SUPR
PacBio Sequencing Analysis for Insect Pheromone Producing Camelina sativa
Dnr:

NAISS 2023/22-1295

Type:

NAISS Small Compute

Principal Investigator:

Kamil Demski

Affiliation:

Sveriges lantbruksuniversitet

Start Date:

2023-12-22

End Date:

2025-01-01

Primary Classification:

40101: Agricultural Science

Webpage:

Allocation

Abstract

Moth larvae are a major threat to global food security as they deplete our produce. Fighting them with pesticides proved to have a negative impact on the environment. Luckily, these pests can also be controlled through moth sex pheromone mating disruption. Unfortunately, chemical synthesis of pheromones is expensive. An alternative is to produce them in plant seeds. Many moth sex pheromones are unusual unsaturated medium-chain fatty alcohols and can be produced in a plant system by transgenic enzymes. Our efforts thus far concentrated on producing fatty acid precursors of the pheromones in plants. In this project we went a step further: by producing the pheromones themselves and then entrapping them in fatty alcohol moieties of seed wax esters (WE). Our target pheromones of choice were 14:1(Z11) and 14:1(E11) (14-carbon-long, monounsaturated compounds) produced by Argyrotaenia velutinana, the red-banded leafroller moth, a pest of many fruits, including apples, consuming plant leaves and fruit alike. We produced seed WE-pheromone-accumulating lines of Camelina sativa, a promising industrial oilcrop, up to the fouth generation. To our surprise different lines, all steming from one initial transformant (T1), produce varying amounts of wax esters and varying amounts of the pheromone precurser compounds, which goes beyond what should be observed due to zygosity. We suspect that these lines might have different genetic backgrounds, maybe because of Camelina's hexaploid nature. We therefore isolated genomic DNA from 10 progenitors from the next generation of the transformant (T2) and sent it for long-read PacBio sequencing. In this project we would like to assemble the estimated genome of the T1 transformant, to be able to find out where the transgene has been inserted, which will help us in downstream analysis of Camelina lines with varying WE content. It is possible that some lines contain multiple inserts of the transgenes. There is a lack of studies directed towards understanding how genetic background might affect the target compound content in transgenic hexaploid Camelina. We want to study it for the lines we developed in this project by linking data obtained from the genome assembly to the abundance of specific inserts in subsequent lines (by qPCR of genomic DNA) and their lipid analysis. This study could prove useful for research on transgenic Camelina and Camelina breeding.