Investigation of heat stress linked RNAi and crop quality determining genes by genome editing technology in barley

Genome editing revolutionizes site-directed mutagenesis in molecular biology. Our lab pioneers barley genome editing, expanding experiments. Using bioinformatics, we'll pinpoint RNA interference factors (DCL, AGO, RDR) in barley. Heat stress-related factors will be identified via gene expression analysis in treated plants, followed by genome editing to understand their role in stress response. We'll also target the SIX-ROWED SPIKE (VRS) gene for enhanced grain characteristics. To bolster reliability, we'll develop a high-fidelity, plant-specific Cas9 construct based on published functional studies.

http://nyilvanos.otka-palyazat.hu/index.php?menuid=930&lang=HU&num=130384

Establishment of genome editing in barley and other crop species for research and crop improvement

The CRISPR/Cas9 system, a swift and precise genome editing tool, is poised for integration into Hungarian research. Our experiments focus on economically vital barley, utilizing established lab techniques to create CRISPR/Cas9-edited plants. Targets include RNA interference components, a key research area, exploring their role in barley development and stress response. CRISPR/Cas9 allows direct genome modifications in local cultivars, expediting trait introduction without traditional breeding delays. This system offers transgenic alternatives, sidestepping typical genetically modified organism concerns. We'll evaluate this aspect by targeting barley genes for trait improvement. Additionally, we'll extend genome editing efforts to wheat, a critical Hungarian crop.

http://nyilvanos.otka-palyazat.hu/index.php menuid=930&num=125300&keyword=125300

Investigation of the regulation and activity of RNA interference executor complexes in model and crop plants

RNA interference (RNAi), a small non-coding RNA (smRNA) system, regulates gene expression, development, stresses, epigenetics, and defense. Micro (mi) RNAs and small interfering (si) RNAs control target RNAs through degradation or repression. The RNA-induced silencing complex (RISC) features ARGONAUTE1 (AGO1), pivotal in siRNA and miRNA pathways. AGO1 regulation, influenced by miR168, exhibits unique aspects compared to canonical miRNA control. Our goal is to explore fine-tuned, tissue-specific AGO1 activity, unveiling specific regulatory processes and its role in development. Utilizing size separation, we'll analyze RISC-bound smRNAs through next-generation sequencing, shedding light on loading regulation. A novel gain-of-function mutant screen will identify factors modulating the RNA interference pathway. Results will be applied to economically crucial crops like pepper and wheat.

http://nyilvanos.otka-palyazat.hu/index.php?menuid=930&lang=HU&num=116602

Genome wide mRNA and small RNA transcriptome profiling and characterization in Capsicum annuum

This study delves into the intricacies of RNA-mediated regulation in plant development, focusing on Capsicum annuum (pepper) cultivars, a significant crop in Hungary. Leveraging deep sequencing technologies, we aim to unveil tissue-specific expression patterns in pepper transcriptomes. Through RNA-seq and sRNA-seq, we will profile mRNA and small RNA, constructing the pepper genome. Comparative bioinformatics will identify known and novel pepper-specific regulatory small RNAs (miRNAs, siRNAs). Promising candidates will undergo thorough investigation using advanced molecular biology techniques. Comparative analyses between ancient and modern cultivars will spotlight key small RNAs or mRNAs shaping economically important traits during pepper development. This research not only uncovers regulatory RNAs in pepper organogenesis but also provides mechanistic insights into small RNA action during plant development broadly.

http://nyilvanos.otka-palyazat.hu/index.php?menuid=930&lang=HU&num=109438