Gene cloning of a thaumatin-like (PR-5) protein of western white pine (Pinus monticola D. Don) and expression studies of members of the PR-5 group |
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| Institution: | 1. Mani L. Bhaumik Institute for Theoretical Physics, Department of Physics and Astronomy, University of California, Los Angeles, CA 90095, USA;2. Department of Mathematics, University of California, Los Angeles, CA 90095, USA;1. Department of Materials Engineering, Indian Institute of Science, Bangalore, 560012, India;2. Department of Chemistry, National Institute of Technology, Durgapur, WB, 713209, India;1. Electrical and Computer Engineering Department, Semnan University, Semnan, Iran;2. Electrical and Computer Engineering Department, University of Tehran, Tehran, Iran;1. College of Biodiversity Conservation and Utilisation, Southwest Forestry University, Kunming, 650224, PR China;2. Institute of Applied Ecology, Chinese Academy of Sciences, Shenyang, 110016, PR China;3. University of Chinese Academy of Sciences, Beijing, 100049, PR China;1. Technical Mechanics Group, Faculty of Mechanical Engineering, Technische Universität Ilmenau, 98684 Ilmenau, Germany;2. Chaif of Magnetofluiddynamics, Measuring and Automation Technology, Technische Universität Dresden, 01062 Dresden, Germany;3. State Scientific Research Institute of Chemistry and Technology of Organoelement Compounds, 111123 Moscow, Russia;1. Plant Reproductive Biology Laboratory, Department of Biochemistry, Cellular and Molecular Biology of Plants, Estación Experimental del Zaidín (CSIC), Profesor Albareda 1, 18008 Granada, Spain;2. Departamento de Biología Molecular y Bioquímica, Universidad de Málaga, Málaga, Spain |
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| Abstract: | To understand the molecular defence response of western white pine to the blister rust pathogen Cronartium ribicola, we have endeavoured to isolate and characterise pathogenesis-related (PR) proteins from western white pine. A full-length cDNA (Pin mTLP) was isolated from a cDNA library constructed from inoculated foliage. BLASTX search results indicated that the sequence shared a high degree of identity with the thaumatin-like proteins (TLPs) of the PR-5 family, and in particular with those of the ‘small-TLP’ subgroup mainly identified in monocots. The mature protein predicted from the cDNA sequence has a molecular mass of 16 kDa and pI of 4.1. Southern blot analysis showed that Pin mTLP is a single-copy gene member of a multi-gene family.Two-dimensional Western blot analysis of total western white pine protein extracts probed with a Douglas-fir anti-TLP antibody detected a major and a minor protein spot of 23 and 17 kDa, respectively. Both 23 and 17-kDa proteins were identified as TLPs by Nanospray MS/MS analysis. The 23-kDa protein was shown to accumulate in canker margins in the bark of infected seedlings indicating that it is locally induced in response to fungal invasion. Both local wounding and methyl jasmonate treatments induced expression of the protein, whereas salicylic acid treatment did not. These results suggest that the 23-kDa TLP plays a role in the molecular defence response of western white pine to wounding and pathogen attack. |
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