转基因水稻愈伤组织中过度表达II型水稻元蛋白酶OsMC4提高内质网应激耐受性

IF 2.2 Q3 GENETICS & HEREDITY Plant Gene Pub Date : 2023-06-01 DOI:10.1016/j.plgene.2023.100421
Nurul Faqihah Mohd Yusof , Nur Fadzliyana Saparin , Zulkifli Ahmad Seman , Zuraida Ab Rahman , Yun Shin Sew , Muhamad Aidilfitri Mohamad Roslan , Nor Aini Abdul Rahman , Noor Azmi Shaharuddin , Patrick Gallois , Zulfazli M. Sobri
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引用次数: 0

摘要

内质网(ER)是一种负责调节植物蛋白质合成的细胞器。高盐度会导致错误折叠蛋白质的积累,导致ER应激反应机制,即未折叠蛋白质反应(UPR)。UPR不能逆转蛋白质错误折叠的作用将激活程序性细胞死亡(PCD)。已知后胱天蛋白酶基因调节植物中的PCD。本研究旨在全面分析水稻叶片中II型水稻异aspase(OsMC)基因在ER和盐度胁迫下的表达模式。在水稻基因组中的五种II型元库蛋白酶中,发现在膜霉素(ER胁迫)和氯化钠(NaCl)(盐度胁迫)处理期间,OsMC4、OsMC5和OsMC8的表达上调。构建了由CaMV35S启动子控制的taqRFP::OsMC4,并将其转化为水稻愈伤组织。过表达taqRFP::OsMC4的转基因水稻愈伤组织表现出内质网应激标记基因、蛋白质二硫化物异构酶(OsPDI)和结合免疫球蛋白(OsBiP)的表达显著变化。该分析的结果提供了初步证据,表明至少有一种II型异天冬酶OsMC4能够降低水稻的ER和盐度胁迫。未来可以在过表达OsMC的转基因水稻中进一步分析OsMC基因在ER和耐盐性中的功能,以提高耐盐性。简单摘要水稻,通常被称为水稻,是世界上消费量最大的作物之一。在对多种生物和非生物因素的反应中,一系列内质网(ER)应激反应调节因子被激活。有证据表明,高盐度会引发植物的内质网应激。本研究旨在确定水稻对ER和盐度胁迫的反应中II型异淀粉酶的基因表达水平,并评估它们如何与水稻愈伤组织中的PCD联系。已经观察到三个异aspase基因OsMC4、OsMC5和OsMC8在膜霉素处理后在水稻叶片中具有显著表达。taqRFP::OsMC4在水稻愈伤组织中的过表达显著降低了胁迫标记OsBiP和OsPDI的表达水平,表明与野生型愈伤组织相比,转基因愈伤组织的胁迫水平相对较低。因此,taqRFP::OsMC4在水稻中的过表达可能增加水稻对ER和盐度胁迫的耐受性。OsMCs家族的这些表达分析为进一步研究OsMCs在ER和盐度胁迫反应中的生物学作用提供了有价值的信息。
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Overexpression of type II rice metacaspase, OsMC4, increases endoplasmic reticulum stress tolerance in transgenic rice calli

The endoplasmic reticulum (ER) is an organelle responsible for regulating protein synthesis in plants. High salinity can lead to the accumulation of misfolded proteins, resulting in an ER stress response mechanism known as the unfolded protein response (UPR). Failure of the UPR to reverse the effect of protein misfolding will activate programmed cell death (PCD). Metacaspase genes are known regulate PCD in plants. This study aims to provide a comprehensive analysis of the expression patterns of type II rice metacaspase (OsMC) genes in response to ER and salinity stress in rice leaf. Among five type II metacaspases in the rice genome, OsMC4, OsMC5, and OsMC8 expressions were found to be upregulated during treatment with tunicamycin (ER stress) and sodium chloride, NaCl (salinity stress). A construct of taqRFP::OsMC4, controlled by the CaMV35S promoter, was generated and transformed into rice calli. The transgenic rice calli overexpressing taqRFP::OsMC4 demonstrated significant changes in the expression of the ER stress-marker genes, protein disulfide isomerase (OsPDI), and binding immunoglobulin protein (OsBiP). The results from this analysis provide preliminary evidence that at least one of the type II metacaspases, OsMC4, is be able to reduce ER and salinity stresses in rice. Further functional analysis of OsMC genes in ER and salinity stress tolerance could be carried out in transgenic rice overexpressing OsMCs in the future to improve stress tolerance.

Simple summary

Oryza sativa, commonly known as rice, is one of the most consumed crops in the world. In response to multiple biotic and abiotic factors, a series of endoplasmic reticulum (ER) stress response regulators are activated. There is evidence that high salinity triggers ER stress in plants. This study aims to determine the level of gene expression among type II metacaspases in rice in response to ER and salinity stress and to assess how they may be linked to PCD in rice calli. Three metacaspase genes, OsMC4, OsMC5, and OsMC8, have been observed to have significant expression post-treatment with tunicamycin in rice leaf. Overexpression of taqRFP::OsMC4 in rice calli significantly reduces the expression level of the stress markers, OsBiP and OsPDI, indicating that the stress level is relatively lower in the transgenic calli compared to the wild-type calli. Therefore, overexpression of taqRFP::OsMC4 in rice may increase rice tolerance towards ER and salinity stress. These expression analyses of the OsMCs family provide valuable information for further functional studies on the biological roles of OsMCs in ER and salinity stress responses.

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来源期刊
Plant Gene
Plant Gene Agricultural and Biological Sciences-Plant Science
CiteScore
4.50
自引率
0.00%
发文量
42
审稿时长
51 days
期刊介绍: Plant Gene publishes papers that focus on the regulation, expression, function and evolution of genes in plants, algae and other photosynthesizing organisms (e.g., cyanobacteria), and plant-associated microorganisms. Plant Gene strives to be a diverse plant journal and topics in multiple fields will be considered for publication. Although not limited to the following, some general topics include: Gene discovery and characterization, Gene regulation in response to environmental stress (e.g., salinity, drought, etc.), Genetic effects of transposable elements, Genetic control of secondary metabolic pathways and metabolic enzymes. Herbal Medicine - regulation and medicinal properties of plant products, Plant hormonal signaling, Plant evolutionary genetics, molecular evolution, population genetics, and phylogenetics, Profiling of plant gene expression and genetic variation, Plant-microbe interactions (e.g., influence of endophytes on gene expression; horizontal gene transfer studies; etc.), Agricultural genetics - biotechnology and crop improvement.
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