a-simple-dry-sectioning-method-for-obtaining-whole-seed-sized-resin

A Simple Dry Sectioning Method for Obtaining Whole-Seed-Sized Resin Section and Its Applications

The morphology, size and quantity of cells, starch granules&#160;and protein bodies in seed determine the weight and quality of seed. They are ...

yangzhou university

Yangzhou University

High-amylose cereal endosperm is rich in heterogeneous starch granules. In this paper, we investigated the morphology, structure and gelatinization properties of high-amylose maize endosperm starch. Starch had individual, aggregate and elongated heterogeneous granules. Most of individual granules were round with small size and had one central hilum. Aggregate and elongated granules consisted of many subgranules with central hila, and had irregular and rod/filamentous shapes, respectively. Iodine stained starch granules showed five types of polarization colors: blue, purple, fuchsia, dark red, and interior dark blue and exterior brown. Most of individual and aggregate granules had the color of dark red, that of elongated granules the color of interior dark blue and exterior brown. Amylose was mainly distributed in the hilum region and the circumference of starch granules. Aggregate and elongated granules had higher amylose content than individual granules. Elongated and individual granules had the highest and the lowest gelatinization resistance among high-amylose maize heterogeneous starch granules, respectively. 

Morphology, structure and gelatinization properties of heterogeneous starch granules from high-amylose maize.

Starch granules from high-amylose cereal mutants or transgenic lines usually have different morphologies. It is not clear whether the structure and spatial distribution of starch granules with different morphologies in endosperm is homogeneous or heterogeneous. In the present study, the structure and spatial distribution in endosperm of morphologically different starch granules from high-amylose transgenic rice line (TRS) were investigated. The TRS endosperm had individual, aggregate, elongated, and interior hollow starch granules. The individual and interior hollow granules had the lowest and the highest amylose content and gelatinization resistance, respectively, among the four types of granules. The individual granules were mainly distributed in the middle of the endosperm; the aggregate granules in the starchy endosperm cells between the subaleurone layer and the middle of the endosperm; the elongated granules in the peripheral starchy endosperm cells adjacent to the subaleurone layer; and the interior hollow granules in the subaleurone layer cells. 

Heterogeneous structure and spatial distribution in endosperm of high-amylose rice starch granules with different morphologies.

Large-, medium-, and small-sized granules were separated from normal and high-amylose maize starches using a glycerol centrifugation method. The different-sized fractions of normal maize starch showed similar molecular weight distribution, crystal structure, long- and short-range ordered structure, and lamellar structure of starch, but the different-sized fractions of high-amylose maize starch showed markedly different structural properties. The amylose content, iodine blue value, amylopectin long branch-chain, and IR ratio of 1045/1022 cm(-1) significantly increased with decrease of granule size, but the amylopectin short branch-chain and branching degree, relative crystallinity, IR ratio of 1022/995 cm(-1), and peak intensity of lamellar structure markedly decreased with decrease of granule size for high-amylose maize starch. The large-sized granules of high-amylose maize starch were A-type crystallinity, native and medium-sized granules of high-amylose maize starch were CA-type crystallinity, and small-sized granules of high-amylose maize starch were C-type crystallinity, indicating that C-type starch might contain A-type starch granules. 

Different structural properties of high-amylose maize starch fractions varying in granule size.

Starches were isolated from rhizomes of Curcuma longa, Canna edulis and Canna indica and bulbs of Lilium lancifolium, and showed a B-type X-ray diffraction pattern. Their physicochemical properties were investigated and compared. These starches showed significantly different granule morphologies and sizes, but all had eccentric hila. The C. longa starch had the lowest content of amylopectin short branch-chain and branching degree and the highest content of amylopectin long branch-chain, and the L. lancifolium starch the highest content of amylopectin short branch-chain and branching degree and the lowest content of amylopectin long branch-chain among the four starches. The L. lancifolium starch had the lowest resistance to gelatinization, and showed the lowest pasting peak, hot and final viscosities, and the C. longa starch had the highest resistance to gelatinization, and showed the highest pasting hot, final and setback viscosities and the lowest pasting breakdown viscosity. The C. longa and L. lancifolium starches possessed very high and low resistance to hydrolysis and digestion, respectively. The above physicochemical properties would be useful for the applications of B-type starches in food and nonfood industries. 

Comparison of physicochemical properties of B-type nontraditional starches from different sources.

Lotus rhizome C-type starch was separated into different size fractions. Starch morphologies changed from irregular to elongated, ellipsoid, oval, and spherical with decreasing granule size. The small- and very-small-sized fractions had a centric hilum, and the other size fractions had an eccentric hilum. The different size fractions all showed C-type crystallinity, pseudoplasticity and shear-thinning rheological properties. The range of amylose content was 25.6 to 26.6%, that of relative crystallinity was 23.9 to 25.8%, that of swelling power was 29.0 to 31.4 g/g, and that of gelatinization enthalpy was 12.4 to 14.2J/g. The very-small-sized fraction had a significantly lower short-range ordered degree and flow behavior index and higher scattering peak intensity, water solubility, gelatinization peak temperature, gelatinization conclusion temperature, consistency coefficient, hydrolysis degrees, and digestion rate than the large-sized fraction. Granule size significantly positively influenced short-range ordered structure and swelling power and negatively influenced scattering peak intensity, water solubility, hydrolysis and digestion of starch (p<0.01).

Effect of granule size on the properties of lotus rhizome C-type starch.

Storage starch in starchy seed influences the seed weight and texture, and determines its applications in food and nonfood industries. Starch granules from different plant sources have significantly different shapes and sizes, and even more the difference exists in the different regions of the same tissue. Therefore, it is very important to in situ investigate the morphology and distribution of starch in the whole seed. However, a simple and rapid method is deficient to prepare the whole section of starchy seed for investigating the morphology and distribution of starch in the whole seeds for a large number of samples.

A simple and rapid method for preparing the whole section of starchy seed to investigate the morphology and distribution of starch in different regions of seed.

Starch, as a main energy storage substance, plays an important role in plant growth and human life. Despite the fact that several enzymes and regulators involved in starch biosynthesis have been identified, the regulating mechanism of starch synthesis is still unclear. In this study, we isolated a rice floury endosperm mutant  from a mutant pool induced by Co. Both total starch content and amylose content in  seeds significantly decreased, and starch thermal and pasting properties changed. Compound starch granules were defected in the floury endosperm of  seeds. Map-based cloning and a complementation test showed that the floury endosperm phenotype was determined by a gene of , which encodes pyruvate orthophosphate dikinase (PPDK, EC 2.7.9.1). Subcellular localization analysis demonstrated that PPDK was localized in chloroplast and cytoplasm, the  highly expressed in leaf and leaf sheath, and the  constitutively expressed with a high expression in developing endosperm. Moreover, the expression of starch synthesis-related genes was also obviously altered in  developing endosperm. The above results indicated that PPDK played an important role in starch metabolism and structure in rice endosperm.

A Novel Mutation of , Encoding Pyruvate Orthophosphate Dikinase, Affects Metabolism and Structure of Starch in the Rice Endosperm.

The accumulation and morphology of starch in the pericarp, embryo and endosperm of normal and waxy maize were investigated using whole sections of complete caryopses. Pericarp starch took the form of compound granules, was distributed in the bottom of caryopses, and degraded from the top to the bottom. Embryo starch mostly took the form of simple granules and accumulated in the scutellum beginning approximately 10 DAP. In the endosperm, starch accumulated longitudinally from the top to the bottom and transversely from the centre to the periphery with caryopsis development. The peripheral endosperm cells synthesized starch faster than did the inner ones. Simple and compound starches were both observed, but the compound starch granules were distributed in the central region of the endosperm. At a late stage of development, compound starch was only observed in the bottom central portion of the endosperm. The pericarp starch of normal maize showed higher amylose content than did the embryo and endosperm starch. The waxy maize pericarp and embryo starches had similar amylose contents, but amylose was hardly detected in the endosperm due to the granule-bound starch synthase I gene mutation. The starches from the endosperm, embryo and pericarp of normal and waxy maize all had A-type crystallinity.

Spatiotemporal accumulation and characteristics of starch in developing maize caryopses.

In cereal seeds, the number, morphology and development of endosperm cells are closely related to grain quality, weight and yield. Endosperm cells differ morphologically in different regions of the seed. Nevertheless, it is important to be able to analyze the morphology of cereal endosperm cells. We established an image processing method to enhance the outlines of endosperm cells. The endosperm cell wall was traced precisely using the "pen tool" in Photoshop software (PS). The tracing was defined as the "work path" and was highlighted using the PS "brush tool." Images of mature rice, maize and wheat endosperm sections stained with different methods were analyzed using this method. Combined with the whole sections of mature and developing cereal kernels, the processed image exhibited clearly the morphology of endosperm cells in any region of endosperm and at any stage of endosperm development. The processed image was more accurate and efficient for analyzing morphological characteristics than the unprocessed image.

An image processing method for investigating the morphology of cereal endosperm cells.

In maize (), kernel weight is an important component of yield that has been selected during domestication. Many genes associated with kernel weight have been identified through mutant analysis. Most are involved in the biogenesis and functional maintenance of organelles or other fundamental cellular activities. However, few quantitative trait loci (QTLs) underlying quantitative variation in kernel weight have been cloned. Here, we characterize a QTL, , associated with maize kernel weight. This QTL encodes a DYW motif pentatricopeptide repeat protein involved in C-to-U editing of , a subunit of the chloroplast NADH dehydrogenase-like complex. In a null  background, C-to-U editing of  was abolished, and photosynthesis was reduced, resulting in less maternal photosynthate available for grain filling. Characterization of  highlights the importance of optimizing photosynthesis for maize grain yield production.

Lingxiao Zhao

yangzhou university

A Simple Dry Sectioning Method for Obtaining Whole-Seed-Sized Resin Section and Its Applications