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.