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

Kód výsledku v IS VaVaI

<a href="https://www.isvavai.cz/riv?ss=detail&h=RIV%2F60077344%3A_____%2F21%3A00559502" target="_blank" >RIV/60077344:_____/21:00559502 - isvavai.cz</a>

Výsledek na webu

<a href="https://www.sciencedirect.com/science/article/pii/S0254629920311911?via%3Dihub" target="_blank" >https://www.sciencedirect.com/science/article/pii/S0254629920311911?via%3Dihub</a>

DOI - Digital Object Identifier

<a href="http://dx.doi.org/10.1016/j.sajb.2020.12.004" target="_blank" >10.1016/j.sajb.2020.12.004</a>

Jazyk výsledku

angličtina

Název v původním jazyce

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

Popis výsledku v původním jazyce

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.

Název v anglickém jazyce

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

Popis výsledku anglicky

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.

Druh

J_imp - Článek v periodiku v databázi Web of Science

CEP obor

—

OECD FORD obor

10610 - Biophysics

Projekt

<a href="/cs/project/EF15_003%2F0000336" target="_blank" >EF15_003/0000336: Kovy, rostliny a lidé</a><br>

Návaznosti

P - Projekt vyzkumu a vyvoje financovany z verejnych zdroju (s odkazem do CEP)<br>I - Institucionalni podpora na dlouhodoby koncepcni rozvoj vyzkumne organizace

Rok uplatnění

2021

Kód důvěrnosti údajů

S - Úplné a pravdivé údaje o projektu nepodléhají ochraně podle zvláštních právních předpisů

Název periodika

South African Journal of Botany

ISSN

0254-6299

e-ISSN

1727-9321

Svazek periodika

138

Číslo periodika v rámci svazku

May 01

Stát vydavatele periodika

ZA - Jihoafrická republika

Počet stran výsledku

7

Strana od-do

50-56

Kód UT WoS článku

000648693200004

EID výsledku v databázi Scopus

2-s2.0-85099249828