ABSTRACT
To gain more insights into the rice base editor (rBE3 and rBE4), we evaluated the mutation efficiency, off-target and inheritance of OsSERK1(D428N) and pi-ta(S918F) genes modified with rBE endonucleases. We predicted and analyzed the putative off-target sites of the sgRNA designed for OsSERK1(D428N) and pi-ta(S918F) by PCR amplification and Sanger sequencing. Then we further characterized the inheritance and stability of targeted base mutations and T-DNA segregation in the progeny of the self-fertilized T0 plants. Analysis of the DNA sequencing data of T0 plants of OsSERK1(D428N) revealed no nucleotide change at any of the four potential off-target sites. For OsSERK1(D428N) and Os08g07774 carry the same sgRNA targeting sites, base substitution at both two loci were detected at a frequency of 41.67%. The targeted base mutations could be transmitted readily to T1 progeny. Furthermore, genetic segregation caused the loss of T-DNA at a frequency between 25.0% and 40.9% in the T1 transgenic plants of OsSERK1(D428N) and pi-ta(S918F). These results demonstrated that the rBE3 and rBE4 systems could mediate specifically targeted base editing in one- or multi-site, and the targeted base editing could be stably inherited to next generation.
ABSTRACT
<p><b>OBJECTIVE</b>To investigate the chemical constituents from the rhizome o fHedychium chrysoleucum.</p><p><b>METHOD</b>The compounds were isolated by column chromatography on silica gel. The structures were identified by NMR, IR and MS analyses.</p><p><b>RESULT</b>Seven compounds were isolated and identified as hedychenone (1), coronarin A (2), yunnancoronarin A (3), coronarin E (4), beta-sitosterol (5), cryptomeridiol (6), and beta-eudesmol (7).</p><p><b>CONCLUSION</b>Compounds 1-7 were obtained from this plant for the first time.</p>