Double T-DNA carrier and its application in cultivating of non selecting sign transgene rice

Abstract

The invention provides a simple and convenient Agrobacterium Tumefacies dual carrier containing double T-DNA structural regions, and a method of using the carrier system to cultivate transgenic rice without resistance selection label. With the help of the principle of co-conversion mediated by root nodule Agrobacterium Tumefacies, the system contains two separate T-DNA structural sections, where the first T-DNA region contains antibiotic resistance selection label gene and the second one contains a universal polyclonal site able to be arbitrarily inserted with destination gene. The double-T-DNA carrier has small molecular weight, easy to clone and after the destination gene and necessary regulation and control series are cloned in the T-DNA region containing the polyclonal site, it realizes rice co-conversion; by selfing, it selects transgenic individual with destination gene but without selectin label gene from the self-bred progeny, thus eliminating the negative effect on transgenic plant commercialized production, etc, possibly caused by selection label gene.

Description

technical field [0001] The invention relates to a method for cultivating transgenic crops without a selection marker for resistance, and belongs to the application technology in the field of biology and modern agricultural technology. More specifically, the present invention relates to a binary vector with two independent T-DNA structural domains, respectively containing a resistance selectable marker gene and a target gene; relates to the use of the double T-DNA binary vector system to generate simultaneously containing selection After the co-transformation of transgenic plants with the marker gene and the target gene, individuals containing only the target gene but no resistance selectable marker gene are selected and bred in the offspring population, thereby eliminating the possible impact on the commercial production of transgenic crops due to the existence of the selectable marker. coming negative impact. Background technique [0002] Since the first transgenic plant e...

AI Technical Summary

Problems solved by technology

Because: (1) Taking marker gene knockout systems based on recombinases, transposons or intrachromosomal recombination as examples, although these systems have been proven in model plants such as tobacco and Arabidopsis, so far there are very few See reports on important food crops such as rice and corn; secondly, the probability of homologous recombination or transposition in rice and other crops is often not high, and these systems still need to be combined with genes containing recombinase or transposase Transgenic plant hybridization and genetic segregation of subsequent generations, so the effect of marker-free transgenic plants that can be screened is still very limited, and the breeding cycle is relatively long

(2) Most of the improved marker gene knockout systems need to combine positive selection markers such as ipt gene as auxiliary markers (such as Endo et al. 2002). Although these technologies have been applied in individual plant varieties, the requirements for receptor materials are relatively low. Extremely strict, especially for most difficult-to-transform maize and rice (especially indica types), it is not suitable to use the ipt gene as a positive selection marker

However, the molecular weight of these double T-DNA vectors is often large and not universal

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