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Acquirement of gymnosperm without external-wall pollen

A gymnosperm and pollen technology, applied in the field of obtaining exine pollen from plants, can solve the problems of lagging biological research, no effective methods and techniques for separating gymnosperm pollen from exine, and achieve great practical application value

Inactive Publication Date: 2006-05-31
INST OF BOTANY CHINESE ACAD OF SCI
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

But so far there is no effective method and technique for isolating the pollen of gymnosperms, which leads to a lag in biological research related to gymnosperms compared to angiosperms.

Method used

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  • Acquirement of gymnosperm without external-wall pollen
  • Acquirement of gymnosperm without external-wall pollen

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0018] Embodiment 1, obtaining the exine wall pollen of white bark loosening

[0019] Include the following steps:

[0020] 1) After drying the mature white bark pine pollen grains at room temperature, store them at -20°C for later use;

[0021] 2) Prepare 0.01% boric acid (H 3 BO 3 ) and 0.01% CaCl 2 15% sucrose solution;

[0022] 3) Use the solution in step 2) to prepare an enzymolysis solution containing 1% cellulase R-10 (Yakult Honsha Co.Ltd) and 0.8% macerozyme R-10 (Yakult Honsha Co.Ltd);

[0023] 4) Put 1-2 mg of pine pollen grains from step 1) at room temperature, put them in 2 mL of the enzymatic hydrolysis solution in step 3), and oscillate the enzymolysis on a shaker at 25° C. at 120 rpm for 2 hours;

[0024] 5) Centrifuge at 1000-2000rpm / min, collect the precipitate, discard the supernatant, replace it with the sucrose solution in step 2) and wash the precipitate 2-3 times with this centrifugal method;

[0025] 6) Place the pollen grains collected in step 5)...

Embodiment 2

[0027] Embodiment 2, obtain the exine wall pollen that green skewer takes off

[0028] 1) After drying the mature green skewer pollen grains at room temperature, store them at -20°C for later use;

[0029] 2) Prepare 0.01% boric acid (H 3 BO 3 ) and 0.03% calcium chloride (CaCl 2 ) of 12% sucrose solution;

[0030] 3) Use the solution in step 2) to prepare an enzymolysis solution containing 1.5% cellulase (Yakult Honsha Co.Ltd) and 1.2% isolated enzyme (Yakult Honsha Co.Ltd);

[0031] 4) After warming 500 μg of green skewer pollen grains from step 1) at room temperature, put it in 20 ml of the enzymatic hydrolysis solution in step 3), and oscillate the enzymatic hydrolysis on a shaking table at 25° C. and 120 rpm for 1.5 hours;

[0032] 5) Centrifuge at 1000-2000rpm / min, collect the precipitate, discard the supernatant, replace it with the sucrose solution in step 2) and wash the precipitate 2-3 times with this centrifugal method;

[0033] 6) Place the pollen grains colle...

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Abstract

A method for preparing the pollen without external wall of gymnosperm includes such steps as enzymolyzing the pollen of gymno sperm in the enzymolyzing liquid containing cellulase (1-2 mass%) and macerozyme (0.5-1.5) for 1.5-2 hr, and slightly pressing the enzymolyzed pollen particles.

Description

technical field [0001] The invention relates to a method for obtaining plant exine pollen, in particular to a method for obtaining gymnosperm exine pollen. Background technique [0002] Pollen grains are considered to be an ideal cell system in the field of biotechnology because of their simple structure, large population size, and ease of in vitro manipulation. The wall of plant pollen grains is usually divided into two layers, namely the inner wall composed of cellulose, pectin, etc. and the outer wall composed of sporopollenin. The inner wall can be removed by enzymatic hydrolysis, while the sporopollenin in the outer wall is lacking. Potent enzymes that degrade it. Exine pollen is a structural unit between intact pollen grain and pollen grain protoplast. It is a unique pollen morphology and physiology research material, which can be used to study the surface structure of the entire pollen The structure and composition of the exgranular wall and the role of the exine in...

Claims

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Application Information

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IPC IPC(8): A01H7/00
Inventor 林金星房克凤
Owner INST OF BOTANY CHINESE ACAD OF SCI
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