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Ultralow-temperature preservation method for grapefruit germplasm pollen

An ultra-low temperature preservation and pollen technology, which is applied in the fields of plant preservation, botanical equipment and methods, and application, can solve the problems of poor sealing and packaging of pollen, insufficient rewetting of recovered pollen, and increased damage to pollen cells by ice crystals. The effect of germination rate

Active Publication Date: 2016-12-07
INST OF CROP SCI CHINESE ACAD OF AGRI SCI
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, the inventors have found through experiments that the ultra-low temperature preservation of pomelo germplasm pollen using this technical solution will lead to ① insufficient rehumidification during ultra-low temperature preservation of revived pollen, which may cause the pollen to fail to germinate in vitro on the medium after thawing; ② pollen If the airtight packaging is not tight, the pollen may be immersed in liquid nitrogen to cause freezing damage and lose pollen germination vigor; Pollen germination activity and other defects, therefore, this method is not suitable for cryopreservation of pomelo germplasm pollen

Method used

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  • Ultralow-temperature preservation method for grapefruit germplasm pollen
  • Ultralow-temperature preservation method for grapefruit germplasm pollen
  • Ultralow-temperature preservation method for grapefruit germplasm pollen

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0049] Example 1 Effects of thawing methods and germination medium on pollen germination rate after cryopreservation

[0050] 1. Experimental materials

[0051] The materials used in the experiment were three pomelo germplasm pollen of "Zuoshi pomelo", "Kui pomelo" and "Beizhuan pomelo" provided by the National Fruit Tree Germplasm Citrus Garden (Chongqing).

[0052] 2. Experimental method and specific steps

[0053] - Pollen drying treatment: the collected anthers are dried under an incandescent lamp (at a temperature of about 30°C) until the water content is 10-20%.

[0054] —Preservation in sealed packaging with liquid nitrogen: After the pollen is properly dried, put it into a 1.8ml cryopreservation tube (Nunc, USA), and put it into liquid nitrogen (CBS S3000-AB SERIES) for storage.

[0055] — Thawing: Take the frozen storage tubes containing pollen out of the liquid nitrogen tank and thaw them in two ways: (1) rinse with tap water at 25°C for about 15 minutes; (2) quick...

Embodiment 2

[0061] Example 2 The effect of rehumidification on the germination rate of grapefruit pollen

[0062] 1. Experimental materials

[0063] The varieties used in the experiment were two pomelo germplasm pollen of "Zuoshiyou" and "Kuiyou" provided by the National Fruit Tree Germplasm Citrus Garden (Chongqing).

[0064] 2. Experimental method and specific steps

[0065] - Pollen drying treatment: the collected anthers are dried under an incandescent lamp (at a temperature of about 30°C) until the water content is 10-20%. Three samples of "left pomelo - water content 10%", "left pomelo - water content 20%" and "kui pomelo - water content 10%" were obtained.

[0066] —Preservation in sealed packaging with liquid nitrogen: After the pollen is properly dried, put it into a 1.8ml cryopreservation tube (Nunc, USA), and put it into liquid nitrogen (CBS S3000-AB SERIES) for storage.

[0067] — Thawing: Take the frozen storage tubes containing pollen out of the liquid nitrogen tank and t...

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Abstract

The invention relates to a plant ultralow-temperature preservation method, and in particular to an ultralow-temperature preservation method for a grapefruit germplasm pollen. The ultralow-temperature preservation method comprises the following steps: (1) drying a pollen, namely, drying the pollen till the water content is 10-20%; (2) performing pollen cryopreservation, namely, sealing and packaging the dried pollen, and putting the pollen in liquid nitrogen for preservation; (3) performing pollen anabiosis, namely, taking out the pollen from the liquid nitrogen, and thawing the pollen for 5-15 minutes at 25-37 DEG C; (4) performing pollen rewetting for 0-3 hours; (5) performing in-vitro germination. By adopting the method, key factors of ultralow-temperature preservation applicable to the grapefruit germplasm pollen, such as the water content range, the thawing condition, the rewetting condition and the in-vitro germination condition, are specified. Due to optimization and restriction on the conditions, the germination rate of the grapefruit germplasm pollen after ultralow-temperature preservation is remarkably increased.

Description

technical field [0001] The invention relates to a method for ultra-low temperature preservation of plants, in particular to a method for ultra-low temperature preservation of pomelo germplasm pollen. Background technique [0002] Pomelo germplasm (Citrus grandis (L.) Osbeck) is an important kind of germplasm resources, which is a citrus plant of Rutaceae, native to Asia. Pomelo fruit is beautiful in shape, delicious in taste, rich in nutrition and high in economic value. In recent years, its development and utilization at home and abroad are in the ascendant. The natural environment of my country's pomelo production areas is superior and the ecological conditions are complex. Coupled with the monoembryonic nature of pomelo, seedlings are prone to variation, and extremely rich germplasm resources have been formed for thousands of years. According to incomplete statistics, there are more than 200 varieties, strains, types and strains of pomelo germplasm resources in China. ...

Claims

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Application Information

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IPC IPC(8): A01N3/00
CPCA01N3/00
Inventor 张金梅陈晓玲卢新雄辛霞尹广鹍何娟娟
Owner INST OF CROP SCI CHINESE ACAD OF AGRI SCI
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