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Cryopreservation method for plant germplasm of Eriobotrya by using combined method of programmed freezing method and vitrification method

A technology of ultra-low temperature preservation and programmed cooling, applied in botany equipment and methods, plant regeneration, gardening methods, etc., can solve the problems of complex components of vitrification protective agents, affecting survival rate, and failure to provide, etc., to achieve stable regeneration survival rate, The method has good repeatability and the effect of overcoming randomness and uncertainty

Active Publication Date: 2016-04-13
SOUTH CHINA AGRI UNIV
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Problems solved by technology

This technical scheme is only applicable to the cultivated loquat varieties in the genus Loquat, and there is no report to test other species of the existing Loquat genus, there is no repeatability between species, and it cannot be repeated; the composition of the vitrification protective agent used in this technical scheme is relatively complicated , and it is necessary to re-add the vitrification protective agent repeatedly. The operation of adding protective agent is relatively complicated, which affects the survival rate. The highest survival rate is 52.3%.
There is no specific technical solution to restore plant regeneration and culture that can be repeated by about 50%

Method used

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  • Cryopreservation method for plant germplasm of Eriobotrya by using combined method of programmed freezing method and vitrification method
  • Cryopreservation method for plant germplasm of Eriobotrya by using combined method of programmed freezing method and vitrification method
  • Cryopreservation method for plant germplasm of Eriobotrya by using combined method of programmed freezing method and vitrification method

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Experimental program
Comparison scheme
Effect test

Embodiment 1

[0051] This embodiment provides a stable, high survival rate, novel program method for cryopreservation of Loquat plants, which is called the programmed cooling vitrification method. The specific processing method flow is as follows:

[0052] S1. Material pretreatment before programmed cooling. Put the stem tip of Loquat (cultivated loquat, wild loquat or tissue-cultured seedlings of loquat interspecific hybrid progeny, take the stem tip, about 0.5cm long, and disinfect it according to the conventional method) in the pre-medium MS+BA1.0mg· L -1 +NAA0.1mg·L -1 +0.1mol·L -1 Sucrose+5%DMSO pre-cultured for 3 days;

[0053] S2. Transfer to PVS (MS solution + 0.4mol L -1 Sucrose+2mol·L -1 Glycerol) loaded at 4°C for 9h, figure 2 Shown is a vitrified loquat shoot tip.

[0054] The treatment conditions of step S2 are based on the long-term results based on the relationship between the loading time of vitrification solution (within 6 hours) and the survival rate of loquat aft...

Embodiment 2

[0075] The experimental results of Example 1, Comparative Example 1, and Comparative Example 2 were compared. The test data were analyzed by variance with SPSS 19.0 software, and the averages were compared by Duncan's new multiple range test. Survival rate (%) = number of surviving shoot tips / number of inoculated shoot tips × 100%. The comparison results are shown in Table 1:

[0076] Table 1

[0077]

[0078] Note: The letters in the table indicate that the mean values ​​are significantly different (P<0.05) through Duncan's new multiple level difference test.

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Abstract

The invention discloses a cryopreservation method for plant germplasm of Eriobotrya by using a combined method of a programmed freezing method and a vitrification method. The programmed freezing method and the vitrification method are scientifically and organically combined. Innovative improvements of key technology, such as a medium formula and a cryoprotectant formula for cryopreservation pretreatment of plant shoot tips of Eriobotrya, a cooling program design for cryopreservation of loquat shoot tips, a plant regeneration medium formula and a cultivation condition after cryopreservation of the plants of Eriobotrya, and the like are carried out. A novel program method for cryopreservation of plants of Eriobotrya with effect stability and high survival rate is innovated, and the survival rate is 90.0-97.50%. The method can be applied to cultivated species, wide species and interspecific hybridized materials of plants of Eriobotrya, and can replace plantation, so that manpower, material resources and land are saved; damages to loquat preservation during plantation due to insect diseases and natural disasters are prevented.

Description

technical field [0001] The invention relates to the technical field of low-temperature preservation of plant germplasm, and more specifically relates to a method for ultra-low temperature preservation of loquat plant germplasm by combining a programmed cooling method and a vitrification method. Background technique [0002] Plant germplasm resources are the most precious natural resources owned by human beings. [0003] Since the 1960s, many countries have increased investment and research efforts in the collection and preservation of plant germplasm resources, especially in countries with developed economies that have established relatively complete field gene banks. The disadvantage is that field germplasm preservation occupies a large area, which consumes a lot of manpower, material and financial resources, and is also vulnerable to various natural disasters, which forces people to explore more practical, safe and economical preservation methods. Until the middle of the ...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): A01H4/00
CPCA01H4/005
Inventor 林顺权刘义存
Owner SOUTH CHINA AGRI UNIV
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