Vitrification cryopreservation method of dendrobium wardianum warner protocorm

A technology of vitrification ultra-low temperature and dendrobium sheaths, which is applied to the preservation of plants, botany equipment and methods, applications, etc., and can solve problems that have not been reported

Inactive Publication Date: 2011-08-10
SHANGHAI JIAO TONG UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

This method has the advantages of simplicity, low cost, and stable heredity of preserved materials, and is of great significance to the preservation of some rare and endangered plant germplasm resources. It has been applied to the orchid family Phaius tankervilleae (Bank ex L'Herit .) Bl.) seeds, bletilla striata (Bletilla striata Maxim.) zygotic embryo, Cymbidium grandiflorum Sw. 'Idol' stem tip, protocorm, vanilla fragrans (Salisb.) Ames 'Andrews'), protocorms of Dendrobium cruentum Rchb.f., and seeds, protocorms and protoplasts of Dendrobium officinale Kimuraet Migo, but the cryopreservation technology of Dendrobium officinale Kimuraet Migo has not been reported yet.

Method used

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  • Vitrification cryopreservation method of dendrobium wardianum warner protocorm
  • Vitrification cryopreservation method of dendrobium wardianum warner protocorm

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0021] The selection state of the protocorm before saving: in 1 / 2MS, 6-BA0.2mg·L -1 , NAA0.5mg·L -1 , Proliferate on 2% sucrose medium for 60 days, and the diameter of a single protocorm after stripping is about 2-3 mm.

[0022] Such as figure 1 As shown, after the protocorm of Dendrobium sclerotii was vitrified and cryopreserved according to the above steps, it turned green and showed vitality after 30 days of recovery culture (left picture), and the protocorm began to proliferate after 90 days (right picture).

Embodiment 2

[0024] Select 4 kinds of osmotic pressure regulators, sucrose, sorbitol, mannitol and glucose, the concentrations are 0.2, 0.4, 0.6, 0.8 mol L -1 , after pretreatment at room temperature for 8 days, the content of 0.8mol L -1 The highest survival rate was 43.5% after sucrose pretreatment.

[0025] Room temperature - hypertonic (25 ℃, 0.8mol L -1 sucrose) and hypotonic-hypertonic (4°C, 0.8mol L -1 Sucrose) pretreatment for 4, 6 and 8 days, after dehydration by loading solution and PVS2, it was concluded that low temperature-hypertonic treatment for 6 days could achieve a higher survival rate, which was 46.6%.

Embodiment 3

[0027] After the protocorm was pretreated by low temperature-hypertonicity, four kinds of vitrification solutions PVS1 (1 / 2MS, 22% glycerol, 15% polyethylene glycol, 15% ethylene glycol, 7% dimethyl Sulfoxide and 0.5mol L -1 sorbitol), PVS2, PVS4 (1 / 2MS, 35% glycerol, 20% ethylene glycol and 0.6mol L -1 sucrose), PVSW (1 / 2MS, 10% proline and 0.6mol L -1 40min after sucrose) treatment and dehydration in the loading solution, the best vitrification solution was screened out to be PVS2.

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Abstract

The invention provides a vitrification cryopreservation method of dendrobium wardianum warner protocorm, relating to the technical field of cryopreservation. The method comprises the following steps of: after preprocessing the protocorm with high permeability at low temperature, transferring the protocorm to a loading liquid; performing dewatering treatment, transferring the protocorm to a freezing tube which is filled with vitrification solution; and rapidly filling with liquid nitrogen for cryopreservation. The method in the invention provides technical guarantee for the preservation of theendangered dendrobium wardianum warner germ plasm resource. The invention provides an effective path for the preservation of the dendrobium plant even the arethusa resource and the cultivation of newspecies by using plant diversity.

Description

technical field [0001] The invention relates to a method in the technical field of low-temperature preservation, in particular to a method for vitrification ultra-low temperature preservation of protocorms of Dendrobium sheaths. Background technique [0002] Dendrobium wardianum Warner has large flowers, bright colors, and a long flowering period. It is a species with high ornamental value in the genus Dendrobium. In addition, fresh or dried stems can be used as medicine, which is beneficial to the stomach, nourishing yin and clearing heat. As a wild species of the second largest genus of Orchidaceae in my country, and a national II-level protected plant, Dendrobium sclerotii has strict requirements on the ecological environment. In a very narrow distribution area, it can only be found in the tropical rainforest at an altitude of 1350-1900m from southeastern to western Yunnan in my country. Among them, Myanmar, Thailand and northeastern India are also distributed. Due to the...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): A01N3/00
Inventor 申晓辉吴元玲张荻任丽
Owner SHANGHAI JIAO TONG UNIV
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