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

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...

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 population of double-strain co-transformation is very heavy, and it is urgent to improve the screening efficiency of transgenic individuals without selection markers, so as to cultivate new germplasm of crops that meet the breeding goals on the basis of efficient and safe transgenic technology

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