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The yellow seed characteristic in Brassica napus L. is desirable because of its association with higher oil content and better quality of oil-extracted meal. YN01-429 is a yellow-seeded canola-quality germplasm developed in Canada arising from several years of research. Seed-coat pigmentation is due to oxidized proanthocyanidins (PA; condensed tannins) derived from phenylpropanoids and malonyl CoA. We found PA accumulation to be most robust in young seed coats (20 d post anthesis; dpa) of a related black-seeded line N89-53 and only very little PA in YN01-429, which also contained much less extractable phenolics. The flavonol content, however, did not show as great a difference between these two lines. Furthermore, sinapine, a product of the general phenylpropanoid metabolism, was present at comparable levels in the embryos of both lines. Dihydroflavonol reductase (DFR) activity that commits phenolics to PA synthesis was lower in YN01-429 seed coats. The results of Southern blot and in silico analyses were indicative of two copies of the DFR gene in B. napus. Both copies were functional in YN01-429, ruling out homeoallelic repression or silencing, but together they showed very low expression levels (17-fold fewer transcripts) relative to DFR activity in N89-53 seed coats. These results collectively suggest that YN01-429 differs in regulatory circuits that impact the PA synthesis branch much more than the flavonol synthesis branch in the seed coats and such circuits do not impinge upon general phenylpropanoid metabolism in the embryos.Le jaune caracte\ub4ristique des grains du Brassica napus L. est de\ub4sirable, compte tenu de son association avec une haute teneur en huile et une meilleure qualite\ub4 de la farine extraite de l\u2019huile. Le YN01-429 constitue un germplasme a` graine jaune de qualite\ub4 canola re\ub4sultant de plusieurs anne\ub4es de recherches. La pigmentation des te\ub4guments est attribue\ub4e a` des proanthocyanidines oxyde\ub4es (PA; tannins condense\ub4s) de\ub4rive\ub4es de phe\ub4nylpropano\u131\ua8des et du CoA du malonyl. Les auteurs ont trouve\ub4 une plus forte accumulation de PA chez les jeunes te\ub4guments (20 jours apre`s l\u2019anthe`se; dpa) d\u2019une ligne\ub4e apparente\ub4e a` grains noirs N89-53 et tre`s peu de PA chez la ligne\ub4e YN01-429 qui contient a` la fois beaucoup moins de phe\ub4nols extractibles. Cependant, la teneur en flavonols ne montre pas autant de diffe\ub4rences entre ces deux ligne\ub4es. De plus, la sinapine, un produit du me\ub4tabolisme ge\ub4ne\ub4ral des phe\ub4nylpropano\u131\ua8des, se retrouve a` des teneurs comparables chez les embryons des deux ligne\ub4es. L\u2019activite\ub4 de la re\ub4ductase du dihydroflavonol (DFR) qui pousse les phe\ub4nols vers la synthe` se de PA, est plus faible dans les te\ub4guments du YN01-429. Les re\ub4sultats du transfert Southern et des analyses in silico indiquent la pre\ub4sence de deux copies du ge`ne DFR chez le B. napus. Les deux copies sont fonctionnelles chez le YN01- 429, ce qui e\ub4limine la re\ub4pression ou le silencing home\ub4oalle\ub4lique, mais ensemble elles ne montrent que de tre`s faibles degre \ub4s d\u2019expression (transcriptions 17 fois infe\ub4rieures) par rapport a` l\u2019activite\ub4 du DFR chez les te\ub4guments du N89-53. Ces re\ub4- sultats pris dans leur ensemble sugge`rent que le YN01-429 diffe`re a` l\u2019e\ub4chelle des circuits qui agissent sur la ligne de synthe`se des PA beaucoup plus que sur la ligne de synthe`se des flavonols dans les te\ub4guments, et de tels circuits n\u2019affectent pas le me\ub4tabolisme ge\ub4ne\ub4ral des phe\ub4nylpropano\u131\ua8des chez les embryons.Peer reviewed: YesNRC publication: Ye

botany

Proanthocyanidin biosynthesis in the seed coat of yellow-seeded, canola quality<i>Brassica napus</i>YN01-429 is constrained at the committed step catalyzed by dihydroflavonol 4-reductaseThis paper is one of a selection of papers published in a Special Issue from the National Research Council of Canada – Plant Biotechnology Institute.