Direct in situ RT-PCR as a useful tool in localisation of abundant and organ-specific genes expressed in Rosa hybrida

• Published in: Acta horticulturae no. 953; pp. 201 - 207
• Main Authors: Jedrzejuk, A, Mibus, H, Serek, M
• Format: Journal Article
• Language: English
• Published: International Society for Horticultural Science 01.01.2012
• Subjects: buds; color; cultivars; flowers; genes; hybrids; nucleic acids; ornamental plants; reverse transcriptase polymerase chain reaction; Rosa
• ISSN: 0567-7572
• DOI: 10.17660/actahortic.2012.953.28

In situ PCR is a technique, which allows detection of specific nucleic acid sequences in particular cells and tissues. In plants, in situ RT-PCR is widely used in localisation of specific genes, including MADS-box, and other function-specific genes of floral buds and other organs. Roses are one of the most economically important ornamental plants, and a number of cultivars have been selected based on flower traits, such as petal form, colour, and number. Of particular interest to floriculture researchers and breeders is flower colour and petal number, which is connected to MADS-box genes expression. In this report, we present a convenient protocol for the localisation of transcript expression in Rosa hybrid. The protocol is more appealing because of a high sensitivity for the in situ RT-PCR reaction and its speed. We demonstrated that a two-step reaction can be completed in two days, and a one-step reaction with fluorescent nucleotide used as a label can be completed in one day. Our results showed that a normal RT-PCR reaction performed directly on tissue showed a high specific expression of examined genes in rose organs.