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Cryopreservation and recovery culture method for medicinal wild rice stem tip

A technology of cryopreservation and medicinal wild rice, which is applied in the field of plant cell engineering, can solve the problems of long material acquisition cycle, unfavorable large-scale preservation of germplasm resources, and difficulty in obtaining a large number of neat and consistent materials, so as to simplify the operation process and processing time , Conducive to large-scale preservation and the effect of controlling endophyte pollution

Inactive Publication Date: 2019-02-26
SHANGHAI AGROBIOLOGICAL GENE CENT
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

For the first time, Lin Tian et al. adopted a series of measures to improve the resistance of the shoot tip and reduce the damage in each step, and adopted liquid nitrogen direct injection instead of relying on the freezing step of the programmed cooling device, and established the ultra-low temperature preservation system of the shoot tip of common wild rice. , and obtained a patent in 2018 (patent number: ZL 201610131944.1), but there are still disadvantages: 1. Since wild rice explants are sampled in the field, endophyte pollution is difficult to control
2. Since the young stem tip is to be separated from the tightly wrapped leaves, the step of stripping the stem tip requires high technical requirements for the operator, and the work intensity is high, and it is easy to cause mechanical damage to the stem tip during the stripping process, which is not conducive to Large-scale conservation of germplasm resources
3. The period from sampling in the field to obtaining materials suitable for freezing is too long, and it is difficult to obtain a large number of neat and consistent materials, which is not conducive to the quantity and quality control of frozen storage

Method used

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  • Cryopreservation and recovery culture method for medicinal wild rice stem tip
  • Cryopreservation and recovery culture method for medicinal wild rice stem tip

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0025] Embodiment 1, the acquisition of aseptic seedlings of wild rice and the detection of germ removal

[0026] Take the stem section of the underground stem with sprouted buds of wild rice, carefully wash away the sediment attached to the root, soak it with 1% carbendazim for 1 hour, and cultivate it in clean water for 10-20 days, take the newly sprouted aerial buds on the ground, and put them in After repeated rinsing under running water for about 1 hour, disinfect with 70% alcohol for 1 min, 0.1% mercuric chloride for 15 min, rinse with sterile water for several times, strip off the shoot tip of about 2 mm under the dissecting microscope, and inoculate it in a plant containing 4 mg·L -1 BA and 0.5 mg·L -1 Cluster shoots were induced on MS medium with NAA. The culture temperature was 25±2°C, and the light was 12 h d -1 , light intensity 36 µmol m -2 the s -1 . After inducing clustered shoots, transfer to a 10 g L -1 Glucose and 5 g L -1Cultivate in the medium of...

Embodiment 2

[0027] Embodiment 2, the clustered bud of wild rice obtains

[0028] Get the tissue culture seedling of embodiment 1 gained, get the aseptic tissue culture seedling of subculture 30 days, strip off 2-3mm stem tip, put into containing TDZ4mgL -1 2-3 mm clustered shoots were induced in about 5-7 days in MS medium.

Embodiment 3

[0029] Embodiment 3, the device fixation of clustered buds of wild rice

[0030] Use tweezers to lightly dip the pre-cultured clustered buds in 2-4% calcium ion-free sodium alginate solution, and place them neatly on a 5mm×20mm metal mesh loading strip, with 10 buds on each loading strip. Dip the loading strip with the shoot tip into a solution containing 0.2% CaCl, 0.4 molL -1 Sucrose and 2moL -1 Glycerol-based MS liquid medium (without NH 4 + ), after 20 min of fixation, place the loading strip on sterile filter paper to absorb excess liquid.

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Abstract

The invention relates to a cryopreservation and plant regeneration method for medicinal wild rice stem tip buds, in particular to a cryopreservation method of the medicinal wild rice stem tip buds, and belongs to the technical field of plant cell engineering. The cryopreservation and plant regeneration method comprises the following steps: 1. field sampling and tissue culture system establishment;2. detection of endophytes; 3. induction of cluster buds; 4. pre-cultivation of the cluster buds; 5. embedding of the cluster buds; 6. loading and vitrification of the cluster buds; 7. freezing, thawing and washing; 8. recovery culture. The invention provides successful cryopreservation for the medicinal wild rice stem tip buds for the first time, and cryopreserved wild rice recovers well. The method has a very good reference value for cryopreservation of wild rice and gramineous plants.

Description

technical field [0001] The invention relates to a method for preserving the stem tip of the medicinal wild rice, in particular to a method for ultra-low temperature preservation of the shoot tip shoot of the medicinal wild rice. It belongs to the technical field of plant cell engineering. Background technique [0002] Medicinal wild rice is one of the three kinds of wild rice native to China. Due to long-term growth under harsh natural conditions, medicinal wild rice has withstood various natural disasters and natural selection from adverse environments. It is a very scarce and important The germplasm resources of wild rice have been in the wild state for a long time, and have undergone various disasters and natural selection of the environment, forming rich variation types, which have strong resistance to rice diseases and insect pests, strong adaptability to the environment, and It has excellent genes such as high quality, high yield, efficient use of nitrogen and phospho...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): A01N3/00A01H4/00
CPCA01H4/001A01H4/005A01N3/00
Inventor 林田李天菲杨华刘鸿艳韩静周莉魏仕伟龙萍罗利军
Owner SHANGHAI AGROBIOLOGICAL GENE CENT
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