Abstract
Agrobacterium-mediated transformation has been widely used in plants. However, the mechanism in plant cells’ response to Agrobacterium infection was very complex. The mechanism of the determinants in host cell remains obscure, especially in barley, which is recalcitrant for Agrobacterium-mediated transformation. In the present study, microspore-derived embryogenic calli (MDEC) from barley elite cultivar were employed as unique subjects to characterize the mechanisms during the Agrobacterium infection process. Hua 30 MDEC can be successfully infected by Agrobacterium. RNA-sequencing at different infection points (0, 2, 6, 12, 24 hpi) was performed. The average expressional intensity of the whole genomics increased from 0 to 2 hpi, and then decreased subsequently. More upregulated than downregulated differentially expressed genes (DEGs) were counted at the same time. GO enrichment analysis showed that protein modification was significantly overrepresented in upregulated DEGs. Chromosome-related biological processes, gene expression and cellular metabolic processes were significantly overrepresented in downregulated DEGs. KEGG analysis showed that plant defense responses, phenylpropanoid biosynthesis and biosynthesis of amino acids were significantly enriched across the infection time course. Nine DEGs related to defense responses were identified. All DEGs were upregulated from 2 to 24 hpi. We speculate that these genes are possibly related to Agrobacterium infection. These findings will provide deep insights into the molecular events occurring during the process of Agrobacterium-mediated transformation.
Highlights
Agrobacterium tumefaciens possesses the ability to integrate self T-DNA into plant genomic [1]
3.1 Infection of Hua 30 microspore-derived embryogenic calli (MDEC) by Agrobacterium Microspores isolated from the anthers of Hua 30 were cultured under controlled aseptic conditions
The results indicated that Hua 30 MDEC can be successfully infected by Agrobacterium strain LBA4404, but the infection efficiency was not high
Summary
Agrobacterium tumefaciens possesses the ability to integrate self T-DNA into plant genomic [1]. The. Agrobacterium tumefaciens-mediated genetic transformation system has been widely used in both basic research and breeding in plants, based on the low copy number of large segments of DNA into host genome DNA and low cost. 2022, vol., no.6 studies, the Agrobacterium infection process involves two steps: the attachment of the bacterium to host cells and the transfer of the DNA-protein complex into the plant cell [3]. The plants have evolved defense systems for the Agrobacterium infection, which may be an important factor influencing the infection efficiency of plant cells [4]. As a gram-negative bacterium, Agrobacterium tumefaciens possesses bacteria-derived compounds known as pathogen-associated molecular patterns (PAMP), which caused immune response termed as PAMP-triggered immunity (PTI) in host cells [5]. The mechanisms in plant cells’ response to Agrobacterium infection were very complex [6,7]; any molecular interactions at these stages can affect the infection efficiency. It is critically important to get more knowledge on the molecular mechanisms involved in the plant-Agrobacterium interaction [8]
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