Changes in fine structure of amylopectin and internal structures of starch granules in developing endosperms and culms caused by starch branching enzyme mutations of japonica rice.

KEY MESSAGE: BEIIb plays a specific role in determining the structure of amylopectin in rice endosperm, whereas BEIIa plays the similar role in the culm where BEIIb is absent. Cereals have three types of starch branching enzymes (BEs), BEI, BEIIa, and BEIIb. It is widely known that BEIIb is specifically expressed in the endosperm and plays a distinct role in the structure of amylopectin because in its absence the amylopectin type changes to the amylose-extender-type (ae-type) or B-type from the wild-type or A-type and this causes the starch crystalline allomorph to the B-type from the wild-type A-type. This study aimed to clarify the role of BEIIa in the culm where BEIIb is not expressed, by using a be2a mutant in comparison with results with be2b and be1 mutants. The results showed that the amylopectin structure exhibited the B-type in the be2a culm compared with the A-type in the wild-type culm. The starch granules from the be2a culm also showed the B-type like allomorph when examined by X-ray diffraction analysis and optical sum frequency generation spectroscopy. Both amylopectin chain-length profile and starch crystalline properties were found to be the A-type at the very early stage of endosperm development at 4-6 days after pollination (DAP) even in the be2b mutant. All these results support a view that in the culm as well as in the endosperm at 4-6 DAP, BEIIa can play the role of BEIIb which has been well documented in maturing endosperm. The possible mechanism as to how BEIIa can play its role is discussed.

On the cluster structure of amylopectin.

KEY MESSAGE: Two opposing models for the amylopectin structure are historically and comprehensively reviewed, which leads us to a better understanding of the specific fine structure of amylopectin. Amylopectin is a highly branched glucan which accounts for approximately 65-85 of starch in most plant tissues. However, its fine structure is still not fully understood due to the limitations of current methodologies. Since the 1940 s, many scientists have attempted to elucidate the distinct structure of amylopectin. One of the most accepted concepts is that amylopectin has a structural element known as "cluster", in which neighboring side chains with a degree of polymerization of ≥ 10 in the region of their non-branched segments form double helices. The double helical structures are arranged in inter- and intra-clusters and are the origin of the distinct physicochemical and crystalline properties of starch granules. Several models of the cluster structure have been proposed by starch scientists worldwide during the progress of analytical methods, whereas no direct evidence so far has been provided. Recently, Bertoft and colleagues proposed a new model designated as "the building block and backbone (BB) model". The BB model sharply contrasts with the cluster model in that the structural element for the BB model is the building block, and that long chains are separately synthesized and positioned from short chains constituting the building block. In the present paper, we conduct the historical review of the cluster concept detailing how and when the concept was established based on experimental results by many scientists. Then, differences between the two opposing concepts are explained and both models are critically discussed, particularly from the point of view of the biochemical regulation of amylopectin biosynthesis.

Contributions of Dexter French (1918-1981) to cycloamylose/cyclodextrin and starch science.

Professor Dexter French (1918-1981) was an American chemist and biochemist at Iowa State College (University in 1959). He devoted his career to advance knowledge of polysaccharides and oligosaccharides, in particular starch, cyclodextrins, and enzymes. Cyclodextrins are oligosaccharides obtained from starch and are typically cage molecules with a hydrophobic cavity that can encapsulate other compounds nowadays the basis for many industrial applications. Since the 1960s, he has been recognized as an outstanding authority in the field of starches and cyclodextrins and has inspired researchers in laboratories around the world. This review, on the fortieth anniversary of his death, commemorates his remarkable contribution to starch and cyclodextrin chemistry. Firstly, we give an overview of his personal life and career. Secondly, we highlight some of the results on starch and cyclodextrins from Professor French and his group. A third part discusses his impact on the modern chemistry of cyclodextrins and starch.

Effects of BEIIb-Deficiency on the Cluster Structure of Amylopectin and the Internal Structure of Starch Granules in Endosperm and Culm of Japonica-Type Rice.

It is known that one of starch branching enzyme (BE) isoforms, BEIIb, plays a specific role not only in the synthesis of distinct amylopectin cluster structure, but also in the formation of the internal structure of starch granules in rice endosperm because in its absence the starch crystalline polymorph changes to the B-type from the typical A-type found in the wild-type (WT) cereal endosperm starch granules. In the present study, to examine the contribution of BEIIb to the amylopectin cluster structure, the chain-length distributions of amylopectin and its phosphorylase-limit dextrins (Φ-LD) from endosperm and culm of a null be2b mutant called amylose-extender (ae) mutant line, EM10, were compared with those of its WT cultivar, Kinmaze, of japonica rice. The results strongly suggest that BEIIb specifically formed new short chains whose branch points were localized in the basal part of the crystalline lamellae and presumably in the intermediate between the crystalline and amorphous lamellae of amylopectin clusters in the WT endosperm, whereas in its absence branch points which were mainly formed by BEI were only located in the amorphous lamellae of amylopectin. These differences in the cluster structure of amylopectin between Kinmaze and EM10 endosperm were considered to be responsible for the differences in the A-type and B-type crystalline structures of starch granules between Kinmaze and EM10, respectively. The changes in internal structure of starch granules caused by BEIIb were analyzed by wide angle X-ray diffraction, small-angle X-ray scattering, solid state 13C NMR, and optical sum frequency generation spectroscopy. It was noted that the size the amylopectin cluster in ae endosperm (approximately 8.24 nm) was significantly smaller than that in WT endosperm (approximately 8.81 nm). Based on the present results, we proposed a model for the cluster structure of amylopectin in WT and ae mutant of rice endosperm. We also hypothesized the role of BEIIa in amylopectin biosynthesis in culm where BEIIb was not expressed and instead BEIIa was the major BE component in WT of rice.