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Key message  Overexpression of BoMYB29 gene up-regulates the aliphatic glucosinolate pathway in  Brassica oleracea  plants increasing the production of the anti-cancer metabolite glucoraphanin, and the toxic and pungent sinigrin.    Abstract  Isothiocyanates, the bio-active hydrolysis products of glucosinolates, naturally produced by several  Brassicaceae  species, play an important role in human health and agriculture. This study aims at correlating the content of aliphatic glucosinolates to the expression of genes involved in their synthesis in  Brassica oleracea , and perform functional analysis of  BoMYB29  gene. To this purpose, three genotypes were used: a sprouting broccoli, a cabbage, and a wild genotype (Winspit), a high glucosinolate containing accession. Winspit showed the highest transcript level of  BoMYB28 ,  BoMYB29  and  BoAOP2  genes, and  BoAOP2  expression was positively correlated with that of the two  MYB  genes. Further analyses of the aliphatic glucosinolates also showed a positive correlation between the expression of  BoAOP2  and the production of sinigrin and gluconapin in Winspit. The Winspit  BoMYB29  CDS was cloned and overexpressed in Winspit and in the DH AG1012 line. Overexpressing Winspit plants produced higher quantities of alkenyl glucosinolates, such as sinigrin. Conversely, the DH AG1012 transformants showed a higher production of methylsulphinylalkyl glucosinolates, including glucoraphanin, and, despite an up-regulation of the aliphatic glucosinolate genes, no increase in alkenyl glucosinolates. The latter may be explained by the absence of a functional  AOP2  gene in DH AG1012. Nevertheless, an extract of DH AG1012 lines overexpressing  BoMYB29  provided a chemoprotective effect on human colon cells. This work exemplifies how the genetic diversity of  B. oleracea  may be used by breeders to select for higher expression of transcription factors for glucosinolate biosynthesis to improve its natural, health-promoting properties.    Electronic supplementary material  The online version of this article (10.1007/s11103-019-00890-2) contains supplementary material, which is available to authorized users.

Overexpression of the MYB29 transcription factor affects aliphatic glucosinolate synthesis in Brassica oleracea