转<em>TaTLP1</em>基因小麦的获得及抗叶锈性分析

doi:10.3969/j.issn.1674-7968.2020.06.002

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Abstract Thaumatin-like proteins (TLPs), which is a kind of pathogenesis-related proteins possessing antifungal activity against many plant pathogens. In recent years, the antifungal function of TLPs has received more and more attention in plant pathology research. Our previous studies have reported that one wheat (Triticum aestivum) TLP gene named TaTLP1. It is related to wheat resistance to Puccinia triticina (Pt), and the expression of TaTLP1 was significantly up-regulated at 96 h after inoculation with Pt. In this study, the full sequence of the TaTLP1 gene was inserted into the expression vector pLGY-02 driven by the strong ubiquitin (Ubi) promoter, which can be highly expressed in monocotyledons. Then the recombinant vector pLGY-TaTLP1 was transformed into susceptible material 'JW' by Agrobacterium-mediated genetic transformation. Total 140 young embryos were infested, and 31 positive lines were obtained with a transformation rate of 22.1%. The transgenic TaTLP1 wheat from T₁~T₄ generations were verified by qRT-PCR, and the positive plants with stable genetic expression were screened out. qRT-PCR analysis confirmed the overexpression of TaTLP1 gene in transgenic positive plants. To further confirm the function of TaTLP1 gene in wheat resistance to Pt, the impact of TaTLP1 gene on transgenic wheat including 4 consecutive generations response to leaf rust pathogen infection was analyzed, the results showed that overexpression of TaTLP1 gene increased the resistance of transgenic plants to leaf rust. In addition, the determination of physiological and biochemical content of homozygous offspring further confirmed the phenotype observations, and proline, β-1,3-glucanase, hydrogen peroxide and peroxidase were also involved in wheat resistance to leaf rust. The results indicated that overexpression of TaTLP1 gene enhanced transgenic plants resistance to wheat leaf rust and played an important role in defense against wheat leaf rust, which provides a theoretical basis for further study on the function of TaTLP1 gene and the molecular mechanism between host-pathogen interaction.

Key words： Wheat leaf rust Thaumatin-like protein Transgene Disease resistance Functional analysis

Received: 17 November 2019

ZTFLH:

S432.1

Corresponding Authors: *, ndwanghaiyan@163.com; ldq@hebau.edu.cn

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	LIANG Fang
	CUI Zhong-Chi
	WANG Hai-Yan
	LIU Da-Qun

Cite this article:

LIANG Fang,CUI Zhong-Chi,WANG Hai-Yan, et al. Generation of TaTLP1 Transgenic Wheat (Triticum aestivum) and Analysis of Its Resistance to Leaf Rust Fungus[J]. 农业生物技术学报, 2020, 28(6): 963-973.

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http://journal05.magtech.org.cn/Jwk_ny/EN/10.3969/j.issn.1674-7968.2020.06.002 OR http://journal05.magtech.org.cn/Jwk_ny/EN/Y2020/V28/I6/963

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