Detection of Cercospora beticola by PCR in amended and naturally infested field soil.

2007 ◽  
Author(s):  
Robert E. Lartey ◽  
TehCan Caesar-TonThat ◽  
Sophia Hanson ◽  
William M. Iversen ◽  
Robert G. Evans
Keyword(s):  
Cercospora Beticola ◽  
Field Soil ◽  
Infested Field

scholarly journals Direct Polymerase Chain Reaction-Based Detection of Cercospora beticola in Field Soils

Plant Disease ◽  
2010 ◽  
Vol 94 (9) ◽  
pp. 1100-1104 ◽  
Author(s):  
R. T. Lartey ◽  
T. C. Caesar-TonThat ◽  
A. W. Lenssen ◽  
J. Eckhoff ◽  
S. L. Hanson ◽  
...  
Keyword(s):  
Polymerase Chain Reaction ◽  
Sugar Beet ◽  
Fragment Size ◽  
Rapid Screening ◽  
Cercospora Beticola ◽  
Field Soil ◽  
Chain Reaction ◽  
Field Soils ◽  
Total Dna ◽  
Polymerase Chain

Cercospora beticola, the causal agent of Cercospora leaf spot of sugar beet, survives as pseudostromata in infected sugar beet residues in the soil. Under optimal conditions, overwintering propagules germinate and produce conidia that are dispersed as primary inoculum to initiate infection in sugar beet. We developed a polymerase chain reaction (PCR) technique for rapid detection of C. beticola in field soils. Total DNA was first isolated from soil amended with C. beticola culture using the PowerSoil DNA Kit. The purified DNA was subjected to PCR in Extract-N-Amp PCR mix with CBACTIN primers over 35 cycles. The amplified products were resolved and compared by electrophoresis in 1% agarose gels. The PCR fragment size of C. beticola from the amended field soil correlated in size with the amplicon from control C. beticola culture DNA extract. Additionally, sample soils were collected from sugar beet fields near Sidney, MT and Foxholm, ND. Total DNA was extracted from the samples and subjected to PCR and resolved as previously described. The amplicons were purified from the gels and subjected to BigDye Terminator Cycle sequencing. All sequences from field soils samples, C. beticola-amended field soil, and pure culture were compared by alignment with a C. beticola actin gene sequence from GenBank. The result of the alignment confirmed the amplicons as products from C. beticola. Rapid screening for the presence of C. beticola in the soil by PCR will improve research capabilities in biological control, disease forecasting, and management of this very important sugar beet pathogen.

scholarly journals CRISPR-Based Isothermal Next-Generation Diagnostic Method for Virus Detection in Sugarbeet

2021 ◽  
Vol 12 ◽  
Author(s):  
Vanitharani Ramachandran ◽  
John J. Weiland ◽  
Melvin D. Bolton
Keyword(s):  
Virus Detection ◽  
Detection Methods ◽  
Target Sequence ◽  
Field Soil ◽  
Human Virus ◽  
Guide Rna ◽  
Highly Sensitive ◽  
Infested Field ◽  
Virus Diagnostics

Rhizomania is a disease of sugarbeet caused by beet necrotic yellow vein virus (BNYVV) that significantly affects sugarbeet yield globally. Accurate and sensitive detection methods for BNYVV in plants and field soil are necessary for growers to make informed decisions on variety selection to manage this disease. A recently developed CRISPR-Cas-based detection method has proven highly sensitive and accurate in human virus diagnostics. Here, we report the development of a CRISPR-Cas12a-based method for detecting BNYVV in the roots of sugarbeet. A critical aspect of this technique is the identification of conditions for isothermal amplification of viral fragments. Toward this end, we have developed a reverse transcription (RT) recombinase polymerase amplification (RPA) for detecting BNYVV in sugarbeet roots. The RT-RPA product was visualized, and its sequence was confirmed. Subsequently, we designed and validated the cutting efficiency of guide RNA targeting BNYVV via in vitro activity assay in the presence of Cas12a. The sensitivity of CRISPR-Cas12a trans reporter-based detection for BNYVV was determined using a serially diluted synthetic BNYVV target sequence. Further, we have validated the developed CRISPR-Cas12a assay for detecting BNYVV in the root-tissue of sugarbeet bait plants reared in BNYVV-infested field soil. The results revealed that BNYVV detection is highly sensitive and specific to the infected roots relative to healthy control roots as measured quantitatively through the reporter signal. To our knowledge, this is the first report establishing isothermal RT-RPA- and CRISPR-based methods for virus diagnostic approaches for detecting BNYVV from rhizomania diseased sugarbeet roots.

Flaming to Reduce Inocula of the Boxwood Blight Pathogen, Cylindrocladium pseudonaviculatum, in Field Soil

2012 ◽  
Vol 13 (1) ◽  
pp. 33 ◽  
Author(s):  
Norman L. Dart ◽  
Sarah M. Arrington ◽  
Sarah M. Weeda
Keyword(s):  
Significant Role ◽  
Plant Tissue ◽  
Field Soil ◽  
Infested Plant ◽  
Infested Field

Infested plant tissue can play a significant role in the epidemiology of boxwood blight in ornamental nursery systems. In an effort to reduce inoculum levels in an infested field in Carroll Co., Virginia, symptomatic plants were removed, collected into a pile, and destroyed by burning using a propane push flamer. The authors investigated whether soil flaming would be effective to reduce viable inocula of C. pseudonaviculatum in the upper layer of soil. Accepted for publication 4 October 2012. Published 26 October 2012.

scholarly journals Response of cucurbit species to Rotylenchulus reniformis

2005 ◽  
Vol 30 (1) ◽  
pp. 85-87 ◽  
Author(s):  
Gustavo R. C. Torres ◽  
Elvira M. R. Pedrosa ◽  
Kércya M. S. Siqueira ◽  
Romero M. Moura
Keyword(s):  
Cucumis Melo ◽  
Citrullus Lanatus ◽  
Control Measures ◽  
Effective Control ◽  
Field Soil ◽  
Rotylenchulus Reniformis ◽  
Nematode Reproduction ◽  
Infested Field ◽  
High Reduction ◽  
Shoot Mass

The state of Rio Grande do Norte is the major melon (Cucumis melo) producer in Brazil and the reniform nematode Rotylenchulus reniformis has become increasingly important due to damages to that crop and the lack of resistant cultivars and effective control measures. Under these circumstances, eight cucurbit genotypes were screened for resistance or tolerance to a population of R. reniformis in naturally infested field soil, under greenhouse conditions. A high reduction in shoot mass was found in all infected genotypes. Watermelon (Citrullus lanatus) cv. Sugar Baby showed the lowest rate of nematode reproduction.

A pythium root rot of muskmelon

1968 ◽  
Vol 46 (10) ◽  
pp. 1165-1171 ◽  
Author(s):  
C. D. McKeen ◽  
H. J. Thorpe
Keyword(s):  
Experimental Data ◽  
Root Rot ◽  
Pythium Ultimum ◽  
Wilt Disease ◽  
Field Soil ◽  
Southern Ontario ◽  
Pythium Root Rot ◽  
High Soil ◽  
Infested Field ◽  
Soil Temperatures

Pythium ultimum was readily isolated from the necrotic roots of young and mature muskmelon plants growing in soil in which root rot had been severe. Muskmelons planted in steamed soils inoculated with P. ultimum developed necrotic roots and aboveground symptoms closely similar to those produced in naturally infested field soil. The fungus was considerably more pathogenic at low than at high soil temperatures. All of seven commercial varieties of muskmelon commonly grown in southern Ontario were moderately to highly susceptible to P. ultimum. Experimental data support the conclusion that P. ultimum probably plays an important role in the "sudden wilt" disease of mature muskmelon plants.

scholarly journals Detection of Soilborne Alternaria radicina and Its Occurrence in California Carrot Fields

Plant Disease ◽  
1998 ◽  
Vol 82 (8) ◽  
pp. 891-895 ◽  
Author(s):  
B. M. Pryor ◽  
R. M. Davis ◽  
R. L. Gilbertson
Keyword(s):  
Selective Medium ◽  
Causal Agent ◽  
Black Rot ◽  
Field Soil ◽  
Positive Correlation ◽  
Organic Debris ◽  
Selective Agar ◽  
Infested Field ◽  
Dry Soil ◽  
Rot Disease

Alternaria radicina, causal agent of black rot disease of carrot, was recovered from soil by plating dilutions on a semi-selective medium, A. radicina semi-selective agar. The efficiency of this soil assay was 93% based on recovery of the fungus from non-infested field soil amended with A. radicina conidia. Soilborne A. radicina was recovered from five of six carrot-growing areas in California, but was only commonly found in the Cuyama Valley, where the fungus was detected in 83% of sampled fields. Over a 3-year period of sampling, A. radicina soil populations in Cuyama Valley fields prior to carrot planting ranged from 0 to 317 CFU/g. There was a positive correlation between A. radicina soil populations in these fields and the incidence of black rot disease at harvest. A. radicina was recovered from dry soil after 4 years of storage, and the fungus survived in this soil as solitary conidia or as conidia associated with organic debris.

scholarly journals Management of Diseases Induced by Soil- Borne Pathogens, Solarization and Biological Control

1998 ◽  
Vol 3 (1) ◽  
pp. 65
Author(s):  
C. Alabouvette
Keyword(s):  
Biological Control ◽  
Population Density ◽  
Plant Pathogens ◽  
Field Soil ◽  
Environment Friendly ◽  
Soil Borne Pathogens ◽  
Beneficial Microorganisms ◽  
Antagonistic Microorganisms ◽  
Infested Field ◽  
Selection Of

Diseases induced by soil-borne plant pathogens are among the most difficult to control. Prophylactic methods aiming at preventing the introduction of pathogens in healthy soils have to be respected, because it is almost impossible to eradicate pathogens from an infested field soil. Even the drastic disinfestation techniques based on the application of biocide molecules such as methylbromide failed to eliminate the pathogens, but are harmful to man and the environment. Growers should prefer new, environment friendly techniques such as solarization and biological disinfestation of soils. These methods induce changes in the microbial balance, reducing the population density of the pathogens and stimulating the activity of some beneficial microorganisms. The study of soils that naturally suppress diseases induced by soil-borne pathogens has led to a better understanding of the interactions between pathogenic and antagonistic microorganisms and has resulted in the selection of bio control agents.

Effects of Host Age on Development of Cavity Spot Disease of Carrots Caused by Pythium coloratum in Western Australia Khaled

1997 ◽  
Vol 45 (4) ◽  
pp. 727 ◽  
Author(s):  
A. El-Tarabily ◽  
Giles E. St J. Hardy ◽  
Krishnapillai Sivasithamparam
Keyword(s):  
Western Australia ◽  
Infested Soil ◽  
Host Age ◽  
Field Soil ◽  
Spot Disease ◽  
Cavity Spot ◽  
Infested Field ◽  
Time Of Infection ◽  
Pythium Coloratum ◽  
Time Of Harvest

Three experiments were conducted with Pythium coloratum Vaartaja, a causal agent of cavity spot disease of carrots in Western Australia, to study the relationships between host age, time of infection and development of cavity spot lesions. Pythium coloratum was isolated frequently from 3-6-week-old asymptomatic roots of carrots grown in soils infested naturally or artificially with the pathogen. Carrots grown in containers of soil artificially infested with P. coloratum, but not those in naturally infested field soil, developed cavity spot lesions after 6 weeks. Early infection of carrot seedlings at or before 3 weeks by P. coloratum in artificially infested soils followed by their transfer to pathogen-free soil was sufficient to cause cavity spot disease at the time of harvest (16 weeks). The disease levels in this treatment were not different from those transferred to P. coloratum-infested soil. There was no significant (P > 0.05) association between carrot age and the ability of P. coloratum to infect the roots and to cause cavity spot lesions at harvest.

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