Accepted_test

Development of genetic models for validation of candidate genes associated with cold stress response in tea plant
by Egorova Anastasiya | Fizikova Anastasiya | Fomin Ivan | Kostina Nina | Gerasimova Sophia | Samarina Lidiia | Malyukova Lyudmila | FRC SSC RAS, ICG SBRAS | FRC SSC RAS, Sirius University of Science and Technology | ICG SBRAS | ICG SBRAS | FRC SSC RAS, ICG SBRAS | FRC SSC RAS,Sirius University of Science and Technology | FRC SSC RAS
Abstract ID: 173
Event: BGRS-abstracts
Sections: [Sym 6] Section “Genomics, genetics and systems biology of plants”

Tea (Camellia sinensis (L.) Kuntze) is a perennial woody plant, one of the most important agricultural crops for human consumption, medicinal and functional purposes, grown in more than 60 countries worldwide. Cold is the main factor limiting the expansion of tea plantations to more northern latitudes and limiting their productivity. Transcriptional analysis of cv. Kolkhida identified candidate genes involved in long-term cold stress responses. The aim of this study is to functionally validate these candidate genes using knockout and overexpression methods.

Two candidate genes (COR413PM1-like and ELIP1) were selected for functional validation. Cas9/gRNA-mediated site-directed mutagenesis was used to knock out candidate genes in cv. Kolkhida. The extraction and transformation of protoplasts from young tea leaves was established and the mutagenic activity of the vectors was demonstrated in protoplasts.

Overexpression of candidate genes was performed in Nicotiana tabacum SR1 plants. The candidate genes were cloned from the cv. Kolkhida and overexpression vectors were constructed. Agrobacterium-mediated genetic transformation of N. tabacum was performed, resulting in ten transgenic lines for each gene. Based on qPCR analysis, five lines with the highest expression for each gene were selected and T1 generations were obtained.

During this work, the development of genetic models was initiated, which will be used to study the functions of candidate genes in response to cold stress.