RNAi-induced silencing in floral tissues of Petunia hybrida by agroinfiltration: a rapid assay for chalcone isomerase gene function analysis.
Variegation in flower color is commonly observed in many plant species and also occurs on Petunia (Petunia hybrida) as an ornamental plant. Variegated plants are of highly valuable in the floricultural market. Agroinﬁltration is an Agrobacterium-mediated transient assay for the analysis of gene function and genetic modiﬁcation in leaves, ﬂowers and fruit tissues of various plants. Transient RNAi-induced silencing by agroinﬁltration has been developed in leaves and fruits of several plant species. Here we report the establishment of a transient hairpin RNAi-induced silencing system for color modiﬁcation assay in ﬂoral tissues of Petunia with different colors. chiRNAi construct was cloned into the pBI121 vector under the control of 35S promoter. Transient RNA silencing of chi in the ﬂoral tissues of Petunia was induced by delivering 530 bp chi hairpin RNAs (hpRNAs) into the petals of ﬂowers using agroinﬁltration. Impaired anthocyanin accumulation and reduction of endogenous mRNAs of the corresponding targets were observed in the inﬁltrated areas of the petals of four colors of Petunia. Silencing of the endogenous chi mRNAs was highly effective in reduction of chi gene and anthocyanin accumulation. This transient silencing system is a prototype for modiﬁcation of the anthocyanin biosynthetic pathway in Petunia through chi gene suppression.
Iranian Academic Center for Education, Culture and Research, Branch of Mashhad Department of Ornamental Plant Biotechnology Mashhad Iran F.keykha@jahrom.um.ac.ir.
F
F
Keykha
Ferdowsi University of Mashhad Department of Plant Breeding and Biotechnology, College of Agriculture Mashhad Iran.
A
A
Bagheri
Ferdowsi University of Mashhad Department of Plant Breeding and Biotechnology, College of Agriculture Mashhad Iran.
N
N
Moshtaghi
Ferdowsi University of Mashhad Department of Biology, Faculty of Sciences Mashhad Iran.
A R
AR
Bahrami
Iranian Academic Center for Education, Culture and Research, Branch of Mashhad Department of Ornamental Plant Biotechnology Mashhad Iran.
A
A
Sharifi
D060054:Agrobacterium / Q000378:metabolism*; D020029:Base Pairing / Q000235:genetics; D035264:Flowers / Q000201:enzymology* / Q000235:genetics*; D018506:Gene Expression Regulation, Plant; D005821:Genetic Techniques; D019753:Intramolecular Lyases / Q000235:genetics*; D010641:Phenotype; D010858:Pigmentation; D034622:RNA Interference; D060888:Real-Time Polymerase Chain Reaction
D019753:Intramolecular Lyases; C019865:chalcone isomerase