ISSN:
1615-6102
Keywords:
Texas cytoplasmic-male-sterility
;
Flow cytometry
;
Image analysis
;
Maize
;
Tapetum
Source:
Springer Online Journal Archives 1860-2000
Topics:
Biology
Notes:
Summary A cytological study of Texas cytoplasmic male sterile (Tcms) and normal (N) anther tapetal protoplasts ofZea mays was undertaken to determine whether there were any differences prior to Tcms male cell abortion not noted in previous published studies. Squash preparations, tapetal protoplast separation via flow cytometry, image analysis, and electron microscopy were utilized. Chemically preserved tapetal protoplasts from both lines were prominently angular in shape and typically smaller than any other cell type in the anthers. The tapetum from both lines consisted of a mixture of uninucleate and binucleate protoplasts. The Tcms tapetum consistently had a higher proportion of binucleate protoplasts during all stages of microsporogenesis prior to abortion. The size of Tcms uniand binucleate tapetal protoplasts was more variable than the N tapetal protoplasts and was largest during the microspore stage when male cells abort. Tapetal nuclear size in both lines was less variable. Uni- and binucleate tapetal protoplasts from each line could be separated from the other anther cells and from each other by filtration and then by flow cytometry, based on intensity of nuclear fluorescence. These results suggest that Tcms uninucleate tapetal protoplasts have a higher level of DNA than N uninucleate tapetal protoplasts. Both fluorescence microscopy and electron microscopy confirmed pure populations of intact uni- and binucleate tapetal protoplasts using flow cytometry. The results from this study indicate that the methodology presented here could be used for a variety of further studies to better understand the cellular and molecular basis of male sterility in maize, and in other taxa, where the tapetum is the primary target that leads to male sterility.
Type of Medium:
Electronic Resource
URL:
http://dx.doi.org/10.1007/BF01378861
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