFSP) Varieties.

This study reports the inherent phytochemical contents in leaves and roots of nine sweet potato varieties from Kenya. Results indicated that vitamin C content varied significantly (P < 0.05) among the sweet potato varieties regardless of the plant part, leaves having significantly (P < 0.05) higher levels than in the roots. Total flavonoids and phenolic compounds differed significantly (P < 0.05) among varieties, higher values were found in leaves than in roots. Flavonoid contents in roots ranged from below detectable limits (Whitesp) to 25.8 mg CE/100 g (SPK031), while in leaves it ranged from 4097 to 7316 mg CE/100 g in SPK4 and Kenspot 5, respectively. Phenolic content was below detectable limits in the roots of whitesp but it was in substantial amounts in orange fleshed varieties. The ß-carotene content was significantly (P < 0.05) higher in leaves (16.43-34.47 mg/100 g dry weight) than in roots (not detected-11.1 mg/100 g dry weight). Total and phytic phosphorus were directly correlated with phytate contents in leaves and the roots. Tannins and soluble oxalates varied significantly (P < 0.05) with variety and plant part being higher in leaves. The current information is important for ration formulations and dietary recommendations utilizing sweet potato leaves and roots. Future studies on effects of processing methods on these phytochemicals are recommended.

Comparative Analysis Delineates the Transcriptional Resistance Mechanisms for Pod Borer Resistance in the Pigeonpea Wild Relative Cajanus scarabaeoides (L.) Thouars.

Insect pests pose a serious threat to global food production. Pod borer (Helicoverpa armigera (Hübner)) is one of the most destructive pests of leguminous crops. The use of host resistance has been an effective, environmentally friendly and sustainable approach for controlling several agricultural pests. The exploitation of natural variations in crop wild relatives could yield pest-resistant crop varieties. In this study, we used a high-throughput transcriptome profiling approach to investigate the defense mechanisms of susceptible cultivated and tolerant wild pigeonpea genotypes against H. armigera infestation. The wild genotype displayed elevated pest-induced gene expression, including the enhanced induction of phytohormone and calcium/calmodulin signaling, transcription factors, plant volatiles and secondary metabolite genes compared to the cultivated control. The biosynthetic and regulatory processes associated with flavonoids, terpenes and glucosinolate secondary metabolites showed higher accumulations in the wild genotype, suggesting the existence of distinct tolerance mechanisms. This study provides insights into the molecular mechanisms underlying insect resistance in the wild pigeonpea genotype. This information highlights the indispensable role of crop wild relatives as a source of crucial genetic resources that could be important in devising strategies for crop improvement with enhanced pest resistance.

Genetic Profiling of Aspergillus Isolates with Varying Aflatoxin Production Potential from Different Maize-Growing Regions of Kenya.

Highly toxigenic strains of Aspergillus flavus have been reported to frequently contaminate maize, causing fatal aflatoxin poisoning in Kenya. To gain insights into the environmental and genetic factors that influence toxigenicity, fungi (n = 218) that were culturally identified as A. flavus were isolated from maize grains samples (n = 120) from three regions of Kenya. The fungi were further characterized to confirm their identities using a PCR-sequence analysis of the internal transcribed spacer (ITS) region of rDNA which also revealed all of them to be A. flavus. A subset of 72 isolates representing ITS sequence-based phylogeny and the agroecological origin of maize samples was constituted for subsequent analysis. The analysis of partial calmodulin gene sequences showed that the subset consisted of A. flavus (87%) and Aspergillus minisclerotigenes (13%). No obvious association was detected between the presence of seven aflatoxin biosynthesis genes and fungal species or region. However, the presence of the aflD and aflS genes showed some association with aflatoxin production. The assessment of toxigenicity showed higher aflatoxin production potential in A. minisclerotigenes isolates. Given that A. minisclerotigenes were mainly observed in maize samples from Eastern Kenya, a known aflatoxin hotspot, we speculate that production of copious aflatoxin is an adaptative trait of this recently discovered species in the region.

Mycotoxin Contamination of Edible Non-Timber Forest Products in Cameroon.

The prevalence and concentrations of three major mycotoxins, total aflatoxin (AFs), fumonisin (F), and zearalenone (ZEN), were determined on seven edible non-timber forest products (ENTFP) in Cameroon. A total of 210 samples consiting of 30 samples from each ENTFP commodity was collected from farmers and local markets in three agroecological zones of Cameroon and analyzed for moisture content and mycotoxins. Mycotoxins were analyzed using commercial enzyme-linked immunosorbent assay (ELISA) kits and results were validated using the VICAM fluorometric method. The European Union regulation of mycotoxins for human consumption (N°1881/2006) was adopted as reference. The moisture content of samples varied from 5.0% to 22.6%. Aflatoxin contamination was detected in 84.3% samples and only 5.7% exceeded the legal limit (10 ppb). Similarly, 53% of samples were contaminated with fumonisin and 5% of samples exceeded the legal limit (1000 ppb). Zearalenone contamination was detected in 92% of samples and 21% of samples exceeded the legal limit (100 ppb). This is the first report on mycotoxin contamination of ENTFP in the Congo Basin forest. The findings of this study will form a basis for educating farmers and other stakeholders of ENTFP values chain on mycotoxins and mycotoxin mitigation measures to produce safe ENTFP for local and international markets.

The effect of different processing methods on nutrient and isoflavone content of soymilk obtained from six varieties of soybean grown in Rwanda.

Soymilk is rich in nutrients and isoflavones, and could greatly promote nutrition and health. However, this product is not widely accepted due to an objectionable beany flavor. Several methods involving heat treatment and soaking in basic solutions prior to soymilk extraction have been reported to reduce the objectionable flavor. However, the effects of such treatments on the nutritional value and isoflavone content of soymilk, and the responses of different soybean varieties to nutrient extraction by these methods is not well studied. The aim of this study was to determine the effect of three processing methods on protein, fat, minerals, and isoflavone content in soymilk from six soybean varieties grown in Rwanda (Peka-6, SB 24, Sc. Sequel, Sc, Squire, and a local variety) to find the best variety and processing method. The first method (M1) involved soaking soybeans in water for 12 hr prior to milk extraction, M2 involved blanching in NaHCO 3 prior to extraction and M3 involved soaking in NaHCO 3 solution for 16 hr and subsequent cooking prior to extraction. M1 resulted in significantly higher nutrient and isoflavone extraction than M2 and M3. Thus, M1 extracted more nutrients and can be recommended for soymilk production. However, where consumers prefer soymilk obtained by M2 or M3, Sc Squire and the local variety may be recommended. Sc. Squire has another advantage of higher isoflavone content than the other varieties. Further comprehensive studies on the sensory acceptability of products made from different varieties by different methods among different consumer categories will be necessary.

Metagenomic Analysis of Plant Virus Occurrence in Common Bean (Phaseolus vulgaris) in Central Kenya.

Two closely related potyviruses, bean common mosaic virus (BCMV) and bean common mosaic necrosis virus (BCMNV), are regarded as major constraints on production of common bean (Phaseolus vulgaris L.) in Eastern and Central Africa, where this crop provides a high proportion of dietary protein as well as other nutritional, agronomic, and economic benefits. Previous studies using antibody-based assays and indicator plants indicated that BCMV and BCMNV are both prevalent in bean fields in the region but these approaches cannot distinguish between these potyviruses or detect other viruses that may threaten the crop. In this study, we utilized next generation shotgun sequencing for a metagenomic examination of viruses present in bean plants growing at two locations in Kenya: the University of Nairobi Research Farm in Nairobi's Kabete district and at sites in Kirinyaga County. RNA was extracted from leaves of bean plants exhibiting apparent viral symptoms and sequenced on the Illumina MiSeq platform. We detected BCMNV, cucumber mosaic virus (CMV), and Phaseolus vulgaris alphaendornaviruses 1 and 2 (PvEV1 and 2), with CMV present in the Kirinyaga samples. The CMV strain detected in this study was most closely related to Asian strains, which suggests that it may be a recent introduction to the region. Surprisingly, and in contrast to previous surveys, BCMV was not detected in plants at either location. Some plants were infected with PvEV1 and 2. The detection of PvEV1 and 2 suggests these seed transmitted viruses may be more prevalent in Eastern African bean germplasm than previously thought.

Brachiaria Grasses (Brachiaria spp.) harbor a diverse bacterial community with multiple attributes beneficial to plant growth and development.

Endophytic and plant-associated bacteria were isolated from plants and rhizoplane soil of naturally grown Brachiaria grasses at International Livestock Research Institute in Nairobi, Kenya. Eighty-four bacterial strains were isolated from leaf tissues, root tissues, and rhizoplane soil on nutrient agar and 869 media. All bacterial strains were identified to the lowest possible taxonomic unit using 16S rDNA primers and were characterized for the production of Indole-3-acetic acid, hydrogen cyanide, and ACC deaminase; phosphate solubilization; siderophore production; antifungal properties; and plant biomass production. The 16S rDNA-based identification grouped these 84 bacterial strains into 3 phyla, 5 classes, 8 orders, 12 families, 16 genera, and 50 unique taxa. The four most frequently isolated genera were Pseudomonas (23), Pantoea (17), Acinetobacter (9), and Enterobacter (8). The functional characterization of these strains revealed that 41 of 84 strains had a minimum of three plant beneficial properties. Inoculation of maize seedlings with Acinetobacter spp., Microbacterium spp., Pectobacterium spp., Pseudomonas spp., and Enterobacter spp. showed positive effects on seedling biomass production. The ability of Brachiaria grasses to host genetically diverse bacteria, many of them with multiple plant growth-promoting attributes, might have contributed to high biomass production and adaptation of Brachiaria grasses to drought and low fertility soils.

Molecular Characterizations of Kenyan Brachiaria Grass Ecotypes with Microsatellite (SSR) Markers.

Brachiaria grass is an emerging forage option for livestock production in Kenya. Kenya lies within the center of diversity for Brachiaria species, thus a high genetic variation in natural populations of Brachiaria is expected. Overgrazing and clearing of natural vegetation for crop production and nonagricultural uses and climate change continue to threaten the natural biodiversity. In this study, we collected 79 Brachiaria ecotypes from different parts of Kenya and examined them for genetic variations and their relatedness with 8 commercial varieties. A total of 120 different alleles were detected by 22 markers in the 79 ecotypes. Markers were highly informative in differentiating ecotypes with average diversity and polymorphic information content of 0.623 and 0.583, respectively. Five subpopulations: International Livestock Research Institute (ILRI), Kitui, Kisii, Alupe, and Kiminini differed in sample size, number of alleles, number of private alleles, diversity index, and percentage polymorphic loci. The contribution of within-the-individual difference to total genetic variation of Kenyan ecotype population was 81%, and the fixation index (F ST = 0.021) and number of migrant per generation (Nm = 11.58) showed low genetic differentiation among the populations. The genetic distance was highest between Alupe and Kisii populations (0.510) and the lowest between ILRI and Kiminini populations (0.307). The unweighted neighbor-joining (NJ) tree showed test ecotypes grouped into three major clusters: ILRI ecotypes were present in all clusters; Kisii and Alupe ecotypes and improved varieties grouped in clusters I and II; and ecotypes from Kitui and Kiminini grouped in cluster I. This study confirms higher genetic diversity in Kenyan ecotypes than eight commercial varieties (Basilisk, Humidicola, Llanero, Marandú, MG4, Mulato II, Piatá and Xaraés) that represent three species and one three-way cross-hybrid Mulato II. There is a need for further collection of local ecotypes and their morphological, agronomical, and genetic characterizations to support Brachiaria grass breeding and conservation programs.

Development of Microsatellite Markers and Analysis of Genetic Diversity and Population Structure of Colletotrichum gloeosporioides from Ethiopia.

Twenty three polymorphic microsatellite markers were developed for citrus plant pathogenic fungus, Colletotrichum gloeosporioides, and were used to analyze genetic diversity and population structure of 163 isolates from four different geographical regions of Ethiopia. These loci produced a total of 118 alleles with an average of 5.13 alleles per microsatellite marker. The polymorphic information content values ranged from 0.104 to 0.597 with an average of 0.371. The average observed heterozygosity across all loci varied from 0.046 to 0.058. The gene diversity among the loci ranged from 0.106 to 0.664. Unweighted Neighbor-joining and population structure analysis grouped these 163 isolates into three major groups. The clusters were not according to the geographic origin of the isolates. Analysis of molecular variance showed 85% of the total variation within populations and only 5% among populations. There was low genetic differentiation in the total populations (FST = 0.049) as evidenced by high level of gene flow estimate (Nm = 4.8 per generation) among populations. The results show that Ethiopian C. gloeosporioides populations are generally characterized by a low level of genetic diversity. The newly developed microsatellite markers were useful in analyzing the genetic diversity and population structure of the C. gloeosporioides populations. Information obtained from this study could be useful as a base to design strategies for better management of leaf and fruit spot disease of citrus in Ethiopia.

Interspecific hybridization and bioactive alkaloid variation increases diversity in endophytic Epichloë species of Bromus laevipes.

Studying geographic variation of microbial mutualists, especially variation in traits related to benefits they provide their host, is critical for understanding how these associations impact key ecological processes. In this study, we investigate the phylogenetic population structure of Epichloë species within Bromus laevipes, a native cool-season bunchgrass found predominantly in California. Phylogenetic classification supported inference of three distinct Epichloë taxa, of which one was nonhybrid and two were interspecific hybrids. Inheritance of mating-type idiomorphs revealed that at least one of the hybrid species arose from independent hybridization events. We further investigated the geographic variation of endophyte-encoded alkaloid genes, which is often associated with key benefits of natural enemy protection for the host. Marker diversity at the ergot alkaloid, loline, indole-diterpene, and peramine loci revealed four alkaloid genotypes across the three identified Epichloë species. Predicted chemotypes were tested using endophyte-infected plant material that represented each endophyte genotype, and 11 of the 13 predicted alkaloids were confirmed. This multifaceted approach combining phylogenetic, genotypic, and chemotypic analyses allowed us to reconstruct the diverse evolutionary histories of Epichloë species present within B. laevipes and highlight the complex and dynamic processes underlying these grass-endophyte symbioses.

Deletion of the fungal gene soft disrupts mutualistic symbiosis between the grass endophyte Epichloë festucae and the host plant.

Hyphal anastomosis, or vegetative hyphal fusion, establishes the interconnection of individual hyphal strands into an integrated network of a fungal mycelium. In contrast to recent advances in the understanding of the molecular basis for hyphal anastomosis, knowledge of the physiological role of hyphal anastomosis in the natural habitats of filamentous fungi is still very limited. To investigate the role of hyphal anastomosis in fungal endophyte-plant interactions, we generated mutant strains lacking the Epichloë festucae soft (so) gene, an ortholog of the hyphal anastomosis gene so in the endophytic fungus E. festucae. The E. festucae Δso mutant strains grew similarly to the wild-type strain in culture but with reduced aerial hyphae and completely lacked hyphal anastomosis. The most striking phenotype of the E. festucae Δso mutant strain was that it failed to establish a mutualistic symbiosis with the tall fescue plant host (Lolium arundinaceum); instead, it killed the host plant within 2 months after the initial infection. Microscopic examination revealed that the death of the tall fescue plant host was associated with the distortion and disorganization of plant cells. This study suggests that hyphal anastomosis may have an important role in the establishment/maintenance of fungal endophyte-host plant mutualistic symbiosis.

Phytophthora aquimorbida sp. nov. and Phytophthora taxon 'aquatilis' recovered from irrigation reservoirs and a stream in Virginia, USA.

Two distinct subgroups (L2 and A(-2)) were recovered from irrigation reservoirs and a stream in Virginia, USA. After molecular, morphological and physiological examinations, the L2 subgroup was named Phytophthora aquimorbida and the A(-2) designated as Phytophthora taxon 'aquatilis'. Both taxa are homothallic. P. aquimorbida is characterized by its noncaducous and nonpapillate sporangia, catenulate and radiating hyphal swellings and thick-walled plerotic oospores formed in globose oogonia mostly in the absence of an antheridium. P. taxon 'aquatilis' produces plerotic oospores in globose oogonia mostly with a paragynous antheridium. It has semi-papillate, caducous sporangia with variable pedicels, but it does not have hyphal swelling. Analyses of ITS, CO1, ß-tubulin and NADH1 sequences revealed that P. aquimorbida is closely related to P. hydropathica, P. irrigata and P. parsiana, and P. taxon 'aquatilis' is related to P. multivesiculata. The optimum temperature for culture growth is 30 and 20 C for P. aquimorbida and P. taxon 'aquatilis' respectively. Both taxa were pathogenic to rhododendron plants and caused root discoloration, pale leaves, wilting, tip necrosis and dieback. Their plant biosecurity risk also is discussed.

Enhancement of switchgrass (Panicum virgatum L.) biomass production under drought conditions by the ectomycorrhizal fungus Sebacina vermifera.

Experiments were conducted to examine the effects of cocultivating the important bioenergy crop switchgrass with the ectomycorrhizal fungus Sebacina vermifera under severe drought conditions. Plants cocultivated with the fungus produced significantly higher biomass and had a higher macronutrient content than uninoculated control plants under both adequately watered and drought conditions.

Prevalence of an intraspecific Neotyphodium hybrid in natural populations of stout wood reed (Cinna arundinacea L.) from eastern North America.

Members of genus Neotyphodium are asexual derivatives of sexual Epichloë species and maintain endophytic relationships with many cool-season grasses. Most Neotyphodium species analyzed so far are interspecific hybrids with combined or partial genomes of two or three ancestral species. In this study we characterized Neotyphodium isolates from Cinna arundinacea, a perennial cool-season grass from eastern North America. A total of 23 isolates grouping into two distinct morphotypes were obtained from five local populations of C. arundinacea. PCR amplification and cloning of translation-elongation factor 1-α (tefA) and ß-tubulin (tubB) genes of 10 isolates comprising both morphotypes (two isolates per location) revealed that all 10 contain two copies of tefA and tubB genes. Surprisingly phylogenetic analysis of mainly non-coding sequence from these genes revealed that both copies in each isolate were inherited from Epichloë typhina ancestors, indicating that the C. arundinacea endophytes arose through intraspecific hybridization between two E. typhina progenitors with extant relatives infecting hosts Poa nemoralis and Poa pratensis. Furthermore the tefA sequences were identical between isolates, as were tubB sequences, despite obvious morphological differences. Profiling of alkaloid biosynthetic genes from these isolates indicated the presence of the peramine biosynthetic gene (perA) and the absence of genes required for biosynthesis of lolines, indole-diterpenes and ergot alkaloids. Thus this endophyte is potentially capable of producing peramine in planta and providing protection to its host from insect pests. The absence of genes for indole-diterpenes and ergot alkaloid biosynthesis makes this endophyte a candidate for agricultural applications. Based on our phylogenetic analysis, alkaloid profiling and description of morphological characteristics, we propose the name Neotyphodium schardlii for these isolates from C. arundinacea, a new member of genus Neotyphodium and the first described to have arisen through intraspecific hybridization.