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Ultralow-temperature preservation method for stem tips of wild rice

A technology of ultra-low temperature preservation and rice strips, which is applied in the field of plant cell engineering, can solve problems such as difficult to obtain materials, damage to shoot tip growth points, and difficult to cultivate, so as to reduce the difficulty of operation, simplify the operation process and processing time, and facilitate large-scale production saved effect

Inactive Publication Date: 2020-02-18
SHANGHAI AGROBIOLOGICAL GENE CENT
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, unlike ordinary wild rice, Oryza sativa is distantly related to cultivated rice, and is a typical difficult-to-cultivate plant, and it is difficult to obtain a large amount of neat and consistent materials in a short period of time.
In addition, although the operation steps are simplified by cutting the buds, the base plate of the buds is often cut off due to careless operation during the cutting process, which will damage the nearby shoot tip growth points and affect the survival rate.

Method used

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  • Ultralow-temperature preservation method for stem tips of wild rice
  • Ultralow-temperature preservation method for stem tips of wild rice

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0023] Embodiment 1, the disinfection of wild rice and the processing of breaking dormancy

[0024] Take wild rice seeds, sterilize with 70% alcohol for 1 min, 0.1% sodium hypochlorite for 15 min, rinse with sterile water several times, and wash with 5% PEG, 200 mgL -1 GA 3 Soak the seeds in MS liquid medium for 2 days at room temperature in the dark.

Embodiment 2

[0025] Embodiment 2, the embryogenic callus induction of wild rice

[0026] Get the processed seed of embodiment 1, inoculate and contain 6-BA0.5mgL -1 , 2,4-D 2mgL -1 The callus was induced in MS medium, cultured at 25±2°C, and cultured in the dark. White granular callus can be seen in about 20 days, which is stripped from the seeds, transferred to a new medium, and subcultured once every 10-20 days;

Embodiment 3

[0027] Embodiment 3, wild rice cluster bud induction and rooting culture

[0028] Get the embryogenic callus that embodiment 2 obtains, put into containing 6-BA0.5mgL -1 , NAA0.1mgL -1 In MS medium, culture temperature is 25±2℃, light is 12 h·d -1 , light intensity 36 µmol m -2 the s -1 , About 10-15 days to induce callus containing 2-3mm clustered shoots. Take clustered shoots of 2-3mm, cut off the base disc callus, and inoculate the buds one by one with NAA0.1mgL -1 In MS medium, new roots of 1-2mm were induced in about 3-5 days.

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Abstract

The invention relates to an ultralow-temperature preservation and plant regeneration method for rooted buds of Oryza meyeriana, in particular to an ultralow-temperature preservation method for the rooted buds of the Oryza meyeriana, and belongs to the technical field of plant cell engineering, the ultralow-temperature preservation and plant regeneration method comprises the following steps: 1, seed dormancy breaking treatment and embryogenic callus induction; 2, inducing of cluster buds; 3, rooting culture of the cluster buds; 4, pre-culturing of the rooted buds; 5, embedding of the rooted buds; 6, loading of the rooted buds and vitrification treatment; 7, freezing, unfreezing and washing; and 8. recovery culture. According to the method, the rooted buds of the Oryza meyeriana are successfully preserved at ultralow temperature for the first time, and the growth recovery condition of the preserved wild rice is good. Good reference values are realized on the ultralow-temperature preservation of wild rice and gramineous plants.

Description

technical field [0001] The invention relates to a method for preserving wild rice verruca, in particular to a method for ultra-low temperature preservation of rooted shoots of wild rice verruca, belonging to the technical field of plant cell engineering. Background technique [0002] Oryza sativa is one of the three species of wild rice native to China. It is an important genetic resource for rice breeding and improvement, and has extremely high scientific research value. The current rapidly deteriorating environment, unfavorable climatic conditions and the rapid expansion of modern human economic activities have led to the loss and deterioration of wild rice habitats, and the sharp reduction of habitats; in addition, wild rice has exposed stigmas, high outcrossing seed setting, and seed dormancy. Strong, strong shattering, low germination rate of biological characteristics. These factors make Oryza verruca species face the danger of extinction, and it is urgent to protect ...

Claims

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Application Information

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IPC IPC(8): A01N3/00A01H4/00
CPCA01H4/001A01H4/005A01H4/008A01N3/00
Inventor 林田阿新祥李天菲聂元元韩静董超周莉腾小英杨华刘鸿艳龙萍罗利军
Owner SHANGHAI AGROBIOLOGICAL GENE CENT
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