Современные методы диагностики некоторых видов трипсов на основе PCR-RFLP и прямого секвенирования

The study reveals diagnosis peculiarities for main Thrips spp. occurring during phytosanitary testing. Morphological diagnostic methods were compared with PCR-RFLP and direct sequencing. High reliability of molecular and genetic methods has been shown. These methods provide for determining the species of thrips larvae and damaged specimens of imagoes.В работе изложены особенности диагностики основных видов трипсов, встречающихся в фитосанитарной экспертизе. Проведено сравнение морфологических методов диагностики с PCR-RFLP и прямым секвенированием. Показана высокая достоверность молекулярно-генетических методов, которые позволяют устанавливать видовую принадлежность личинок трипсов, а также поврежденных имаго

Excavation of an early 17th-century glassmaking site at Glasshouse, Shinrone, Co. Offaly, Ireland

An archaeological research excavation was conducted in the area immediately surrounding an upstanding glassmaking furnace near Shinrone, Co. Offaly, Ireland. It dates to the early to mid 17th century and was built and operated by French Huguenots, probably de Hennezells (de Hennezel/Henzeys/Hensie) who had settled in this region as part of the Crown plantation of King’s County (now Co. Offaly). This furnace, which employed wood rather than coal as a fuel, is a very rare survival, with no other upstanding examples known in Ireland, Britain or the Lorraine region of France where the form probably originated

Контроль достоверности результатов фитосанитарной экспертизы при использовании молекулярных методов диагностики

An internal PCR control system based on a mouse gene fragment cloned into a vector was developed. The created internal control system allowed for choosing a method for DNA extraction from bio-baits which doesn't inhibit the amplification process when Phytophthora sp. is being diagnosed. During the process of diagnosing the pathogen of the ring rot of potato, it was shown that the developed internal control in real-time PCR turned out to be more informative than the mitochondrial gene of a host plant used for that purpose.Разработана система внутреннего контроля (ВК) прохождения ПЦР на основе фрагмента гена мыши, встроенного в вектор. Созданная система ВК позволила выбрать метод выделения ДНК из биоприманок, не ингибирующий процесс амплификации при диагностике Phytophthora sp. При диагностике возбудителя кольцевой гнили картофеля было показано, что разработанный ВК в режиме ПЦР «в реальном времени» оказался более информативным, чем при использовании в качестве ВК митохондриального гена растения-хозяина

Видовое разнообразие фитопатогенных бактерий рода Xanthomonas, поражающих растения семейства мятликовые в России

We reviewed the published data on a composition of plant pathogenic bacteria of the genus Xanthomonas damaging plant of family Poaceae (cereals) in Russia together with our new data obtained from surveys of cereal seeds from different regions of Russia. We assayed the xanthomonads on cereal seeds from different regions and have identified for the first time pathogens similar to species X. hortorum, X. campestris, X. cynarae, X. pisi, and X. gardeneri, which have not been reported yet on the cereal crops

Видовое разнообразие фитопатогенных бактерий рода Xanthomonas, поражающих растения семейства мятликовые в России

We reviewed the published data on a composition of plant pathogenic bacteria of the genus Xanthomonas damaging plant of family Poaceae (cereals) in Russia together with our new data obtained from surveys of cereal seeds from different regions of Russia. We assayed the xanthomonads on cereal seeds from different regions and have identified for the first time pathogens similar to species X. hortorum, X. campestris, X. cynarae, X. pisi, and X. gardeneri, which have not been reported yet on the cereal crops

First report of Agrobacterium vitis causing crown galls of wine grape in Russia

Symptoms of crown gall were observed in 9 of 15 surveyed vineyards located along Black Sea coast of Russia. Sampled tumor tissue was placed in a mortar and pestle for maceration. Serial dilutions of the resulting suspension were plated onto RS medium described by Roy and Sasser (1983). Isolation plates were incubated for 5 days at 28°C until bacteria developed. Colonies were consistent with morphology expected of Agrobacterium spp. On RS medium (opaque red center, translucent margin, mucoid) were purified on yeast dextrose calcium carbonate agar (YDC). Sixty-nine putative Agrobacterium isolates were confirmed by PCR with consensus primers virD2A/2C from the virD2 gene (Haas et al. 1995). Isolates—identified by PCR and producing tumors on indicator plants (carrot, red beet, sunflower) and on grapevine plants in 30 days after needle prick inoculation—were subjected to additional biochemical and physiological tests for Agrobacterium spp. (Moore et al. 2001). The tests included evaluation of 3-ketolactose production, alkaline reaction in litmus milk, growth on 2% and 5% NaCl, growth at 36°C, acid production from erythritol and melezitose, and alkali production from malonic acid and l-tartaric acid. Bacteria reisolated from inoculated grapevine plants were similar to original isolates in PCR test and 3-ketolactose production. The PGF/PGR primers amplifying the chromosomal polygalacturonase gene pehA (Szegedi and Bottka 2002) were used to identify A. vitis isolates and differentiate them from A. tumefaciens. Based on PCR, 18 of 69 tested isolates belonged to A. vitis and showed results of biochemical tests consisted with this species. In addition, for nine isolates, DNA sequence analysis of the housekeeping genes dnaK and trpE confirmed the isolates as A. vitis (Aujoulat et al. 2011). Sequences were deposited in GenBank as Accessions Nos. KT831413 to KT831421 for the dnaK gene and KT831422 to KT831430 for the trpE gene. Sequences were compared with corresponding genes of sequenced strain Agrobacterium vitis S4 (Accession No. CP000633.1). BLAST analysis revealed 99% homology for dnaK and 100% homology for trpE gene. This is the first documented Russian record of Agrobacterium vitis. © 2016, American Phytopathological Society. All rights reserved

First report of Agrobacterium vitis causing crown galls of wine grape in Russia

Symptoms of crown gall were observed in 9 of 15 surveyed vineyards located along Black Sea coast of Russia. Sampled tumor tissue was placed in a mortar and pestle for maceration. Serial dilutions of the resulting suspension were plated onto RS medium described by Roy and Sasser (1983). Isolation plates were incubated for 5 days at 28°C until bacteria developed. Colonies were consistent with morphology expected of Agrobacterium spp. On RS medium (opaque red center, translucent margin, mucoid) were purified on yeast dextrose calcium carbonate agar (YDC). Sixty-nine putative Agrobacterium isolates were confirmed by PCR with consensus primers virD2A/2C from the virD2 gene (Haas et al. 1995). Isolates—identified by PCR and producing tumors on indicator plants (carrot, red beet, sunflower) and on grapevine plants in 30 days after needle prick inoculation—were subjected to additional biochemical and physiological tests for Agrobacterium spp. (Moore et al. 2001). The tests included evaluation of 3-ketolactose production, alkaline reaction in litmus milk, growth on 2% and 5% NaCl, growth at 36°C, acid production from erythritol and melezitose, and alkali production from malonic acid and l-tartaric acid. Bacteria reisolated from inoculated grapevine plants were similar to original isolates in PCR test and 3-ketolactose production. The PGF/PGR primers amplifying the chromosomal polygalacturonase gene pehA (Szegedi and Bottka 2002) were used to identify A. vitis isolates and differentiate them from A. tumefaciens. Based on PCR, 18 of 69 tested isolates belonged to A. vitis and showed results of biochemical tests consisted with this species. In addition, for nine isolates, DNA sequence analysis of the housekeeping genes dnaK and trpE confirmed the isolates as A. vitis (Aujoulat et al. 2011). Sequences were deposited in GenBank as Accessions Nos. KT831413 to KT831421 for the dnaK gene and KT831422 to KT831430 for the trpE gene. Sequences were compared with corresponding genes of sequenced strain Agrobacterium vitis S4 (Accession No. CP000633.1). BLAST analysis revealed 99% homology for dnaK and 100% homology for trpE gene. This is the first documented Russian record of Agrobacterium vitis. © 2016, American Phytopathological Society. All rights reserved