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Clone and application of gene INDETERMINATE1 for regulating and controlling plant height of gramineous plants

A technology for regulating Grass and Beaceae, applied in the field of plant genetic engineering, can solve problems such as ID1 gene that no research has paid attention to

Inactive Publication Date: 2015-10-07
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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  • Clone and application of gene INDETERMINATE1 for regulating and controlling plant height of gramineous plants
  • Clone and application of gene INDETERMINATE1 for regulating and controlling plant height of gramineous plants
  • Clone and application of gene INDETERMINATE1 for regulating and controlling plant height of gramineous plants

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

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

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

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

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

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

[0054] The amino acid seque...

Embodiment 2

[0083] Example 2: BdID1 complements rice rid1 mutants

[0084] Using the constructed 35S::BdID1 overexpression vector to complement the rice rid1 mutant: Afterwards, the isocomplementation experiment of the rice rid1 mutant was carried out according to the existing technology, and the specific process will not be described in detail. "RID1, encoding a Cys2 / His2-type zinc finger transcript ion factor, acts as a master switch from vegetable to floral development in rice." in Proceedings of the Academy of Sciences.

[0085] The results showed that the transgenic plants could 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 a dwarf phenotype (such as image 3 A), using P1 and P2, P2 and P3 (primer sequences as follows) and BdID1 bidirectional primer verification found that the transgenic seedlings that restored the heading phenotype were indeed complementary transgenic positive s...

Embodiment 3

[0090] Example 3: Obtaining of Brachypodium distachyon and Wheat ID1 Overexpression Transgenic Plants

[0091] Utilize the method mediated by Agrobacterium to infect the callus of Brachypodium distachyon to obtain transgenic seedlings, the specific implementation method is as follows:

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

[0093] (2) Collect the bacteria on the culture medium into 20ml infection solution, shake it at 28°C, measure the concentration, and make the OD 600 Values ​​between 0.6-1.0, A 600 It is 0.06-0.1. Collect about 300 pieces of calluses induced by young embryos of Brachypodium distachyda tha...

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Abstract

The invention relates to the technical field of plant biology, in particular to a clone and application of gene INDETERMINATE1 for regulating and controlling a plant height of gramineous plants. The inventor separated and cloned homologous ID1 genes from Brachypodium distachyon and wheat, and named the genes as BdID1 and TaID1, the inventor connected full length cDNA of the BdID1 and the TaID1 of A and B genomes to an expression vector initiated by an over expression vector, infected and converted Brachypodium distachyon with agrobacterium, and found that the genes enable Brachypodium distachyon to generate a dwarfing character.

Description

technical field [0001] The invention relates to the technical field of plant genetic engineering, and provides the cloning and application of a gene INDETERMINATE1 (ID1) regulating the plant height of gramineous plants. Background technique [0002] The agricultural production in the Huang-Huai-Hai region of my country is mainly the rotation of wheat and corn. Since the production of maize tends to be directly broadcast by machine after wheat harvest and harvested by machine after maturity, it is urgent to breed wheat and maize varieties with short growth period and lodging resistance. Moderate early flowering can shorten the growth period of wheat and maize and improve the adaptability of varieties, so it is an important agronomic trait of crops. The dwarfing of crop plants can significantly increase the lodging resistance of crops. During the Green Revolution in the 1960s, gibberellin-related dwarf genes were used to breed dwarf and lodging-resistant wheat and rice variet...

Claims

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Application Information

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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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