Specific and sensitive detection of the conifer pathogen Gremmeniella abietina by nested PCR

BACKGROUND: Gremmeniella abietina (Lagerb.) Morelet is an ascomycete fungus that causes stem canker and shoot dieback in many conifer species. The fungus is widespread and causes severe damage to forest plantations in Europe, North America and Asia. To facilitate early diagnosis and improve measures to control the spread of the disease, rapid, specific and sensitive detection methods for G. abietina in conifer hosts are needed. RESULTS: We designed two pairs of specific primers for G. abietina based on the 18S rDNA sequence variation pattern. These primers were validated against a wide range of fungi and 14 potential conifer hosts. Based on these specific primers, two nested PCR systems were developed. The first system employed universal fungal primers to enrich the fungal DNA targets in the first round, followed by a second round selective amplification of the pathogen. The other system employed G. abietina-specific primers in both PCR steps. Both approaches can detect the presence of G. abietina in composite samples with high sensitivity, as little as 7.5 fg G. abietina DNA in the host genomic background. CONCLUSION: The methods described here are rapid and can be applied directly to a wide range of conifer species, without the need for fungal isolation and cultivation. Therefore, it represents a promising alternative to disease inspection in forest nurseries, plantations and quarantine control facilities

Optically-Nonactive Assorted Helices Array with Interchangeable Magnetic/Electric Resonance

We report here the designing of optically-nonactive metamaterial by assembling metallic helices with different chirality. With linearly polarized incident light, pure electric or magnetic resonance can be selectively realized, which leads to negative permittivity or negative permeability accordingly. Further, we show that pure electric or magnetic resonance can be interchanged at the same frequency band by merely changing the polarization of incident light for 90 degrees. This design demonstrates a unique approach to construct metamaterial.Comment: 15 pages, 4 figure