Indica rice classified control gene PHR1 and use thereof

A rice and amino acid technology, applied in the fields of application, genetic engineering, plant gene improvement, etc., can solve problems such as unseen cloned genes

Inactive Publication Date: 2006-05-31
INST OF GENETICS & DEVELOPMENTAL BIOLOGY CHINESE ACAD OF SCI
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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  • Indica rice classified control gene PHR1 and use thereof
  • Indica rice classified control gene PHR1 and use thereof
  • Indica rice classified control gene PHR1 and use thereof

Examples

Experimental program
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Effect test

Embodiment 1

[0037] Example 1 Cloning of Rice Phenol Staining Reaction Control Gene PHR1

[0038] 1. Rice material

[0039] Rice (Oryza sativa ssp.) dyeable indica variety Minghui63 (Minghui63) and non-dyeable japonica variety Chunjiang06 (Chunjiang06).

[0040] 2. Analyze and target groups

[0041] The homozygous indica variety Minghui 63 was crossed with the japonica variety Chunjiang 06, F 1 A total of 8000 F 2 Individuals, and 2,506 individuals were selected as the positioning group. Take about 2 grams of young leaves from each plant at the seedling stage to extract DNA.

[0042] 3. Localization of the PHR1 gene by SSR, STS, and CAPS markers

[0043] The improved CTAB (Cetyltrimethyl Ammonium Bromide) method [11] was used to extract genomic DNA for gene mapping from rice leaves. About 100mg of rice leaves were taken, frozen in liquid nitrogen, ground into powder in a small mortar with a diameter of 5em, transferred to a 1.5ml centrifuge tube to extract DNA, and the obtained DNA p...

Embodiment 2

[0053] Example 2 Functional Complementation and Transgenic Research of Rice Phenol Staining Reaction Control Gene PHR1

[0054] Primers were designed according to the sequence of the indica rice 93-11 PHR1 gene, and the primers PH1F, PH1R, PH2F and PH2R (see Appendix 1 for the sequence) were used to divide into two sections of high-fidelity PCR (see Table 1 for the sequence, pre-denatured at 94°C for 5min, 94°C for 1min, 60 ℃ 1min, 72℃ 1min, 35 cycles, 72℃ extension for 10mins and sequencing with ABI3730 DNA sequencer (ABI company), select the clones with completely correct sequence and use the common BstBI site to connect them into a 4.2kb fragment, including the initial The full-length sequence of 1,422 bases upstream of the codon ATG and 454 bases after the stop codon TGA was cloned into the binary vector pCAMBIA1300 (purchased from CAMIA Company), and the plasmid pCAMPHR1 ( Figure 4 ). The plasmid was transferred into Agrobacterium tumefaciens strain EHA105 (purchased fr...

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Abstract

A long-grained rice classified controlling gene PHR1 and its use are disclosed. It is cloned from long-grained rice 93-11 and approved gene for controlling long-grained classification and named PHR1. Transfer-gene complementary examination proofs that PHR1 is gene for differentiating long-grained rice classification. It can obtain transfer-gene rice with various anti-insect abilities by inhibiting or excess expressing PHR1 to regulate plant resistance by gene engineering technology.

Description

technical field [0001] The present invention relates to the field of plant genetic engineering, more specifically, the present invention relates to the protein encoded by the rice phenol dyeing reaction control gene PHR1 and its functional analogue, the nucleotide sequence encoding it, the vector containing the nucleotide and the vector containing the vector In addition, the present invention also relates to a method for controlling plant phenol dyeing reaction and a method for improving plant resistance to disease and pest breeding. Background technique [0002] During the long evolutionary history of cultivated rice, approximately 80,000-100,000 cultivars have been produced. These cultivars are divided into two types according to the difference in fertility after crossing with each other, one is Indica (Indica) and the other is Japonica (Japonica) ]. However, classification by fertility is a heavy workload and is not easy to be used. In 1962, Oka[3] discovered through a...

Claims

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Application Information

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IPC IPC(8): C07K14/415C12N15/29C12N15/63C12N15/82
Inventor 李家洋钱前于彦春曾大力周奕华
Owner INST OF GENETICS & DEVELOPMENTAL BIOLOGY CHINESE ACAD OF SCI
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