Looking for breakthrough ideas for innovation challenges? Try Patsnap Eureka!

Method for appraising quality of plant callus

A callus and quality identification technology, applied in the field of plant tissue culture, can solve problems such as inability to distinguish callus, and achieve the effects of accurate judgment and simple operation.

Inactive Publication Date: 2013-01-09
WUHAN UNIV
View PDF0 Cites 0 Cited by
  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

The existing callus judgment methods cannot distinguish callus with different physiological functions. Therefore, a physiological and biochemical method that can transform callus quality research from qualitative judgment to quantitative analysis and more accurately distinguish callus quality status is very much needed. The application of this analysis method can not only judge whether the callus is suitable for genetic transformation, but also help to study the influence of medium components on the quality of callus, the difference of gene expression and expression of callus in different states. epigenetic mechanism

Method used

the structure of the environmentally friendly knitted fabric provided by the present invention; figure 2 Flow chart of the yarn wrapping machine for environmentally friendly knitted fabrics and storage devices; image 3 Is the parameter map of the yarn covering machine
View more

Image

Smart Image Click on the blue labels to locate them in the text.
Viewing Examples
Smart Image
  • Method for appraising quality of plant callus
  • Method for appraising quality of plant callus
  • Method for appraising quality of plant callus

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0017] Example 1 Analysis of Transgenic Efficiency Using Relative Fluorescence Values

[0018] 1.1 Material description

[0019] 10 kinds of calli were selected for the transgenic experiment: ZH11-1, ZH11-2, 9311-1, 9311-2, 9311-3, Njx74-1, Njx74-2, YTA-1, YTA-2, YTA-3 . Among them, ZH11-1, 9311-1, Njx74-1, and YTA-1 are calli induced and subcultured on LY medium, and ZH11-2, 9311-3, and YTA-3 are induced and subcultured on N6 medium. Subcultured calli, 9311-2, Njx74-2, YTA-2 are calli induced on LY medium but subcultured on N6 medium. Among them, LY medium was purchased from Wuhan Luoying Biotechnology Co., Ltd. (LY medium kit includes LY medium, LY-A reagent, LY-B reagent, etc.).

[0020] Transgenic strain: EHA105

[0021] Transgenic plasmid: pCAMBIA1301

[0022] Fluorescence detection: Take 0.5g of various calli and add them to 2ml of 1M NaCl solution, grind until homogenized, centrifuge at 14,000g for 5min, resuspend the supernatant and precipitate with 60% (W / V) su...

Embodiment 2

[0100] Example 2 Using Relative Fluorescence Values ​​to Analyze Changes in Callus Quality During Culture

[0101] Materials and methods

[0102] Experimental materials: japonica rice material Zhonghua 11 (ZH11), indica rice material Nanjing Xian 74 (NJX74).

[0103] experimental method:

[0104] Induction: Induction and subculture were carried out on LY medium (purchased from Wuhan Luoying Biotechnology Co., Ltd.) to obtain embryogenic callus. Induction method, first remove the glumes of mature embryos, then soak in 75% ethanol for 1 min, 0.75% mercuric chloride aqueous solution for 15 min, wash 5-6 times with sterile water, and inoculate on the induction medium. The induction medium is LY medium supplemented with 0.5g / L glutamine, 0.8g / L acid hydrolyzed casein, 30g / L sucrose, 2.5g / L phytogel, and 100μl / L LY-A; to induce growth Conditions: Induction at 33 °C. Starting from the 10th day, samples were taken and analyzed on the tenth, fourteenth, seventeenth and twen...

Embodiment 3

[0110] Example 3 Analyzing the Effects of Different Mediums with Relative Fluorescence Values

[0111] Materials and methods

[0112] One japonica variety ZH11 was selected. Four indica varieties MH63, YTB, NJX74, 9311 were selected.

[0113] ZH11-L, MH63-L, YTB-L, NJX74-L, 9311-L are the corresponding varieties of calli induced and subcultured on LY medium. ZH11-N, MH63-N, YTB-N, NJX74-N, 9311-N were induced and subcultured on N6 medium for the corresponding varieties of calli. The culture condition was 30°C, and samples were taken for analysis 12 days after induction for 30 days. Analysis method is with embodiment 1.

[0114] result

[0115] Table 2 The culture results of LY medium and N6 medium on callus of different varieties

[0116] MH63-L MH63-N YTB-L YTB-N NJX74-L NJX74-N ZH11-L ZH11-N 9311-L 9311-N RUF-1 a 1767.84 2624.26 905.58 1792.5 1458.64 1754.49 875.394 775.494 1355.69 3341.35 RUF-2 a 1784.96 2560...

the structure of the environmentally friendly knitted fabric provided by the present invention; figure 2 Flow chart of the yarn wrapping machine for environmentally friendly knitted fabrics and storage devices; image 3 Is the parameter map of the yarn covering machine
Login to View More

PUM

PropertyMeasurementUnit
wavelengthaaaaaaaaaa
Login to View More

Abstract

The invention relates to a method for appraising quality of plant callus. In the method, the quality of the callus is appraised by detecting relative fluorescence intensity of the callus. In the invention, physiological indices of the callus are adopted to judge the quality of the callus for the first time, so that the judgment on the quality of the callus is advanced from subjective empirical qualitative research of conventional surface observation to physiological and biochemical quantitative analysis of an instrument. The method is easy to operate, and the quality of the callus can be accurately judged.

Description

technical field [0001] The invention relates to the technical field of plant tissue culture, specifically the detection and application of plant callus. technical background [0002] At present, our judgment on the quality of the callus state is based on observation, that is, through the observation of the morphology of the callus surface, or through the observation of the section of the callus. The simple and convenient observation of tissue morphology requires a lot of experience, and the judgment results of different individuals are quite different. Sometimes the surface state does not necessarily represent the physiological state, so misjudgment occurs. For example, we found that the appearance of indica rice variety G44 is It conforms to the characteristics of embryogenic callus but the transformation rate is extremely low. Although slice observation can be more accurately judged from the cell level, the callus is not uniform, but is composed of cells of different qual...

Claims

the structure of the environmentally friendly knitted fabric provided by the present invention; figure 2 Flow chart of the yarn wrapping machine for environmentally friendly knitted fabrics and storage devices; image 3 Is the parameter map of the yarn covering machine
Login to View More

Application Information

Patent Timeline
no application Login to View More
Patent Type & Authority Patents(China)
IPC IPC(8): C12Q1/02G01N21/64C12Q1/68
Inventor 李阳李阳生丁博
Owner WUHAN UNIV
Who we serve
  • R&D Engineer
  • R&D Manager
  • IP Professional
Why Patsnap Eureka
  • Industry Leading Data Capabilities
  • Powerful AI technology
  • Patent DNA Extraction
Social media
Patsnap Eureka Blog
Learn More
PatSnap group products

Browse by: Latest US Patents, China's latest patents, Technical Efficacy Thesaurus, Application Domain, Technology Topic, Popular Technical Reports.

© 2024 PatSnap. All rights reserved.Legal|Privacy policy|Modern Slavery Act Transparency Statement|Sitemap|About US| Contact US: help@patsnap.com