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Anti-insect anti-herbicide corn conversion event and creation method and detection method thereof

A transformation event, corn technology, applied in the field of plant biotechnology and plant breeding, can solve problems such as long residual period, no selectivity for weeding, and inability to be directly used in the growing period of crops

Active Publication Date: 2017-08-25
先正达集团股份有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0006] At present, the widely used selective herbicides are applied in a large amount and have a long residual period, which is likely to affect the normal growth of the following crops
Glyphosate and glufosinate-ammonium and other herbicides have the characteristics of high efficiency, low toxicity, easy degradation, and no residue, but they have no selectivity for weeding and cannot be directly used in the growth period of crops

Method used

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  • Anti-insect anti-herbicide corn conversion event and creation method and detection method thereof
  • Anti-insect anti-herbicide corn conversion event and creation method and detection method thereof
  • Anti-insect anti-herbicide corn conversion event and creation method and detection method thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0076] Example 1 Obtaining and Molecular Detection of Maize Transformation Event M00595A006a

[0077] The immature embryos of maize inbred line Xiang 249 with a size of about 1mm were stripped and pollinated about 10 days ago, infected with Agrobacterium containing the constructed transformation vector pZHZH35005, co-cultured for 3 days, and rested for 6 days. Next, the culture medium containing 5 mg / L and 8 mg / L bialafosine was used for one round of screening respectively. The screened resistant calli were differentiated into seedlings, and after rooting, they were transferred to soil for growth and samples were taken for molecular detection.

[0078] The results of molecular testing showed that the RQ value of exogenous gene cry1C of T0 transformed seedlings with sample number 2334 was 0.62, while the RQ value of the wild-type control sample was 0, and the Ct value of skeleton element Ori amplification was 30, so the transformation Seedlings are positive, low-copy, high-q...

Embodiment 2

[0119] Example 2 Breeding and Field Performance of Maize Transformation Event M00595A006a

[0120] As described in Example 1, the recipient material of the transformation event in this example is the maize inbred line Xiang 249. After obtaining the transformation event of the T0 generation (single-copy insertion and no vector skeleton contamination), planting management, the flowering period is based on the The transformation event was the female parent, and the excellent backbone wild-type maize inbred line Zheng 58 was the male parent, and the F1 seeds of the T1 generation were obtained by crossing. Afterwards, the T1 generation seeds were germinated, sprayed with glufosinate, and the glufosinate-resistant and sensitive plants were selected in line with the Mendelian genetic law, and samples were taken to detect the genotype, and the target single plant was selected for backcrossing with Zheng 58 as the male parent. , to harvest the BC1F1 seeds of the T2 generation; after ...

Embodiment 3

[0137] Example 3 Isolation of left (5') and right (3') flanking sequences of maize transformation event M00595A006a

[0138] In general, DNA primer sequences and methods for detecting transgenic plants are simple and consistent, and these detection methods usually focus on frequently used gene expression elements, such as promoters, terminators, and marker genes, because for many transformation vectors, the coding sequence Zones are interchangeable. Therefore, these methods cannot be used to distinguish between constructs that differ only in their coding sequence, or between different transformation events, especially those generated using the same transformation vector, unless they are chromosomally adjacent to the inserted heterologous DNA. The sequence of the DNA, the "flanking DNA", is known.

[0139] To this end, in this example, the 5' flanking sequence of maize transformation event 00595A006a was isolated and identified by Adaptor-PCR.

[0140] After genomic DNA ext...

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Abstract

The invention provides an anti-insect anti-herbicide corn conversion event and a creation method and a detection method thereof. The invention provides the creation method of the anti-insect anti-herbicide corn conversion event, and meanwhile provides nucleic acid molecules which are inserted into a maize genome and side wing nucleic acid molecules thereof, the detection method of the corn conversion event is built based on the nucleic acid molecules and the side wing nucleic acid molecules thereof, and the method can be applied to corn breeding.

Description

technical field [0001] This application relates to the fields of plant biotechnology and plant breeding. Specifically, the present application relates to insect-resistant and herbicide-resistant nucleic acid molecules, plant transformation events created by them, and methods for their creation and detection. Background technique [0002] As the crop with the largest planting area in China, corn has been more than self-sufficient for a long time, but since 2010, the import volume has increased year by year. [0003] The corn borer (Ostrinia nubilalis), commonly known as the corn borer, is one of the major biological disasters that cause perennial corn yield reduction, seriously affecting the yield and quality of corn. China is a frequent and re-occurring area of ​​Asian corn borer (Ostrinia furnacalis), which occurs almost once every two years. In normal years, the corn borer will reduce its yield by 10%-15%, and the annual yield reduction of Ostrinia furnacalis can reach mo...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N15/54C12N15/31C12N15/82C12N5/10C12N1/21C12N15/84C12N15/11C12Q1/68A01H5/00A01H1/02
CPCC12N15/8277C12Q1/6895C12N15/8286C12Q2600/13
Inventor 邱龙杨倩倩安吉翠聂东明王勇佘秋明许洁婷孙出吴晶晶刘博林马崇烈章旺根
Owner 先正达集团股份有限公司
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