TALE protein mediated overexpression of embryogenesis related marker genes in wheat microspores

0559502 - BC 2022 RIV ZA eng J - Journal Article
Sinha, Rakesh Kumar - Jiang, F. - Eudes, F.
TALE protein mediated overexpression of embryogenesis related marker genes in wheat microspores.
South African Journal of Botany. Roč. 138, May 01 (2021), s. 50-56. ISSN 0254-6299. E-ISSN 1727-9321
R&D Projects: GA MŠMT EF15_003/0000336
Institutional support: RVO:60077344
Keywords : Microspore: RKD ECA1 TPD1 * TALE protein * Embryogenesis
OECD category: Biophysics
Impact factor: 3.111, year: 2021
Method of publishing: Open access
https://www.sciencedirect.com/science/article/pii/S0254629920311911?via%3Dihub

Stress treatment of microspore embryogenesis leads to the reprogramming of male gametophyte development pathways, which generates haploid green plants. However, low green plant production efficiency is a major hindrance to the adoption of the double haploid (DH) production platform for wheat breeding. The aim of the current study was to induce the expression of microspore embryogenesis-associated genes by overexpressing the RKD transcription factor using Transcription activator-like effector (TALE) proteins. The TALE protein can modulate the gene expression by recognizing and binding host plant promoter sequences. Taking into consideration, we have cloned and analysed RKD promoters from wheat and triticale and used them for the custom synthesis of TaRKD-TALE protein. The isolated and purified protein was conjugated with R9 (Cys (Npys)-(D-Arg)9) cargo peptides and successfully transduced into wheat microspores. The expression of early-culture abundant (ECA1), RWP-RK domain-containing proteins (RKD1) and Tapetum determinant 1 (TPD1) marker genes in wheat linked to microspore embryogenesis were studied. The expression of all three (ECA1, TPD1 and RKD1) embryogenesis-related marker genes in the TALE transduced microspore was significantly higher than in the control. Microspore culture transduced by R9-TaRKD-TALE protein yielded a significantly higher number of embryo-like structures (ELS) and the total green plants in wheat cultivars AC Fielder. Conversely, the microspore cell death has not been significantly affected by the transduction of R9TaRKD-TALE, while showing a higher recovery at 120 h of microspore culture. This work will be helpful to exploit the TALE protein to improve our understanding of DH production and other pathways-related genes.
Permanent Link: https://hdl.handle.net/11104/0332780