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In-vitro culturing method of gymnospermae pollen and its special culturing base

A technology for in vitro culturing of pollen and gymnosperms is applied in the field of an in vitro culturing method of plant pollen and its special medium, and can solve the problems that there is no effective method and technology for culturing gymnosperm pollen in vitro, and the lag of biological research angiosperms and the like.

Inactive Publication Date: 2009-01-28
INST OF BOTANY CHINESE ACAD OF SCI
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Whether the pollen can germinate normally and whether the pollen tube can grow normally is an important prerequisite for the above-mentioned research work, but so far there is no effective method and technology for cultivating gymnosperm pollen in vitro, which leads to biological problems related to gymnosperms. Research lags relative to angiosperms

Method used

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  • In-vitro culturing method of gymnospermae pollen and its special culturing base
  • In-vitro culturing method of gymnospermae pollen and its special culturing base
  • In-vitro culturing method of gymnospermae pollen and its special culturing base

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Embodiment 1

[0024] The in vitro culture of embodiment 1, green stalk and white stalk pollen

[0025] 1. Preparation medium

[0026] Get sucrose 120g, boric acid (H 3 BO 3 ) 0.2g and calcium chloride (CaCl 2 ) 0.3g, and dilute to 1000mL with water to obtain the in vitro culture medium of gymnosperm pollen.

[0027] 2. In vitro culture of green stem and white stem pollen

[0028] Using the method of the present invention to carry out in vitro culture of the pollen of the green stalk and the white stalk, comprising the following steps:

[0029] 1) After drying the pollen grains of mature green stems and white stems at room temperature, store them at -20°C for later use;

[0030] 2) Take 5 mg of pollen grains from step 1), hydrate at room temperature for 30 minutes, place in 5 mL of medium prepared in step 1, and shake and culture at 25°C and 120 rpm for 18 hours;

[0031] 3) Centrifuge at 1000rpm to collect the precipitate, discard the supernatant, wash the precipitate twice with 12% s...

Embodiment 2

[0033] The in vitro culture of embodiment 2, white bark pine pollen

[0034] 1. Preparation medium

[0035] Take 150 g of sucrose, 0.1 g of boric acid and 0.1 g of calcium chloride, and dilute to 1000 mL with water to obtain an in vitro culture medium for gymnosperm pollen.

[0036] 2. In vitro culture of pine pollen

[0037] Carry out in vitro culture with the method of the present invention to white bark pine pollen, comprise the following steps:

[0038] 1) After drying the mature white bark pine pollen grains at room temperature, store them at -20°C for later use;

[0039] 2) Take 5 mg of pollen grains from step 1), hydrate them at room temperature for 20 minutes, place them in 4 mL of medium prepared in step 1, and shake and culture them for 72 hours at 27°C and 100 rpm in the dark;

[0040] 3) Centrifuge at 500rpm to collect the precipitate, discard the supernatant, wash the precipitate 3 times with 15% sucrose solution and centrifuge, and obtain the pollen tube of Pi...

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Abstract

The invention discloses gymnosperm isolated culture method and the special using culture medium. The culture medium receipt is that: cane sugar 120-150g, boric acid 0.1-0.2g and calcium chloride 0.1-0.3g. They are settled to 1000 ml by water. The culturing method is that the gymnosperm pollen is hydrated for 20-30 minutes under room temperature, then it is cultured in the said medium for 6-72 hours, finally the pollen tube is got after pollen sprouting. The essential environment is supplied for pollen sprouting and growth. The culture by this invention is not only fast and easy, but also good in culturing result. A great deal of gymnosperm pollen tube with vitality can be got, and there is great practical application value.

Description

technical field [0001] The invention relates to an in vitro culture method of plant pollen and a special culture medium thereof, in particular to an in vitro culture method of gymnosperm pollen and a special culture medium thereof. Background technique [0002] Pollen germination is the process in which the volume of pollen increases after absorbing water, and the internal turgor pressure increases, so that the inner wall of pollen protrudes outward along the germination hole, forming a pollen tube. Initiating pollen germination is a complex process, and the basic conditions for germination require the participation of sugar sources, boric acid and mineral elements (such as exogenous Ca). At present, the pollen of various angiosperms has been successfully cultured in vitro. For example, the in vitro culture technology of lily and tobacco pollen has been quite mature, while the research on the in vitro culture of gymnosperm pollen is less and the technology is relatively back...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12N5/04C12N3/00
Inventor 林金星田翠婷吕洪飞
Owner INST OF BOTANY CHINESE ACAD OF SCI
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