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Rice plant height and grain shape control gene tud1 and its application

A rice and biolistic technology, applied in application, genetic engineering, plant genetic improvement, etc., can solve problems such as adverse pleiotropic effects, reduction of plant height and internode length, complex dwarf gene expression, etc., and achieve strong operability. Effect

Active Publication Date: 2011-11-30
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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  • Rice plant height and grain shape control gene tud1 and its application
  • Rice plant height and grain shape control gene tud1 and its application
  • Rice plant height and grain shape control gene tud1 and its application

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0043] Example 1: Obtaining Candidate Gene of Rice TUD1

[0044] 1. Rice material

[0045] The short-stalked 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), the other three etc. The single recessive spontaneous mutants tud1-2, tud1-3, and tud1-4 were derived from the breeding materials D506 (Minghui 63 / Zhui 100), H7788 (Tetepu), ZH3 (Miyang 23), respectively. This line is also indica rice background ( figure 1 )

[0046] 2. Analyze and locate groups

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

[0048] 3. Locate the TUD1 gene through SSR and STS markers

[0049] The rapid extraction method of rice trace DNA is u...

Embodiment 2

[0056] Example 2: Functional complementation verification of TUD1 gene

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

[0058] The plasmid was transferred into AgroBacterium tumefaciens strain EHA105 (prepared in our laboratory, refer to "Plant Genetic Engineering", Wang Guanlin, Fang Hongyun, Science Press, 2004, 2nd edition, e...

Embodiment 3

[0066] Example 3: Expression of TUD1 protein in wild-type TUD1 plants, tud1-3 mutant plants, and TUD1 transgenic plants

[0067] In order to verify the expression of TUD1 protein in wild-type TUD1 plants, tud1-3 mutant plants, and transgenic plants with TUD1 gene, we cloned the full-length coding sequence of TUD1 in wild-type TUD1 plants, tud1-3 mutant plants, and transgenic plants with TUD1 gene Into the protein expression vector pGEX-6P1 (Amersham Biosciences). The PCR amplification primers are 5'-CGGGATCCATGCCGCAGTACCAGGAGC-3' (underlined is EcoR I restriction site) and 5'-CAGTCGACTCACTGGATTGTCTTCGTAA-3' (underlined is Sal I restriction site). For cloning methods, 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 transgenic plants with TUD1 gene was identified by sequencing. Aft...

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Abstract

The invention discloses a gene and its encoded protein for controlling the plant height and seed size of rice. The invention also discloses a vector comprising the gene and its fragments, and a host cell. The invention also discloses the regulation and control of plant height and seed size in plants. A seed-sized method comprising transforming a plant cell with an expression vector comprising said gene, and growing the transformed plant cell into a plant. The gene of the invention has important theoretical significance in revealing the formation mechanism of rice plant height and seed size, and has strong operability for rice molecular plant type and yield design breeding, shaping suitable rice yield factors, and effectively increasing rice yield.

Description

Invention field [0001] The invention belongs to the field of plant genetic engineering. Specifically, the present invention relates to a rice plant height and grain shape gene TUD1 (TAIHU Dwarf1), the protein encoded by the gene and its functional analogues, and a vector containing the nucleotide sequence of the TUD1 gene and containing the gene The gene nucleotide sequence or the host cell of the vector; in addition, the present invention also relates to a method of using the vector to transform plant cells to change plant height and seed size. Background technique [0002] Rice plant height is an important factor that constitutes rice plant type. Proper plant height is an important guarantee for high yield of rice. The "green revolution" caused by breeding using the rice dwarf mutant gene-sd1 is a clear proof. (1，2) (This article uses Arabic numerals to indicate the cited documents, the same below). The ecological and production conditions of the main rice-producing areas in ...

Claims

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Application Information

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Patent Type & Authority Patents(China)
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