Eos 700d digital camera

Images of the tassels and anthers were captured with a Canon EOS 700D digital camera (Canon, Tokyo, Japan) and a SZX2-ILLB stereomicroscope (Olympus, Tokyo, Japan) respectively. Routine analysis of I₂-KI staining and SEM (scanning electron microscopy) were performed as described previously [44 (link)]. SEM images of anther and pollen grain were detected with a HITACHI S-3400N scanning electron microscope (HITACHI, Tokyo, Japan).

Tassels and spikelets were photographed using a Canon EOS 700D digital camera (Canon, Japan). To evaluate the pollen viability, anthers and pollen grains were stained with 1% I₂‐KI and photographed under an Olympus SZ51 microscope (Olympus, Japan). For transverse section analysis, the spikelet was fixed in 3:1 ethanol: acetic acid. Fixed samples were dehydrated through an ethanol series and embedded in epoxy resin. Semi‐thin sections were obtained using an Ultracut E Ultramicrotome (Reichert), stained with 0.1% toluidine blue O and observed under an Olympus BX61 microscope (Olympus, Japan).

Paraformaldehyde-fixed GA muscles were embedded in paraffin and sliced into 5-μm-thick sections, followed by H&E staining to examine the morphology of muscle tissues. Five fields per section were digitally captured at 200× magnification using an Olympus BX43 Upright microscope (Waltham, MA, USA) and a Canon EOS 700D digital camera (Tokyo, Japan).

Samples of decaying wood were collected from October 2019 to November 2020 in forests and nature reserves of Guizhou, Hainan and Yunnan Provinces in China. The specimens were observed with a stereomicroscope while microscopic images of the samples were taken using a Nikon ECLIPSE Ni compound microscope, with a Canon EOS 700D digital camera. Measurements were taken with Tarosoft (R) Image Frame Work (v.0.9.7). More than 30 asci and ascospores were measured for each specimen examined. Photoplates were arranged and improved by using Adobe Photoshop CS6 software. Isolations of fungi were made by single spore isolation (Chomnunti et al. 2014 (link)) and germinated spores were transferred to potato dextrose agar (PDA) medium for purification. The specimens were deposited at the Herbarium of Cryptogams, Kunming Institute of Botany Academia Sinica (KUN-HKAS) and Herbarium of Guizhou Medical University (GMB). Strains of the new genus and new species are maintained in the Guizhou Medical University Collection Centre (GMBC).

Agrobacterium tumefaciens was transformed with the individual constructs described above and grown overnight in the YEP medium containing the appropriate antibiotics. A. tumefaciens cultures were pelleted and then resuspended individually in an infiltration buffer [10 mM MgCl₂, 10 mM 2-(N-Morpholino) ethane sulfonic acid (MES), and 100 μM acetosyringone]. Individual A. tumefaciens cultures were adjusted to OD₆₀₀ = 1.2, or as indicated otherwise, and then mixed with the infiltration buffer by 1 to 1 volume for 3 h in dark conditions. The mixed A. tumefaciens suspension co-infiltrated into leaves of 7–8 leaf stage N. benthamiana plants using 1 mL needleless syringes. The agro-infiltrated leaves were examined for GFP expression under a hand-held 100 W, long-wave UV lamp (UV Products, Upland, CA, USA). The infiltrated leaves were photographed with a Canon EOS 700 D digital camera (Canon, Taiwan, China) with an Y48 yellow filter.