BIOLOGY 201L9 Molecular Biology Lab

Question:

Herbicide resistance refers to the ability of weeds to reproduce and survive in herbicide environments that are fatal to wild species.

Erigeron bonariensis, also known as hairy fleabane, is an agricultural weed found in California’s Central Valley. It has developed resistance to RoundUp(r), an herbicide that kills wild weeds.

E. canadensis is a close relative to E.

Bonariensis is a non-target site, reduced translocation of herbicide. This prevents the chemical’s spread around the plant.

The target gene EPSPS is thought to be responsible for resistance in Erigeron canadensis. This combination of ABC transporter genes M10, M11 may also play a role.

This study will use quantitative PCR (qPCR), to determine if these candidate genes play a role in the glyphosate resistance of wild Erigeron bonariensis populations.

Samples of the weed were taken from plants from 10 populations wild-collected in the Central Valley, and two control populations of Erigeron Bonariensis.

To determine percent resistance, each wild-collected population was analyzed for its response to glyphosate.

RNA was extracted from the leaves and used to synthesize cDNA.

For the E, quantitative PCR primers were created.

Bonariensis orthologues to the E. canadensis gene EPSPS, ABC M10 and ABC M11 were analysed using qPCR. The pre- and post-spraying expression of each gene (relatively to the housekeeping gene Actin) was also analyzed.

This experiment will look at gene expression patterns in ABC transporter genetics (M10, M11), to determine if these genes are upregulated by Glyphosate application. It also establishes their role in glyphosate resistant genes.

Future RNA-Seq analysis using Illumina HiSeq could reveal additional genes that are differentially affected in resistant populations.

The genetic basis of herbicide resistance can be determined. This will give fundamental information about parallel evolution under strong selection pressures and may help to suggest other methods for controlling this weed in the field.

Answer:

None

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