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Application of corn nuclear factor gene ZmNF-YA1 in plant adverse resistance transformation

A technology of plant stress resistance and nuclear factor, applied in the field of bioengineering breeding of crops

Active Publication Date: 2018-05-04
SHANDONG UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

There are few reports on the specific application of maize nuclear factor gene ZmNF-YA1 in plant stress resistance transformation

Method used

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Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0031] Example 1: Transducing the ZmNF-YA1 gene to create a drought-tolerant and salt-tolerant inbred line of maize

[0032] 1) The establishment of the receptor system is based on the backbone inbred lines used in agricultural production in my country, such as Zheng 58, Chang 7-2, DH4866, etc. The seeds are soaked in 70% ethanol for 8 minutes, then soaked in 0.1% mercuric chloride for 8-12 minutes, and then washed 3-5 times with sterile water. Shake the seeds constantly during sterilization to ensure complete surface sterilization. After sterilization, the seeds are placed in a sterile Erlenmeyer flask to germinate, and a small amount of sterile water is put into the bottle for 1-2 days under dark conditions (25-28° C.). After the seeds germinated (lubai), they were placed on the modified MS medium to germinate in the dark. When the germ elongates to 3-4 cm, the coleoptile and 2-3 young leaves are peeled off to expose the growth cone at the tip of the stem.

[0033]2) Maiz...

Embodiment 2

[0038] Example 2: Transducing the ZmNF-YA1 gene to create a heat-resistant inbred line in maize

[0039] 1. Obtaining of transgenic maize plants

[0040] Seed germination, plasmid construction and genetic transformation of corn elite inbred lines are the same as in Example 1. After the transformed plant grows 3 leaves, spray herbicide (Hoechst Schering AgrEvo GmbH, containing herbicide glufosinateammonium) aqueous solution, the concentration is 9.6ml-10.8ml It is advisable for the plants to shed droplets. Untransformed control plants stopped growing after 4 days after spraying and started to die after 9 days. After the transformed plants were sprayed, some individual changes were similar to those of the control plants, while others continued to grow without obvious changes. When the surviving plants grow to 5 leaves, they are planted in the field, bagged and self-seeded.

[0041] 2. Homozygous transgenic plants and progeny analysis

[0042] Use 10.8ml for T1 generation...

Embodiment 3

[0046] Example 3: Transforming the ZmNF-YA1 gene to create drought- and salt-tolerant bluegrass

[0047] Using shoot tips of different varieties of bluegrass (Poa pratensis L), such as prize, new song Laide, rugby-A, midnight, etc., as test materials, induce callus expansion at the base, cut the expanded tissue, and place it in the subculture Cluster buds were induced on the clump and clump medium, and the latter continued to proliferate on the subculture and clump medium to provide the recipient material for the transgene. Using the Agrobacterium-mediated method to introduce foreign genes into recipient cells, and obtain transformed cells and plants through selection. In Agrobacterium-mediated genetic transformation, the use of surfactant Silwet L-77 treatment and reduced pressure treatment effectively increased the transformation frequency, and established an efficient transgenic technology system for Poa annua with less genotype restriction. The specific operation is as fo...

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PUM

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Abstract

The invention discloses application of a corn nuclear factor gene ZmNF-YA1 in plant adverse resistance transformation. According to the application, the ZmNF-YA1 gene is cloned out from corns, the gene is recombined into a plant expression carrier in a positive or antisense mode or an RNAi structure mode, and the ZmNF-YA1 gene is guided into a plant by a transgenic technology; by means of detecting transgenic expression and measuring adverse resistance of the plant, transgenic plants and descendants capable of improving or reducing adverse resistance are screened out, and novel germplasm withapplication value in plant breeding is created out. The application disclosed by the invention utilizes the ZmNF-YA1 gene to participate the plant adverse resistance adjustment through gene expressionregulation, so that important significance in breeding high-yield transgenic crops is achieved.

Description

technical field [0001] The invention belongs to the field of bioengineering breeding of crops, in particular, it relates to the application of a corn nuclear factor gene ZmNF-YA1 in the transformation of plant stress resistance; that is, a scheme for changing the stress resistance of corn by constructing a transgene overexpression structure and a transgene pathway and use. Background technique [0002] NF-Y (nuclear factor-y) is a ubiquitous transcription factor complex in eukaryotes, which consists of three different subunits (NF-YA / CBF-B, NF-YB / CBF-A and NF- YC / CBF-C), and regulate the expression of the target gene by acting on the promoters of other regulatory factors. The complete NF-Y transcription complex binds to the CCAAT motif in the promoter of the target gene to regulate the transcription of the target gene. Among them, NF-YA is a DNA sequence-specific subunit, which binds to the core pentamer nucleotide CCAAT motif (motif) in the gene promoter region. The NF-Y...

Claims

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Application Information

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IPC IPC(8): C07K14/415C12N15/29C12N15/82A01H5/00A01H6/46
CPCC12N15/8271C12N15/8273C07K14/415
Inventor 张举仁王保海王洁敏李朝霞
Owner SHANDONG UNIV
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