Transgene silencing and transgene-derived siRNA production in tobacco plants homozygous for an introduced AtMYB90 construct.

PloS One
Jeff VeltenChristopher I Cazzonelli

Abstract

Transgenic tobacco (Nicotiana tabacum) lines were engineered to ectopically over-express AtMYB90 (PAP2), an R2-R3 Myb gene associated with regulation of anthocyanin production in Arabidopsis thaliana. Independently transformed transgenic lines, Myb27 and Myb237, accumulated large quantities of anthocyanin, generating a dark purple phenotype in nearly all tissues. After self-fertilization, some progeny of the Myb27 line displayed an unexpected pigmentation pattern, with most leaves displaying large sectors of dramatically reduced anthocyanin production. The green-sectored 27Hmo plants were all found to be homozygous for the transgene and, despite a doubled transgene dosage, to have reduced levels of AtMYB90 mRNA. The observed reduction in anthocyanin pigmentation and AtMYB90 mRNA was phenotypically identical to the patterns seen in leaves systemically silenced for the AtMYB90 transgene, and was associated with the presence of AtMYB90-derived siRNA homologous to both strands of a portion of the AtMYB90 transcribed region. Activation of transgene silencing in the Myb27 line was triggered when the 35S::AtMYB90 transgene dosage was doubled, in both Myb27 homozygotes, and in plants containing one copy of each of the independently seg...Continue Reading

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Citations

Dec 27, 2015·Plant Science : an International Journal of Experimental Plant Biology·Amandeep MittalChristopher D Rock
Oct 7, 2015·Frontiers in Plant Science·Sarma RajeevkumarRamalingam Sathishkumar
Dec 11, 2020·Nature Communications·Mohammed D AljohaniChristian Frøkjær-Jensen

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Methods Mentioned

BETA
transgenic
PCR
RNA assay
electrophoresis

Software Mentioned

smRNA
SAS
BLAST
SOLiD
BioEdit
NextGENe®

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