Protoplasts from the calli were generated as previously described [69 ], then protoplast suspensions were dropped onto microscope slides to observe the plastid modes. Light microscopy of various orange tissues of Hongkong kumquats was performed using a frozen sectioning technique with a Leica CM1900 (Leica, Germany). An optical microscope (BX61, Olympus) equipped with a DP70 camera was used in tandem with a differential interference contrast (DIC) technique.
Transmission electron microscopy (TEM) analysis was performed according to Cao et al. [28 (link)]. Samples were prepared using a normal fixation process with 2.5% glutaraldehyde adjusted to pH 7.4, and a 0.1 M phosphate buffer with 2% OsO4. The preparations were dehydrated and embedded in epoxy resin and SPI-812, respectively. Ultrathin sections obtained with a Leica UC6 ultramicrotome were stained with uranyl acetate and subsequently with lead citrate. Image recording was performed with a HITACHI H-7650 transmission electron microscope at 80 KV and a Gatan 832 CCD camera.
Starch granule morphology was examined with a scanning electron microscope (SEM). The samples were mounted on studs, sputter coated with gold (Balzers, JFC-1600), and examined under a JSM-6390LV SEM (JEOL, Japan).
Transmission electron microscopy (TEM) analysis was performed according to Cao et al. [28 (link)]. Samples were prepared using a normal fixation process with 2.5% glutaraldehyde adjusted to pH 7.4, and a 0.1 M phosphate buffer with 2% OsO4. The preparations were dehydrated and embedded in epoxy resin and SPI-812, respectively. Ultrathin sections obtained with a Leica UC6 ultramicrotome were stained with uranyl acetate and subsequently with lead citrate. Image recording was performed with a HITACHI H-7650 transmission electron microscope at 80 KV and a Gatan 832 CCD camera.
Starch granule morphology was examined with a scanning electron microscope (SEM). The samples were mounted on studs, sputter coated with gold (Balzers, JFC-1600), and examined under a JSM-6390LV SEM (JEOL, Japan).
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