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Plant resistance associated protein MYB, as well as coding gene and application thereof

A technology of plant resistance and related proteins, applied in the direction of plant gene improvement, application, plant peptides, etc., can solve the problems of cloning, expression characteristics and function research that have not been reported, and achieve improved drought resistance, disease resistance and drought resistance sexual effect

Inactive Publication Date: 2011-04-13
INST OF CROP SCI CHINESE ACAD OF AGRI SCI
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

The close relatives of wheat, E. intermedium, are resistant to wheat rust, powdery mildew, viral disease, sheath blight, drought and other adversities (Liang Hongxia, Lu Yan, LiuHongxia, Wang Fengde, Xin Zhiyong, Zhang Zengyan(Correspondance).2008. A novelactivator-type ERF of Thinopyrum intermedium, TiERF1, positively regulates defense responses. Journal of Experimental Botany.59: 3111-3120), however, there is no research on the cloning, expression characteristics and function of the Thinopyrum intermedium resistance-related protein MYB gene to report

Method used

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  • Plant resistance associated protein MYB, as well as coding gene and application thereof
  • Plant resistance associated protein MYB, as well as coding gene and application thereof
  • Plant resistance associated protein MYB, as well as coding gene and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0042] Example 1. Discovery of plant resistance-related protein TiMYB1 and its coding gene TiMYB1

[0043] According to the sequence of wheat TaPIMP1 gene (GenBank Accession NO.EF587267.1), a primer pair was designed: MYB-U: 5'-ACTCGCGTACGTCTTCCTGA-3' and MYB-L: 5'-GCGC TCTAGTTAAGTTCATCGTC-3'.

[0044] The leaves of the 4-5 leaf stage seedlings of Echinopsis intermedia (Z1146) inoculated with Bipolaris sorokiniana were taken, treated with liquid nitrogen, and the total RNA of the leaves was extracted according to the instructions of the Invitrogen TRIZOL Reagent total RNA extraction reagent. According to the procedures of Invitrogen's first-strand cDNA synthesis kit, the extracted RNA samples were reverse-transcribed to synthesize first-strand cDNA, which was used as a template for gene cloning. Using cDNA as a template, PCR amplification was performed with the designed primer pair; the PCR amplification system was: 1 μL of 10×GC buffer, 2 μL of cDNA (50 ng), 2 μL of 2.5 mM dN...

Embodiment 2

[0046] Example 2, Induced expression analysis of TiMYB1 gene

[0047] 1. Induced expression analysis of wheat root rot fungus

[0048] The leaves of the 4-5 stage seedlings of Thinium intermedia (Z1146) were rubbed and inoculated with the mycelium block of root rot pathogenic fungus (Helminium umbilicale), and the mycelium block of root rot pathogenic fungus was placed on the stem of Thinium intermedia. Leaf sheath; using the leaves of Thiopyrum intermedium that were not inoculated with root rot pathogenic bacteria as the control (0h), the leaves of Thinium intermedium were taken 12, 24, 48, 72, and 96 h after inoculation, and stored at -80°C after quick-freezing in liquid nitrogen. ℃ ultra-low temperature refrigerator for later use.

[0049] Total RNA (approximately 5 μg total RNA per sample) was extracted from leaves at each treatment time, and reverse-transcribed into cDNA according to the procedure of the first-strand cDNA synthesis kit from Invitrogen. The concentration...

Embodiment 3

[0059] Embodiment 3, the acquisition of transgenic wheat and identification of disease resistance

[0060] 1. Construction of recombinant expression vector

[0061] 1. Use Bipolaris sorokiniana to inoculate the leaves of Thinium intermedia (Z1146), extract RNA after 12 hours, reverse transcribe it into cDNA; use cDNA as template, and use primers composed of TiM-DF and TiM--SAR Perform PCR amplification to obtain the PCR amplification product (TiMYB1 gene carrying DarI and SacI sites).

[0062] TiM-DF: 5'-GG TTTAAA ATGGACATGGACAAGG-3' (Dar I enzyme recognition site is underlined);

[0063] TiM--SAR: 5'-TT GAGCTC GCGCTCTAGTTAAGTT-3' (SacI enzyme recognition site is underlined).

[0064] PCR reaction program: pre-denaturation at 95°C for 3min; then 95°C for 30s, 58°C for 1min, 72°C for 1min, 5 cycles; 95°C for 30s, 56°C for 1min, 72°C for 1min, 20 cycles; 95°C for 30s, 60°C 1min, 1min at 72°C, 15 cycles; finally fill in the end at 72°C for 10min.

[0065] 2. The PCR ampli...

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Abstract

The invention discloses a plant resistance associated protein MYB as well as a coding gene and an application thereof. The protein provided in the invention is shown in (a) or (b) below, wherein (a) shows the protein consisting of the amino acid sequence shown in the sequence 1 of a sequence table, and (b) shows the plant resistance associated protein derived from the sequence 1 with the substitution and / or the deletion and / or the addition of one or more amino acid residues on the amino acid sequence of sequence 1. The introduction of the coding gene of the plant resistance associated proteinin the plant can enhance the disease resistance and the drought resistance of the plant. The protein and the cultivating method of the transgenic plant with the enhancement of the disease resistance and the drought resistance using the coding gene of the protein provided by the invention have important theoretical and practical significance, and can play an important role in the hereditary improvement of the plant.

Description

technical field [0001] The invention relates to a plant resistance-related protein MYB and its coding gene and application. Background technique [0002] Wheat (Triticuma aestivum) is one of the four major crops that humans rely on for survival. More than one-third of the world's population uses wheat as a staple food. The yield and quality of wheat directly affect human survival and quality of life. With the changes of farming systems, fertilizer and water conditions, and climate conditions, diseases have become one of the important factors restricting high and stable yields of wheat. In recent years, the main wheat diseases that have occurred in my country include powdery mildew, scab, sheath blight, stripe rust, root rot and yellow dwarf. For example, from 2008 to 2009, wheat head blight was severe (level 4) or above in most of Hubei, south of Huaihe River in Jiangsu, mid-late ripening wheat areas along Huaihe River in Anhui and south of it, middle and lower reaches of Y...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C07K14/415C12N15/29C12N15/63C12N5/10C12N1/15C12N1/19C12N1/21C12N15/11A01H5/00
Inventor 张增艳周贤尧周淼平刘红霞刘欣董娜路妍辛志勇
Owner INST OF CROP SCI CHINESE ACAD OF AGRI SCI
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