Structural features of five types of maize starch granule subgroups sorted by flow cytometry.

Subgroups of starch granules from five maize phenotypes including waxy-, normal-, popcorn-, sweet corn- and high-amylose maize were sorted by flow cytometry (FC) utilizing the side scatter channel (SSC) and forward scatter channel (FSC). SSC and FSC mainly reflecting internal object complexity, and object size, respectively. Subgroups with higher FSC signal always showed higher SSC signal, indicating larger granules exhibited higher internal structural complexity. Wide-angle and small-angle X-ray scattering analysis showed that the subgroups showing high SSC signal intensity also had high lamellar scattering intensity, and low crystallinity. Vibrational transitions of bonds analyzed by Fourier Transform Infrared Spectroscopy (FT-IR) showed that the subgroups of maize starches, except sweet corn starch, with high SSC signal had high intensities at 1045 and 1022 cm-1. Hence, our data demonstrate that the structural complexity detected by the SSC signal is mainly associated with lamellar and crystalline features of starch granules.

The relationship between the expression pattern of starch biosynthesis enzymes and molecular structure of high amylose maize starch.

Two high amylose (HAM) inbred lines with apparent amylose contents of 55 % and 62 %, respectively, were selected to explore the relationship between molecular structure and gene expression of starch-synthase involved enzymes. GPC analysis of debranched starches showed that the HAM starches (HAMSs) had shorter amylose chains and longer amylopectin chains than normal maize starch (NMS). FACE analysis showed that these HAMSs had a higher content of amylopectin chains of DP > 21. Quantitative Real-Time PCR analysis showed that the HAM lines had specifically low expression of the starch branching enzyme IIb (SBEIIb), and the starch synthase IIIa (SSIIIa) homologue, and high expression of the isoamylase 2 (ISA2), potentially suppressing the generation of amylopectin molecules through deficient branching and excessive debranching process, thereby increasing the relative amylose content. A high expression of GBSS1 was potentially associated with increased short amylose chain lengths in HAMSs.