Tissue integrity and RNA quality of laser microdissected phloem of potato.
The phloem is an important conduit for the transport of signaling molecules including RNA. Phloem sap has served as a source of RNA to profile uncontaminated phloem transcriptomes but its collection is difficult in many species. Laser capture microdissection techniques offer a valuable alternative for isolating RNA from specific vascular cells. In potato (Solanum tuberosum L.), there are seven BEL1-like transcription factors expressed throughout the plant with diverse functions. The RNA of one of these, StBEL5, moves through the phloem from the leaf to stolon tips to regulate tuber formation. In this study, the presence of several BEL RNAs and one Knotted1-like RNA was determined in phloem cells collected by laser microdissection coupled to laser pressure catapulting (LMPC). Three fixatives were compared for their effect on cell morphology and RNA quality in transverse sections of stems of potato. For optimum tissue integrity and quality of RNA from potato stem sections, the best results were achieved using ethanol acetate as the fixative. In addition, the RT-PCR results demonstrated the presence of six out of seven of the StBEL RNAs and a potato Knox RNA in phloem cells.
Yu
Yueyue
Y
Molecular, Cellular, and Developmental Biology Major, Iowa State University, Ames, IA 50011, USA.
Lashbrook
C C
CC
Hannapel
David J
DJ
D018506:Gene Expression Regulation, Plant; D007834:Lasers; D042282:Microdissection; D052585:Phloem / Q000166:cytology / Q000235:genetics* / Q000378:metabolism*; D018547:Plant Stems / Q000166:cytology; D012333:RNA, Messenger / Q000235:genetics / Q000378:metabolism; D018749:RNA, Plant / Q000378:metabolism*; D011198:Solanum tuberosum / Q000166:cytology / Q000235:genetics* / Q000378:metabolism*
D012333:RNA, Messenger; D018749:RNA, Plant