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Cloning and application of a gene indeterminate1 regulating plant height in Gramineae

A grass and gene technology, applied in the field of plant genetic engineering, can solve the problem that no research has paid attention to ID1 gene and so on

Inactive Publication Date: 2018-01-05
SHANDONG AGRICULTURAL UNIVERSITY
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Therefore, from an evolutionary point of view, it is speculated that the function and mechanism of the ID1 gene in temperate grasses may have a certain degree of specificity, but so far no research has focused on the ID1 gene in temperate grasses

Method used

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  • Cloning and application of a gene indeterminate1 regulating plant height in Gramineae
  • Cloning and application of a gene indeterminate1 regulating plant height in Gramineae
  • Cloning and application of a gene indeterminate1 regulating plant height in Gramineae

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0049] Example 1: Sequence analysis, cloning and vector construction of BdID1 and wheat TaID1 genes

[0050] Through bioinformatics analysis, the inventors found the ID1 homologous gene of Brachypodium biscuitae as shown in SEQ ID NO.1, and the ID1 homologous gene of wheat, and cloned the ID1 gene of Brachypodium biscuitae as shown in SEQ. ID No. 2 and the ID1 gene on the AB genome of wheat are shown in SEQ ID NO. 4 and SEQ ID NO. 6. The BdID1 (BRADI3G26910) gene of Brachypodium dilatatum is located on the third chromosome, and ID1 of wheat is located on 6AS, 6BS, and 6DS (named TaID1_6AS, TaID1_6BS, TaID1_6DS, respectively).

[0051] The open reading frame of BdID1 is 1290 bp, and the code 429aa is shown in SEQ ID NO.3;

[0052] The open reading frame of TaID1-6AS is 1257bp, and the coding 418aa is shown in SEQ ID NO.5;

[0053] The open reading frame of TaID1-6BS is 1230bp, and the code 409aa is shown in SEQ ID NO.7.

[0054] The amino acid sequence encoded by the gene BdID1 was sea...

Embodiment 2

[0083] Example 2: BdID1 complements rice rid1 mutant

[0084] Using the constructed 35S::BdID1 overexpression vector to complement the rice rid1 mutant: Afterwards, the rice rid1 mutant was subjected to an iso-complementation experiment according to the existing technology. The specific process will not be repeated. For details, please refer to Mr. Wu Changyin’s 2008 publication in the United States "RID1, encoding a Cys2 / His2-type zinc fingertranscript ion factor, acts as a master switch from vegetat ive to floral development in rice." on the Academy of Sciences newspaper.

[0085] The results show that the transgenic plants can restore heading (such as image 3 A), the heading date at the population level is slightly later than that of the wild type (such as image 3 B), and some plants have dwarfed phenotypes (such as image 3 A), using P1 and P2, P2 and P3 (the primer sequence is as follows) and the two-way primer verification of BdID1 found that the transgenic seedlings that re...

Embodiment 3

[0090] Example 3: Obtaining of transgenic plants with overexpression of Brachypodium biscuit and wheat ID1

[0091] The Agrobacterium-mediated method was used to infect Brachypodium ergatum callus to obtain transgenic seedlings. The specific implementation methods are as follows:

[0092] (1) Three days before infection, pick Agrobacterium (a single colony of Agrobacterium carrying recombinant plasmid) and inoculate it in YEP medium containing 50mg / L spectinomycin, shake overnight at 28°C, 200rpm, and take 1ml the next day The activated bacteria were plated on MGL medium containing acetosyringone and incubated for two days in the dark before they were ready for infection.

[0093] (2) Collect the bacterial cells on the culture medium into 20ml infection solution, shake them at 28℃, measure the concentration, and make the OD 600 The value is between 0.6-1.0, A 600 Is 0.06-0.1. Collect 300 pieces of callus induced by young embryos of Brachypodium dilatum that have grown for 6-7 weeks,...

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Abstract

The present invention relates to the field of plant biotechnology, in particular to the cloning and application of a gene INDETERMINATE1 (ID1) regulating the plant height of gramineous plants. The inventors isolated and cloned a homologous ID1 gene from Brachypodium distachyon and wheat, and incorporated They were named BdID1 and TaID1. The inventors connected the full-length cDNA of BdID1 and TaID1 of the A and B genomes to the expression vector initiated by the overexpression vector, and transformed Brachypodium distachyon by Agrobacterium infection, and found that the above-mentioned genes can make the two Brachypodium tachypodium appeared dwarf traits.

Description

Technical field [0001] The present invention relates to the technical field of plant genetic engineering, and provides the cloning and application of a gene INDETERMINATE1 (ID1) regulating plant height of gramineous plants. Background technique [0002] The agricultural production in the Huanghuaihai region of my country is mainly the rotation of wheat and corn. As the production of corn tends to be direct seeding after wheat harvest and harvested after ripening, there is an urgent need for breeding wheat and corn varieties with short growth period and lodging tolerance. Moderate early flowering can shorten the growth period of wheat and corn and improve the adaptability of varieties, so it is an important crop agronomic traits. Crop plant dwarfing can significantly increase the lodging tolerance of crops. The Green Revolution in the 1960s used gibberellin-related dwarf genes to bred dwarf and lodging-resistant wheat and rice varieties, leading to worldwide Grain production has...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12N15/29C07K14/415A01H5/00
Inventor 安海龙王也李奇别小敏赵翔宇王付祥
Owner SHANDONG AGRICULTURAL UNIVERSITY
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