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Gene editing of authentic Brassica rapa flavonol synthase 1 generates dihydroflavonol-accumulating Chinese cabbage

Horticulture Research 2023年第12期10卷 58-69页

作者： Sangkyu Park Hyo Lee Jaeeun Song Chan Ju lim Jinpyo Oh Sang Hoon Lee Saet Buyl Lee Jong-Yeol Lee sunhyung lim Jin A.Kim Beom-Gi Kim Metabolic Engineering Division National Institute of Agricultural SciencesRural Development AdministrationJeonju54874South Korea Institute of Biotechnology and Breeding Asiaseed Inc.Icheon17414South Korea Food and Nutrition Division National Institute of Agricultural SciencesRural Development AdministrationWanju55365South Korea Department of Horticultural Biotechnology School of BiotechnologyHankyong National UniversityAnseong17579South Korea

Flavonols are the major class of flavonoids of green Chinese cabbage(Brassica rapa ***).The *** genome harbors seven flavonol synthase genes(BrFLSs),but they have not been functionally ***,transcriptome analysis showed four BrFLSs mainly expressed in Chinese *** them,only BrFLS1 showed major FLS activity and additional flavanone 3β-hydroxylase(F3H)activity,while BrFLS2 and BrFLS3.1 exhibited only marginal F3H *** generated BrFLS1-knockout(BrFLS1-KO)Chinese cabbages using CRISPR/Cas9-mediated genome editing and obtained transgene-free homozygous plants without off-target mutation in the T1 generation,which were further advanced to the T2 generation showing normal ***-ESI-QTOF-MS analysis revealed that flavonol glycosides were dramatically decreased in the T2 plants,while dihydroflavonol glycosides accumulated concomitantly to levels corresponding to the reduced levels of *** PCR analysis revealed that the early steps of phenylpropanoid and flavonoid biosynthetic pathway were upregulated in the BrFLS1-KO *** accordance,total phenolic contents were slightly enhanced in the BrFLS1-KO plants,which suggests a negative role of flavonols in phenylpropanoid and flavonoid biosynthesis in Chinese *** surveys revealed that the BrFLS1-KO Chinese cabbages showed normal head formation and reproductive phenotypes,but subtle morphological changes in their heads were *** addition,their seedlings were susceptible to osmotic stress compared to the controls,suggesting that flavonols play a positive role for osmotic stress tolerance in *** *** this study,we showed that CRISPR/Cas9-mediated BrFLS1-KO successfully generated a valuable breeding resource of Chinese cabbage with distinctive metabolic traits and that CRISPR/Cas9 can be efficiently applied in functional Chinese cabbage breeding.

关键词： steps slightly phenolic

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