Method for determining pollen number in rice anther

Abstract

The invention relates to a method for determining the pollen number in a rice anther. The method is characterized by: making 168 grids with a size of 2280 micrometers*3048 micrometers by metal cautery lines at the center of a reticle; dripping 1-2 drops of an I2-KI solution with a concentration of 0.1%-0.3% on the reticle, squeezing an anther by a tweezer repeatedly at different places in the I2-KI solution till the pollen in the anther is dispersed completely, then adding 1-2 drops of an I2-KI solution with a concentration of 1% into the original solution to dye the pollen into a deep color; distributing the pollen sample in the grids of the reticle uniformly; photographing the grids in the reticle completely one by one in order to record them by a digital microscope; and employing image analysis software to carry out differential counting on the photographed pollen in terms of active pollen, inactive pollen and total pollen. The method provided in the invention can accurately determine the number of active pollen and inactive pollen in the anther, and has the characteristics of convenient determination, reliable data, and high determination efficiency.

Description

Technical field: [0001] The invention relates to a method for measuring the number of pollen in rice anthers, which belongs to the technical field of rice crop planting research and application. Background technique: [0002] The average number of pollen in each anther of rice is about 800-2500 grains. The pollen number is one of the important reproductive performance indicators of rice varieties; Therefore, pollen count has also become an important characteristic index to measure the response of varieties to stress. In addition, in the seed production of hybrid rice, the effective pollen yield of the male parent is also the technical basis for determining the row ratio of the parent parent in the seed production field. Therefore, rice research and production often involve the determination of the number of rice pollen per drug. At present, there are three main methods for measuring the total number of rice pollen per drug: (1) manual counting method: the anther to be meas...

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Problems solved by technology

[0003] The shortcomings of the above method are: method (1) due to the large pollen amount of rice, under a sufficiently large resolution, a microscopic field of view cannot fully display the sample to be tested, and multi-field observations are often used. Therefore, the traditional The method can only choose the sub-field sampling method, which makes the workload of manual counting very large, the accuracy of counting is low, and the amount of anthers measured at one time is very limited.

The problem with method (2) is that it is difficult to accurately prepare a suspension liquid that can evenly distribute the pollen, because the specific gravity of the liquid must match the quality of the pollen, and it is difficult to measure the quality of a single grain of pollen, and the quality of pollen of different varieties There may be large differences; at the same time, the determination of this method is mainly through the determination of the number of pollen in a certain field of view and then converted according to its volume to obtain the total number of pollen of each drug, there is a certain error

Method (3) Since the scanner does not have a microscopic function, and the pollen is very small, there is a large error in counting the scanned pollen photos; in addition, there is also impurity interference during the scanning process, and the inactivated pollen is in the iodine-potassium iodide solution. Lighter in color, can also make accurate counting difficult

Therefore, the above-mentioned method has obvious deficiencies in the measurement accuracy and measurement efficiency, and cannot meet the requirements of rice research and production practice.

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