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Insect and herbicide resistant maize transformation event

A technology for transforming events and corn, applied in the field of plant biology, can solve the problems of not being able to directly use the crop growth period, long residual period, and easily affecting the normal growth of the next crop.

Active Publication Date: 2019-08-20
CHINA NAT SEED GRP
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

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
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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  • Insect and herbicide resistant maize transformation event
  • Insect and herbicide resistant maize transformation event
  • Insect and herbicide resistant maize transformation event

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0106] Embodiment 1 vector construction

[0107] 1. Synthesis of insect resistance genes

[0108] The insect-resistant gene cry1Ab / cry1Ac ZM that is expressed in plants and produces insect-resistant effects disclosed by the applicant in PCT international application WO2017012577A1 is used. The gene is based on the fused and transformed N-terminal 608 amino acid sequences of Cry1Ab and Cry1Ac, uses plant-favored codons to replace the coding sequence, and replaces the rich genes in the DNA sequence that cause plant transcription instability. Containing AT sequences and commonly used restriction enzyme sites for correction and elimination. At the same time, a 67-nucleotide Ωsequence and a 3-nucleotide (ACC) Kozaksequence were added at the 5' end to enhance the translation efficiency of eukaryotic genes. A 135bp polyA sequence was added to the 3' end. The protein encoded by this gene contains three functional segments, in which the two N-terminal functional regions are highly h...

Embodiment 2

[0122]The acquisition of embodiment 2 transgenic corn

[0123] Transgenic maize was obtained using Agrobacterium-mediated genetic transformation method.

[0124] The plasmid DNA of the vector pZHZH35006 was transformed into Agrobacterium EHA105 by electric shock method, and it was identified for later use.

[0125] Transformation is carried out by using the high-efficiency transgenic method of the backbone inbred line of maize disclosed by the applicant in the Chinese invention patent application CN104745622A.

[0126] Specifically, immature embryos with a length of about 1.5 mm were used for transformation after selfing of the maize inbred line Xiang 249. Collect the immature embryos of about 200 ears as a batch, place them in EP tubes, suck out the suspension, add Agrobacterium liquid containing 200 μM acetosyringone, co-cultivate for 5 minutes, and then transfer the immature embryos to the co-culture medium. Incubate in the dark for 3 days. The immature embryos after dar...

Embodiment 3

[0137] Example 3 Resistance identification of transgenic maize

[0138] T 0 The generation plants are selfed, and the resulting seeds are T 1 generation seeds. Will T 1 Generation seeds were sown in greenhouses to obtain T 1 Substitute plants. Repeat the above process until T 4 generation seeds.

[0139] to T 1 to T 3 Transgenic positive detection, herbicide tolerance analysis, insect resistance identification, and agronomic traits analysis were carried out on the generation plants. Select transgene-positive plants from each generation of plants, which have the characteristics of insect resistance, herbicide tolerance, and excellent agronomic performance, and enter the next generation of screening.

[0140] 1. Positive test

[0141] The detection method and steps are the same as described in Example 2.

[0142] 2. Identification of herbicide tolerance traits

[0143] Sow the selfed seeds of the positive plants detected in step 1 in a greenhouse, identify the herbic...

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Abstract

The application provides an insect and herbicide resistant maize transformation event, and provides a related creation method, detection method and application. In this application, a maize inbred line Xiang 249 is used as a receptor, and agrobacterium-mediated genetic transformation is performed to obtain a maize plant into which an exogenous gene insert is inserted at a specific genomic site, wherein the exogenous gene insert comprises the following three genes: an insect resistant gene, a glufosinate resistant gene and a glyphosate resistant gene. In the transformation event obtained in theapplication, the inserted exogenous gene is located at a non-functional site of the maize genome, and does not affect the expression of other genes of the recipient plant itself, so the transgenic maize plants maintain good agronomic traits while obtaining insect and herbicide resistance.

Description

technical field [0001] This application relates to the field of plant biotechnology, in particular to a creation method, detection method and application of an insect-resistant and herbicide-resistant corn transformation event. Background technique [0002] Corn is an important feed and industrial raw material crop. As the crop with the largest planting area in China, it 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, commonly known as the corn borer, is one of the important biological disasters that cause perennial corn yield reduction, seriously affecting the yield and quality of corn, including Asian corn borer (Ostrinia furnacalis) and European corn borer (Ostrinian ubilalis). China is the frequent and re-incidence area of ​​Asian corn borer (Ostrinia furnacalis), which occurs on a large scale almost every two years. In general years, corn is harmed by corn borer, and the yield is...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N15/82C12N15/11
CPCC12N15/8277C12N15/8275C12N15/8286C12Q1/6895C12Q2600/13C12N15/11C12N5/10Y02A40/146C12N15/8274
Inventor 刘博林佘秋明谭超许洁婷王绪霞田裴秀子聂东明韩宇马崇烈章旺根
Owner CHINA NAT SEED GRP
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