10.6084/m9.figshare.8948696.v1 Ni Cai Ni Cai Feng Wang Feng Wang Xiangqun Nong Xiangqun Nong Guangjun Wang Guangjun Wang Mark McNeill Mark McNeill Guangchun Cao Guangchun Cao Kun Hao Kun Hao Shaofang Liu Shaofang Liu Zehua Zhang Zehua Zhang Visualising confirmation of the endophytic relationship of <i>Metarhizium anisopliae</i> with maize roots using molecular tools and fluorescent labelling Taylor & Francis Group 2019 Endophyte entomopathogenic fungus Zea mays egfp-labelling PCR laser scanning confocal microscopy (LSCM) 2019-07-18 05:56:57 Dataset https://tandf.figshare.com/articles/dataset/Visualising_confirmation_of_the_endophytic_relationship_of_i_Metarhizium_anisopliae_i_with_maize_roots_using_molecular_tools_and_fluorescent_labelling/8948696 <p><i>Metarhizium anisopliae</i>, an entomopathogenic fungus, is found to contribute to plant nutrient uptake and condidered as a plant endophyte. However, there is very little visual evidence that demonstratrates the endophytic relationship. Since the incidence of <i>Metarhizium</i> within host plants may be ephemeral or of limited distribution within host tissue, visualisation is reliant on both presence and the ability to detect the fungus even when present at low levels. We inoculated maize seeds with an <i>egfp</i>-labelled isolate of <i>M. anisopliae</i>, and then detected the <i>egfp</i> fragment by qualitative and quantitative PCR, and examined fluorescence of mycelia in the inoculated roots by laser scanning confocal microscopy (LSCM). The optimised PCR qualitatively identified the presence of <i>egfp</i>-labelled <i>M. anisopliae</i> at a 1:3000 <i>M. anisopliae</i>-maize DNA ratio. Root samplings of inoculated plants at intervals over 24 days were discovered <i>egfp</i>-labelled fragments with 14-d roots having the highest proportion with amplification. The <i>egfp</i>-label concentrations ranged very low in 2.976 × 10<sup>−7.42∼−7.02</sup> ng/μL, with a rise process in 4–11 days after <i>M. anisopliae</i> treatment. Under LSCM, fluorescent mycelia in roots were identified, and the clear images were captured to show hyphal development extending along a longitudinal line of root cells and even forming a hyphal mass in the intercellular space. There were many hyphae at the branch junctions of the lateral roots. The methodology used to detect the fungus will be important for exploring the multi-trophic function of <i>M. anisopliae</i> and co-evolution in plants.</p>