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Screening method of allogenic material for promoting vitrification ultra-low temperature storage

A technology of vitrification ultra-low temperature and screening methods, which is applied in the fields of botany equipment and methods, plant preservation, application, etc., can solve the problems of inability to survive, slow recovery of growth, and restriction of exogenous substances, so as to improve recovery growth rate, Reproducible, wide-ranging effects

Inactive Publication Date: 2012-12-19
SHANGHAI JIAO TONG UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0005] At home and abroad, research on cryopreservation technology has been carried out on different explants of more than 200 kinds of plants. However, some plants (such as tropical plants or plants with high water content) have a low survival rate or cannot survive at all after recovery. Source substances are beneficial to improve the survival rate of cryopreservation, but how to screen out some of the more effective exogenous substances from the many exogenous substances that can be added, generally in vitro tissue screening of exogenous substances, and the method of activity detection of tissues or cells after cryopreservation is not direct enough , and there is a certain error, and the recovery of growth is slow to observe directly, and many materials for cryopreservation are rare and endangered plants, and it is impossible to use a large amount of material consumption to screen effective exogenous substances to establish a cryopreservation system. The above reasons greatly restrict The efficiency of effective exogenous substance screening and cryopreservation system establishment

Method used

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  • Screening method of allogenic material for promoting vitrification ultra-low temperature storage
  • Screening method of allogenic material for promoting vitrification ultra-low temperature storage
  • Screening method of allogenic material for promoting vitrification ultra-low temperature storage

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0049] 1. Experimental materials

[0050] Subject: Arabidopsis thaliana ecotype Col-0.

[0051] Experimental reagents:

[0052] 1) Loading solution: MS culture solution containing 2M glycerol and 0.4M sucrose;

[0053] 2) Vitrification solution: MS culture solution containing 30% w / v glycerol, 15% w / v ethylene glycol, 15% w / v dimethyl sulfoxide and 0.4M sucrose;

[0054] 3) Washing solution: MS culture solution supplemented with 1.2M sucrose and MS culture solution supplemented with 1.0M sucrose;

[0055] 4) MS solid medium: 1L medium contains 1900mg KNO 3 , 1650mg NH 4 NO 3 , 170mg KH 2 PO 4 , 370mg MgSO 4 ·7H 2 O, 440 mg CaCl 2 2H 2 O, 37.3 mg Na 2 - EDTA, 27.8mg FeSO 4 ·7H 2 O, 100mg inositol, 0.5mg niacin, 0.5mg pyridoxine hydrochloride, 0.1mg thiamine hydrochloride, 2mg glycine, 0.83mg KI, 6.2mg H 3 BO 3 , 22.3 mg MnSO 4 4H 2 O, 8.6mg ZnSO 4 ·7H 2 O, 0.25 mg Na 2 MoO 4 2H 2 O, 0.025 mg CuSO 4 ·5H 2 O, 0.025 mg CoCl 2 ·6H 2 0, 30g sucrose, 10g ag...

Embodiment 2

[0071] Example 2 Verification experiment of exogenous substances promoting vitrification cryopreservation

[0072] 1. Subjects

[0073] 1) Embryogenic callus of Agapanthus praecox ssp.orientalis, protocorm of Dendrobium wardianum.

[0074] 2) Exogenous substances: 1 mM VC, 1 μM ABA, 10 mM GB and 0.16 mM GSH.

[0075] 3) 1 / 2MS solid medium is a solid medium in which the macroelements in MS solid medium components are halved and the rest of the components remain unchanged.

[0076] 4) 1 / 2 MS culture solution is a culture solution in which the macroelements in the MS culture solution are halved and the rest of the components remain unchanged.

[0077] 2. Experimental method

[0078] 2.1 Verification experiment of exogenous substances promoting the vitrification of protocorms of Dendrobium sclerotii

[0079] 2.1.1 Preparation of Protocorm of Dendrobium spp.

[0080] Acquisition of protocorms of Dendrobium maxima: Uncracked capsules 6 months after pollination (mature) were obt...

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Abstract

The invention relates to the field of vitrification ultra-low temperature storage, in particular to a screening method of an allogenic material for promoting vitrification ultra-low temperature storage; an arabidopsis thaliana seedling is adopted as an experimental object; the allogenic material is added into vitrification solution for the vitrification ultra-low temperature storage of the arabidopsis thaliana seedling; the arabidopsis thaliana seedling which is not added with the allogenic material is adopted as a contrast; and the effect of the allogenic material is evaluated by comparing the recovery rate. According to the screening method of the allogenic material for promoting vitrification ultra-low temperature storage, the result is accurate, the repeatability is strong, the scope of application is wide, batch screening can be carried out, the efficiency is high, an effective approach is provided for screening the allogenic material, technical guarantee is provided for a vitrification ultra-low temperature storage technology, and a guiding role is played to the storage of plant species resources.

Description

technical field [0001] The invention relates to the field of vitrification cryopreservation, in particular to a screening method for exogenous substances that promote vitrification cryopreservation. Background technique [0002] Germplasm resources are the general term for the genetic material that determines the species and transmits its rich genetic information from parents to offspring. It is the material basis for species evolution, genetic research and plant breeding. Realizing the preservation of germplasm resources is the strategic demand of my country's social, ecological and economic development. [0003] Cryopreservation is a modern in vitro preservation technology for germplasm resources developed in the 1970s. Usually stored in liquid nitrogen, the substance metabolism and growth activities in the preserved material cells are almost completely stopped, and they are in a relatively stable biological state, achieving the purpose of long-term preservation of germpl...

Claims

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Application Information

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IPC IPC(8): A01N3/00
Inventor 申晓辉任丽张荻
Owner SHANGHAI JIAO TONG UNIV
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