Smz 168

Table S1 in the supplemental material lists the strains used in this study. C. neoformans strains were cultured and maintained in yeast extract-peptone-dextrose (YPD) medium. Melanin production was assessed on Niger seed medium (70 g Niger seed, 20 g Bacto agar per liter) containing different glucose concentrations. Cells were incubated at 37°C and photographed daily for 1 to 3 days by microscope (SMZ-168; Motic) at ×10 magnification. For the urease assay, equal cell numbers (5 × 10⁴ cells) were spotted onto Christensen’s agar medium. Plates were incubated for 2 to 3 days at 30°C and photographed.

In each 800 m² plot (field plots: 20 m × 40 m; plant spacing: 40 cm; row spacing: 80 cm) of the tomato fields in each landscape, the first sampling started 10 days after tomato transplanting between Jun. and Nov. in 2018 and 2019. Leaves with B. tabaci nymphs and adults were collected every 10 days using the fixed five points sampling method until the end of the tomato growing season. In each sampling point, five tomato plants were sampled (but avoiding the plants closest to any edge to minimize edge effects). On each tomato plant, five leaves of similar age at the upper, middle and lower positions were examined, giving a total of 375 leaves observed per plot. Adults were counted in situ; afterwards, the leaves were cut, individually placed in plastic bags and marked, then brought to the laboratory to count nymphs under a dissecting microscope (20 × magnification, MOTIC, SMZ-168). The area of each leaf was measured by a leaf area measuring instrument (Yaxin-1242), from which standardized density data (no. of individuals per 100 cm² leaf surface) were obtained. The tomato growth period was recorded at each survey. This was divided into seedling, anthesis, fruit expansion and harvest periods [29 (link),30 (link)].

Dipterocarpus alatus leaf litter was collected from Doi Inthanon National Park, Chiang Mai, Thailand. The samples were stored in separate paper bags, brought back to the laboratory, and pressed in between newspaper sheets. After two days, leaf samples were placed in a sterile moisture chamber to favor the emergence of saprobic fungal taxa. The specimens were examined using a dissecting microscope (Motic SMZ-168, Wetzlar, Germany) and a light microscope (Nikon ECLIPSE 80i, Tokyo, Japan). Single-spore isolation was carried out in potato dextrose agar (PDA) plates as described by Senanayake et al. [38 (link)]. The obtained cultures were incubated at 25 °C for 2–3 weeks with frequent observation to detect and record the sporulation and colony characteristics.

Once the S. furcifera adults emerged, individuals from each treatment group were identified to sex and wing form. Fifteen pairs of macropterous females and males were randomly selected to mate and oviposit in numbered glass tubes with rice seedlings (Male NZMF x female NZMF and male GMF x Female GMF). Since each S. furcifera female adult mates multiple times with male adults, a new male adult was added to the cage if the original died before the female. Fecundity was measured by dissecting rice stems every day under a stereomicroscope (MOTIC SMZ-168) until death of the given female individual and counting number of eggs laid per female. Another 40 newly-emerged macropterous females from each treatment group were transferred to large beakers with rice seedlings, maintained continuously under their corresponding magnetic field treatments, and checked for mortality daily until death. The remaining macropterous female adults on the 1st, 4th, 8th day after emergence from each treatment group meeting the requirements of the molecular experiments were individually transferred into 1.5ml clear microtubes (Axygen MCT-150-C) at and stored in a -80°C freezer (Thermo Scientific Forma 702, USA) for the gene expression experiments (90 individuals for each sampling time). Fresh rice seedlings were provided every 3rd day.

Embryos at one-day post-fertilization (dpf) were microinjected individually using a pneumatic picopump (PV830; World Precision Instruments, Sarasota, FL, USA) equipped with a magnetic manipulator (MM33; Kantec, Bensenville, IL, USA) with a pulled microcapillary pipette-using device (PC-10; Narishigen, Tokyo, Japan). Injection of each rHDL alone (16 μg of apoA-I) or co-injection with CML (500 ng) was performed at the same position in the yolk to minimize bias, as described previously [43 (link),59 (link)]. After the injection, the live embryos were observed under a stereomicroscope (Motic SMZ 168; Hong Kong) and photographed (20× magnification) using a Motic cam2300 CCD camera. At 24 h post-injection, each live embryo was compared after removing the chorion to compare the developmental stage at higher magnification (50×).