Intraspecific variation in RFLP patterns and morphological studies on Steinernema feltiae and S. kraussei (Rhabditida: Steinernematidae) from Hokkaido, Japan

AbstractSteinernema feltiae and S. kraussei were isolated from Hokkaido, Japan. This is the first record of S. kraussei and the first definitive record of S. feltiae from Japan. The morphological variation of the infective juveniles and the first generation males of Japanese isolates of both species are reported. Intraspecific variation in the PCR-RFLP analysis of the ITS region of ribosomal DNA was observed in both species. The Japanese isolates of S. feltiae showed different RFLP patterns from European isolates with Dde I and Hinf I restriction digests. The Japanese isolate of S. kraussei also showed an RFLP variation from the UK and Russian isolates upon Dde I restriction digestion. Moreover, in each isolate of S. kraussei, some intra-population variations were observed with some restriction digestions. The intraspecific variation in the ITS region of the rDNA could be used as a molecular marker to distinguish the Japanese isolates from European isolates of S. feltiae if the latter was introduced as a biological insecticide.

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Steinernema apuliae sp. n. (Rhabditida: Steinernematidae): a new entomopathogenic nematode from southern Italy

Zootaxa ◽

10.11646/zootaxa.460.1.1 ◽

2004 ◽

Vol 460 (1) ◽

pp. 1 ◽

Cited By ~ 11

Author(s):

ORESTE TRIGGIANI ◽

ZDENEK MRÁ»EK ◽

ALEX REID

Keyword(s):

First Generation ◽

Southern Italy ◽

Entomopathogenic Nematode ◽

Rflp Analysis ◽

Tail Length ◽

Its Region ◽

The Body ◽

Steinernema Glaseri ◽

Pcr Rflp ◽

H 41

Steinernema apuliae sp. n. has been found in soil samples collected along a saltpan border habitat in southern Italy characterized by a salted silt soil. This species belongs to the long-IJ nematode group represented by Steinernema glaseri (Steiner, 1929) and Steinernema arenarium (Artyukhovsky, 1967) among others. However, it differs from these taxa in some morphometric values such as V%, H%. Females possess asymmetrical, oblique slit vulva, slant vagina and small flap in the vulval opening. These characteristics are more distinct in second generation females which is different from most other steinernematids; the vulva position is behind the mid-body about 57% to 61% of the body length. First-generation females have a conical-like tip bearing 2 to 3 papilla-like protuberances. Male mucron is absent in both generations. Lightly brown spicules have bluntly pointed tip and elongated manubrium. Third-stage infective juveniles are on average over 1000 m long; the position of the excretory pore is posterior (D% 66) and the hyaline layer is less than half the tail length (H% 41 42). Lateral fields are formed by 8 equally distributed ridges. S. apuliae differs from S. glaseri and S. arenarium and is separated by PCR-RFLP analysis of the ITS region. There were no positive cross-breedings among these species.

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Characterization of Dryas octopetala ectomycorrhizas from limestone karst vegetation, western Ireland

Canadian Journal of Botany ◽

10.1139/b02-082 ◽

2002 ◽

Vol 80 (9) ◽

pp. 970-982 ◽

Cited By ~ 20

Author(s):

Thomas J Harrington ◽

Derek T Mitchell

Keyword(s):

Rflp Analysis ◽

Its Region ◽

Fungal Species ◽

Morphological Evidence ◽

Soil Core ◽

Cenococcum Geophilum ◽

Dryas Octopetala ◽

Pcr Rflp ◽

Provisional Identification ◽

Western Ireland

The principal ectomycorrhizas of Dryas octopetala L. from a treeless grass-heath in the Burren, western Ireland, were characterized using morphotyping and molecular methods (PCR-RFLP analysis of ITS-rDNA and sequencing of the ITS region). Twenty-one distinct morphotypes are described. Six of these (Cortinarius atrovirens Kalchbr., Cortinarius caesiocanescens (Mos.) Kühn. & Romagn., Cortinarius calochrous (Mos.) Nezd., Cortinarius odorifer Britz., Cortinarius mussivus Fr., and Tricholoma myomyces (Scop.) Quél.) were distinguished by tracing rhizomorph connections between mycorrhizas and basidiomes. The ectomycorrhizas of Cenococcum geophilum Fr., Craterellus lutescens (Pers.:Fr.) Fr., and Hebeloma sinapizans (Paulet:Fr.) Gill were identified based on molecular and morphological evidence. The ectomycorrhizas of Cortinarius brunneus (Pers.:Fr.) Fr., Cortinarius infractus (Pers.:Fr.) Fr., Hydnum repandum L., and Hebeloma circinans Quél. were distinguished provisionally, because they were consistently found in soil core samples containing basidiomes of a particular fungal species. The provisional identification of Lactarius sanguifluus (Paulet) Fr., and Russula delica Fr. ectomycorrhizas was also based on anatomical evidence, particularly the presence of lacticifers and cystidia, respectively. Six morphotypes could not be assigned to a specific fungal taxon and, therefore, were named "Dryadirhiza" + a characterizing epithet (D. aerea, D. cerina, D. fulgens, D. nigra, D. rugosa, and D. truncata). It is concluded that Dryas octopetala forms ectomycorrhizal associations in the Burren with woodland fungal species.Key words: ectomycorrhizas, Dryas octopetala, morphotyping, ITS-RFLP, mountain avens.

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Determination of Types of Enterocytozoon bieneusiStrains Isolated from Patients with Intestinal Microsporidiosis

Journal of Clinical Microbiology ◽

10.1128/jcm.36.7.1882-1885.1998 ◽

1998 ◽

Vol 36 (7) ◽

pp. 1882-1885 ◽

Cited By ~ 40

Author(s):

Olivier Liguory ◽

Felicia David ◽

Claudine Sarfati ◽

Francis Derouin ◽

Jean-Michel Molina

Keyword(s):

Rflp Analysis ◽

Its Region ◽

Restriction Enzymes ◽

Epidemiological Studies ◽

Enterocytozoon Bieneusi ◽

Type I ◽

Pcr Rflp ◽

Intestinal Microsporidiosis ◽

Rflp Typing

To determine the types of Enterocytozoon bieneusistrains associated with intestinal microsporidiosis, we developed a rapid and efficient approach for typing parasites obtained from stool specimens by PCR-restriction fragment length polymorphism (PCR-RFLP). Typing was based on DNA polymorphism of the ribosomal DNA internal transcribed spacer (ITS) region of E. bieneusi. RFLPs generated with two restriction enzymes (NlaIII andFnu4HI) in PCR-amplified ITS products were used to classify strains into different lineages. This approach was successfully used to differentiate 78 strains that had been obtained from the stools of 65 patients with intestinal microsporidiosis. Among the 78 strains, we found four genetically unrelated lineages, showing the genetic diversity of E. bieneusi. Type I strains of E. bieneusi were found in a majority of the samples, accounting for 51 (78%) of the 65 microsporidiosis cases. In contrast, type II, III, and IV strains were found in only 8 (12%), 3 (5%), and 3 (5%) cases, respectively. All strains of E. bieneusi we have tested so far fall into one of four different lineages, and this study shows that human intestinal microsporidiosis is most often associated with type I strains. PCR-RFLP analysis of the ITS region of E. bieneusishould be useful for epidemiological studies.

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Sequence and PCR–RFLP analysis of the internal transcribed spacers of the rDNA repeat unit in isolates of Cryptosporidium from different hosts

Parasitology ◽

10.1017/s0031182098003412 ◽

1999 ◽

Vol 118 (1) ◽

pp. 49-58 ◽

Cited By ~ 56

Author(s):

U. M. MORGAN ◽

P. DEPLAZES ◽

D. A. FORBES ◽

F. SPANO ◽

H. HERTZBERG ◽

...

Keyword(s):

Neospora Caninum ◽

Repeat Unit ◽

Rflp Analysis ◽

Internal Transcribed Spacers ◽

Sequence Information ◽

Rdna Sequences ◽

Pcr Rflp ◽

Rdna Repeat Unit ◽

The Uk ◽

Epidemiology Studies

The Cryptosporidium ITS1, 5·8S and ITS2 rDNA regions from a number of Cryptosporidium isolates from different hosts and geographical areas were cloned and sequenced in order to investigate the extent of sequence heterogeneity between human and cattle-derived isolates from different geographical locations and also between isolates of Cryptosporidium from different hosts such as cats, pigs, mice and a koala. Calf-derived isolates from different continents were virtually identical as were human-derived isolates from the UK and Australia. Genetic differences between Cryptosporidium isolates were extensive and were in fact greater than the level of nucleotide divergence between Toxoplasma gondii and Neospora caninum rDNA sequences. Based on the sequence information derived from this study, PCR–RFLP of the ITS1 region was undertaken in order to directly amplify and genotype Cryptosporidium isolates from different hosts. This PCR–RFLP approach can now be used for molecular epidemiology studies, circumventing the need for costly sequencing and allowing a wider range of genetically different isolates to be examined.

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Rapid detection and quantification of yeast species during spontaneous wine fermentation by PCR-RFLP analysis of the rDNA ITS region

Journal of Applied Microbiology ◽

10.1046/j.1365-2672.1999.00600.x ◽

1999 ◽

Vol 87 (6) ◽

pp. 949-956 ◽

Cited By ~ 113

Author(s):

L. Granchi ◽

M. Bosco ◽

A. Messini ◽

M. Vincenzini

Keyword(s):

Rapid Detection ◽

Yeast Species ◽

Rflp Analysis ◽

Its Region ◽

Wine Fermentation ◽

Rdna Its ◽

Pcr Rflp ◽

Rdna Its Region ◽

Detection And Quantification

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Characterization of Ascaris from Ecuador and Zanzibar

Journal of Helminthology ◽

10.1017/s0022149x14000431 ◽

2014 ◽

Vol 89 (4) ◽

pp. 512-515 ◽

Cited By ~ 5

Author(s):

A.M. Sparks ◽

M. Betson ◽

G. Oviedo ◽

C. Sandoval ◽

P.J. Cooper ◽

...

Keyword(s):

Fragment Length Polymorphism ◽

Rflp Analysis ◽

Its Region ◽

Restriction Enzymes ◽

Zoonotic Transmission ◽

Pcr Rflp ◽

Double Digestion ◽

Polymerase Chain ◽

Shed Light

AbstractTo shed light on the epidemiology of ascariasis in Ecuador and Zanzibar, 177 adult worms retrieved by chemo-expulsion from either people or pigs were collected, measured and subjected to polymerase chain reaction–restriction fragment length polymorphism (PCR-RFLP) analysis of the ribosomal internal transcribed spacer (ITS) region. Upon double digestion with RsaI and HaeIII, PCR-RFLP analysis revealed the presence of A. lumbricoides in people and A. suum in pigs in Ecuador. In contrast, while there are no pigs on Zanzibar, of the 56 worms obtained from people, one was genotyped as A. suum. No additional genetic variation was detected upon further PCR-RFLP analysis with several other restriction enzymes. Upon measurement, worm mass and length differed by location and by species, A. suum being lighter and longer. While there is no evidence to suggest zoonotic transmission in Ecuador, an enduring historical signature of previous zoonotic transmission remains on Zanzibar.

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An RFLP-Based Technique for Identifying Fungi in the Sooty Blotch and Flyspeck Complex on Apple

Plant Disease ◽

10.1094/pdis-92-5-0794 ◽

2008 ◽

Vol 92 (5) ◽

pp. 794-799 ◽

Cited By ~ 6

Author(s):

K. B. Duttweiler ◽

G. Y. Sun ◽

J. C. Batzer ◽

T. C. Harrington ◽

M. L. Gleason

Keyword(s):

Agar Plate ◽

Pcr Amplification ◽

Rflp Analysis ◽

Its Region ◽

Morphological Description ◽

Sooty Blotch ◽

Pcr Rflp ◽

Sooty Blotch And Flyspeck ◽

Rflp Assay

A restriction fragment length polymorphism (RFLP)-based technique was developed to identify members of the sooty blotch and flyspeck (SBFS) disease complex on apple because these fungi are difficult to identify using agar-plate isolation and morphological description. The method includes polymerase chain reaction (PCR) amplification of the internal transcribed spacer (ITS) region of ribosomal DNA (rDNA) using a fungal-specific forward primer (ITS1-F) and an SBFS-specific reverse primer (Myc1-R), followed by digestion of the PCR product by the HaeIII restriction enzyme. When applied to previously identified isolates of 24 SBFS-causing species in nine genera, the PCR-RFLP assay produced 14 unique banding patterns. Different genera never shared the same RFLP pattern. To evaluate performance in vivo, the technique was applied to DNA extracted directly from SBFS colonies on apple fruit from three Iowa orchards. The primers amplified the rDNA of only SBFS fungi, with the exception of a Cladosporium sp.; however, its RFLP banding pattern was distinct from those of SBFS fungi. The majority (60%) of SBFS colonies in the in vivo trial were identified to genus by RFLP analysis. The PCR-RFLP assay greatly streamlined the identification process by minimizing the need for culturing, indicating its value as a tool for field studies of the SBFS complex.

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First report of entomopathogenic nematode Steinernema feltiae (Rhabditida: Steinernematidae) from Croatia

Helminthologia ◽

10.2478/helm-2018-0024 ◽

2018 ◽

Vol 55 (3) ◽

pp. 256-260 ◽

Cited By ~ 2

Author(s):

I. Majić ◽

A. Sarajlić ◽

T. Lakatos ◽

T. Tóth ◽

E. Raspudić ◽

...

Keyword(s):

Entomopathogenic Nematodes ◽

Agricultural Land ◽

Entomopathogenic Nematode ◽

Its Region ◽

Original Description ◽

First Record ◽

Steinernema Feltiae ◽

Morphological Identification ◽

Evolutionary Analysis ◽

Different Strains

Summary A survey of entomopathogenic nematodes was conducted in Croatia between 2016 and 2017. The steinernematids were recovered in two out of 100 soil samples from agricultural land characterized as loamy soils with acidic reaction. Molecular and morphological identification was used to distinguish the nematodes. The isolates were identified as two different strains conspecific with Steinernema feltiae. The variations in morphometrical characteristics of infective juveniles (IJs) and males were observed among Croatian strains and with the original description. The analysis of ITS region revealed the greatest similarity of Croatian strains with Slovenian B30 and English A2 strains, which together comprised a monophyletic group in evolutionary analysis. This is the first record of steinernematids, namely S. feltiae in Croatia.

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Genetic Sequence Variation in the ITS Region of Nine Rubus Genotypes

HortScience ◽

10.21273/hortsci.39.4.754a ◽

2004 ◽

Vol 39 (4) ◽

pp. 754A-754 ◽

Cited By ~ 1

Author(s):

Eric Stafne* ◽

John Clark ◽

Allen Szalanski

Keyword(s):

Genetic Variation ◽

Rflp Analysis ◽

Small Variation ◽

Its Region ◽

Genetic Distances ◽

Nuclear Ribosomal Dna ◽

Internal Transcribed Spacer Region ◽

Pcr Rflp ◽

Restriction Sites ◽

Polymerase Chain

In this study, the nuclear ribosomal DNA internal transcribed spacer region (ITS) of six Rubus cultivars were sequenced, then compared with sequences of three Rubus species in Genbank. DNA sequencing revealed little genetic variation among blackberry cultivars, but ably revealed distinctions between blackberry and red raspberry genotypes. Analysis by maximum-parsimony and pairwise genetic distances confirmed the small variation among blackberry cultivars. The resulting sequences were analyzed for useful restriction sites and polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP) analysis was conducted on a total of six cultivars to establish genetic variation. Digests were difficult to interpret due to heterogeneity at restriction sites.

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