Leek Proliferation: A New Phytoplasma Disease in the Czech Republic and Italy

During the summer 1996, twelve of twenty-eight leek plants located in a garden near eské Budjovice, South Bohemia exhibited symptoms typical of diseases associated with phytoplasmas. In summer 1998 similar symptoms were detected in leek plants in a field used for seed production located in Romagna, North Italy. In both cases the plants were established in the spring of the previous year. Plants showed flower abnormalities: stamen elongation, anther sterility, pistil proliferation, as well as poor, if any, seed production. Phytoplasma-like structures were detected by scanning and transmission electron microscopy in phloem sieve elements in the Czech diseased plants, but not in healthy ones. Nested-PCR amplifications of extracted DNA with phytoplasma-specific oligonucleotide primer pairs confirmed the presence of phytoplasmas in these plants at low concentrations. Restriction fragment length polymorphism analyses of amplified ribosomal sequences allowed the identification of detected phytoplasmas: all the samples from the Czech Republic contained aster yellows related phytoplasmas (16SrI-B) while in the Italian samples aster yellows related phytoplasmas (16SrI-B) together with stolbur related phytoplasmas (16SrXII-A) were identified. This is the first report of detection and identification of a phytoplasma disease of leek in the Czech Republic and Italy.     

Comparison of phytoplasmas infecting winter oilseed rape in the Czech Republic with Italian Brassica phytoplasmas and their relationship to the aster yellows group

Winter oilseed rape grown in several areas in South Bohemia showed symptoms of stunting, leaf reddening and extensive malformation of floral parts. Phytoplasmas were consistently observed by using electron microscopy only in phloem tissue of symptomatic plants. DNA isolated from infected and healthy control plants was used in PCR experiments. Primer pairs R16F2/R2, P1/P7 and rpF2/R2, amplifying, respectively, 16S rDNA, 16S rDNA plus spacer region and the beginning of the 23S and ribosomal protein gene L22 specific for phytoplasmas, were used. According to RFLP and sequence analyses of PCR products, the phytoplasma from rape was classified in the aster yellows phytoplasma group, subgroup 16SrI-B. The PCR products from the Czech phytoplasma-infected rape also had RFLP profiles identical to those of phytoplasma strains from Italian Brassica . This first molecular characterization of phytoplasmas infecting rape compared with strains from Brassica does not, however, clearly indicate differences among isolates of the same 16SrI-B subgroup. Further studies on other chromosomal DNA portions could help the research on host specificity or on geographical distribution of these phytoplasmas.     

Novel galactolipids from the leaves of Ipomoea batatas L.: characterization by liquid chromatography coupled with electrospray ionization-quadrupole time-of-flight tandem mass spectrometry

Sixteen novel and ten known galactolipids have been isolated and characterized from the leaves of Ipomoea batatas L. (sweet potato) using an analytical method based on high-performance liquid chromatography coupled with electrospray ionization-quadrupole time-of-flight tandem mass spectrometry. Using this technique, the structures and regiochemistries of the fatty acyl groups and the positions of the double bonds on the acyl chains were determined. Sugar moieties were identified by analysis of one- and two-dimensional nuclear magnetic resonance spectra. The positions of the double bonds of polyunsaturated fatty acids were confirmed, and in some cases their geometries determined, by gas chromatography-mass spectrometry. This is the first report of galactolipids in the leaves of sweet potato.     

An oligonucleotide microarray-based assay for identification of phytoplasma 16S ribosomal groups

A method using consensus PCR followed by oligonucleotide microarray hybridization was developed for identification of phytoplasma 16Sr ribosomal groups. The array consisted of 21– to 33-nt-long oligonucleotides which were designed to hybridize to individual 16Sr groups. Two oligonucleotides were designed to detect all phytoplasma groups. The array could efficiently identify samples from 16SrI, 16SrII, 16SrIII, 16SrV, 16SrVI, 16SrVII, 16SrIX, 16SrX and 16SrXII ribosomal groups. This microarray-based test represents a rapid method for detection of phytoplasmas in unknown samples and for identification of most 16Sr groups.     

Note: Molecular identification of ‘Candidatus phytoplasma asteris’ inducing histological anomalies inSilene nicaeensis

Numerous plants ofSilene nicaeensis having symptoms resembling those associated with the presence of phytoplasmas were observed in an extensive coastal area in the south of Italy. Microscopic observation showed histological abnormalities in the organization of tissues in symptomatic plants, and molecular tests, including PCR/RFLP analyses and nucleic acid sequencing, revealed the presence of phytoplasmas belonging to the aster yellows group (‘Candidatus phytoplasma asteris’). This is the first report of phytoplasma infection inS. nicaeensis, a wild species that colonizes the Calabrian coast. http://www.phytoparasitica.org posting June 12, 2008     

Molecular and biologic characterization of a phytoplasma associated with <Emphasis Type="Italic">Brassica campestris</Emphasis> phyllody disease in Punjab province,Pakistan

A phytoplasma-associated disease was identified in Brassica campestris (sarson) plants during a survey conducted in Punjab province of Pakistan in 2014–2016. The symptomatic plants showed characteristic symptoms of phyllody and witches’ broom. Phytoplasma presence was detected by polymerase chain reaction on 16S ribosomal and tuf DNAs, followed by RFLP analysis and sequence comparison of the 16S rRNA and tuf genes. The phytoplasma detected was classified in a new ribosomal subgroup designed 16SrIX-H. The phytoplasma presence in phloem tissues of symptomatic sarson samples was also confirmed through light microscopy and transmission studies to healthy plants through dodder and the leafhopper Orosius albicinctus. This is the first report of identification of 16SrIX-H subgroup phytoplasma associated with sarson and its natural vector in Pakistan.     

Diverse phytoplasmas associated with maize bushy stunt disease in Peru

European Journal of Plant Pathology - Corn plants showing symptoms of midribs chlorosis, and leaf reddening, short internodes, ear proliferation, and plant growth reduction were collected in Peru...     

LC/ESI/MS method for the quantitative detection of guazatine residues in cereals

Guazatine is a fungicide used in agriculture to control a wide range of seed-borne diseases of cereals and other vegetable foods. In this work, a LC-ESI-MS method was developed for the quantitative detection of guazatine residues in maize and hard wheat. Quantitative data were determined for the residues of the main diamines, triamines, and tetramines that cover more than 87% of the total contents of the mixture. The mean recoveries from the fortified cereals at 0.050 mg/kg ranged from 81 to 86%, with the coefficients of variation (CVs) ranging from 0.9 to 5.5% (n = 5). At 0.025 mg/kg, the recoveries ranged from 78 to 87%, with the CVs ranging from 0.8 to 6.3% (n = 5). The limits of quantification have been estimated to be 0.010, 0.004, 0.002, 0.002, 0.005, and 0.002 mg/kg, respectively, for GN, GG, GNG, GGN, GGG, and GGGG in maize and hard wheat (S/N ratio >10).