scholarly journals Widespread Occurrence of Pythium arrhenomanes Pathogenic to Rice Seedlings Around Japanese Rice Fields

Plant Disease ◽  
2015 ◽  
Vol 99 (12) ◽  
pp. 1823-1831 ◽  
Author(s):  
Takeshi Toda ◽  
Akinori Iwasa ◽  
Shinichi Fuji ◽  
Hiromitsu Furuya
Keyword(s):  
Rice Seedling ◽  
Rice Fields ◽  
Rice Seedlings ◽  
Specific Primers ◽  
Widespread Occurrence ◽  
Pythium Arrhenomanes ◽  
Akita Prefecture ◽  
Polymerase Chain ◽  
Species Specific ◽  
Northern Japan

In Japan, rice seedlings grown in nurseries and used for transplanting are subject to a damping-off disease caused by Pythium spp. In this study, 148 isolates of Pythium spp. were obtained from rice seedlings in 39 locations of northern Japan. Among the isolates, 137 were identified as Pythium arrhenomanes using polymerase chain reaction (PCR) with species-specific primers, DNA sequencing analyses of the internal transcribed regions of ribosomal DNA, and the morphologies of oogonia, antheridia, oospores, and zoosporangia. Inoculation tests showed that the isolates identified as P. arrhenomanes were pathogenic to rice seedlings and parasitic to southern crabgrass with only minor damage. P. arrhenomanes was reisolated from the roots of both rice seedlings and southern crabgrass. Poaceae weeds, hosts of Pythium spp., grow in and around nurseries and in ridges surrounding rice fields. To detect Pythium spp., 188 Poaceae weeds were collected from 37 locations in Akita Prefecture. P. arrhenomanes was frequently detected in 164 weed roots from all locations by PCR using species-specific primers. Thus, we determined that P. arrhenomanes exists in and around rice seedling nurseries and rice fields, and that it is much more widely distributed than previously recognized in northern Japan.

scholarly journals Identification and Characterization of Benomyl-Resistant and -Sensitive Populations of Colletotrichum gloeosporioides from Statice (Limonium spp.)

2006 ◽  
Vol 96 (5) ◽  
pp. 542-548 ◽  
Author(s):  
Marcel Maymon ◽  
Aida Zveibil ◽  
Shimon Pivonia ◽  
Dror Minz ◽  
Stanley Freeman
Keyword(s):  
Colletotrichum Gloeosporioides ◽  
Sequence Analyses ◽  
Specific Primers ◽  
Tubulin Genes ◽  
Colletotrichum Spp ◽  
Polymerase Chain ◽  
Species Specific ◽  
Identification And Characterization ◽  
Propagation Material

Sixty-four isolates of Colletotrichum gloeosporioides were isolated from infected Limonium spp. cultivated in 12 different locations in Israel. All isolates were identified as belonging to the C. gloeosporioides complex by species-specific primers. Of these isolates, 46 were resistant to benomyl at 10 μg/ml and 18 were sensitive to this concentration of fungicide. Based on arbitrarily primed polymerase chain reaction of all isolates and internal transcribed spacer-1 sequence analyses of 12 selected isolates, the benomyl-resistant and -sensitive populations belong to two distinct genotypes. Sequence analyses of the β-tubulin genes, TUB1 and TUB2, of five sensitive and five resistant representative isolates of C. gloeosporioides from Limonium spp. revealed that the benomyl-resistant isolates had an alanine substitute instead of a glutamic acid at position 198 in TUB2. All data suggest that the resistant and sensitive genotypes are two independent and separate populations. Because all Limonium plant propagation material is imported from various geographic regions worldwide, and benomyl is not applied to this crop or for the control of Colletotrichum spp. in Israel, it is presumed that plants are bearing quiescent infections from the points of origin prior to arrival.

Microsporidial infections due toEncephalitozoon intestinalisin non-HIV-infected patients with chronic diarrhoea

2011 ◽  
Vol 140 (10) ◽  
pp. 1773-1779 ◽  
Author(s):  
J. YAKOOB ◽  
Z. ABBAS ◽  
M. ASIM BEG ◽  
W. JAFRI ◽  
S. NAZ ◽  
...  
Keyword(s):  
Hepatocellular Carcinoma ◽  
Stool Examination ◽  
Chronic Diarrhoea ◽  
Healthy Controls ◽  
Chain Reaction ◽  
Specific Primers ◽  
Specific Pcr ◽  
Stool Samples ◽  
Polymerase Chain ◽  
Species Specific

SUMMARYWe determined the prevalence of microsporidiaEnterocytozoon(Ent.)bieneusiandEncephalitozoon(E.)intestinalisinfection in patients with chronic diarrhoea and hepatocellular carcinoma (HCC). A total of 330 stool samples were examined from 171 (52%) patients with chronic diarrhoea, 18 (5%) with HCC while 141 (43%) were controls. Stool microscopy, polymerase chain reaction (PCR) with species-specific primers forEnt. bieneusiandE. intestinalisand sequencing were carried out. Microsporidia were found by trichrome staining in 11/330 (3%) andE. intestinalisby PCR in 13/330 (4%) whileEnt. bieneusiwas not detected. PCR forE. intestinaliswas positive in 8/171 (5%) stool samples from patients with chronic diarrhoea, 2/141 (1·4%) samples from healthy controls and in 3/18 (17%) samples from patients with HCC. In the chronic diarrhoea group,E. intestinaliswas positive in 4/171 (2·3%) (P=0·69) stool samples compared to 2/18 (11%) (P=0·06) in the HCC group and 2/141 (1·4%) from healthy controls.E. intestinalisinfection was significantly associated with chronic diarrhoea and HCC in these patients who were negative for HIV. Stool examination with trichrome or species-specific PCR for microsporidia may help establish the cause of chronic diarrhoea.

Detection of cow milk paneer in mixed/buffalo milk paneer through conventional species specific Polymerase Chain Reaction

2016 ◽  
Author(s):  
Tanmay Hazra ◽  
Vivek Sharma ◽  
Rekha Sharma ◽  
S. De ◽  
Sumit Arora ◽  
...  
Keyword(s):  
Buffalo Milk ◽  
Cow Milk ◽  
Market Demand ◽  
Chain Reaction ◽  
Specific Primers ◽  
Mt Dna ◽  
Milk Adulteration ◽  
D Loop ◽  
Polymerase Chain ◽  
Species Specific

Due to higher market demand of buffalo milk paneer, lower price cow milk is often adulterated with higher cost buffalo milk for preparation of paneer. Till date no rapid technique is available in market to ensure that paneer is made from buffalo milk. Currently a PCR based method has been developed to authenticate the buffalo milk paneer. DNA was isolated from paneer by DNeasy Mericon food kit. A set of bovine specific primers (P1) targeting D-loop (displacement loop) of mt- DNA was selected and standardized to amplify cow DNA resulted 126bp amplicon. Using this PCR based approach even upto 1% level of cow milk adulteration in buffalo milk paneer could be detected.

scholarly journals Detection of Pythium ultimum Using Polymerase Chain Reaction with Species-Specific Primers

Plant Disease ◽  
1997 ◽  
Vol 81 (10) ◽  
pp. 1155-1160 ◽  
Author(s):  
K. Kageyama ◽  
A. Ohyama ◽  
M. Hyakumachi
Keyword(s):  
Polymerase Chain Reaction ◽  
Internal Transcribed Spacer ◽  
Pcr Amplification ◽  
Its Region ◽  
Pythium Ultimum ◽  
Pythium Species ◽  
Chain Reaction ◽  
Specific Primers ◽  
Polymerase Chain ◽  
Species Specific

This study was conducted to sequence the rDNA internal transcribed spacer (ITS) region of Pythium ultimum and Pythium group HS, design species-specific primers for polymerase chain reaction (PCR), and detect P. ultimum from diseased seedlings using PCR. The sequence of the ITS region of P. ultimum was identical with that of Pythium group HS. The results support the reports that the HS group is an asexual strain of P. ultimum. Using PCR, the primer pair K1+K3, designed on portions of the sequence of the ITS region, amplified isolates of P. ultimum and the HS group but not isolates of 20 other Pythium species. DNA extracts from damped-off seedlings were not amplified, but a 10-fold dilution of the extracts with Tris-EDTA (TE) buffer diluted the inhibitors and allowed PCR amplification. The primer pair used detected P. ultimum from a single diseased seedling.

Persistence of human mitochondrial DNA throughout the development to the blastocyst of mouse zygotes microinjected with human mitochondria

Zygote ◽  
1999 ◽  
Vol 7 (4) ◽  
pp. 279-283 ◽  
Author(s):  
V.B. Vasilyev ◽  
V.A. Sokolova ◽  
A.V. Sorokin ◽  
M.G. Bass ◽  
N.I. Arbuzova ◽  
...  
Keyword(s):  
Mitochondrial Dna ◽  
Blastocyst Stage ◽  
Preimplantation Embryos ◽  
Hepg2 Cell Line ◽  
Chain Reaction ◽  
Specific Primers ◽  
Human Mitochondrial Dna ◽  
Mouse Zygotes ◽  
Polymerase Chain ◽  
Species Specific

The conditions for transfer of human mitochondria into fertilised mouse ova were elaborated. Species-specific primers were designed to discriminate human mitochondrial DNA (mtDNA) and the endogenous mtDNA in the preimplantation embryos. Human mitochondria isolated from the HepG2 cell line were microinjected into murine zygotes, and the latter cultured for 96 h to the blastocyst stage. The polymerase chain reaction allowed the detection of human mtDNA at every stage of embryo cleavage. In some cases a clear disparity in distribution of human mtDNA among blastomeres was evident.

scholarly journals DNA-Dependent Detection of the Grapevine Fungal Endophytes Aureobasidium pullulans and Epicoccum nigrum

Plant Disease ◽  
2009 ◽  
Vol 93 (10) ◽  
pp. 993-998 ◽  
Author(s):  
M. Martini ◽  
R. Musetti ◽  
S. Grisan ◽  
R. Polizzotto ◽  
S. Borselli ◽  
...  
Keyword(s):  
Aureobasidium Pullulans ◽  
Potential Role ◽  
Fungal Endophytes ◽  
Specific Primers ◽  
Variable Regions ◽  
Epicoccum Nigrum ◽  
Polymerase Chain ◽  
Species Specific ◽  
Reference Strains ◽  
Its1 And Its2

Aureobasidium pullulans and Epicoccum nigrum are frequently reported as endophytes of various crops, including grapevine (Vitis vinifera). Because of their potential role as biological control agents against grapevine pathogens, we examined the occurrence of A. pullulans and E. nigrum in two grapevine varieties (Merlot and Prosecco) in Italian vineyards where spontaneous recovery from phytoplasma disease is recurrent. Species-specific primers for A. pullulans and two genetically distinct strains of E. nigrum were designed in variable regions of ITS1 and ITS2. Primer specificity was confirmed by polymerase chain reaction using purified DNA from other fungal endophytes that are usually encountered during isolation attempts from grapevine tissues and from several other strains of A. pullulans and E. nigrum isolated from other sources. In order to determine the occurrence of the two endophytes in grapevine plants, DNA was extracted from shoots of 44 grapevines collected in six vineyards from different localities of northeast Italy. Both endophytes were detected and their identity was confirmed by restriction fragment length polymorphism (RFLP) patterns obtained from reference strains. RFLP analyses confirmed the presence of two E. nigrum strains belonging to different RFLP groups in grapevine. The molecular methods described allowed a sensitive, specific, and reliable identification of the two endophytes in grapevine.

scholarly journals Curly Top Survey in the Western United States

2008 ◽  
Vol 98 (11) ◽  
pp. 1212-1217 ◽  
Author(s):  
C. A. Strausbaugh ◽  
W. M. Wintermantel ◽  
A. M. Gillen ◽  
I. A. Eujayl
Keyword(s):  
United States ◽  
Host Resistance ◽  
Field Trials ◽  
Western United States ◽  
Management Options ◽  
Specific Primers ◽  
Beet Curly Top Virus ◽  
Polymerase Chain ◽  
Species Specific ◽  
Polymerase Chain Reaction Primers

Curly top in sugar beet continues to be a challenging disease to control in the western United States. To aid in development of host resistance and management options, the curtovirus species composition was investigated by sampling 246 commercial fields along with nursery and field trials in the western United States. DNA was isolated from leaf samples and the species were identified using species-specific polymerase chain reaction primers for the C1 gene. Amplicons from 79 isolates were also sequenced to confirm identifications. Beet severe curly top virus (BSCTV) and Beet mild curly top virus (BMCTV) were widely distributed throughout the western United States, while only a few isolates of Beet curly top virus (BCTV) were found. In phylogenetic analysis, BSCTV, BMCTV, and BCTV isolates formed distinct groups in the dendrogram. Seven isolates not amplifiable with species-specific primers did amplify with curly top coat protein primers, indicating novel curtovirus species or strains may be present. Given the wide host range of the viruses responsible for curly top, frequent co-infections, and genetic diversity within and among species, establishing better host resistance, and controlling curly top will continue to be a challenge.

scholarly journals Morphological and Molecular Identification of Fusarium oxysporum Sch. Isolated From Guava Wilt in Bangladesh

2012 ◽  
Vol 41 (1) ◽  
pp. 49-54 ◽  
Author(s):  
M Zakir Hussain ◽  
MA Rahman ◽  
Mohammad Nurul Islam ◽  
MA Latif ◽  
MA Bashar
Keyword(s):  
Fusarium Oxysporum ◽  
Psidium Guajava ◽  
18S Rrna Gene ◽  
Rrna Gene ◽  
Chain Reaction ◽  
Species Identity ◽  
Specific Primers ◽  
Polymerase Chain ◽  
Serious Disease ◽  
Species Specific

Wilt of guava plants (Psidium guajava L.) is a serious disease in Bangladesh. Sixteen isolates of Fusarium oxysporum Sch. were collected from the root and stem fragments of guava plants growing in six districts of Bangladesh. Species identity was based on the colony character, nature of conidiogenous cell, morphology of microconidia, macroconidia and chlamydospores. Eleven isolates were confirmed as F. oxysporum through polymerase chain reaction (PCR) using species specific primers designed from the conserved regions of 18S rRNA gene. DOI: http://dx.doi.org/10.3329/bjb.v41i1.11082 Bangladesh J. Bot. 41(1): 49-54, 2012 (June)

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