A genotyping protocol for multiple tissue types from the polyploid tree species Sequoia sempervirens (Cupressaceae)

Area of Science:

• Botany
• Genetics
• Ecology

Background:

• Asexual reproduction in plants like coast redwood (Sequoia sempervirens) complicates clonal identification due to genetic variations within lineages.
• Somatic mutations, null alleles, and scoring errors can lead to non-identical genotypes from the same clonal lineage.

Purpose of the Study:

• To develop and validate a clonal identification protocol for coast redwood that is resilient to common sources of error.
• To enable accurate clonal lineage identification in hexaploid asexually reproducing species.

Main Methods:

• Utilized microsatellite data from six primers (four published, two new) with a modified scoring protocol.
• Employed Bruvo genetic distances for clone identification.
• Assessed protocol effectiveness through simulations and genotyping of paired tissue samples from the same trees.

Main Results:

• Simulations confirmed the protocol's accuracy in identifying clonal lineages.
• The protocol correctly identified multiple tissue samples from the same coast redwood trees as clones.
• Observed variations in genetic distances between certain tissue type pairs.

Conclusions:

• The developed protocol accurately identifies coast redwood clones, supporting future ecological studies.
• This methodology is applicable to the study of clonal organisms beyond coast redwoods.