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碘-碘化钾染色法鉴定水稻花粉育性   

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Original research article

A brief version of this protocol appeared in:
Rice Protocol e-book

实验原理:成熟花粉粒含有淀粉粒,遇碘变蓝。
实验目的:鉴定水稻花粉的育性。

关键词: 水稻, 碘-碘化钾, 染色法, 花粉, 育性

材料与试剂

  1. 离心管
  2. 载玻片
  3. 盖玻片
  4. 75%乙醇

  5. 碘化钾
  6. 1%碘-碘化钾溶液 (见溶液配方)

仪器设备

  1. 镊子
  2. 烧杯
  3. 显微镜

实验步骤

  1. 花药采集:水稻抽穗时,取稻穗第二天开花的颖花放于装有75%乙醇的离心管内,一般要求取稻穗的上、中、下部位,花药长度超过颖壳长度的2/3视为成熟花药。
  2. 镜检:取稻穗上、中、下各一朵颖花的花药放在载玻片上,加1~2滴1%碘-碘化 钾溶液,然后用镊子将花药夹碎,使花粉粒充分释放,盖上盖玻片,用镊子轻压下,静置2-3 min。将其于10倍显微镜下观察。随机观测一个视野,每个视野花 粉量多于200粒。
    注:如果花药不能当天检测,要放入4 °C或 -20 °C冰箱。

结果与分析

显微镜下观测结果:
可育花粉粒呈圆形、大而饱满、深蓝色、着色均匀;不育花粉包括无花粉型、典败、圆败和染败共四种类型。无花粉型指显微镜下观测,无花粉或仅留残余花粉壁;典败指显微镜下观测花粉不染色,形状不规则状,如三管形、多边形等;圆败指花粉粒外观圆形,无染色淀粉粒;染败指大多数花粉形态正常,但着色较浅或着色不均匀,也有部分花粉深染色,但粒形明显异于正常可育花粉粒。

溶液配方

  1. 1%碘-碘化钾溶液
    称取1克碘放入盛有8 g碘化钾及20 ml水的烧杯中。待碘全部溶解后,将溶液移入100 ml棕色容量瓶中,稀释至刻度,摇匀。
    注:1%碘-碘化钾溶液要避光储存于棕色瓶中。
Copyright: © 2018 The Authors; exclusive licensee Bio-protocol LLC.
引用格式:高冠军, 范优荣, 李一博. (2018). 碘-碘化钾染色法鉴定水稻花粉育性. Bio-101: e1010139. DOI: 10.21769/BioProtoc.1010139.
How to cite: Gao, G. J., Fan, Y. R. and Li, Y. B. (2018). Identification of Rice Pollen Fertility by Iodine-potassium Iodide Staining. Bio-101: e1010139. DOI: 10.21769/BioProtoc.1010139.
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