Epsilon Archive for Student Projects

Abstract

The Norway spruce (Picea abies) stands as one of Sweden’s most valuable natural resources, yet its slow reproductive cycle poses challenges for swift breeding in response to climate change, spanning 20-25 years to reach reproductive maturity. The discovery of the natural early cone-setting variant,
acrocona, offers insights into the genetic mechanisms governing cone development timing. Notably, the SQUAMOSA-PROMOTER BINDING PROTEIN-like-1 (PaSPL1) gene has been linked to this trait through a mutation in a micro-RNA-regulated region.
This study focused on creating yeast libraries containing potential transcription factors (TFs) of PaSPL1, intended for Yeast 1-hybrid mating experiments. These libraries will be complemented by yeast libraries containing PaSPL1 promoters from genotypes with irregular cone-setting phenotypes and will help elucidate the regulatory network of PaSPL1. As acrocona only produces early female
cones, we also aimed to develop entry clones carrying putative B-class (“male-forming”) genes DEFICIENT AGAMOUS LIKE12 (DAL12) and DAL13, that will be used for transforming acrocona calli and evaluation of their role in cone development. Lastly, protocols for stable calli transformation, alongside novel methods for transient transformation of seedlings with RUBY vectors, were established and evaluated.