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NAC transcription factor gene TaNACs in wheat as well as expression vector and application thereof

A technology of transcription factor and expression vector, applied in the field of genetic engineering, can solve problems such as rapid virulence variation, outbreak of wheat powdery mildew, agricultural production disaster, etc., and achieve the effect of improving resistance

Active Publication Date: 2015-05-20
NANJING AGRICULTURAL UNIVERSITY
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Due to the large number of physiological races of wheat powdery mildew and rapid virulence variation, once a new toxic race is produced or the population of the race changes, it will lead to a large outbreak of wheat powdery mildew and bring disastrous consequences to agricultural production

Method used

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  • NAC transcription factor gene TaNACs in wheat as well as expression vector and application thereof
  • NAC transcription factor gene TaNACs in wheat as well as expression vector and application thereof
  • NAC transcription factor gene TaNACs in wheat as well as expression vector and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0017] Example 1 Cloning of a NAC transcription factor gene TaNACs in Nannong 9918 containing Pm21 gene

[0018] Nannong 9918 is a wheat variety containing broad-spectrum powdery mildew resistance gene Pm21 bred by the Institute of Cytogenetics of Nanjing Agricultural University (publicly known and public), and Yangmai 158 is a popularized variety of high-susceptibility wheat powdery mildew bred by the Institute of Agricultural Sciences in Lixiahe District, Jiangsu (publicly known and public). In order to screen the genes expressed by powdery mildew in Nannong 9918, high-throughput sequencing was used to obtain the digital gene expression profiles of the stripe rust-resistant wheat Nannong 9918 and the stripe rust-susceptible wheat Yangmai 158, and on this basis to screen the resistant genes. Differentially expressed genes in sensitive materials. The specific process is as follows: the seeds of the powdery mildew resistant wheat Nannong 9918 and the stripe rust susceptible wh...

Embodiment 2

[0020] Example 2 Construction of TaNACs Gene Transient Expression Vector

[0021] Using the TaNACs gene cDNA cloned in Nannong 9918 induced by powdery mildew as a template, the primer pair TaNACs-BamHI-F (cgcGGATCCATGAGCGGCGGACAGGAGCT, SEQ ID NO.5) and TaNACs-KpnI-R (cggGGTACCTTAGAACGGCTTGCCCCAGT, SEQ ID NO.6) were used as templates Perform PCR amplification and recover the amplified fragment. The amplified product was double digested with BamHI and KpnI, and the digested product was inserted into the vector pBI220 (Jefferson RA, Kavanagh TA, Bevan MW. GUS fusions: beta-glucuronidase as a sensitive and versatile gene fusion marker in higher plants.EMBO J.1987,6:3901-3907.), put TaNACs at the multiple cloning site behind the 35S promoter. Thus, the target gene TaNACs was cloned to the downstream of the strong promoter 35S, and the expression vector pBI220:TaNACs( figure 1 ). It was verified by sequencing that the vector was constructed successfully.

Embodiment 3

[0022] Example 3 Transfer of TaNACs gene into wheat leaves by transient expression method

[0023] The transient expression method is a reliable and rapid method for identifying gene functions (Schweizer, Pokorny et al. A Transient Assay System for the Functional Assessment of Defense-Related Genes in Wheat Molecular Plant-Microbe Interactions.1999,12:647-654. ). In this study, the transient expression method was used to wrap the plasmid DNA on the outer layer of metal particles, and the metal particles and genes were bombarded into the epidermal cells of wheat leaves with the help of a gene gun, and then the haustoria index of bombarded TaNACs cells was compared with that of non-bombarded TaNACs cells. Bacteria haustoria index, to clarify whether the target gene has powdery mildew resistance function.

[0024] The procedure for encapsulating carrier DNA and metal particles is as follows:

[0025] Preparation of tungsten powder: Weigh 30mg of tungsten powder into a 1.5ml epp...

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Abstract

The invention belongs to the field of gene engineering and discloses a NAC transcription factor gene TaNACs in wheat as well as an expression vector and application thereof. A cDNA sequence with NAC transcription factor gene TaNACs is SEQ ID No.1, and the coded amino acid sequence is SEQ ID No.2. The gene comes from common wheat (Triticum asetivum L.) Nannong 9918. The expression of TaNACs is enhanced under the induction of powdery mildew in the powdery mildew-resistant wheat variety Nannong 9918, and the expression level is much higher than the expression level in a susceptible wheat variety Yangmai 158. The susceptible wheat variety Yangmai 158 is transformed with the gene through transient expression, and the result indicates that overexpression of TaNACs can reduce the haustorium index of Yangmai 158. Therefore, TaNACs is expected to be applied to gene engineering breeding; and the powdery mildew resistance of wheat is expected to be improved by introducing TaNACs into a wheat variety susceptible to powdery mildew.

Description

technical field [0001] The invention belongs to the field of genetic engineering and discloses a NAC transcription factor gene TaNACs in wheat, its expression carrier and application. Background technique [0002] The whole growth period of wheat (Triticum aestivum) is harmed by various diseases and insect pests, among which wheat powdery mildew is a fungal disease caused by wheat powdery mildew (Blurmeria graminis f.sp.tritici, Bgt). Because wheat powdery mildew has many physiological races and rapid virulence variation, once a new toxic race is produced or the population of the race changes, it will lead to a large outbreak of wheat powdery mildew and bring disastrous consequences to agricultural production. The discovery and utilization of broad-spectrum and long-lasting disease resistance genes are of great significance to the control of powdery mildew. [0003] The broad-spectrum powdery mildew resistance gene Pm21 from Haynaldia villosa (2n=2x=14, VV) is located on th...

Claims

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Application Information

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IPC IPC(8): C12N15/29C07K14/415C12N15/82A01H5/00
Inventor 曹爱忠钱晨胡平李若晨邢莉萍周渭皓王慧王秀娥王海燕肖进陈佩度张瑞奇袁春霞
Owner NANJING AGRICULTURAL UNIVERSITY
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