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Small-interfering RNAs from natural antisense transcripts derived from a cellulose synthase gene modulate cell wall biosynthesis in barley

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Small-interfering RNAs (siRNAs) from natural cis-antisense pairs derived from the 3'-coding region of the barley (Hordeum vulgare) CesA6 cellulose synthase gene substantially increase in abundance during leaf elongation. Strand-specific RT-PCR confirmed the presence of an antisense transcript of HvCesA6 that extends greater-than-or-equal1230 bp from the 3' end of the CesA-coding sequence. The increases in abundance of the CesA6 antisense transcript and the 21-nt and 24-nt siRNAs derived from the transcript are coincident with the down-regulation of primary wall CesAs, several Csl genes, and GT8 glycosyl transferase genes, and are correlated with the reduction in rates of cellulose and (1 [rightward arrow] 3),(1 [rightward arrow] 4)-β-D-glucan synthesis. Virus induced gene silencing using unique target sequences derived from HvCesA genes attenuated expression not only of the HvCesA6 gene, but also of numerous nontarget Csls and the distantly related GT8 genes and reduced the incorporation of D-¹⁴C-Glc into cellulose and into mixed-linkage (1 [rightward arrow] 3),(1 [rightward arrow] 4)-β-D-glucans of the developing leaves. Unique target sequences for CslF and CslH conversely silenced the same genes and lowered rates of cellulose and (1 [rightward arrow] 3),(1 [rightward arrow] 4)-β-D-glucan synthesis. Our results indicate that the expression of individual members of the CesA/Csl superfamily and glycosyl transferases share common regulatory control points, and siRNAs from natural cis-antisense pairs derived from the CesA/Csl superfamily could function in this global regulation of cell-wall synthesis.
Held, Michael A. , Penning, Bryan , Brandt, Amanda S. , Kessans, Sarah A. , Yong, Weidong , Scofield, Steven R. , Carpita, Nicholas C.
Hordeum vulgare , barley , genes , hexosyltransferases , gene expression regulation , gene silencing , antisense RNA , cell walls , leaves , leaf development , cellulose , beta-glucans , carbohydrate metabolism , small interfering RNA
p. 20534-20539.
Includes references
Proceedings of the National Academy of Sciences of the United States of America 2008 Dec. 23, v. 105, no. 51
National Academy of Sciences
Journal Articles, USDA Authors, Peer-Reviewed
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