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Low-temperature over-summering preservation and cultivation method of brassica napus L microspore culture embryoid

A technology for microspore culture and Brassica napus, which is applied in horticultural methods, botanical equipment and methods, horticulture, etc., can solve problems such as the difficulty of oversummering seedlings, and achieve the effects of low cost, simple method, and improved efficiency

Inactive Publication Date: 2014-11-19
贵州省油料研究所
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0004] The purpose of the present invention is to provide a low-temperature oversummer preservation and cultivation method for brassica napus microspores to culture embryoid bodies, so as to solve the problem in the prior art that it is difficult to culture brassica napus microspores or seedlings to survive summer

Method used

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  • Low-temperature over-summering preservation and cultivation method of brassica napus L microspore culture embryoid
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  • Low-temperature over-summering preservation and cultivation method of brassica napus L microspore culture embryoid

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Embodiment 1

[0020] Embodiment 1 of the present invention: a low-temperature over-summer preservation culture method for culturing embryoid bodies of Brassica napus microspores. After culturing free microspores in NLN-13 medium for 26 days, an embryoid body with an embryo age of 26 days was obtained. The obtained embryoid bodies were placed in new NLN-13 medium, and the pH of the new NLN-13 medium was reduced to 5.5. The embryoid bodies were placed in a glass petri dish with a diameter of 6 cm, and each dish was stored for 150~ 200 embryoid bodies; then they will be stored in a refrigerator at 3°C. After 60 days of storage, they need to be taken out of the refrigerator at 3°C ​​and transferred to an incubator at 8°C for 24 hours, and then replaced with new NLN-13 (PH 5.5) liquid culture medium; replace it and put it back in the incubator at 8°C for 24 hours, then store it in the refrigerator at 3°C ​​for 120 days; during this period, replace the NLN-10 medium every 60 days, The method is t...

Embodiment 2

[0021] Embodiment 2 of the present invention: Brassica napus microspore culture embryoid body low-temperature over-summer preservation culture method, after the free microspores are cultured in NLN-13 medium for 25 days, the embryoid body with an embryo age of 25 days is obtained, The obtained embryoid bodies were placed in new NLN-13 medium, and the pH of the new NLN-13 medium was reduced to 5.5. The embryoid bodies were placed in a glass petri dish with a diameter of 6 cm, and each dish was stored for 150~ 200 embryoid bodies; and then stored in a refrigerator at 3°C ​​for 60 days.

Embodiment 3

[0022] Embodiment 3 of the present invention: a low-temperature over-summer preservation culture method for culturing embryoid bodies of Brassica napus microspores, after culturing free microspores in NLN-13 medium for 28 days, to obtain embryoid bodies with an embryo age of 28 days, The obtained embryoid bodies were placed in new NLN-13 medium, and the pH of the new NLN-13 medium was reduced to 5.5. The embryoid bodies were placed in a glass petri dish with a diameter of 6 cm, and each dish was stored for 150~ 200 embryoid bodies; then they will be stored in a refrigerator at 3°C. After 60 days of storage, they need to be taken out of the refrigerator at 3°C ​​and transferred to an incubator at 8°C for 24 hours, and then replaced with new NLN-13 (PH 5.5) liquid culture medium; replace it and put it back in the incubator at 8°C for 24 hours, and then store it in the refrigerator at 3°C ​​for 90 days; The method is the same as above, and the pH of the NLN-10 medium is 5.5.

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Abstract

The invention discloses a low-temperature over-summering preservation and cultivation method of brassica napus L microspore culture embryoid. The method comprises the following steps: culturing free microspores in an NLN-13 culture medium for 25-28 days to obtain an embryoid at the age of 25-28 days, and arranging the obtained embryoid in a novel NLN-13 culture medium; and preserving the embryoid in a refrigerator at the temperature of 3 DEG C for 60 days; performing transition culture on the embryoid in an incubator, replacing the culture medium with an NLN-10 shallow liquid culture medium, culturing in the incubator again, and preserving the embryoid in the refrigerator at the temperature of 3 DEG C for 120-150 days, wherein the culture medium is replaced with a NLN-10 culture medium once per 60 days in the period of preservation. According to the method, the free microspores of brassica napus L are cultivated to obtain the embryoid at a proper age, which is then preserved at a low temperature, the transfer time is greatly saved, lots of transfer expenditures and manpower resources are saved, and the method has significance for innovating germplasm resources, particularly excellent and specific germplasm resources.

Description

technical field [0001] The invention relates to the field of agricultural planting, in particular to a low-temperature over-summer preservation and culture method for culturing embryoid bodies of brassica napus microspores. Background technique [0002] Brassica napus Brassica napus L is the species with the highest grain yield among the three types of oilseed rape (Chinese cabbage-type rape, mustard-type rape, and Brassica napus). It is currently planted in large quantities in the upper reaches of the Yangtze River in my country and is the most important oil-bearing crop in my country. [0003] Under normal circumstances, after the microspores of Brassica napus produce embryos, the embryoid bodies are inoculated on MS solid medium, and need to be subcultured every 30-45 days in a culture bottle. From March to October, at least 4 subcultures are required. Pollution in summer can cause a large amount of seedling loss, and it is also a large waste of time, labor and financia...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): A01H4/00
Inventor 李超林茂
Owner 贵州省油料研究所
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