Detection of pollen viability and stigma pollinability

The pollination process begins with anther dehiscence and dispersal of mature pollen. Pollen grains carrying male gametes or their precursors are exposed to dry conditions and must reach a suitable receptive stigma when they are viable in order to complete the pollination process, and flowers with suitable stigmas for pollen receptivity are in the stigmatic pollinization period. The combination of the length of time that pollen remains viable and the length of the stigma pollination period profoundly affects the success of pollination in plants, especially heterogamous pollinated plants. Therefore, the study of pollen viability and stigma pollinability of plants can provide important theoretical guidance for artificial cross-breeding.

Operation method

Detection of pollen viability and stigma pollinability

Materials and Instruments

Material:
Arabidopsis thaliana in full bloom.
Apparatus:
① Microscope
① Microscope ② Slide
③ Concave slide
④ Coverslip
⑤ Straight-tipped tweezers
⑥ Dissecting needle
⑦ Filter paper
⑧ Petri dish (60 mm)
Reagents:
① Distilled water
② Triphenyltetrazolium chloride
Sucrose
Sucrose ④ Benzidine
⑤ Hydrogen peroxide

Move

The basic process of pollen viability and stigma pollination test can be divided into the following steps:

The pollen viability test was performed using the TTC (2,3,5-triphenyltetrazolium chloride) method. The pollen viability is determined by using 0.5% TTC sucrose solution to make the viable pollen turn red while the non-viable or abortive pollen does not show red color. The method was as follows: the pollen scattered at different times was scattered on the slide, 0.5% TTC sucrose solution was added dropwise, the coverslip was covered quickly, and placed into a petri dish with wet filter paper, and the petri dish was placed in the dark at 37 ℃ for 2 h. The proportion of red pollen in the 3-5 fields of view in the center of the coverslip was counted, and the average value was calculated by randomly selecting 6 florets for the determination.

The data were processed by Excel and analyzed for significance by SPSS, and then the results were presented in the form of graphs.

The stigmatic pollination was determined by the benzidine-hydrogen peroxide method. During the blooming period, flowers were collected at noon every day at different days after blooming, and the stigmas were put into concave slides, and benzidine-hydrogen peroxide reaction solution (1% benzidine: 3% hydrogen peroxide: water = 4:11:22, v/v) was added drop by drop, then covered with a coverslip and observed under a microscope quickly. If the column head is recognizable, the reaction solution around the column head appears blue with a large number of bubbles. By comparing the amount and size of the bubbles, the strength of the column head can be measured.

The results are analyzed and presented in a graphical format in Table 32-1.

Source: Huang Xiumei et al. 2008.

Note: "+" indicates that the stigma is fertile; "++" indicates that the stigma is strongly fertile: "+/-" indicates that part of the stigma is fertile and part of the stigma is not fertile.

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