| Autor: |
Prabhakaran, Narayanasamy, Kumar, Aundy, Sheoran, Neelam, Singh, Vaibhav Kumar, Nallathambi, Perumal |
| Zdroj: |
Journal of Plant Diseases and Protection; April 2021, Vol. 128 Issue: 2 p457-469, 13p |
| Abstrakt: |
Fluorescence microscopy and qPCR-based pathogen tracking tools were developed to elucidate the growth and development of Puccinia graminisf. sp. triticion leaves of nonhost plant (rice: Oryza sativaL.). Stem rust race-Puccinia graminisf. sp. tritici40A was identified using Internal Transcribed Spacer (ITS) sequences and gene sequences coding for hypothetical protein (PGTG_08233_hypothetical protein, HP). A reliable qPCR-based quantitation assay was developed for stem rust fungus exploiting HP primers that yielded 377-bp amplicon in conventional PCR and a forma specialis-specific 146 bp in qPCR with a detection sensitivity of 250-fg genomic DNA. The fungal cell wallN-acetyl-glucosamine was green fluorescence labelled and visualized on propidium iodide-stained nonhost leaf. This technique along with scanning electron microscopy allowed imaging of various developmental structures of Puccinia graminisf. sp. tritici40A on plant epiphytic and endophytic niches. The microscopy coupled with qPCR-based pathogen load estimation revealed that the nonhost (rice) and host (wheat) phyllosphere surface supported uredospore germination, germ tube formation, hyphal elongation, epiphytic growth, stomatal entry and endophytic growth of fungus in an identical manner. However, the nonhost plant did not show any sign of rusting caused by uredospore production, which instead displayed induced H2O2accumulation, on leaf. Development of qPCR-based Pucciniaquantitation and green fluorescent tag-based qualitative assessment of Pucciniproliferation will facilitate nonhost resistance phenotyping not only in rice but also in other nonhost plants of stem rust pathogen. |
| Databáze: |
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