Genetic Differentiation of Phytoplasma Isolates Determined by a DNA Heteroduplex Mobility Assay.

Zhong, Qihong; Hiruki, C.

doi:10.2183/pjab.70.127

Cited by 10 publications

(6 citation statements)

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“…HMA has a higher sensitivity for detecting DNA mutations than any other technique currently available, except exhaustive DNA sequencing (Delwart et al ., 1993, 1994; Wang & Hiruki, 1999, 2000a, 2001b). In phytoplasma studies, HMA has been suggested as an accurate means for identifying and classifying phytoplasmas, while other methods such as RFLP do not readily differentiate very closely related phytoplasmas at the subgroup level (Zhong & Hiruki, 1994; Ceranic‐Zagorac & Hiruki, 1996; Wang & Hiruki, 1999, 2000a, 2001a,b).…”

Section: Introductionmentioning

confidence: 99%

Distinctions between phytoplasmas at the subgroup level detected by heteroduplex mobility assay

Wang

Hiruki

2005

Plant Pathology

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A total of 62 phytoplasma isolates were collected from North America, Europe and Asia and analysed by heteroduplex mobility assay (HMA) of the 16/23S spacer region amplified by the polymerase chain reaction. The results revealed wide genetic diversity among the phytoplasmas studied and a number of new phytoplasma strains were identified from known or new plant hosts in Alberta, Canada. Two distinctive subgroups were revealed by HMA in phytoplasmas associated with canola yellows, Chinese aster yellows, dandelion yellows and monarda yellows. In Alberta, two subgroups of the aster yellows group of phytoplasmas, I-A and I-B, were prevalent in naturally infected field crops and ornamentals in open gardens. The results indicated that HMA is a simple, but rapid and accurate, alternative method for the detection and estimation of genetic divergence of phytoplasmas when finer molecular characterization of phytoplasmas is required at the subgroup level.

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Section: Introductionmentioning

confidence: 99%

Distinctions between phytoplasmas at the subgroup level detected by heteroduplex mobility assay

Wang

Hiruki

2005

Plant Pathology

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“…However, the technique of RFLP analysis does not necessarily detect all subtle DNA‐based differences among strains, and does not give a reliable estimation of phylogenetic distance (Seemüller et al ., 1998), while DNA sequencing is expensive and time‐consuming when several samples must be analysed. In contrast, heteroduplex mobility assay (HMA) provides a comparative means of detecting mutations in phytoplasma DNA sequences (Zong & Hiruki, 1994; Ceranic‐Zagorac & Hiruki, 1996; Cousin et al ., 1998; Wang & Hiruki, 1999; Wang & Hiruki, 2000) – it is simple to perform, and can yield a good estimation of phylogenetic distance (Delwart et al ., 1993).…”

Section: Introductionmentioning

confidence: 99%

Phylogenetic relationships among Flavescence dorée strains and related phytoplasmas determined by heteroduplex mobility assay and sequence of ribosomal and Nonribosomal DNA

et al. 2003

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Heteroduplex mobility assay (HMA) and DNA sequencing were performed on Flavescence dorée (FD) phytoplasma strains and related phytoplasmas belonging to the elm yellows group. Part of the ribosomal RNA gene operon and a nonribosomal DNA region were utilized for phylogenetic analyses. Two FD strains, FD92 and FD-D, detected in France and Italy, respectively, were identical in both DNA fragments, confirming previous results. Other FD strains were all very similar and most closely resembled ALY, an Italian alder phytoplasma. Phytoplasmas associated with German Palatinate grapevine yellows were shown to form a distinct subcluster, also different from the elm yellows phytoplasma subcluster. Strain disparities revealed by HMA and sequence data were mostly in agreement, highlighting the utility of HMA in differentiation and classification of phytoplasmas belonging to the same ribosomal RNA group.

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“…In addition, heteroduplex mobilities are proportional to the degree of genetic distance between the two DNA strands (Delwart et al., 1995). Hence, this method has been used for identification, phylogenetic and epidemiological studies of several organisms including viruses (Delwart et al., 1993; Lin et al., 2000), phytoplasma (Zhong and Hiruki, 1994; Cousin et al., 1998), protozoan parasites (Leoni et al., 2002), bacteria (Jensen and Hubner, 1996; Garrec et al., 2003) and fungi (Kumeda and Asao, 2001; Ramos et al., 2001, 2006).…”

Section: Introductionmentioning

confidence: 99%

Heteroduplex Mobility Assay for Identification and Phylogenetic Analysis of Anthracnose Fungi

Huang

Yeh²,

Tzeng

2010

Journal of Phytopathology

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Colletotrichum spp. are casual agents of anthracnose on various economically important crops. To cope with the pitfalls of identifying the fungi by morphotaxonomic criteria, the application of heteroduplex mobility assay (HMA) of internal transcribed spacer (ITS) regions as a biochemical tool was explored. The ITS regions of 29 Colletotrichum isolates including Colletotrichum gloeosporioides, Colletotrichum acutatum, Colletotrichum musae, Colletotrichum graminicola, Colletotrichum capsici, Colletotrichum dematium, Colletotrichum lindemuthianum and three unidentified species of Colletotrichum, were PCR amplified. Comparison of the ITS sequences from 15 Colletotrichum isolates revealed a greater DNA divergence within ITS1 region than that within ITS2. The DNA distance and sequence identity within intra-species ranged from 0.0 to 1.1% and from 98.9 to 100%, respectively; whereas those within inter-species ranged from 1.46 to 13.43% and 90.02 to 98.56%, respectively. From the correlation of DNA distance and relative heteroduplex mobility observed among 15 reference isolates, a formula for estimation of distances of a tested DNA sequence was developed for estimation of DNA distances of a compared strain. The phylogenetic analysis of ITS regions of 29 Colletotrichum isolates using DNA distance inferred from relative heteroduplex mobility divided them into 5 distinctive species groups, namely CG, CA, CC, CM and CL, similar to that assembled based on DNA sequences analysis. Our results show that HMA of ITS regions is a relatively rapid and convenient method for species-specific identification of Colletotrichum spp. The potential use of the established techniques for identification of anthracnose and even other fungal diseases are discussed

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Genetic Differentiation of Phytoplasma Isolates Determined by a DNA Heteroduplex Mobility Assay.

Cited by 10 publications

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Distinctions between phytoplasmas at the subgroup level detected by heteroduplex mobility assay

Distinctions between phytoplasmas at the subgroup level detected by heteroduplex mobility assay

Phylogenetic relationships among Flavescence dorée strains and related phytoplasmas determined by heteroduplex mobility assay and sequence of ribosomal and Nonribosomal DNA

Heteroduplex Mobility Assay for Identification and Phylogenetic Analysis of Anthracnose Fungi

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