Potential of Wheat dwarf virus (Geminiviridae: Mastrevirus) Truncated Promoter for Improvement of Transgene Expression in Rice

IF 1.1 Q3 AGRONOMY Agricultural Research Pub Date : 2024-01-29 DOI:10.1007/s40003-023-00689-0

Marzieh Taghi-Malekshahi, Khalil Alami-Saeid, Esmaeel Ghasemi-Goojani, Mohamad Hamed Ghodoum Parizipour

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Abstract

Wheat dwarf virus (WDV) is a monocotyledonous plant-infecting virus that causes economical loss to cereal cultivations annually. The objective of this study was to use Cauliflower mosaic virus 35S promoter in combination with WDV large intergenic region (LIR) in rice (Oryza sativa L.) plant. Herein, a partial sequence from the genome of an Iranian isolate of WDV was used to synthesize a promoter for improvement of transgene expression in rice cultivars including Champa, Hashemi and Najafi. The promoter was cloned into pCAMBIA1304 and transferred into rice cells using in planta method. PCR, hygromycin resistance and in vivo expression of green fluorescent protein (mGFP5) were used for evaluating the efficiency of the synthesized promoter. The results showed that the WDV LIR can act as an efficient promoter for transgene expression in rice plant. Moreover, a significant difference was found between transformation rates in different rice cultivars; thus, Champa and Najafi cultivars showed the highest and lowest rate of transformation, respectively. WDV LIR was used to design a novel promoter for gene expression in rice cells. PCR assay, phenotypic experiment (antibiotic selection and GFP expression) and nucleotide sequencing showed that LIR is a promising source for the improvement of transgene expression within cells of monocotyledonous plants.

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小麦矮小病毒（Geminiviridae：Mastrevirus）截短启动子在改善水稻转基因表达方面的潜力

小麦矮缩病毒（WDV）是一种单子叶植物感染病毒，每年都会给谷物种植造成经济损失。本研究的目的是在水稻（Oryza sativa L.）植株中结合使用花椰菜花叶病毒 35S 启动子和 WDV 大基因间区（LIR）。在此，研究人员利用伊朗分离的 WDV 基因组的部分序列合成了一个启动子，用于改善 Champa、Hashemi 和 Najafi 等水稻栽培品种的转基因表达。启动子被克隆到 pCAMBIA1304 中，并通过植物体内法转移到水稻细胞中。利用 PCR、百菌清抗性和绿色荧光蛋白（mGFP5）的体内表达来评估合成启动子的效率。结果表明，WDV LIR 可作为转基因表达的高效启动子。此外，不同水稻栽培品种的转化率存在显著差异，其中占婆和纳杰菲的转化率分别最高和最低。利用 WDV LIR 设计了一种新型启动子，用于水稻细胞中的基因表达。PCR 检测、表型实验（抗生素选择和 GFP 表达）和核苷酸测序表明，LIR 是改善单子叶植物细胞内转基因表达的一个很有前景的来源。

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来源期刊

Agricultural Research AGRONOMY-

CiteScore

3.80

自引率

0.00%

发文量

期刊介绍： The main objective of this initiative is to promote agricultural research and development. The journal will publish high quality original research papers and critical reviews on emerging fields and concepts for providing future directions. The publications will include both applied and basic research covering the following disciplines of agricultural sciences: Genetic resources, genetics and breeding, biotechnology, physiology, biochemistry, management of biotic and abiotic stresses, and nutrition of field crops, horticultural crops, livestock and fishes; agricultural meteorology, environmental sciences, forestry and agro forestry, agronomy, soils and soil management, microbiology, water management, agricultural engineering and technology, agricultural policy, agricultural economics, food nutrition, agricultural statistics, and extension research; impact of climate change and the emerging technologies on agriculture, and the role of agricultural research and innovation for development.