Detection and differentiation of phytoplasmas in Australia

1997 ◽  
Vol 48 (5) ◽  
pp. 535 ◽  
Author(s):  
R. I. Davis ◽  
B. Schneider ◽  
K. S. Gibb
Keyword(s):  
Sweet Potato ◽  
Restriction Site ◽  
Fragment Length Polymorphism ◽  
Aster Yellows ◽  
Site Analysis ◽  
Blot Hybridisation ◽  
Total Dna ◽  
Polymerase Chain ◽  
Southern Blot Hybridisation ◽  
Polymorphism Pattern

In a polymerase chain reaction (PCR) diagnostic test, phytoplasma (formerly known as plant-pathogenic mycoplasma-like organism or MLO) ribosomal DNA was detected in total DNA extracts prepared from 56 out of 63 plants collected from geographically diverse locations across Australia. The list of phytoplasma hosts consisted of 38 different species in 16 different families. Restriction site analysis of the PCR-amplified DNA accessions was used to divide the phytoplasmas into 2 groups. The majority of the tomato big bud group and sweet potato little leaf group phytoplasmas were closely related to a phytoplasma originally obtained from Crotalaria in Thailand, which is a member of the faba bean phyllody strain cluster. In contrast, phytoplasmas associated with Australian grapevine yellows and papaya dieback were most similar to members of the aster yellows strain cluster. Twelve phytoplasmas were compared by Southern blot hybridisation with DNA cloned from the sweet potato little leaf phytoplasma strain V4. The restriction fragment length polymorphism pattern of all phytoplasmas compared was identical except for 2 sweet potato little leaf phytoplasmas.

scholarly journals Molecular Characterization and Classification of Phytoplasmas Associated with Canola Yellows and a New Phytoplasma Strain Associated with Dandelions

Plant Disease ◽  
2001 ◽  
Vol 85 (1) ◽  
pp. 76-79 ◽  
Author(s):  
Keri Wang ◽  
Chuji Hiruki
Keyword(s):  
Fragment Length Polymorphism ◽  
Rflp Analysis ◽  
Universal Primers ◽  
Dna Fragments ◽  
Chain Reaction ◽  
Aster Yellows ◽  
Distinct Subgroup ◽  
16S Ribosomal Dna ◽  
Polymerase Chain

DNA isolated from symptomatic canola (Brassica napus, Brassica rapa) and dandelion (Taraxacum officinale) was used to amplify 16S ribosomal DNA fragments by polymerase chain reaction using two pairs of universal primers P1/P6 and R16F2n/R2. Restriction fragment length polymorphism (RFLP) analysis of the amplified DNA fragments using endonucleases AluI, HhaI, HpaII, MseI, RsaI, and Sau 3AI revealed two distinct types of phytoplasmas in canola with similar symptoms. One had the same RFLP profiles as the phytoplasmas in subgroup 16SrI-A, whereas the other one had RFLP profiles similar to those of phytoplasmas in subgroup 16SrI-B. Phytoplasmas were detected in symptomatic dandelion plants that were collected from canola and alfalfa fields where severe alfalfa witches'-broom occurred. Comparative studies indicated that two different phytoplasmas were associated with the dandelion plants. One was identified as a member of subgroup 16SrI-A, whereas another one was classified as a member of a distinct subgroup in the aster yellows group on the basis of the unique RFLP patterns.

scholarly journals Molecular and Symptom Analyses of Phytoplasma Strains from Lettuce Reveal a Diverse Population

2004 ◽  
Vol 94 (8) ◽  
pp. 842-849 ◽  
Author(s):  
Jianhua Zhang ◽  
Saskia A. Hogenhout ◽  
Lowell R. Nault ◽  
Casey W. Hoy ◽  
Sally A. Miller
Keyword(s):  
Fragment Length Polymorphism ◽  
Phylogenetic Analyses ◽  
The Other ◽  
16S Rrna Genes ◽  
Rrna Genes ◽  
Aster Yellows ◽  
Pcr Rflp ◽  
Horizontal Growth ◽  
Polymerase Chain ◽  
Leaf Distortion

Epidemics of aster yellows in lettuce in Ohio are caused by at least seven distinct phytoplasma strains in the aster yellows (AY) group. Five of the strains are newly reported: AY-BW, AY-WB, AY-BD3, AY-SS, and AY-SG. All seven strains were characterized based on symptoms in aster and lettuce, and by polymerase chain reaction (PCR). Strain AY-BD2 (formerly ‘Bolt’) causes yellowing and leaf distortion in lettuce and bolting in aster, whereas strain AY-S (formerly ‘Severe’) causes stunting, leaf clustering, and phyllody. Strain AY-WB causes yellowing and wilting in lettuce and witches'-broom in aster. Strain AY-SG induces horizontal growth in lettuce and aster plants. Strain AY-BW causes chlorosis of emerging leaves and abnormally upright growth of leaf petioles. AY-SS causes symptoms similar to those caused by AY-S but has a different PCR-restriction fragment length polymorphism (RFLP) banding pattern. Strains AY-BD2 and AY-BD-3 cause mild leaf and stem distortion in lettuce but are differentiated by PCR-RFLP. All phytoplasma strains collected from lettuce in Ohio belong to the 16SrI group. AY-WB belongs to the 16SrI-A subgroup and the other six belong to the 16SrI-B subgroup. Five of the seven strains were distinguished from each other by primer typing. The results of phylogenetic analyses of sequences of the 16S rRNA genes were basically consistent with the classification based on PCR-RFLP, in which AY-WB clustered with phytoplasmas of the 16rIA subgroup and the other Ohio lettuce strains clustered with phytoplasmas in the 16SrI-B subgroup.

An intracellular symbiont and other microbiota associated with field-collected populations of sawflies (Hymenoptera: Symphyta)

2008 ◽  
Vol 54 (9) ◽  
pp. 758-768 ◽  
Author(s):  
Robert I. Graham ◽  
Viviane Zahner ◽  
Christopher J. Lucarotti
Keyword(s):  
Fragment Length Polymorphism ◽  
Denaturing Gradient Gel Electrophoresis ◽  
Rrna Gene ◽  
Chain Reaction ◽  
Intracellular Symbiont ◽  
Mountain Ash ◽  
Pcr Rflp ◽  
Gradient Gel Electrophoresis ◽  
Total Dna ◽  
Polymerase Chain

Six species of sawfly (Hymenoptera: Symphyta) from four taxonomic families (Agridae, Diprionidae, Pamphiliidae, and Tenthredinidae) were collected from locations across Canada and surveyed for their associated microbiota. Total DNA was extracted from individual insects, and polymerase chain reaction (PCR) was used to amplify the conserved 16S rRNA gene from microbiota. Denaturing gradient gel electrophoresis (DGGE) and restriction fragment length polymorphism (RFLP) were undertaken to separate bacterial clones associated with the host insect. Sequencing of the PCR–DGGE and PCR–RFLP products revealed a dominance of α- and γ-Proteobacteria, with most sequences showing high similarity to bacteria previously identified from other insect species and environmental samples. Additionally, a strain of the bacterial endosymbiont Wolbachia and a Wolbachia bacteriophage were identified from the mountain ash sawfly ( Pristiphora geniculata ).

MUTATION IN THE FMO3 LOCUS AND ITS DISTRIBUTION IN A GROUP OF AYRSHIRE BREEDING BULLS USED IN FARMS OF THE KRASNODAR TERRITORY

2021 ◽  
pp. 10-15
Author(s):  
Н.В. КОВАЛЮК ◽  
Е.В. ШИРЯЕВА ◽  
Л.И. ЯКУШЕВА ◽  
Ю.Ю. ШАХНАЗАРОВА
Keyword(s):  
Polymerase Chain Reaction ◽  
Mutant Allele ◽  
Restriction Site ◽  
Fragment Length Polymorphism ◽  
Test System ◽  
Frequency Of Occurrence ◽  
Wild Type ◽  
Chain Reaction ◽  
Polymerase Chain ◽  
Restriction Fragment Length

Рыбный привкус в коровьем молоке вызван наличием нонсенс-мутации (g.39523051C>T) в гене бычьего FMO3. Нами разработана тест-система для выявления FMO3- полиморфизма, основанная на полимеразной цепной реакции с последующим анализом полиморфизма длин фрагментов рестрикции с использованием эндонуклеазы TaqI. Фрагменты, которые амплифицировались с участка гена FMO3 «дикого» типа, расщеплялись эндонуклеазой TaqI  на 2 фрагмента: 136 и 99 пн. Фрагменты, амплифицированные с мутантного аллеля, сайта рестрикции не имели (их размер составлял  235 пн). Определена частота встречаемости носителей мутации в отечественной субпопуляции айрширского скота. Установлено, что среди айрширских быков-производителей (n=45), принадлежащих различным отечественным и зарубежным племорганизациям, частота встречаемости носителей мутации в гене FMO составила 9%. Учитывая, что выявленные носители мутации интенсивно используются и могут передавать эту аномалию значительному числу дочерей, генотипирование по локусу FMO3 должно стать обязательным для быков-производителей и групп быкопроизводящих коров племенных айрширских хозяйств. The fishy taste in cow's milk is caused by the presence of a nonsense mutation (g.39523051C>T) in the bovine FMO3 gene. We have developed a test system for detecting FMO3 polymorphism based on a polymerase chain reaction followed by analysis of restriction fragment length polymorphism using TaqI endonuclease. Fragments that were amplified from the wild-type FMO3 gene site were cleaved by TaqI endonuclease into 2 fragments: 136 and 99 bp. The fragments amplified from the mutant allele did not have a restriction site (their size was — 235 bp). The frequency of occurrence of mutation carriers in the domestic subpopulation of Ayrshire cattle was determined. It was found that among Ayrshire bulls (n=45) belonging to various domestic and foreign breeding organizations, the frequency of occurrence of carriers of the mutation in the FMO gene was 9%. Given that the identified carriers of the mutation are intensively used and can transmit this anomaly to a significant number of daughters, genotyping by the FMO3 locus should become mandatory for breeding bulls and groups of bull-producing cows of breeding Ayrshire farms.

scholarly journals Survey of mutations in prolificacy genes in Santa Ines and Morada Nova sheep

2017 ◽  
Vol 69 (4) ◽  
pp. 1047-1053
Author(s):  
G.M.L. Holanda ◽  
J.C. Oliveira ◽  
D.M.F. Silva ◽  
S.S.N. Rocha ◽  
V. Pandolfi ◽  
...  
Keyword(s):  
Restriction Site ◽  
Fragment Length Polymorphism ◽  
Nucleotide Sequencing ◽  
Chain Reaction ◽  
Specific Primers ◽  
Pcr Rflp ◽  
Pcr Products ◽  
Polymerase Chain ◽  
Santa Inês ◽  
Restriction Fragment Length

ABSTRACT Polymorphisms in the BMP-15 gene related to Galway (FecXG) and Inverdale (FecXI) and in the BMPR-1B gene known as Booroola (FecB) mutations were investigated using the Polymerase Chain Reaction - Restriction Fragment Length Polymorphism (PCR-RFLP) method, on sheep from the breeds Santa Inês (n= 574) and Morada Nova (n=282). DNA was extracted and amplified through PCR with specific primers that introduced a restriction site in association with the mutation. The PCR products were submitted to endonucleases. The experiment found no FecXG and FecXI mutations. Six samples of animals with multiple offspring/birth history presented polymorphism for FecB similar to control samples, but this pattern was not confirmed by nucleotide sequencing. Although the absence of these mutations in the studied breeds, other factors related to prolificacy should be investigated to explain the inherent prolificity mechanisms.

scholarly journals Detection and Differentiation of Phytoplasmas Associated with Diseases of Strawberry in Florida

Plant Disease ◽  
1997 ◽  
Vol 81 (2) ◽  
pp. 230-230 ◽  
Author(s):  
N. A. Harrison ◽  
D. E. Legard ◽  
R. DiBonito ◽  
P. A. Richardson
Keyword(s):  
Ribosomal Protein ◽  
Ribosomal Rna ◽  
Fragment Length Polymorphism ◽  
Gene Products ◽  
Aster Yellows ◽  
Central Florida ◽  
Phytoplasma Infection ◽  
Polymerase Chain ◽  
Subgroup Member ◽  
Clover Phyllody

Strawberry (Fragaria × ananassa Duchesne) plants with symptoms suggestive of phytoplasmal disease were identified in commercial fields and a breeder's plot in west central Florida during the 1995 to 1996 winter growing season. Affected plants were all conspicuously stunted and unproductive. Primary symptoms on cvs. Rosa Linda and Carlsbad and on a breeder's accession resembled those of strawberry green petal (SGP). Plants displayed sparse clusters of virescent flowers with enlarged sepals and phylloid receptacles that failed to develop fully into fleshy structures or redden on ripening. Symptoms on cv. Oso Grande were more typical of multiplier disease and included a proliferation of branch crowns producing numerous small leaves with spindly petioles. Oso Grande and Carlsbad originated as transplants from a nursery in Montreal, Canada, whereas Rosa Linda transplants were from Nova Scotia. Plants were assessed for phytoplasma infection by polymerase chain reaction with total DNAs from leaves and petioles as template and phytoplasma-specific ribosomal RNA primers P1 and P7 (3), or mollicute-specific ribosomal protein (rp) gene primers rpF1 and rpR4 (2). Amplification of a 1.8-kb rDNA or 1.2-kb rp gene product, respectively, confirmed infection of Rosa Linda (7 of 7 plants), Carlsbad (3 of 7), Oso Grande (4 of 4), and a single breeder's accession. No products were amplified from DNAs of healthy plants. Restriction fragment length polymorphism patterns of rDNA digested with AluI, EcoRI, HaeIII, HhaI, HpaII, KpnI, ScaI, or Tru9I endonucleases, or of rp gene products digested with AluI, DraI, RsaI, TaqI, or Tru9I, revealed no differences among phytoplasma strains affecting both Rosa Linda and Carlsbad. Collectively, patterns were comparable to those of clover phyllody and SGP phytoplasmas, two Canadian strains previously classified as members of phytoplasma 16S rRNA (rr)-ribosomal protein (rp) group 16S rI, subgroup C (16S rI-C (rr-rp)) (1). Similarly, no differences were evident among phytoplasmas associated with all four diseased Oso Grande plants. Both rDNA and rp fragment profiles associated with this cultivar were characteristic of strains such as tomato big bud and eastern aster yellows delineated as 16S rI-A (rr-rp) subgroup members (1). However, AluI rDNA and TaqI rp fragment patterns were unique, identifying Oso Grande-infecting strains as representatives of a new subgroup within the larger 16S rI (rr-rp) group. Cumulative rDNA and rp fragment profiles of the phytoplasma associated with the breeder's accession matched those of the Mexican periwinkle virescence phytoplasma, identifying this strain as a 16S rI-I (rr-rp) subgroup member (1) and a second possible etiological agent of SGP. This is the first report of phytoplasmas infecting strawberry in Florida. References: (1) D. E. Gundersen et al. Int. J. Syst. Bacteriol. 46:64, 1996. (2) P.-O. Lim and B. B. Sears. J. Bacteriol. 174:2602, 1993. (3) C. D. Smart et al. Appl. Environ. Microbiol. 62:2988, 1996.

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