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Block-Surface Staining for Differentiation of Starch and Cell Walls in Wheat Endosperm

 

作者: GlennGregory M.,   PittsMarvin J.,   LiaoKe,   IrvingDelilah W.,  

 

期刊: Biotechnic&Histochemistry  (Taylor Available online 1992)
卷期: Volume 67, issue 2  

页码: 88-97

 

ISSN:1052-0295

 

年代: 1992

 

DOI:10.3109/10520299209110014

 

出版商: Taylor&Francis

 

关键词: Triticum aestivumL;block surface staining;wheat classification

 

数据来源: Taylor

 

摘要:

A staining technique for differentiating starch granules and cell walls was developed for computer-assisted studies of starch granule distribution in cells of wheat [Triticum aestivumL.] caryopses. Blocks of embedded caryopses were sectioned, exposing the endosperm tissue, and stained with iodine potassium iodide (IKI) and Calcofluor White. Excessive tissue hydration during staining was avoided by using stains prepared in 80% ethanol and using short staining times. The IKI quenched background fluorescence which facilitated the use of higher concentrations of Calcofluor White. Cell wall definition was improved with the IKI-Calcofluor staining combination compared to Calcofluor alone. The high contrast between darkly stained starch granules and fluorescent cell walls permitted computer assisted analysis of data from selected hard and soft wheat varieties. The ratio of starch granule area to cell area was similar for both wheat classes. The starch granule sizes ranged from 2.1μm3to 22,000μm3with approximately 90% of the granules measuring less than 752μm3(ca. 11μm in diameter). Hard wheat samples had a greater number of small starch granules and a lower mean starch granule area compared to the soft wheat varieties tested. The starch size distribution curve was bimodal for both the hard and soft wheat varieties. Three-dimensional starch size distribution was measured for four cells near the central cheek region of a single caryopsis. The percentage of small granules was higher at the ends than at the mid-section of the cells.

 

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