Anther Development and Pollen Viability Analysis

Photographs of the fresh anthers at each sequential stage were obtained from XN1376 and XN1376-CIMS with a Nikon E995 digital camera (Nikon, Tokyo, Japan) fixed firmly to a Motic K400 dissecting microscope (Preiser Scientific, Louisville, KY, United States). Pollen grains were also analyzed via 1% iodine–potassium iodide (1% KI–I₂) staining to determine the viability of the mature pollen, as previously described [65 (link)]. Scanning electron microscopy was used to characterize the surface characteristics of the anthers. Before the experiment, fresh anthers and pollen grains were fixed in 4% glutaraldehyde and then treated with an alcohol gradient, dried, and broken in sequence. Finally, the anthers and pollen grains were mounted on a stub with colloidal silver and photographed using a JSM-6360LV scanning electron microscope (JEOL, Tokyo, Japan) [66 (link)].

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Zhang L., Wang C., Yu Y., Zhang Y., Song Y., Li Z., Wang S., Zhang Y., Guo X., Liu D., Li Z., Ma S., Zheng J., Zhao H, & Zhang G. (2020). Cyclin-Dependent Kinase Inhibitor Gene TaICK1 acts as a Potential Contributor to Wheat Male Sterility induced by a Chemical Hybridizing Agent. International Journal of Molecular Sciences, 21(7), 2468.

Publication 2020

JSM-6360LV scanning electron microscope

Alcohol Cims Colloidal silver Glutaraldehyde Iodine potassium iodide Microscope Pollen grains Scanning electron microscope

Corresponding Organization : Northwest A&F University

Other organizations : Henan Institute of Science and Technology, Yangtze University

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Variable analysis

independent variables

Genotype (XN1376 and XN1376-CIMS)

dependent variables

Characteristics of fresh anthers at each sequential stage
Viability of mature pollen grains
Surface characteristics of anthers

control variables

Nikon E995 digital camera fixed firmly to a Motic K400 dissecting microscope
1% iodine–potassium iodide (1% KI–I2) staining for pollen viability analysis
Scanning electron microscopy for anther and pollen grain surface characterization
Fixation of fresh anthers and pollen grains in 4% glutaraldehyde, treatment with alcohol gradient, drying, and mounting on stub with colloidal silver

positive controls

None specified

negative controls

None specified

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