Some zoosporic fungi of New Zealand

Summary In further studies of the zoosporic fungi of New Zealand nine additional species were isolated on various substrata from soil. These include Rhizophydium pythii de Wildemann, R. condylosum Karling, Rhizophlyctis oceanis Karling, R. ingoldii Sparrow, R. boneysi Sparrow, Rhizophlyctis sp., Rhizidium reniformis sp. nov., Chytriomyces rotoruaensis sp. nov., Sparrowia parasitica Willoughby, and Aphanomycopsis punctatus Karling. Rhizidium reniformis is characterized by predominantly reniform, appendiculate zoosporangia and small zoospores which emerge slowly in a columnar mass. This usually floats away from the zoosporangium and explands, and after a while the zoospores swarm collectively in a vesicle. Chytriomyces rotoruaensis resembles R. reniformis by the structure and appearance of its thallus and behavior of the zoospores after discharge, but differs by the presence of a thin inconspicuous operculum and the development of smooth hyaline resting spores with coarsely granular content. Rhizophlyctis ingoldii, Sparrowia parasitica, and R. boneysi, known previously only from England and Hawaii, respectively, occurred abundantly in New Zealand.This study has been supported by a grant from the National Science Foundation, Washington, D.C.     

POLYPHASIC CHARACTERIZATION OF THREE STRAINS OF ANABAENA RENIFORMIS AND APHANIZOMENON APHANIZOMENOIDES (CYANOBACTERIA) AND THEIR RECLASSIFICATION TO SPHAEROSPERMUM GEN. NOV. (INCL. ANABAENA KISSELEVIANA)1

Occurrences of rare cyanobacteria Anabaena reniformis Lemmerm. and Aphanizomenon aphanizomenoides (Forti) Horecká et Komárek were recently detected at several localities in the Czech Republic. Two monoclonal strains of An. reniformis and one strain of Aph. aphanizomenoides were isolated from distant localities and different sampling years. They were characterized by a combination of morphological, genetic, and biochemical approaches. For the first time, partial 16S rRNA gene sequences were obtained for these morphospecies. Based on this gene, all of these strains clustered separately from other planktonic Anabaena and Aphanizomenon strains. They appeared in a cluster with Cylindrospermopsis Seenaya et Subba Raju and Raphidiopsis F. E. Fritsch et M. F. Rich, clustered closely together with two An. kisseleviana Elenkin strains available from GenBank. A new generic entity was defined (Sphaerospermum gen. nov., with the type species S. reniforme, based on the traditional species An. reniformis). These results contribute significantly to the knowledge base about genetic heterogeneity among planktonic Anabaena–like and Aphanizomenon–like morphospecies. Accordingly, the subgenus Dolichospermum, previously proposed for the group of planktonic Anabaena, should be revaluated. Secondary metabolite profiles of the An. reniformis and Aph. aphanizomenoides strains differed considerably from 17 other planktonic Anabaena strains of eight morphospecies isolated from Czech water bodies. Production of puwainaphycin A was found in both of the An. reniformis strains. Despite the relatively short phylogenetic distance from Cylidrospermopsis, the production of cylindrospermopsin was not detected in any of our strains.     

Root-Parasitic Nematodes Enhance Soil Microbial Activities and Nitrogen Mineralization

Obligate root-parasitic nematodes can affect soil microbes positively by enhancing C and nutrient leakage from roots but negatively by restricting total root growth. However, it is unclear how the resulting changes in C availability affect soil microbial activities and N cycling. In a microplot experiment, effects of root-parasitic reniform nematodes (Rotylenchulus reniformis) on soil microbial biomass and activities were examined in six different soils planted with cotton. Rotylenchulus reniformis was introduced at 900 nematodes kg^–1 soil in May 2000 prior to seeding cotton. In 2001, soil samples were collected in May before cotton was seeded and in November at the final harvest. Extractable C and N were consistently higher in the R. reniformis treatments than in the non-nematode controls across the six different soils. Nematode inoculation significantly reduced microbial biomass C, but increased microbial biomass N, leading to marked decreases in microbial biomass C:N ratios. Soil microbial respiration and net N mineralization rates were also consistently higher in the nematode treatments than in the controls. However, soil types did not have a significant impact on the effects of nematodes on these microbial parameters. These findings indicate that nematode infection of plant roots may enhance microbial activities and the turnover of soil microbial biomass, facilitating soil N cycling. The present study provides the first evidence about the direct role of root-feeding nematodes in enhancing soil N mineralization.     

Functional C‐TERMINALLY ENCODED PEPTIDE (CEP) plant hormone domains evolved de novo in the plant parasite Rotylenchulus reniformis

Sedentary plant‐parasitic nematodes (PPNs) induce and maintain an intimate relationship with their host, stimulating cells adjacent to root vascular tissue to re‐differentiate into unique and metabolically active ‘feeding sites’. The interaction between PPNs and their host is mediated by nematode effectors. We describe the discovery of a large and diverse family of effector genes, encoding C‐TERMINALLY ENCODED PEPTIDE (CEP) plant hormone mimics (RrCEPs), in the syncytia‐forming plant parasite Rotylenchulus reniformis. The particular attributes of RrCEPs distinguish them from all other CEPs, regardless of origin. Together with the distant phylogenetic relationship of R. reniformis to the only other CEP‐encoding nematode genus identified to date (Meloidogyne), this suggests that CEPs probably evolved de novo in R. reniformis. We have characterized the first member of this large gene family (RrCEP1), demonstrating its significant up‐regulation during the plant–nematode interaction and expression in the effector‐producing pharyngeal gland cell. All internal CEP domains of multi‐domain RrCEPs are followed by di‐basic residues, suggesting a mechanism for cleavage. A synthetic peptide corresponding to RrCEP1 domain 1 is biologically active and capable of up‐regulating plant nitrate transporter (AtNRT2.1) expression, whilst simultaneously reducing primary root elongation. When a non‐CEP‐containing, syncytia‐forming PPN species (Heterodera schachtii) infects Arabidopsis in a CEP‐rich environment, a smaller feeding site is produced. We hypothesize that CEPs of R. reniformis represent a two‐fold adaptation to sustained biotrophy in this species: (i) increasing host nitrate uptake, whilst (ii) limiting the size of the syncytial feeding site produced.     

Molecular Characterization of a Nonfibrillar Collagen from the Marine Sponge <Emphasis Type="Italic">Chondrosia reniformis</Emphasis> Nardo 1847 and Positive Effects of Soluble Silicates on Its Expression

We report here the complete cDNA sequence of a nonfibrillar collagen (COLch) isolated from the marine sponge Chondrosia reniformis, Nardo 1847 using a PCR approach. COLch cDNA consists of 2,563 nucleotides and includes a 5′ untranslated region (UTR) of 136 nucleotides, a 3′ UTR of 198 nucleotides, and an open reading frame encoding for a protein of 743 amino acids with an estimated M _r of 72.12 kDa. The phylogenetic analysis on the deduced amino acid sequence of C-terminal end shows that the isolated sequence belongs to the short-chain spongin-like collagen subfamily, a nonfibrillar group of invertebrate collagens similar to type IV collagen. In situ hybridization analysis shows higher expression of COLch mRNA in the cortical part than in the inner part of the sponge. Therefore, COLch seems to be involved in the formation of C. reniformis ectosome, where it could play a key role in the attachment to the rocky substrata and in the selective sediment incorporation typical of these organisms. qPCR analysis of COLch mRNA level, performed on C. reniformis tissue culture models (fragmorphs), also demonstrates that this matrix protein is directly involved in sponge healing processes and that soluble silicates positively regulate its expression. These findings confirm the essential role of silicon in the fibrogenesis process also in lower invertebrates, and they should give a tool for a sustainable production of marine collagen in sponge mariculture.     

Note Sistematiche ed Osservazioni Fitosociologiche Sulle Laminariales del Mediterraneo Occidentale

Abstract

Laminariales from western Mediterranean Sea: taxonomical and phytosociological study. — Laminariales found in the Mediterranean Sea are: Laminaria rodriguezii Bornet, L. ochroleuca De la Pylaie, Saccorhiza polyschides Batters, Phyllaria reniformis Rostafinski, Ph. purpurascens Rostafinski. By diving technique L. rodriguezii and Ph. reniformis was collected off isles Ustica, Stromboli, Pianosa and Montecristo (Thyrrenean Sea) from 50 to 90 mts depth in the coralligenous biocoenose; L. ochroleuca and Ph. purpurascens between Scilla and Ganzirri in the straits of Messina from 45 to 85 mts depth in facies of deep rheophilous biocoenoses; S. polyschides and Ph. reniformis near Villa S. Giovanni and Messina in the facies with Ulva-Corallina-Dictyopteris of photophilous algae biocoenose. Algal vegetation associated with these species of Laminariales in the straits of Messina is characterized by Atlantic-temperate elements (Ulva olivascens, Halurus equisetifolius, Cryptopleura ramosa, Giffordia hincksiae, etc…).     

Effect of Controlled Cold Storage on Recovery of Rotylenchulus reniformis from Naturally Infested Soil

Rotylenchulus reniformis is rapidly becoming the most economically important pest associated with cotton in the southeastern United States. Incentive programs have been implemented to support sampling of production fields to determine the presence and abundance of R. reniformis. These sampling programs have dramatically increased the number of soils samples submitted to nematology laboratories during autumn. The large numbers of samples overwhelm most labs and require placement in cold storage until extraction. Therefore, the objective of this study was to examine the length of time soils infested with R. reniformis can be stored before nematode extraction without compromising the accuracy of estimates of population densities. A sandy loam and a silty loam were the two cotton production soils used in this study. Rotylenchulus reniformis numbers decreased 61%during the first 180 days of storage in both soils. Rotylenchulus reniformis numbers from the initial sampling through 180 days decreased as a linear function. The decline of R. reniformis numbers during storage was estimated as 0.28% of the population lost daily from the maximum population through 180 days. The diminution of nematode numbers from 180 through 1,080 days in storage continued, but at a slower rate. Numbers of R. reniformis declined to less than 89%, 93%, and 99% of the initial population within 360, 720, and 1,080 days, respectively, of storage. The reduction of R. reniformis numbers over 180 days can be adjusted, allowing a more accurate estimation of R. reniformis levels in soil samples stored at 4 °C.     

Reproductive biology and pollination of Utricularia reniformis A.St.‐Hil. (Lentibulariaceae)

Utricularia reniformis is an endemic Brazilian carnivorous plant, most common in high‐altitude grasslands. Knowledge of the reproductive biology of U. reniformis is essential for planning conservation strategies, but it is currently poorly understood. Thus, we studied the floral morphology, floral biology, breeding system and pollination of this species. U. reniformis produces and stores nectar in the flower spur, a classic feature of bee‐pollinated flowers, and we recorded Xylocopa sp. and Bombus sp. as pollinators. Moreover, although it is self‐compatible it is an obligate animal‐pollinated species, as the sensitive stigma avoids self‐pollination. However, in natural conditions reproductive success is low due to the rarity of visits from pollinators. We suggest that the low reproductive success caused by a deficit of pollinators may affect gene flow, causing loss of genetic diversity in U. reniformis.     

Hypothesized Kinetic Models for Describing the Growth of Globular and Encrusting Demosponges

The marine sponges Dysidea avara and Chondrosia reniformis (globular forms) were cultured in the laboratory on a diet of viable Phaeodactylum tricornutum cells and dissolved nutrients (algae and fish powders). Our growth data were combined with literature data for Pseudosuberites andrewsi (a globular sponge) and for the encrusting sponges Oscarella lobularis, Hemimycale columella, and Crambe crambe. The suitability of three growth models—linear, exponential, and radial accretive—for describing the growth of globular and encrusting sponges was assessed. Radial accretive growth was determined to be the best model to describe growth of both encrusting and globular sponges. Average growth rates of 0.051 ± 0.016 and 0.019 ± 0.003 mm/day (calculated as the increase of the radius of the sponge per day) were obtained experimentally for D. avara and C. reniformis, respectively.     

Two Simple Methods for the Collection of Individual Life Stages of Reniform Nematode,Rotylenchulus reniformis

The sedentary semi-endoparasitic nematode Rotylenchulus reniformis, the reniform nematode, is a serious pest of cotton and soybean in the United States. In recent years, interest in the molecular biology of the interaction between R. reniformis and its plant hosts has increased; however, the unusual life cycle of R. reniformis presents a unique set of challenges to researchers who wish to study the developmental expression of a particular nematode gene or evaluate life stage–specific effects of a specific treatment such as RNA-interference or a potential nematicide. In this report, we describe a simple method to collect R. reniformis juvenile and vermiform adult life stages under in vitro conditions and a second method to collect viable parasitic sedentary females from host plant roots. Rotylenchulus reniformis eggs were hatched over a Baermann funnel and the resultant second-stage juveniles incubated in petri plates containing sterile water at 30°C. Nematode development was monitored through the appearance of fourth-stage juveniles and specific time-points at which each developmental stage predominated were determined. Viable parasitic sedentary females were collected from infected roots using a second method that combined blending, sieving, and sucrose flotation. Rotylenchulus reniformis life stages collected with these methods can be used for nucleic acid or protein extraction or other experimental purposes that rely on life stage–specific data.     

Studies on the pathogenicity of three host races of Rotylenchulus reniformis on castor

Three races (race 2, 3 and 4) of Rotylenchulus reniformis were identified using host differential from 20 populations collected from Aligarh, India. Pathogenicity tests were conducted using 250, 500, 1000, 2000, 4000 and 8000 immature females/kg soil of three races of R. reniformis on castor. The threshold inoculum levels of Race-2, Race-3 and Race-4 on castor were 500, 1000 and 2000 immature females/kg soil, respectively. Out of these three races present in Aligarh district, Race-2 was found more pathogenic followed by Race-3 and Race-4. Infected plants with all the three races of reniform nematode showed stunting, necrosis, leaf shedding and growth reduction at and above the inoculum threshold level of each race. Reduction in castor growth was directly proportional to the inoculum level of R.reniformis. The rate of nematode multiplication was density dependent.     

Biology of Pseudopotamilla reniformis (Müller 1771) in the White Sea,with description of asexual reproduction

The tube-dwelling polychaete Pseudopotamilla reniformis (Sabellidae) forms dense and complex aggregations of flexible tubes on hard substrates in the subtidal zone of the White Sea. No sexual reproduction was observed in this study and recruitment appeared to be due to asexual reproduction by architomy in winter, from October to March. The posterior part of the abdomen undergoes spontaneous fission into from 2 to 4 fragments and depending on their position, the fragments regenerate their anterior ends or both anterior and posterior ends. Regeneration in P. reniformis takes place via a combination of epimorphosis (replacement of missing parts by cell proliferation and the growth of new tissue) and morphallaxis (the remodelling of pre-existing structures without cell proliferation). The morphogenetic events during regenerative restoration include de novo formation of branchial crown, formation of thoracic segments and restoration of the posterior end. Asexual reproduction appears to play a crucial role for formation of P. reniformis aggregations and is very important for the population in the White Sea, at the margin of the species’ range.     

Nematicides Enhance Growth and Yield of Rotylenchulus Reniformis Resistant Cotton Genotypes

Rotylenchulus reniformis resistant LONREN-1×FM966 breeding lines developed at Auburn University have demonstrated that the nematode resistance is accompanied by severe stunting, limited growth, and low yields. The objectives of this study were to evaluate the effects of applying nematicides to selected LONREN breeding lines on R. reniformis nematode populations, plant stunting, and yield. Four resistant breeding lines from the LONREN-1×FM966 cross, one susceptible line from the LONREN-1×FM966 cross, as well as LONREN-1, BARBREN-713, and the susceptible cultivar DP393 were evaluated with and without nematicides in the presence of R. reniformis. In the greenhouse, nematicides increased plant height across all genotypes compared with no nematicide. Rotylenchulus reniformis populations were 50% lower in the resistant lines compared with the susceptible lines at 45 days after planting (DAP). In microplot and field trials, the phenotypic stunting of all genotypes was reduced by aldicarb with increases in plant heights at 30 and 75 DAP. Increases in yields were evident across all genotypes treated with aldicarb. In all three trial environments, BARBREN-713 outperformed the LONREN-derived lines as well as ‘DP393’ in seed cotton yields, while having significantly lower R. reniformis egg densities than the susceptible genotypes.     

The Effect of Soil Texture and Irrigation on Rotylenchulus reniformis and Cotton

The reniform nematode, Rotylenchulus reniformis, is the most damaging nematode pathogen of cotton in Alabama. Soil texture is currently being explored as a basis for the development of economic thresholds and management zones within a field. Trials to determine the reproductive potential of R. reniformis as influenced by soil type were conducted in microplot and greenhouse settings during 2008 to 2010. Population density of R. reniformis was significantly influenced by soil texture and exhibited a general decrease with increasing median soil particle size (MSPS). As the MSPS of a soil increased from 0.04 mm in clay soil to > 0.30 mm in very fine sandy loam and sandy loam soils, R. reniformis numbers decreased. The R. reniformis population densities on all soil types were also greater with irrigation. Early season cotton development was significantly affected by increasing R. reniformis Pi, with plant shoot-weight-to-root-weight ratios increasing at low R. reniformis Pi and declining with increasing R. reniformis Pi. Plant height was increased by irrigation throughout the growing season. The results suggests that R. reniformis will reach higher population densities in soils with smaller MSPS; however, the reduction in yield or plant growth very well may be no greater than in a soil that is less preferential to the nematode.     

Characterization of an Indonesian Isolate of Paecilomyces reniformis

An entomopathogenic fungus (IndGH 96), identified as Paecilomyces reniformis, was isolated from long-horned grasshoppers (Orthoptera: Tettigoniidae) in Sulawesi, Indonesia. The phenotypic and molecular data identified the IndGH 96 as a P. reniformis. We present the first comprehensive characterization of this species using morphological features, sequencing of the ITS1-5.8s-ITS2 region, D1/D2 region of 28S of rDNA, and a portion of the tubulin gene, and laboratory bioassays. Distinguishing features include a hyphal body stage during vegetative growth and the production of distinctly curved, light-green conidia. High dosage bioassays showed that IndGH 96 was infectious to both long-horned and short-horned grasshoppers but not to the house cricket, Acheta domestica, or to the lepidopterans velvetbean caterpillar, Anticarsia gemmatalis or fall armyworm, Spodoptera frugiperda. Phenotypic and genetic analyses suggest that IndGH 96 and other isolates of P. reniformis are more closely related to Nomuraea rileyi than to other species of Paecilomyces.     

Control of root-knot nematode,Meloidogyne incognita and reniform nematode,Rotylenchulus reniformis on pigeonpea and linseed

Abstract

Pigeonpea (Cajanus cajan) and linseed (Linum usitatissimum) are susceptible to Meloidogyne incognita and Rotylenchulus reniformis nematodes. Reduction in different growth parameters (length and weight of plant, number of pods), bulk density of pigeonpea stem, oil content of linseed, chlorophyll content of leaf and water absorption of roots caused by M. incognita and R. reniformis were statistically significant. Similar effects were also observed in plants raised from seeds soaked in different concentrations of water soluble fractions (WSF) of rice polish and pyridoxine solutions, however, the reductions were of a comparatively lesser extent. Higher concentrations of the solutions were more effective when compared to lower ones and pyridoxine was more beneficial than WSF for improving plant growth and reducing disease incidence.     

Effects of the Integration of Sunn Hemp and Soil Solarization on Plant-Parasitic and Free-Living Nematodes

Sunn hemp (SH), Crotolaria juncea, is known to suppress Rotylenchulus reniformis and weeds while enhancing free-living nematodes involved in nutrient cycling. Field trials were conducted in 2009 (Trial I) and 2010 (Trial II) to examine if SH cover cropping could suppress R. reniformis and weeds while enhancing free-living nematodes if integrated with soil solarization (SOL). Cover cropping of SH, soil solarization, and SH followed by SOL (SHSOL) were compared to weedy fallow control (C). Rotylenchulus reniformis population was suppressed by SHSOL at the end of cover cropping or solarization period (Pi) in Trial I, but not in Trial II. However, SOL and SHSOL did not suppress R. reniformis compared to SH in either trial. SH enhanced abundance of bacterivores and suppressed the % herbivores only at Pi in Trial II. At termination of the experiment, SH resulted in a higher enrichment index indicating greater soil nutrient availability, and a higher structure index indicating a less disturbed nematode community compared to C. SOL suppressed bacterivores and fungivores only in Trial II but not in Trial I. On the other hand, SHSOL enhanced bacterivores and fungivores only at Pi in Trial I. Weeds were suppressed by SH, SOL and SHSOL throughout the experiment. SHSOL suppressed R. reniformis and enhanced free-living nematodes better than SOL, and suppressed weeds better than SH.     

Accelerated Movement of Nematodes from Soil in Baermann Funnels with Temperature Gradients

Baermann funnels were modified to eliminate or reverse the small temperature gradient (1-2 C/cm) across the soil layer that normally results from water evaporation. Effects of modifications on extraction efficiency were examined at various ambient temperatures and after overnight adaptation of three nematode species at 20 and 30 C. Extraction of Meloidogyne incognita from sandy loam, Tylenchulus semipenetrans from sandy clay loam, and Rotylenchulus reniformis from silt was greatly accelerated simply by covering funnels to prevent evaporation. In most cases, covering increased the nematodes extracted by 10-100 times after 5.5-48 hours. Faster and more efficient extraction of R. reniformis occurred over a wide range of ambient temperature (18-29 C). Effects of ambient temperature and temperature gradient direction on Baermann funnel extraction of R. reniformis were partly inconsistent with the behavior of R. reniformis in agar. Nematodes in agar moved toward cold at some ambient temperatures and toward heat at other temperatures. They always appeared to move toward cold on Baermann funnels. Differences were not attributable to blockage of gas exchange by covers. In agar and in funnels, the patterns of response to ambient temperature were shifted in the direction of the storage temperature.     

Tolerance to reniform nematode (Rotylenchulus reniformis) race A in pigeonpea (Cajanus cajan) genotypes

The reniform nematode (Rotylenchulus reniformis) is an important pathogen of pigeonpea (Cajanus cajan). Forty‐six medium maturity (mature in 151–200 days at Patancheru, India) pigeonpea genotypes were evaluated for resistance and tolerance to the reniform nematode in greenhouse and field tests, over the period 1990–97. Each genotype was screened for number of nematode egg masses on a 1 (no egg mass = highly resistant) to 9 (> 50 egg masses = highly susceptible) scale. Plant biomass production in carbofurantreated plots was compared with that in non‐treated plots in a field naturally infested with R. reniformis. Pigeonpea genotypes C 11, ICPL 87119 and ICPL 270 were used as nematode susceptible checks. Genotypes with good plant growth, both in nematode‐free and nematode‐infested plots, were identified as tolerant and evaluated for plant growth and yield for at least three years. All the tested genotypes were susceptible (7 and 9 egg mass score). Single‐plant‐selections, based on plant vigour and yield, were made from genotypes showing tolerance to nematode infection. The level of tolerance was enhanced by plant‐to‐progeny row selection for plant vigour and seed yield in a nematode‐sick field for at least three years. The most promising nematode tolerant genotypes produced significantly greater yield and biomass than the locally grown pigeonpea cultivars in fields naturally infested with R. reniformis at two locations. Pigeonpea landraces are considered to be the most likely sources of tolerance to the nematode. These reniform nematode tolerant lines represent new germplasm and they are available in the genebank of pigeonpea at ICRISAT bearing accession numbers ICP 16329, ICP 16330, ICP 16331, ICP 16332, and ICP 16333.