不同固定液对小麦小孢子发育和花粉育性检测的影响

为了明确FAA和卡诺氏固定液对小麦小孢子花粉染色效果的差异,以温敏雄性不育系BNS366及其近等基因系郑麦366为材料,采用I_2-KI法、醋酸洋红法和DAPI法染色观察了小孢子发育过程,并统计了花粉可育率和自交结实率。结果表明,采用FAA固定液固定、I_2-KI法染色,小孢子内淀粉积累区域染色较深,单核期和二核期醋酸洋红法与DAPI法染色细胞核清晰程度均较高;采用卡诺氏固定液固定、三核期和开花期醋酸洋红法染色,细胞核清晰度较高,二核期与三核期DAPI法细胞图像略偏红。采用两种固定液固定、I_2-KI法和DAPI法染色,以及采用卡诺氏固定液固定、醋酸洋红法染色,花粉可育率均较高;采用FAA固定液固定、醋酸洋红法染色,花粉可育率偏低并与自交结实率一致。因此,从获得小孢子高清晰细胞图像角度考虑,对于I_2-KI、单核期和二核期醋酸洋红及DAPI染色法来说,采用FAA固定液较好;对于三核期和开花期醋酸洋红染色法来说,采用卡诺氏固定液较好。

Effects of Different Fixative on Microspore Development and Pollen Fertility Detection of Wheat

In order to clarify the differences in the staining effects of the microspores after the fixation of FAA and carnoys fixatives,the thermo-sensitive male-sterile wheat line BNS 366 and its near-isogenic line Zhengmai366 were used as the materials,and the microspore development process was investigated by I₂-KI method,acetate carmine method and DAPI method. Moreover,the pollen fertility at flowering stage and the selfing seedset rate were measured. The results showed that for the microspores fixed with FAA fixative,the starch accumulation area in microspore was heavily stained by I₂-KI,and the nuclei were lightly stained by acetate carmine and DAPI in uninucleate and binucleate periods. When fixed with carnoys fixative,the nuclei were lightly stained by acetate carmine in trinucleate and flowering periods,and the images were reddish in uninucleate and trinucleate periods when the wheat microspores were stained by DAPI. Pollen fertility was higher when two kinds of fixatives combined with I₂-KI or DAPI staining and acetate carmine staining after fixed with carnoys fixatives were used,Pollen fertility was low and the self-fertility rate was consistent with that of self-pollination by FAA fixation and acetate carmine staining. Therefore,to obtain high-definition cell images of microscopes,FAA fixative is advisable for I₂-KI method,and for acetate carmine method in uninucleate and binucleate period,and for DAPI method; as for carnoys fixative,it is advisable for acetate carmine method in trinucleate and flowering period.