期刊
JOURNAL OF EXPERIMENTAL BOTANY
卷 72, 期 22, 页码 7808-7825出版社
OXFORD UNIV PRESS
DOI: 10.1093/jxb/erab365
关键词
Barley; brassinosteroids; cell reprogramming; cork oak; glycogen synthase kinase; microspore embryogenesis; small molecule inhibitors; somatic embryogenesis; rapeseed
资金
- Spanish National Agency of Research (Agencia Estatal de Investigacion, AEI)
- European Regional Development Fund (ERDF/FEDER) [AGL2017-82447-R, PID2020-113018RB-I00, SAF2016-76693-R, PID2019105600RB-I00]
The use of synthetic small molecule inhibitors of mammalian GSK-3 beta has been shown to improve in vitro embryogenesis efficiency in plants, promoting somatic embryogenesis by activating the BR pathway.
Plant in vitro regeneration systems, such as somatic embryogenesis, are essential in breeding; they permit propagation of elite genotypes, production of doubled-haploids, and regeneration of whole plants from gene editing or transformation events. However, in many crop and forest species, somatic embryogenesis is highly inefficient. We report a new strategy to improve in vitro embryogenesis using synthetic small molecule inhibitors of mammalian glycogen synthase kinase 3 beta (GSK-3 beta), never used in plants. These inhibitors increased in vitro embryo production in three different systems and species, microspore embryogenesis of Brassica napus and Hordeum vulgare, and somatic embryogenesis of Quercus suber. TDZD-8, a representative compound of the molecules tested, inhibited GSK-3 activity in microspore cultures, and increased expression of embryogenesis genes FUS3, LEC2, and AGL15. Plant GSK-3 kinase BIN2 is a master regulator of brassinosteroid (BR) signalling. During microspore embryogenesis, BR biosynthesis and signalling genes CPD, GSK-3-BIN2, BES1, and BZR1 were up-regulated and the BAS1 catabolic gene was repressed, indicating activation of the BR pathway. TDZD-8 increased expression of BR signalling elements, mimicking BR effects. The findings support that the small molecule inhibitors promoted somatic embryogenesis by activating the BR pathway, opening up the way for new strategies using GSK-3 beta inhibitors that could be extended to other species.
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