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Schwenkbier L.,Institute of Photonic Technology | Schwenkbier L.,Friedrich - Schiller University of Jena | Pollok S.,Institute of Photonic Technology | Pollok S.,Friedrich - Schiller University of Jena | And 9 more authors.
Analytical Methods | Year: 2015

Rapid detection and accurate identification of plant pathogens in the field is an ongoing challenge. In this study, we report for the first time on the development of a helicase-dependent isothermal amplification (HDA) in combination with on-chip hybridization for the detection of selected Phytophthora species. The HDA approach allows efficient amplification of the yeast GTP-binding protein (Ypt1) target gene region at one constant temperature in a miniaturized heating device. The assay's specificity was determined by on-chip DNA hybridization and subsequent silver nanoparticle deposition. The silver deposits serve as stable endpoint signals that enable the visual as well as the electrical readout. Our promising results point to the direction of a near future on-site application of the combined techniques for a reliable detection of Phytophthora species. © 2014 The Royal Society of Chemistry.

Julich S.,Institute of Photonic Technology | Riedel M.,Julius Kuehn Institute Federal Research Center for Cultivated Plants | Kielpinski M.,Institute of Photonic Technology | Urban M.,Institute of Photonic Technology | And 8 more authors.
Biosensors and Bioelectronics | Year: 2011

A lab-on-a-chip system for rapid nucleic acid-based analysis was developed that can be applied for diagnosis of selected Phytophthora species as a first example for use in plant pathology. All necessary polymerase chain reaction process (PCR) and hybridization steps can be performed consecutively within a single chip consisting of two components, an inflexible and a flexible one, with integrated microchannels and microchambers. Data from the microarray is collected from a simple electrical measurement that is based on elementary silver deposition by enzymatical catalyzation. Temperatures in the PCR and in the hybridization zone are managed by two independent Peltier elements. The chip will be integrated in a compact portable system with a pump and power supply for use on site. The specificity of the lab-on-a-chip system could be demonstrated for the tested five Phytophthora species. The two Pythium species gave signals below the threshold. The results of the electrical detection of the microarray correspond to the values obtained with the control method (optical grey scale analysis). © 2011 Elsevier B.V.

Miedaner T.,University of Hohenheim | Mirdita V.,University of Hohenheim | Rodemann B.,Julius Kuehn Institute Federal Research Center for Cultivated Plants | Drobeck T.,Bundessortenamt | Rentel D.,Bundessortenamt
Plant Breeding | Year: 2010

Ergot caused by Claviceps purpurea (Fr.) Tul. is a serious threat to rye (Secale cereale L.) cultivation. It produces sclerotia containing a mixture of alkaloids toxic to animals and humans. For improving ergot resistance in rye a reliable field testing with a minimal disturbance by pollen transfer between plots is indispensable. We evaluated 85 rye cultivars of four types of cultivar (population, synthetic, and hybrid cultivars, hybrids blended with 10% population rye) under inoculation in 3 years (2005, 2006 and 2007) and analysed ergot incidence (% of affected heads) and ergot severity (% of sclerotia in grain by weight) at nine and ten environments (location × year combinations), respectively. Cultivar types were separated by border plots and each entry was additionally surrounded by four border plots of similar size (3-8 m2) in a chess-board design. Disease level ranged from 10-27% affected heads and 0.7-2.6% sclerotia in grain in individual environments. Both, cultivar type and cultivars showed significant (P = 0.01) variation for both traits, interactions with environment were also significant (P = 0.01). Population cultivars had less than half the disease level than the other types of cultivar for both traits. Synthetic, hybrid, and blended hybrid cultivars did on average not differ significantly from each other. Blending had an ergot-reducing effect only for the more susceptible entries. The best two hybrid cultivars showed an ergot reaction similar to some population cultivars. However, the best population cultivar still had only half the amount of sclerotia in grain than the best hybrid cultivar (0.37% vs. 0.89%). Ergot incidence and ergot severity were highly (P = 0.01) correlated. We conclude that a good genotypic differentiation among rye cultivars is achievable by field inoculation of Claviceps purpurea across years with this design and testing within national listing trials is feasible. © 2009 Blackwell Verlag GmbH.

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