Cornerstone: A Collection of Scholarly and Creative Works for Minnesota State University, Mankato
Abstract
Lipoxygenases (LOXs) are enzymes which catalyze the peroxidation of polyunsaturated fatty acids. They function in development, growth and response to pathogenic attack including mechanical wounding. There are multiple isozymes of LOX in soybean. It has been reported that each isozyme has a distinct function in different stages of growth. Studies have shown that the expression of some isoforms of LOX is enhanced after wounding or treatment with jasmonic acid (JA) or methyl jasmonate (MJ), a mediator of plant defense mechanism. This project investigated the relationship between wounding, MJ treatment and expression of LOX isoenzymes including LOX10 using Quantitative Polymerase Chain Reaction. Soybean plants were raised in a growth chamber. When the plants reached the bifoliate stage, two sets of experimental plants were used. One set was wounded by crimping the leaves with a clamp. The other set was not wounded. Both sets of experimental plants were sealed in an aquarium containing MJ vapor. Control plants were sealed in an aquarium with no MJ. Leaves were harvested at 0, 3, 6, and 24 hours after treatment. RNA was isolated from the samples using an RNeasy Plant Minikit from Qiagen. RNA quantities were estimated from the absorbance at 260nm. A High-Capacity cDNA Reverse Transcription kit from Applied Biosystems was used to make cDNA copies of the mRNAs. Relative quantities of the LOX mRNA were measured by qPCR on a Plus One Real-Time PCR System from Applied Biosystems. SYBR green was used to detect the PCR products
