The yellow seed trait is preferred by breeders for its potential to improve the seed quality and commercial value of Brassica napus.In the present study,we produced yellow seed mutants using a CRISPR/Cas9 system when ...The yellow seed trait is preferred by breeders for its potential to improve the seed quality and commercial value of Brassica napus.In the present study,we produced yellow seed mutants using a CRISPR/Cas9 system when the two BnPAP2 homologs were knocked out.Histochemical staining of the seed coat demonstrated that proanthocyanidin accumulation was significantly reduced in the pap2 double mutants and decreased specifically in the endothelial and palisade layer cells of the seed coat.Transcriptomic and metabolite profiling analysis suggested that disruption of the BnPAP2 genes could reduce the expression of structural and regulated genes in the phenylpropanoid and flavonoid biosynthetic pathways.The broad suppression of these genes might hinder proanthocyanidin accumulation during seed development,and thereby causing the yellow seed trait in B.napus.These results indicate that BnPAP2 might play a vital role in the regulatory network controlling proanthocyanidin accumulation.展开更多
Brassica napus is an important cash crop broadly grown for the vegetable and oil values.Yellow-seeded B.napus is preferred by breeders due to its improved oil and protein quality,less pigments and lignin compared with...Brassica napus is an important cash crop broadly grown for the vegetable and oil values.Yellow-seeded B.napus is preferred by breeders due to its improved oil and protein quality,less pigments and lignin compared with the blackseeded counterpart.This study compared the differences in flavonoid and fatty acid contents between yellow rapeseed from the progenies of B.napus-Sinapis alba somatic hybrids and the black-seeded counterpart using RNA-seq analysis.Through HPLC-PDA-ESI(-)/MSanalysis,it was found that phenylpropanoids and flavonoids(i.e.,isorhamnetin,epicatechin,kaempferol,and other derivatives)in yellow seed were significantly lower than those in black seed.The fatty acid(FA)content in yellow rapeseed was higher than that in black rapeseed due to the variation of C16:0,C18:0,C18:1,C18:2,and C18:3 contents.RNA-seq analysis of seeds at four and five weeks after flowering(WAF)indicated that differentially expressed genes(DEGs)between black and yellow rapeseeds were enriched in flavonoid and FA biosynthesis,including BnTT3,BnTT4,BnTT18,and BnFAD2.Also,genes related to FA biosynthesis,desaturation and elongation(FAD3,LEC1,FUS3,and LPAT2)in yellow seed were up-regulated compared to those in black seed,while genes involved in beta-oxidation cycle(AIM1 and KAT2)of yellow seed were down-regulated compared to those in black seed.The DEGs related to the variation of flavonoids,phenylpropanoids,and FAs would help improve the knowledge of yellow seed character in B.napus and promote rapeseed improvement.展开更多
14 yellow-seeded rapeseed lines (Brassia napus L.) from different genetic sources were used to analyze diversity of testa pigments content, oil and protein content, and RAPD markers. The results showed that the anthoc...14 yellow-seeded rapeseed lines (Brassia napus L.) from different genetic sources were used to analyze diversity of testa pigments content, oil and protein content, and RAPD markers. The results showed that the anthocyanin and melanin were the most important pigments in testa and their content were responsible for the variation in seed color ranging from orange to black yellow, 14 yellow-seeded lines could be classified into 3 groups: high anthocyanin content group with anthocyanin content over 2. 54 mg g-1 DW, the seed color was light yellow or orange; low pigments content group with low content of anthocyanin and melanin, the testa was transparent and the seed color was light yellow, greenish yellow or twany; high melanin content group with melanin content over 178. 4U(A290nm) , the testa was black, the seed color was black yellow. Oil content changed from 36.2% to 45. 5%, protein content from 21.1% to 27.7% , and the correlation analysis revealed that the oil content is highly significantly negatively correlated with the protein content. The cluster analysis showed that the extensive genetic variation existed among 14 yellow-seeded lines by using unweighted paired group method with arithmetic average (UPGMA) based on RAPD markers which were amplified with decamer primers, the genetic similarity among them ranged from 0. 25 to 0.909, and 14 yellow-seeded lines could put into 2 clusters corresponding to genome difference.展开更多
Yellow-seeded rapeseed (Brassica napus L.) is a new kind of breeding resources with yellow color, increased oil and protein content and less unwanted crude fiber content due to the thinner and transparent testa compar...Yellow-seeded rapeseed (Brassica napus L.) is a new kind of breeding resources with yellow color, increased oil and protein content and less unwanted crude fiber content due to the thinner and transparent testa compared with traditional black or brown-seeded rapeseed. To analyze the longevity of the yellow-seeded rapeseed during storage, the physiological differences between the yellow and black-seeded near-isogenic lines were studied by artificial ageing method. The testa rate, anthocyanin content and melanin content of yellow-seeded rapeseeds decreased by 20.1, 25.2, and 80.4% respectively than black-seeded rapeseed. During artificial ageing, the yellow-seeded rapeseed showed significantly different effect of ageing compared with the black-seeded, as demonstrated by faster deterioration with lower germination percentage, seed vigour index, reducing sugar and soluble protein contents than the black-seeded, as well as a drastic increase in electrical conductivity, malnodialdehyde (MDA) content and a rapid decrease in activities of antioxidant enzymes catalase (CAT) and superoxide dismutase (SOD). The results suggested that the transparent testa of the yellow-seeded rapeseed lost some abilities to protect the embryo against adverse environmental conditions and thus led to a poor storability.展开更多
基金supported by the National Natural Science Foundation of China(31971980,U19A2029)The science and technology innovation Program of Hunan Province,China(2023RC1077)+1 种基金the Agricultural Science and Technology Innovation Foundation of Hunan,China(2022CX55)the Scientific Research Fund of Hunan Provincial Science and Technology Department,China(2021JC0007)。
文摘The yellow seed trait is preferred by breeders for its potential to improve the seed quality and commercial value of Brassica napus.In the present study,we produced yellow seed mutants using a CRISPR/Cas9 system when the two BnPAP2 homologs were knocked out.Histochemical staining of the seed coat demonstrated that proanthocyanidin accumulation was significantly reduced in the pap2 double mutants and decreased specifically in the endothelial and palisade layer cells of the seed coat.Transcriptomic and metabolite profiling analysis suggested that disruption of the BnPAP2 genes could reduce the expression of structural and regulated genes in the phenylpropanoid and flavonoid biosynthetic pathways.The broad suppression of these genes might hinder proanthocyanidin accumulation during seed development,and thereby causing the yellow seed trait in B.napus.These results indicate that BnPAP2 might play a vital role in the regulatory network controlling proanthocyanidin accumulation.
基金funded by the National Natural Science Foundation of China(U20A2028 and 31972963)the Open Funds of the Key Laboratory of Plant Functional Genomics of the Ministry of Education,China(ML201804)+2 种基金the Project of Special Funding for Crop Science Discipline Development,China(yzuxk202006)the Priority Academic Program Development of Jiangsu Higher Education Institutions,Chinathe Yangzhou University for Excellent Talent Support Program,China。
文摘Brassica napus is an important cash crop broadly grown for the vegetable and oil values.Yellow-seeded B.napus is preferred by breeders due to its improved oil and protein quality,less pigments and lignin compared with the blackseeded counterpart.This study compared the differences in flavonoid and fatty acid contents between yellow rapeseed from the progenies of B.napus-Sinapis alba somatic hybrids and the black-seeded counterpart using RNA-seq analysis.Through HPLC-PDA-ESI(-)/MSanalysis,it was found that phenylpropanoids and flavonoids(i.e.,isorhamnetin,epicatechin,kaempferol,and other derivatives)in yellow seed were significantly lower than those in black seed.The fatty acid(FA)content in yellow rapeseed was higher than that in black rapeseed due to the variation of C16:0,C18:0,C18:1,C18:2,and C18:3 contents.RNA-seq analysis of seeds at four and five weeks after flowering(WAF)indicated that differentially expressed genes(DEGs)between black and yellow rapeseeds were enriched in flavonoid and FA biosynthesis,including BnTT3,BnTT4,BnTT18,and BnFAD2.Also,genes related to FA biosynthesis,desaturation and elongation(FAD3,LEC1,FUS3,and LPAT2)in yellow seed were up-regulated compared to those in black seed,while genes involved in beta-oxidation cycle(AIM1 and KAT2)of yellow seed were down-regulated compared to those in black seed.The DEGs related to the variation of flavonoids,phenylpropanoids,and FAs would help improve the knowledge of yellow seed character in B.napus and promote rapeseed improvement.
文摘14 yellow-seeded rapeseed lines (Brassia napus L.) from different genetic sources were used to analyze diversity of testa pigments content, oil and protein content, and RAPD markers. The results showed that the anthocyanin and melanin were the most important pigments in testa and their content were responsible for the variation in seed color ranging from orange to black yellow, 14 yellow-seeded lines could be classified into 3 groups: high anthocyanin content group with anthocyanin content over 2. 54 mg g-1 DW, the seed color was light yellow or orange; low pigments content group with low content of anthocyanin and melanin, the testa was transparent and the seed color was light yellow, greenish yellow or twany; high melanin content group with melanin content over 178. 4U(A290nm) , the testa was black, the seed color was black yellow. Oil content changed from 36.2% to 45. 5%, protein content from 21.1% to 27.7% , and the correlation analysis revealed that the oil content is highly significantly negatively correlated with the protein content. The cluster analysis showed that the extensive genetic variation existed among 14 yellow-seeded lines by using unweighted paired group method with arithmetic average (UPGMA) based on RAPD markers which were amplified with decamer primers, the genetic similarity among them ranged from 0. 25 to 0.909, and 14 yellow-seeded lines could put into 2 clusters corresponding to genome difference.
基金The authors thank 863 Project of the Ministry of Sci-ence and Technology of P.R.China(AA2003207150)948 Project of the Ministry of Agricutureof P.R.China(2003-Q04)for financial support.
文摘Yellow-seeded rapeseed (Brassica napus L.) is a new kind of breeding resources with yellow color, increased oil and protein content and less unwanted crude fiber content due to the thinner and transparent testa compared with traditional black or brown-seeded rapeseed. To analyze the longevity of the yellow-seeded rapeseed during storage, the physiological differences between the yellow and black-seeded near-isogenic lines were studied by artificial ageing method. The testa rate, anthocyanin content and melanin content of yellow-seeded rapeseeds decreased by 20.1, 25.2, and 80.4% respectively than black-seeded rapeseed. During artificial ageing, the yellow-seeded rapeseed showed significantly different effect of ageing compared with the black-seeded, as demonstrated by faster deterioration with lower germination percentage, seed vigour index, reducing sugar and soluble protein contents than the black-seeded, as well as a drastic increase in electrical conductivity, malnodialdehyde (MDA) content and a rapid decrease in activities of antioxidant enzymes catalase (CAT) and superoxide dismutase (SOD). The results suggested that the transparent testa of the yellow-seeded rapeseed lost some abilities to protect the embryo against adverse environmental conditions and thus led to a poor storability.
