Method for converting disease-resistance genes of rice and obtaining transgenic descendants without selective markers

A technology without selectable markers and disease-resistant genes, applied in biochemical equipment and methods, horticultural methods, botanical equipment and methods, etc., can solve the problems of heavy analysis work and difficult screening of transgenic individuals without selectable markers, and achieve improved screening Efficiency and workload reduction effects

Inactive Publication Date: 2012-10-03
ZHEJIANG ACADEMY OF AGRICULTURE SCIENCES
View PDF1 Cites 10 Cited by
  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

In addition, in the genome of about 50% of the co-transformed plants, the T-DNA of the marker gene and the target gene are linked, and it is difficult to obtain isolated transgenic individuals without selection markers from their progeny populations
Therefore, the analysis of the progeny pop

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 converting disease-resistance genes of rice and obtaining transgenic descendants without selective markers
  • Method for converting disease-resistance genes of rice and obtaining transgenic descendants without selective markers
  • Method for converting disease-resistance genes of rice and obtaining transgenic descendants without selective markers

Examples

Experimental program
Comparison scheme
Effect test

Embodiment

[0021] Example: Taking the transformation of rice broad-spectrum rice blast resistance gene Pi9 as an example, through an improved double-strain (double-vector) co-transformation system, a rice germplasm material without a selection marker for the disease-resistant gene was created.

[0022] Vector construction: co-transformation vector system includes marker gene vector (pRB01) and target gene vector (pLJ42), such as figure 1 . The green fluorescent protein (GFP) gene controlled by the maize ubiquitin promoter Ubi was cloned into the HindIII and EcoRI restriction sites of the Agrobacterium transformation vector pCAMBIA1300 (Genbank AF234296) to generate the marker gene vector pRB01, which also carries the cauliflower mosaic virus Hygromycin phosphotransferase gene (HPT) controlled by the CaMV35S promoter. HPT is used as a hygromycin resistance selection marker to screen transformed rice cells, and GFP gene is used as a visible genetic marker to track transformed rice cells, ...

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

No PUM Login to view more

Abstract

The invention relates to a method for converting disease-resistance genes of rice and obtaining transgenic descendants without selective markers. The method comprises the following steps: first, cloning green fluorescent protein (GFP) and hygromycin phosphotransferase (HPT) on a marker gene carrier, placing a target gene in other T-DNA carrier, mixing strains of agrobacterium tumefaciens carrying the marker gene carrier and strains of agrobacterium tumefaciens carrying the target gene carrier and converting callus of the rice, performing PCR (polymerase chain reaction) detection on the disease-resistance genes of the target gene carrier by a specific primer, and screening to obtain co-transformation plants (T0); then, screening the marker gene plants with positive GFP rapidly and massively by means of a desk lamp fluorescence detector in segregative generations (T1 or T2) of the co-transformation plant for removing, and performing PCR detection for disease-resistance genes to the plants with negative GFP so as to obtain the individuals without selective markers of the transgenic disease-resistance genes. The method can be applied to transgenic breeding without selective markers of rice blast-resistant genes or other functional genes of the rice, and enhances the disease resistance of the rice or improves other agronomic traits.

Description

technical field [0001] The invention relates to a rice transgenic technology, in particular to a method for transforming rice disease resistance genes and obtaining transgenic offspring without selection markers. Background technique [0002] Breeding disease-resistant varieties is one of the main goals of crop breeding. As an important supplement to traditional breeding methods, transgenic technology can avoid cumbersome processes such as hybridization and multi-generation selection, transform the disease-resistant gene cloned from the plant genome into susceptible varieties, and improve the disease resistance of recipient varieties in a short period of time . In terms of the breeding application of rice disease resistance genes, with the improvement of rice genomics and map-based cloning technology, a series of rice disease resistance genes have been isolated and studied one after another, providing rich genetic resources for transgenic rice research. [0003] Breeding d...

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
IPC IPC(8): C12N15/82C12Q1/68A01H4/00A01H5/00
Inventor 瞿绍洪李军王歆赵建华何海燕刘中来
Owner ZHEJIANG ACADEMY OF AGRICULTURE SCIENCES
Who we serve
  • R&D Engineer
  • R&D Manager
  • IP Professional
Why Eureka
  • Industry Leading Data Capabilities
  • Powerful AI technology
  • Patent DNA Extraction
Social media
Try Eureka
PatSnap group products

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

© 2024 PatSnap. All rights reserved.Legal|Privacy policy|Modern Slavery Act Transparency Statement|Sitemap