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Gene TUD1 for controlling rice height and grain shape and application thereof

A gene and rice technology, applied in the fields of application, genetic engineering, plant genetic improvement, etc., can solve problems such as adverse pleiotropic effects, limitations, and complex dwarf gene expression, and achieve strong operability

Active Publication Date: 2009-09-30
INST OF GENETICS & DEVELOPMENTAL BIOLOGY CHINESE ACAD OF SCI +1
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Among the dwarf genes, the vast majority are dwarf genes without breeding or economic value, because they have adverse effects on one or two of the three factors of panicle number, grain number and grain weight that constitute rice yield. effect, so that so far, the use of indica rice dwarf genes is still limited to sd-1
However, the expression of dwarf genes is quite complicated, and may have the characteristics of "one cause and multiple effects", that is, in addition to reducing plant height and internode length, it may also affect plant growth vigor, plant type, and seed shape. For example, the sd-1 gene has the effect of promoting early tillering and tillering number at the same time (4，5)

Method used

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  • Gene TUD1 for controlling rice height and grain shape and application thereof
  • Gene TUD1 for controlling rice height and grain shape and application thereof
  • Gene TUD1 for controlling rice height and grain shape and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0043] Embodiment 1: the acquisition of rice TUD1 candidate gene

[0044] 1. Rice material

[0045] The dwarf and small-grained rice (Oryza sativa ssp.) used for positioning is a single recessive spontaneous mutant, and its original wild type is an indica variety—Minghui 63 (IR36 / Gui 630), and the other three etc. The single-recessive spontaneous mutants tud1-2, tud1-3, and tud1-4 came from breeding materials D506 (Minghui 63 / Zihui 100), H7788 (Tetep), ZH3 (Miryang 23), respectively. This line is also an indica rice background ( figure 1 )

[0046] 2. Analyze and target groups

[0047] The homozygous dwarf indica line tud1 was crossed with the japonica variety Nipponbare, F 1 A total of 7500 F 2 Individuals, and 1,680 individuals were selected as the positioning group. Take about 2 grams of young leaves from each plant at the seedling stage to extract DNA.

[0048] 3. Localization of TUD1 gene by SSR and STS markers

[0049] Genomic DNA for gene localization was extra...

Embodiment 2

[0056] Example 2: Verification of functional complementation of TUD1 gene

[0057] Use two restriction enzymes, SmaI and SalI, to excise a 4,662kb fragment with the full length of the TUD1 part from the BAC clone OsJNBa0084L08 (purchased from Shanghai Southern Gene Center), and then use SalI and PstI as enzyme linker-specific primers to amplify DNA fragments of 1,773 kb were ligated into a 6,429 kb fragment, including 3,169 bases upstream of the start codon ATG and a full-length sequence of 1,880 bases after the stop codon TGA, cloned into the binary vector pCAMBIA1300( Purchased from CAMBIA Company), obtained the plasmid pCAMBIA1300-TUD1 ( Figure 5 ) (for cloning and identification methods, see Molecular Cloning Experiment Guide (Third Edition) (Chinese translation) translated by Huang Peitang et al., Science Press, published in September 2002 ").

[0058] The plasmid was transferred into the Agrobacterium tumefaciens strain EHA105 (prepared in this laboratory, referring to...

Embodiment 3

[0066] Embodiment 3: TUD1 protein expression of wild-type TUD1 plant, tud1-3 mutant plant, transgenic TUD1 gene plant

[0067] In order to verify the expression of TUD1 protein in wild-type TUD1 plants, tud1-3 mutant plants, and TUD1-transgenic plants, we cloned the full-length coding sequence of TUD1 from wild-type TUD1 plants, tud1-3 mutant plants, and TUD1-transgenic plants. into the protein expression vector pGEX-6P1 (Amersham Biosciences). PCR amplification primers were 5'-CGGGATCCATGCCGCAGTACCAGGAGC-3' (the underlined EcoRI restriction site) and 5'-CAGTCGACTCACTGGATTGTCTTCGTAA-3' (the underlined Sal I restriction site). For the cloning method, see "Molecular Cloning Experiment Guide" (Third Edition) (Chinese translation) translated by Huang Peitang et al., Science Press, published in September 2002". The full-length coding sequence of TUD1 in wild-type TUD1 plants, tud1-3 mutant plants, and TUD1 transgenic plants was identified by sequencing. After being correctly fused...

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Abstract

The invention discloses a gene for controlling the rice height and the seed size and encoding protein thereof, discloses a vector including the gene and a fragment thereof and a host cell and also discloses a method for adjusting the rice height and the seed size in plants, and the method comprises the following steps: the plant cell is transformed by using the expression vector including the gene; and the transformed plant cell is cultivated into a plant. The gene has important theoretical significance on the aspect of disclosing the formation mechanism of the rice height and the seed size and also has extremely high operability on typing the rice molecular plants, breeding by yield design, molding the proper rice yield factor and effectively improving the rice yield.

Description

field of invention [0001] The invention belongs to the field of plant genetic engineering. Specifically, the present invention relates to a gene TUD1 (TAIHU Dwarf1) for controlling plant height and grain shape of rice, the protein encoded by the gene and its functional analogue, as well as the vector containing the nucleotide sequence of the TUD1 gene and the vector containing the TUD1 gene nucleotide sequence. The nucleotide sequence of the gene or the host cell of the vector; in addition, the present invention also relates to a method for transforming plant cells with the vector to change plant height and seed size. Background technique [0002] Rice plant height is an important factor for rice plant architecture. Appropriate plant height is an important guarantee for high rice yield. The "green revolution" caused by the use of rice dwarf mutant gene - sd1 for breeding is a clear proof (1，2) (This paper uses Arabic numerals to indicate the cited documents, the same below...

Claims

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Application Information

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IPC IPC(8): C07K14/415C12N15/29C12N15/82C12N1/21C12N5/10A01H1/00A01H5/00C12R1/19
Inventor 薛勇彪钱前胡兴明郭龙彪
Owner INST OF GENETICS & DEVELOPMENTAL BIOLOGY CHINESE ACAD OF SCI
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