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What is RNA interference (RNAi) and how this can be relevant to our food?

A Seminar by Nobel Laureate, Dr. Andrew Fire









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    Wood transcriptome profiling identifies critical pathway genes of secondary wall biosynthesis and novel regulators for vascular cambium development in populus
    XV World Forestry Congress, 2-6 May 2022
    2022
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    Wood, the most abundant biomass on Earth, is composed of secondary xylem differentiated from vascular cambium. However, the underlying molecular mechanisms of wood formation remain largely unclear. To gain insight into wood formation, we performed a series of wood-forming tissue-specific transcriptome analyses from a hybrid poplar (Populus alba × P. glandulosa, clone BH) using RNA-seq. Together with shoot apex and leaf tissue, cambium and xylem tissues were isolated from vertical stem segments representing a gradient of secondary growth developmental stages (i.e., immature, intermediate, and mature stem). In a comparative transcriptome analysis of the ‘developing xylem’ and ‘leaf’ tissue, we could identify critical players catalyzing each biosynthetic step of secondary wall components (e.g., cellulose, xylan, and lignin). Several candidate genes involved in the initiation of vascular cambium formation were found via a co-expression network analysis using abundantly expressed genes in the ‘intermediate stem-derived cambium’ tissue. We found that transgenic Arabidopsis plants overexpressing the PtrHAM4-1, a GRAS family transcription factor, resulted in a significant increase of vascular cambium development. This phenotype was successfully reproduced in the transgenic poplars overexpressing the PtrHAM4-1. Taken together, our results may serve as a springboard for further research to unravel the molecular mechanism of wood formation, one of the most important biological processes on this planet. Keywords: Genetic resources, Research ID: 3622616
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    Characterization of formate dehydrogenase from Trametes versicor for formate production from CO2 gas
    XV World Forestry Congress, 2-6 May 2022
    2022
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    Enzymatic CO2 reduction has been reported as a promising approach to greenhouse gas fixation. There are few biocatalysts capable of biological CO2 fixation, e.g. pyruvate decarboxylase, carbonic anhydrase, and formate dehydrogenase. Formate dehydrogenase(FDH) can reduce CO2 to formate without organic chemicals, and formate can be sequentially reduced to formaldehyde and methanol by coupling aldehyde dehydrogenase and alcohol dehydrogenase reactions. Therefore, FDH has been widely adopted inCO2 reducing reaction.
    In this study, we researched CO2-reducing activities of FDH derived from wood rot fungi, Trametes versicolor. Microbial species, T. versicolor, were separated in National institute of Forest Science. The fungi were grown in a potato dextrose agar medium. For the transformation, the host, Shizosaccaromyces pombe of wild type, was obtained from Bioneer(Korea). Total RNA was extracted with Hybrid-R (GeneAll) from T. versicolor mycelium. cDNAs were synthezied by AccuPower (Bioneer) as recommended by the manufacturer. For sub-cloning, PCR of fragments (1.2 kb) were amplified with pairs of TvFDH ORF primers(Foword: 5’-ATGCTCGCCGGCATCT-3’, Reverse: 5’-TCACTTGCGCTGGCCA-3’). The amplified fragments were sub-cloned into a pCR2.1-TOPO (Invitrogen, Carlsbad, CA, USA) vector and the plasmid DNA was sequenced. The amplified fragments were digested by Not 1 and Bam 1 and then inserted into a pSLF272 vector (Bioneer, Korea). The recombinant plasmid was amplified in the Escherichia coli DH5α, and then extracted by mini prep kit. The recombinant plasmids were chemically (lithium acetate method) transformed into yeast S. pombe. Ligation products, transformants, were named as pSLF272-TvFDH. Selection of transformants were performed in the minimal medium, EMM. Expression and purification of the TvFDHs is being performed to obtain pure enzymes. Keywords: Climate change, Research, Genetic resources, Agriculture, Innovation ID: 3604933
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    Wood transcriptome analysis of Pinus densiflora identifies genes critical for secondary cell wall formation and NAC transcription factors involved in tracheid formation
    XV World Forestry Congress, 2-6 May 2022
    2022
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    Although conifers have significant ecological and economic value, information on transcriptional regulation of wood formation in conifers is still limited. Here, to gain insight into secondary cell wall (SCW) biosynthesis and tracheid formation in conifers, we performed wood tissue-specific transcriptome analyses of Pinus densiflora (Korean red pine) using RNA sequencing. In addition, to obtain full-length transcriptome information, PacBio single molecule real-time (SMRT) iso-sequencing was carried out using RNAs from 28 tissues of P. densiflora. Subsequent comparative tissue-specific transcriptome analysis successfully pinpointed critical genes encoding key proteins involved in biosynthesis of the major secondary wall components (cellulose, galactoglucomannan, xylan, and lignin). Furthermore, we predicted a total of 62 NAC (NAM, ATAF1/2 and CUC2) family transcription factor members and identified seven PdeNAC genes preferentially expressed in developing xylem tissues in P. densiflora. Protoplast-based transcriptional activation analysis found that four PdeNAC genes, homologous to VND, NST and SND/ANAC075, upregulated GUS activity driven by an SCW-specific cellulose synthase promoter. Consistently, transient overexpression of the four PdeNACs induced xylem vessel cell-like SCW deposition in both tobacco (Nicotiana benthamiana) and Arabidopsis leaves. Taken together, our data provide a foundation for further research to unravel transcriptional regulation of wood formation in conifers, especially SCW formation and tracheid differentiation. Keywords: Research, Genetic resources ID: 3622610

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