Rapid visco analyzer rva techmaster

Nipponbare (Oryza sativa, ssp. japonica) and all the transgenic plants used in this study were grown in the experimental field and greenhouse of China National Rice Research Institute. The thousand‐grain‐weight, seed length width and chalkiness of WT and mutant lines were examined by a seed phenotyping system (Wangsheng, Hangzhou, China).
Total starch and amylose contents of brown seedswere measured with a starch assay kits Megazyme K‐TSTA and K‐AMYL (Megazyme, Ireland, UK). The content is expressed as the percentage of total sample weight on an oven‐dry basis. The total amylose, crude fiber, lipid and protein contents in the grains were measured following the previous report (Kang et al., 2005). RVA analysis was done on a Rapid Visco Analyzer (RVA Techmaster, Newport Scientific, Narrabeen, Australia) as described by Sun et al., 2017. DSC assay was conducted on a differential scanning calorimeter DSC1 STARe system (Mettler Toledo, Switzerland) by following Sun et al., 2017.

The total starch content of the rice powder was measured using a starch assay kit (Megazyme, Wicklow, Ireland; http://www.megazyme.com). Amylose content was determined following the method depicted by Liu et al. (2009). Lipid and protein contents in the grains were measured according to the method described by Kang et al. (2005). Content of FA was determined by gas chromatography‐mass spectrometer (GC‐MS, Li et al., 2006). To determine the starch pasting properties, 3 g of milled rice power (0.5 mm or less, 14% moisture basis) was transferred into a container with 25 mL of distilled water. The sample was mixed and measured with a Rapid Visco Analyzer (RVA Techmaster; Newport Scientific, Narrabeen, Australia), applying the protocol described by the manufacturer. To determine the chain length distribution of amylopectin, 5 mg of rice flour was digested with Pseudomonas amyloderamosa isoamylase (Megazyme), then analysed using the capillary electrophoresis (PA800 plus pharmaceutical analysis system; Beckman Coulter, http://www.beckmancoulter.com). The swelling and gelatinization properties of endosperm starch in urea solution were measured according to the method described by Nishi et al. (2001). All parameters related to physicochemical properties included three biological replications.