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A kind of rapid culture method of rice suspension cell line

A technology of suspension cells and culture methods, which is applied in the field of rapid cultivation of rice suspension cell lines, can solve the problems of long cycle of rice suspension cell lines, achieve the effects of shortening the test period, maintaining consistency, and increasing the proliferation speed

Inactive Publication Date: 2019-09-20
云南省农业科学院生物技术与种质资源研究所
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AI Technical Summary

Problems solved by technology

[0006] The purpose of the present invention is to solve the problem that the cycle of obtaining rice suspension cell lines is too long in the prior art, and to provide a method for quickly establishing rice suspension cell lines

Method used

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Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0017] Example 1: Establishment of Cultivated Rice 02428 Suspension Cell Line

[0018] In this example, immature or mature embryos of the cultivated rice variety "02428" were used as test materials, and a suspension cell line was rapidly obtained by using the method of the present invention. The implementation process is as follows:

[0019] 1) Selection of explants: The explants were selected from immature embryos or mature embryos of 02428 material independently cultivated in the greenhouse in our laboratory, and the selected embryos of 02428 were placed in air-permeable mesh bags and stored in a refrigerator at 4°C for future use. ;

[0020] 2) Sterilization of explants: Take out the 02428 immature or mature embryos stored in the refrigerator at 4°C, peel off the glumes, put them into a sterilized 100mL triangular flask and soak them in 75% alcohol by volume for 1 minute Carry out surface disinfection, shake the Erlenmeyer flask, rinse 5 times with sterile distilled water...

Embodiment 2

[0024] Embodiment two: the establishment of the suspension cell line of wild rice verruca

[0025] In this example, O. verruculata seed embryos collected in the wild in Yuanjiang County, Yuxi City, Yunnan Province were used as test materials, and a suspension cell line of Oryza verruca was rapidly obtained by using the method of the present invention. The implementation process is the same as in the first embodiment, except for the following points:

[0026] 1) Selection of explants: For explants, the mature embryos of wild rice verruca collected by our research group in Yuanjiang County, Yuxi City, Yunnan Province were selected, and the selected mature embryos were packed in kraft paper envelopes, and brought back to the laboratory and placed at 4 °C Store in the refrigerator for later use;

[0027] 2) Sterilization of explants: Take out the mature embryos of Oryza sativa preserved in a refrigerator at 4°C, peel off the glumes, put them into a sterilized 100mL triangular fla...

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Abstract

The invention relates to a method for rapidly culturing a rice suspension cell line, and belongs to the biotechnical field. The method for rapidly culturing the rice suspension cell line includes the following steps: selection of an explant, sterilization of the explant, callus induction, callus subculture, and suspension cell culture. A suspension cell culture medium and a suspension cell subculture treatment technology are improved. During the subculture treatment of suspension cells, the suspension cultured cells are sequentially screened by sterilized screens with the aperture sizes being about 10 meshes, 20 meshes and 40 meshes after the third-time subculture using a liquid medium to filter out large-granule cell clusters and only reserve cell clusters with uniform size, and the good embryogenic suspension cell line is established through the method in 1.5-2 months. The method has the advantages of low cost, simplicity in operation, shortening of the test cycle, and reduction of the suspension cell line contamination risk, is a new efficient and rapid method, and provides a platform for the relevant study of the rice suspension cell line used as an experimental material.

Description

technical field [0001] The invention relates to a method for rapidly cultivating rice suspension cell lines, which belongs to the field of biological technology. Background technique [0002] Rice is one of the most important food crops in the world, the staple food of more than 50% of the world's population, and also one of the model plants for genetics and genome research. The wild rice verruca is mainly distributed in Yunnan Province and Hainan Province of my country. Because wild rice has been in the wild state for a long time, it has withstood natural selection of various disasters and adverse environments, and has strong stress resistance. It is a natural gene pool and maintains favorable genes that cultivated rice does not have or has disappeared. [0003] Suspension cell lines have a unified source and a consistent genetic background, and there is no influence of individual differences in plant research; the growth conditions are easy to control, the physiological s...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): A01H4/00
Inventor 陈越程在全付坚王波钟巧芳肖素勤陈玲柯学张敦宇殷富有王玲仙余腾琼蒋聪李娥贤黄兴奇
Owner 云南省农业科学院生物技术与种质资源研究所
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