Department of Mycology and Plant Pathology, Institute of Agricultural Sciences, Banaras Hindu University, Varanasi, UP, 221005, India.
Agharkar Research Institute, G. G. Agarkar Road, Pune, 411004, India.
Genes Genomics. 2019 Sep;41(9):1027-1043. doi: 10.1007/s13258-019-00821-x. Epub 2019 May 28.
Membrane-bound NADPH oxidases (Nicotinamide adenine ainucleotide phosphate oxidase) also called respiratory burst oxidase homologs (Rboh) play an essential role in ROS production under normal as well as environmental stress conditions in plants.
To identify and study respiratory burst homologs (Rboh) from the wheat genome as well as characterize their role in various biological and molecular processes along with expression in response to biotic and abiotic stresses.
The Rboh homologs in the wheat genome were predicted based on data processing, alignment of sequences and phylogenetic analysis of sequences in numerous plant species and wheat. The conserved motifs were known followed by domain design study. The 3-D structure prediction and similarity modeling were administered for NADPH enzyme domain. Gene ontology and a functional study were done in addition to expression analysis of Triticum aestivum respiratory burst oxidase (TaRboh) gene family in response to biotic as well as abiotic stress.
Phylogenetic analysis of Rboh gene family members among seven plant species including wheat, classified the family into four subfamilies. Rboh genes are mainly involved in various biological processes such as Response to oxidative stress, Superoxide anion generation, Hydrogen peroxide biosynthetic process. Among the molecular functions, calcium ion binding, peroxidase activity, oxidoreductase activity, superoxide-generating NADPH oxidase activity are essential. Enzyme annotation of the family and superfamily revealed that it encodes to five structural clusters and coding to enzymes NAD(P)H oxidase (HO-forming) (EC:1.6.3.1), Ferric-chelate reductase (NADH) (EC: 1.16.1.7), Peroxidase (EC: 1.11.1.7), Ribose-phosphate diphosphokinase (EC: 2.7.6.1). The enzymes contain six membrane-spanning domains, two hemes, and conserved motifs associated with NADPH, EF-hand and FAD binding. The outcomes additionally reflect a distinct role of this enzyme in different molecular functions which are responsible for the stress signaling. Further, the transcripts of TaRboh found expressed in various plant parts such as stem, leaves, spike, seed, and roots. We also observed expression of these gene family members under drought/combination of drought + heat and important wheat pathogens such as Puccinia striformis, Blumeria graminis f.sp. tritici, Fusarium graminiarum, F. pseudograminiarum, and Zymoseptoria tritici.
The investigation demonstrated that identified respiratory burst homologs (Rboh) in T. aestivum were involved in pathogen activated ROS production and have regulatory functions in cell death and defense responses.
膜结合 NADPH 氧化酶(烟酰胺腺嘌呤二核苷酸磷酸氧化酶)也称为呼吸爆发氧化酶同源物(Rboh),在植物正常和环境胁迫条件下的 ROS 产生中发挥重要作用。
从小麦基因组中鉴定和研究呼吸爆发同源物(Rboh),并研究其在各种生物学和分子过程中的作用,以及对生物和非生物胁迫的表达。
基于数据处理、序列比对和众多植物物种和小麦序列的系统发育分析,预测小麦基因组中的 Rboh 同源物。随后进行保守基序的研究。对 NADPH 酶结构域进行 3D 结构预测和相似性建模。进行基因本体论和功能研究,以及对小麦呼吸爆发氧化酶(TaRboh)基因家族对生物和非生物胁迫的表达分析。
对包括小麦在内的七种植物物种的 Rboh 基因家族成员进行系统发育分析,将该家族分为四个亚家族。Rboh 基因主要参与各种生物学过程,如对氧化应激的反应、超氧阴离子的产生、过氧化氢的生物合成过程。在分子功能方面,钙结合、过氧化物酶活性、氧化还原酶活性、生成超氧化物的 NADPH 氧化酶活性是必不可少的。家族和超家族的酶注释表明,它编码五个结构簇,并编码 NAD(P)H 氧化酶(HO 形成)(EC:1.6.3.1)、三价铁螯合还原酶(NADH)(EC:1.16.1.7)、过氧化物酶(EC:1.11.1.7)、核糖磷酸二磷酸激酶(EC:2.7.6.1)。这些酶包含六个跨膜结构域、两个血红素和与 NADPH、EF 手和 FAD 结合相关的保守基序。结果还反映了该酶在不同分子功能中的独特作用,这些功能负责应激信号转导。此外,还观察到 TaRboh 的转录本在茎、叶、穗、种子和根等各种植物部位表达。我们还观察到这些基因家族成员在干旱/干旱+热组合和重要小麦病原体如小麦条锈菌、叶锈菌、禾谷镰刀菌、禾谷镰孢菌和小麦纹枯病菌等下的表达。
研究表明,鉴定的小麦呼吸爆发同源物(Rboh)参与了病原体激活的 ROS 产生,并在细胞死亡和防御反应中具有调节功能。