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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.

Yellow seed breeding is an effective method to improve oil yield and quality in rapeseed (Brassica napus L.). However, naturally occurring yellow-seeded genotypes have not been identified in B. napus. Mustard (Brassica juncea L.) has some natural, yellow-seeded germplasms, yet the molecular mechanism underlying this trait remains unclear. In this study, a BC9 population derived from the cross of yellow seed mustard "Wuqi" and brown seed mustard "Wugong" was used to analyze the candidate genes controlling the yellow seed color of B. juncea. Subsequently, yellow-seeded (BY) and brown-seeded (BB) bulks were constructed in the BC9 population and subjected to bulked segregant RNA sequencing (BSR-Seq). A total of 511 differentially expressed genes (DEGs) were identified between the brown and yellow seed bulks. Enrichment analysis revealed that these DEGs were involved in the phenylpropanoid biosynthetic process and flavonoid biosynthetic process, including key genes such as 4CL, C4H, LDOX/TT18, PAL1, PAL2, PAL4, TT10, TT12, TT4, TT8, BAN, DFR/TT3, F3H/TT6, TT19, and CHI/TT5. In addition, 111,540 credible single-nucleotide polymorphisms (SNPs) and 86,319 INDELs were obtained and used for quantitative trait locus (QTL) identification. Subsequently, two significant QTLs on chromosome A09, namely, qSCA09-3 and qSCA09-7, were identified by G' analysis, and five DEGs (BjuA09PAL2, BjuA09TT5, BjuA09TT6, BjuA09TT4, BjuA09TT3) involved in the flavonoid pathway were identified as hub genes based on the protein-to-protein network. Among these five genes, only BjuA09PAL2 and BjuA09F3H had SNPs between BY and BB bulks. Interestingly, the majority of SNPs in BjuA09PAL2 were consistent with the SNPs identified between the high-quality assembled B. juncea reference genome "T84-66" (brown-seed) and "AU213" (yellow-seed). Therefore, BjuA09PAL2, which encodes phenylalanine lyase, was considered as the candidate gene associated with yellow seed color of B. juncea. The identification of a novel gene associated with the yellow seed coloration of B. juncea through this study may play a significant role in enhancing yellow seed breeding in rapeseed. [ABSTRACT FROM AUTHOR]

Yellow seeds are desirable in rapeseed breeding because of their higher oil content and better nutritional quality than black seeds. However, the underlying genes and formation mechanism of yellow seeds remain unclear. Here, a novel yellow-seeded rapeseed line (Huangaizao, HAZ) was crossed with a black-seeded rapeseed line (Zhongshuang11, ZS11) to construct a mapping population of 196 F2 individuals, based on which, a high-density genetic linkage map was constructed. This map, comprising 4174 bin markers, was 1618.33 cM in length and had an average distance of 0.39 cM between its adjacent markers. To assess the seed color of the F2 population, three methods (imaging, spectrophotometry, and visual scoring) were used and a common major quantitative trait locus (QTL) on chromosome A09, explaining 10.91–21.83% of the phenotypic variance, was detected. Another minor QTL, accounting for 6.19–6.69% of the phenotypic variance, was detected on chromosome C03, only by means of imaging and spectrophotometry. Furthermore, a dynamic analysis of the differential expressions between the parental lines showed that flavonoid biosynthesis-related genes were down-regulated in the yellow seed coats at 25 and 35 days after flowering. A coexpression network between the differentially expressed genes identified 17 candidate genes for the QTL intervals, including a flavonoid structure gene, novel4557 (BnaC03.TT4), and two transcription factor genes, namely, BnaA09G0616800ZS (BnaA09.NFYA8) and BnaC03G0060200ZS (BnaC03.NAC083), that may regulate flavonoid biosynthesis. Our study lays a foundation for further identifying the genes responsible for and understanding the regulatory mechanism of yellow seed formation in Brassica napus. [ABSTRACT FROM AUTHOR]

Yellow seed breeding is an effective method to improve oil yield and quality in rapeseed (Brassica napus L.). However, naturally occurring yellow-seeded genotypes have not been identified in B. napus. Mustard (Brassica juncea L.) has some natural, yellow-seeded germplasms, yet the molecular mechanism underlying this trait remains unclear. In this study, a BC9 population derived from the cross of yellow seed mustard “Wuqi” and brown seed mustard “Wugong” was used to analyze the candidate genes controlling the yellow seed color of B. juncea. Subsequently, yellow-seeded (BY) and brown-seeded (BB) bulks were constructed in the BC9 population and subjected to bulked segregant RNA sequencing (BSR-Seq). A total of 511 differentially expressed genes (DEGs) were identified between the brown and yellow seed bulks. Enrichment analysis revealed that these DEGs were involved in the phenylpropanoid biosynthetic process and flavonoid biosynthetic process, including key genes such as 4CL, C4H, LDOX/TT18, PAL1, PAL2, PAL4, TT10, TT12, TT4, TT8, BAN, DFR/TT3, F3H/TT6, TT19, and CHI/TT5. In addition, 111,540 credible single-nucleotide polymorphisms (SNPs) and 86,319 INDELs were obtained and used for quantitative trait locus (QTL) identification. Subsequently, two significant QTLs on chromosome A09, namely, qSCA09-3 and qSCA09-7, were identified by G’ analysis, and five DEGs (BjuA09PAL2, BjuA09TT5, BjuA09TT6, BjuA09TT4, BjuA09TT3) involved in the flavonoid pathway were identified as hub genes based on the protein-to-protein network. Among these five genes, only BjuA09PAL2 and BjuA09F3H had SNPs between BY and BB bulks. Interestingly, the majority of SNPs in BjuA09PAL2 were consistent with the SNPs identified between the high-quality assembled B. juncea reference genome “T84-66” (brown-seed) and “AU213” (yellow-seed). Therefore, BjuA09PAL2, which encodes phenylalanine lyase, was considered as the candidate gene associated with yellow seed color of B. juncea. The identification of a novel gene associated with the yellow seed coloration of B. juncea through this study may play a significant role in enhancing yellow seed breeding in rapeseed.

Abstract Objective The brown seed coat colour of flax (Linum ustiatissimum) results from proanthocyanidin synthesis and accumulation. Glutathione S-transferases (GSTs), such as the TT19 protein in Arabidopsis, have been implicated in the transport of anthocyanidins during the synthesis of the brown proanthocyanidins. This study fine mapped the g allele responsible for yellow seed colour in S95407 and identified it as a putative mutated GST. Results We developed a Recombinant Inbred Line population with 320 lines descended from a cross between CDC Bethune (brown seed coat) and S95407 (yellow seed) and used molecular markers to fine map the G gene on Chromosome 6 (Chr 6). We used Next Generation Sequencing (NGS) to identify a putative GST was identified in this region and Sanger sequenced the gene from CDC Bethune, S95407 and other yellow seeded genotypes. The putative GST from S95407 had 13 SNPs encoding, including four non-synonymous amino acid changes, compared to the CDC Bethune reference sequence and the other genotypes. The GST encoded by Lus10019895 is a lambda-GST in contrast to the Arabidopsis TT19 which is a phi-GST.

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Yellow seeds are desirable in rapeseed breeding because of their higher oil content and better nutritional quality than black seeds. However, the underlying genes and formation mechanism of yellow seeds remain unclear. Here, a novel yellow-seeded rapeseed line (Huangaizao, HAZ) was crossed with a black-seeded rapeseed line (Zhongshuang11, ZS11) to construct a mapping population of 196 F2 individuals, based on which, a high-density genetic linkage map was constructed. This map, comprising 4174 bin markers, was 1618.33 cM in length and had an average distance of 0.39 cM between its adjacent markers. To assess the seed color of the F2 population, three methods (imaging, spectrophotometry, and visual scoring) were used and a common major quantitative trait locus (QTL) on chromosome A09, explaining 10.91–21.83% of the phenotypic variance, was detected. Another minor QTL, accounting for 6.19–6.69% of the phenotypic variance, was detected on chromosome C03, only by means of imaging and spectrophotometry. Furthermore, a dynamic analysis of the differential expressions between the parental lines showed that flavonoid biosynthesis-related genes were down-regulated in the yellow seed coats at 25 and 35 days after flowering. A coexpression network between the differentially expressed genes identified 17 candidate genes for the QTL intervals, including a flavonoid structure gene, novel4557 (BnaC03.TT4), and two transcription factor genes, namely, BnaA09G0616800ZS (BnaA09.NFYA8) and BnaC03G0060200ZS (BnaC03.NAC083), that may regulate flavonoid biosynthesis. Our study lays a foundation for further identifying the genes responsible for and understanding the regulatory mechanism of yellow seed formation in Brassica napus.

Objective: The brown seed coat colour of flax (Linum ustiatissimum) results from proanthocyanidin synthesis and accumulation. Glutathione S-transferases (GSTs), such as the TT19 protein in Arabidopsis, have been implicated in the transport of anthocyanidins during the synthesis of the brown proanthocyanidins. This study fine mapped the g allele responsible for yellow seed colour in S95407 and identified it as a putative mutated GST. Results: We developed a Recombinant Inbred Line population with 320 lines descended from a cross between CDC Bethune (brown seed coat) and S95407 (yellow seed) and used molecular markers to fine map the G gene on Chromosome 6 (Chr 6). We used Next Generation Sequencing (NGS) to identify a putative GST was identified in this region and Sanger sequenced the gene from CDC Bethune, S95407 and other yellow seeded genotypes. The putative GST from S95407 had 13 SNPs encoding, including four non-synonymous amino acid changes, compared to the CDC Bethune reference sequence and the other genotypes. The GST encoded by Lus10019895 is a lambda-GST in contrast to the Arabidopsis TT19 which is a phi-GST. [ABSTRACT FROM AUTHOR]

Introduction. Rapeseed is a facultative autogamous plant; its percentage of cross-pollination is 10–50%, so rapeseed breeding engages the line and population breeding methods as well as the development of hybrids and all innovative research approaches.Material and methods. Among the main techniques used for the development of parent material and, eventually, cultivars at the Pustovoit Institute (VNIIMK) is the pedigree method (individual selection from intraspecific and interspecific hybrid populations, combined with inbreeding). Physical and chemical mutagenesis is also employed in rapeseed breeding at VNIIMK. The material with yellow-colored seed coat has been obtained by exposing rapeseed seeds to various doses of gamma rays (50, 100 and 150 kR). Seed treatment with chemical mutagens is primarily aimed at changing the fatty acid composition of oil.Results. Breeding achievements for winter and spring rapeseed, registered by VNIIMK in the State Register of Breeding Achievements of the Russian Federation over the past 15 years, are highly productive line cultivars with a seed yield of 4–5 t/ha for winter rapeseed, and 2–3 t/ha for spring rapeseed. Promising experimental interline hybrids, significantly exceeding the reference in seed yield (by 1.61–2.26 t/ha), have been developed. Yellowseeded lines, which exceed the black-seeded reference cv. ‘Tavrion’ in seed yield by 0.08–0.15 t/ha, have been selected. We developed the material with an oleic acid level of 78.9–80.5%.

Yellow seed is one desirable trait with great potential to improve seed oil quality and yield. The present study surveys the redundant role of BnTTG1 genes in the proanthocyanidins (PA) biosynthesis, oil content and abiotic stress resistance. Stable yellow seed mutants were generated after mutating BnTTG1 by CRISPR/Cas9 genome editing system. Yellow seed phenotype could be obtained only when both functional homologues of BnTTG1 were simultaneously knocked out. Homozygous mutants of BnTTG1 homologues showed decreased thickness and PA accumulation in seed coat. Transcriptome and qRT-PCR analysis indicated that BnTTG1 mutation inhibited the expression of genes involved in phenylpropanoid and flavonoid biosynthetic pathways. Increased seed oil content and alteration of fatty acid (FA) composition were observed in homozygous mutants of BnTTG1 with enriched expression of genes involved in FA biosynthesis pathway. In addition, target mutation of BnTTG1 accelerated seed germination rate under salt and cold stresses. Enhanced seed germination capacity in BnTTG1 mutants was correlated with the change of expression level of ABA responsive genes. Overall, this study elucidated the redundant role of BnTTG1 in regulating seed coat color and established an efficient approach for generating yellow-seeded oilseed rape genetic resources with increase oil content, modified FA composition and resistance to multiple abiotic stresses. • Yellow seed mutants were generated after mutation of two BnTTG1 copies by CRISPR/Cas9. • BnTTG1 mutants Increased seed oil content and modified FA composition. • BnTTG1 accelerated seed germination rate under salt and cold stresses. • BnTTG1 mutants increase abiotic stresses was correlated with the ABA signaling. [ABSTRACT FROM AUTHOR]

Yellow seed coat color is a desirable characteristic in rapeseed (Brassica napus), as it is associated with higher oil content and higher quality of meal. Alternative splicing (AS) is a vital post-transcriptional regulatory process contributing to plant cell differentiation and organ development. To identify novel transcripts and differences at the isoform level that are associated with seed color in B. napus, we compared 31 RNA-seq libraries of yellow- and black-seeded B. napus at five different developmental stages. AS events in the different samples were highly similar, and intron retention accounted for a large proportion of the observed AS pattern. AS mainly occurred in the early and middle stage of seed development. Weighted gene co-expression network analysis (WGCNA) identified 23 co-expression modules composed of differentially spliced genes, and we picked out two of the modules whose functions were highly associated with seed color. In the two modules, we found candidate DAS (differentially alternative splicing) genes related to the flavonoid pathway, such as TT8 (BnaC09g24870D), TT5 (BnaA09g34840D and BnaC08g26020D), TT12 (BnaC06g17050D and BnaA07g18120D), AHA10 (BnaA08g23220D and BnaC08g17280D), CHI (BnaC09g50050D), BAN (BnaA03g60670D) and DFR (BnaC09g17150D). Gene BnaC03g23650D, encoding RNA-binding family protein, was also identified. The splicing of the candidate genes identified in this study might be used to develop stable, yellow-seeded B. napus. This study provides insight into the formation of seed coat color in B. napus.

Soybean [Glycine max (L.) Merr.] is an important source for protein, oil, carbohydrates, isoflavones, and many other nutrients to humans and animals. Lipoxygenase protein is the main antinutritional factor in mature soybean seed. Soybean cultivar, "Choseon" with yellow medium seed size and free of lipoxygenase 2,3 protein was developed. "Choseon" was selected from the population derived from the cross between cultivar, "Jinpum 2" and germplasm, PI408155. "Choseon" has purple flowers, tawny pubescence, a determinate growth habit, and light yellow pods at maturity. The seed of "Choseon" has yellow hilum and yellow seed coat color. Seed protein and oil content on a dry weight basis were 34.2 and 16.9%, respectively. "Choseon" has shown a resistant reaction to soybean necrosis, soybean mosaic virus, Cercospora leaf spot and blight, black root rot, pod and stem blight, and bacterial pustule. "Choseon" matured in 5 October with plant height of 51 cm and a 100-seed weight of 15.1 g. Average yield of "Choseon" was 2.66 (Ton/ha) on the regional yield trials of 2013 at 4 locations. "Choseon" has been registered as a soybean cultivar (registration number: 5985, registration date: April 11, 2016) by Korea Seed & Variety Service, Republic of Korea. [ABSTRACT FROM AUTHOR]

Brassica napus L. is rich in phenolic components and it has natural antioxidant characteristics which are important to human health. In the present study, the total phenolic and flavonoid contents of developing seeds of yellow- and black-seeded B. napus were compared. Both phenolic and flavonoid contents were significantly higher at 5 weeks after flowering (WAF) in black seeds (6.44 ± 0.97 mg EE/g phenolics and 3.78 ± 0.05 mg EE/g flavonoids) than yellow seeds (2.80 ± 0.13 mg/g phenolics and 0.83 ± 0.01 mg/g flavonoids). HPLC–DAD–ESI/MS analysis revealed different content of 56 phenolic components between yellow and black-seeded B. napus, including kaempferol-3-O-glucoside, isorhamnetin-3-O-glucoside, quercetin-3-O-sophoroside, procyanidin B2 ([DP 2]), which were significantly reduced in yellow seeds compared with black seeds. Applying the 2,2-diphenyl-1-picrylhydrazyl (DPPH) and 2,2′-azinobis-(3-ethylbenzothiazoline-6-sulfonic acid) (ABTS) radical assay, we found maximum clearance of DPPH and ABTS in the late developmental stages of yellow and black seeds. Additionally, the ferric reducing antioxidant power (FRAP) value maximized at 5 WAF in black seeds (432.52 ± 69.98 μmol Fe (II)/g DW) and 6 WAF in yellow seeds (274.08 ± 2.40 μmol Fe (II)/g DW). Generally, antioxidant ability was significantly reduced in yellow-seeded B. napus compared to black rapeseed, and positive correlations between antioxidation and flavonoid content were found in both yellow- and black-seeded B. napus.

DNA methylation is a crucial modification process implicated in the epigenetic regulation of gene expression, which may be responsible for agronomic trait variation. However, the epigenetic regulation of the yellow- and black-seeded characters of Brassica napus has yet to be reported. In this study, methylation-sensitive amplification polymorphism (MSAP) analysis was performed to investigate the target regions of seed coat variation in B. napus. The yellow-seeded rapeseed line derived from the somatic hybrids of B. napus- Sinapis alba and the black-seeded rapeseed line were used as experimental materials. Extensive methylation changes were observed between yellow- and black-seeded B. napus. In particular, 10 % of demethylation and 5 % of hypermethylation were detected in yellow rapeseeds compared with black rapeseeds. Nonetheless, this variation was barely identified in different developmental stages. The relative expression levels of polymorphic fragments in MSAP profiles were also analyzed. The gene expression of the demethylated fragments in the yellow rapeseeds was upregulated. For instance, bHLH, a transcription factor regulating flavonoid biosynthesis, was upregulated at 3-5 weeks after flowering of yellow seeds. Therefore, epigenetic changes among rapeseed lines with different seed colors help elucidate the formation of the yellow seed character. [ABSTRACT FROM AUTHOR]

Abstracts To determine the effect of genotype and the environment on tocopherol (T) and plastochromanol-8 (PC-8) contents in winter oilseed rape ( Brassica napus L.), a three-year study was performed. The plant material consisted of 25 doubled haploid (DH) lines derived from the F 1 hybrid obtained from the cross between two DH lines (Z × H), (Z-114- yellow seeds and H 2 -26- black seeds). Quantitative contents of α-T, β-T, γ-T, δ-T, and PC-8 were determined by HPLC. Total tocopherol content ranged between 40.62 and 51.20 mg/100 g and PC-8 5.43 – 8.92 mg/100 g. The α-T/γ-T ratio ranged from 0.77 to 1.23. The heritability in the broad sense was examined for the following components: PC-8, α-T, β-T, γ-T, δ-T, α/γ-T, and γ-T contents in oil. Heritability levels were 0.60, 0.58, 0.53, 0.50, and 0.43, respectively. This suggest the superiority of the effect of genotype on tocopherol contents. The h 2 values for δ-T and β-T were 0.36 and 0.32, which demonstrates a great influence of environmental conditions. Among all analyzed genotypes, eleven were selected as satisfactory and stable regarding tocopherol or PC-8 contents. [ABSTRACT FROM AUTHOR]

Soybean is the main source of protein and oil for human and animal nutrition. However, antinutritional factors in the raw mature soybean are exist. Lipoxygenase protein is the main antinutritional factor in mature soybean seed. A new soybean cultivar, ‘Jinyang' with yellow seed coat and free of lipoxygenase 1,2,3 protein was developed. It was selected from the population derived from the cross between ‘Jinpumkong2ho’ and GS301. ‘Jinyang' has purple flowers, tawny pubescence, a determinate growth habit, oval leaflet shape, and light brown pods at maturity. The seed has yellow hilum and yellow seed coat color. Seed protein and oil content on a dry weight basis were 33.5% and 16.6%, respectively. It has shown a resistant reaction to soybean necrosis, soybean mosaic virus, cercospora leaf spot and blight, black root rot, pod and stem blight, and soybean pod borer. ‘Jinyang' matured in 1 October with plant height of 65cm and a 100-seed weight of 21 g. Average yield of ‘Jinyang' was 300 - 330 kg/10a in 2010 - 2012. ‘Jinyang' has been registered as a new soybean cultivar (registration number: 4279, registration date: Dec. 28, 2012) by Korea Seed & Variety Service, Republic of Korea. [ABSTRACT FROM AUTHOR]

Rape Dahuang is a special landrace of Brassica rapa L. which originated from Qinghai - Tibetan Plateau with bright yellow seeds. Previous results showed the yellow - seeded trait in Dahuang was controlled by a recessive gene (Brsc1), which was mapped on A09 in B. rapa. For more specific markers linked to Brsc1, BC1 population and F2 population were constructed from the cross of Dahuang and 09 A - 126 ( brown - seeded, B. rapa) . Amplified fragment length polymorphism (AFLP) technology combined with bulked segregant analysis (BSA) was conducted by screening 256 pairs of AFLP primers in BC1 population, 5 markers tightly linked to Brsc1 were obtained. 5 specific AFLP fragments were cloned, sequenced and identified, and compared to that of synteny in B. rapa genome, showing homology to A09 of B. rapa. The AFLP markers were converted into 5 SCAR (sequence characterized amplified region) markers for F2 population. One co - dominant marker was detected. [ABSTRACT FROM AUTHOR]

Breeders have focused on yellow-seeded Brassica napus (rapeseed) for its better quality compared with the black-seeded variety. Moreover, flavonoids have been associated with this kind of rapeseed. In this study, we applied lipid chromatography-electrospray ionization mass spectrometry (LC-ESI-MSn) to compare flavonoids in developing seeds of natural black-seeded B. napus and yellow-seeded introgression lines selected from progenies of B. napus-Sinapis alba somatic hybrids. Aside from the most abundant phenolic compounds (sinapine and sinapic acid) and 1, 2-disinapoylglucose, 16 different flavonoids were identified and quantified, including (-)-epicatechin, five monocharged oligomers of (-)-epicatechin ([DP 2]−, [DP 3]−, [DP 4] [DP 2]−B2 and [DP 2]−B5), quercetin, kaempferol, isorhamnetin-dihexoside, kaempferol-sinapoyl-trihexoside, isorhamnetin-sinapoyl-trihexoside, isorhamnetin-hexoside-sulfate, and isorhamnetin-3-O-glucoside. Most of the flavonoids accumulated with seed development, whereas some rapidly decreased during maturation. The content of these flavonoids were lower in the yellow-seeded materials than in the black seeds. In addition, variations of insoluble procyanidin oligomers and soluble phenolic acids were observed among both rapeseed varieties. Transcriptome changes of genes participating in the flavonoid pathway were discovered by quantitative reverse transcription polymerase chain reaction analysis. Consistent with flavonoid changes identified by high performance liquid chromatography analysis, the expression of most genes in the flavonoid biosynthetic pathway was also downregulated. [ABSTRACT FROM AUTHOR]

BACKGROUND Yellow-seeded Brassica napus is preferred by breeders for economic reasons, and seed coat is the most obvious comparable character between yellow and black rapeseed. RESULTS Owing to an incomplete understanding of chemical components in yellow rapeseed coat, here we optimized a technique based on attenuated total reflectance-Fourier transform infrared ( FT-IR) microspectroscopy to screen differences in chemical compositions of Brassica napus, Sinapis alba and four progeny lines with different yellow seed color derived from somatic hybrids between B. napus and S. alba. Nuclear magnetic resonance ( NMR) analysis of seed coat from B. napus and S. alba represented two specific peaks representing components of lignin in B. napus, which were absent in the progenies. Also, the intensity of peaks related to fiber were stronger in B. napus than S. alba and yellow seed lines, indicating the differences of fiber and lignin contents in yellow-seeded lines and parent lines. Differences in the lipid, protein and carbohydrate features were identified between B. napus and S. alba, some of which were specifically inherited in progenies of the somatic hybrids. CONCLUSION FT-IR spectrometry and NMR provide a rapid, direct bioanalytical technique for exploring the changes in molecular chemistry within the canola seed coat and for selecting rapeseed materials with high quality, which can be further used for breeding or nutrition of human and animals. © 2012 Society of Chemical Industry [ABSTRACT FROM AUTHOR]

Polymerase chain reaction and genomic in situ hybridization techniques were conducted to demonstrate the genomic introgressions in somatic hybrid progenies between Brassica napus and Sinapis alba. With minisatellite core sequence 33.6 as primer, an S. alba-specific band (288 bp) was amplified in yellow seed progenies. No hybridization signals were found using the genomic DNA of S. alba as probe. In addition, degenerate primers were used for the detection of related genes based on the genes related to flavonoid biosynthesis of the model plant Arabidopsis thaliana. Sequencing results show that flavonoid pathway genes are highly conserved in A. thaliana, S. alba and B. napus, and present subtle differences because of genetic evolution. A specific band (1,672 bp) consistent with S. alba was characterized in the yellow seed lines. This study demonstrates that the new yellow seed germplasms, derived from backcrossed and self-crossed progenies of B. napus- S. alba hybrids, are stable and homozygous introgression lines with yellow seed color, and differ from existing yellow seed materials. [ABSTRACT FROM AUTHOR]

Agronomic traits, yield and quality of yellow seed lines, obtained from somatic hybrids between Sinapis alba and Brassica napus, were analyzed. Among them two lines(D244 - 18 and D244 - 6) had higher application potential. The genetic model in D244 - 18 line showed that 2 pairs of independent genes controlled the yellow seed trait. The yellow seed phenotype was presented only when both pairs of genes were recessive. Its different genetic pattern made it a new type of yellow seed rape. [ABSTRACT FROM AUTHOR]

A number of variant phenotypes, such as long siliques, high density of pods, increased seed number per pod and yellow seed color resembling Sinapis alba, were selected from intergeneric somatic hybrids between Brassica napus and Sinapis alba through successive backcrosses. Resistance to Sclerotinia sclerotiorum among 24 BCF4 lines was also tested by inoculation with mycelia, and four lines showed stronger resistance than the resistant rapeseed variety ‘Zhongshuang 9’. Comparative anatomy studies on mature seed revealed that the seed coat pigments were mainly distributed in the palisade layer, which is considerably thinner in S. alba compared to B. napus. The area index of protein bodies in cotyledon cells was highest in S. alba, lowest in B. napus and intermediate in the intergeneric progenies. This study demonstrates that wide hybridization can enable exploitation of valuable trait diversity from S. alba for broadening the genetic basis for rapeseed breeding. [ABSTRACT FROM AUTHOR]

Two rapeseed (Brassica napus L.) phenylalanine ammonia-lyase 2 genes (BnPAL2-1 and BnPAL2-2) were isolated. The full-length cDNAs of BnPAL2-1 and BnPAL2-2 are 2422 bp and 2421 bp, with corresponding genomic sequences of 2735 bp and 2742 bp, respectively. Clues from NCBI BLAST, multi-alignment, the phylogenetic tree, conserved domains, preferential amino acids, and tertiary structures all indicated that they are orthologs of Arabidopsis thaliana PAL2 (AtPAL2). The purified His-tagged BnPAL2-1 and BnPAL2-2 proteins, expressed in Escherichia coli, were both active for PAL catabolism, but BnPAL2-2 was much more active than BnPAL2-1. Southern blot analysis showed that the BnPAL2 subfamily might contain 2-6 members. Transcripts of both BnPAL2-1 and BnPAL2-2 were detectable in all tested organs, but BnPAL2-2 was more organ-specific than BnPAL2-1. Transcription of BnPAL2-1 was most abundant in buds, with sequentially decreased abundance in stems, roots, flowers, seeds of all stages, silique pericarps and leaves, and was weak in hypocotyls and cotyledons. Expression of BnPAL2-2 was most intensive in roots, declining sequentially in buds, stems, flowers, and 30D seeds, and was weak in all other organs. Transcription of both genes in yellow-seeded rapeseed line L2's most reproductive organs, including developing seeds, was lower than that of black-seeded line L1. These results, combined with our previous investigations on BnPAL1 genes, demonstrate that most Brassicaceae species contain orthologs of all 4 AtPAL genes. Both PAL1 and PAL2 were most probably triplicated in Brassiceae ancestor and simultaneous down-regulation of all/most PAL paralogs contributes to the formation of the yellow seed trait in L2. [ABSTRACT FROM AUTHOR]

Two yellow-seeded white-petalled Brassica napus F7 inbred lines, developed from interspecific crosses, containing 26–28% erucic acid and more than 40 μmol glucosinolates (GLS)/g seed were crossed with two black/dark brown seeded B. napus varieties of double low quality and 287 doubled haploid (DH) lines were produced. The segregation in the DH lines indicated that three to four gene loci are involved in the determination of seed colour, and yellow seeds are formed when all alleles in all loci are in the homozygous recessive state. A dominant gene governed white petal colour and is linked with an erucic acid allele that, in the homozygous condition, produces 26–28% erucic acid. Four gene loci are involved in the control of total GLS content where low GLS was due to the presence of recessive alleles in the homozygous condition in all loci. From the DH breeding population a yellow-seeded, yellow-petalled, zero erucic acid line was obtained. This line was further crossed with conventional B. napus varieties of double low quality and, following pedigree selection, a yellow seeded B. napus of double low quality was obtained. The yellow seeds had higher oil plus protein content and lower fibre content than black seeds. A reduction of the concentration of chromogenic substances was found in the transparent seed coat of the yellow-seeded B. napus . [ABSTRACT FROM AUTHOR]

Yellow-seeded Brassica napus was developed from interspecific crosses between yellow-seeded Brassica rapa var. `yellow sarson' (AA), black-seeded Brassica alboglabra (CC), yellow-seeded Brassica carinata (BBCC) and black-seeded B. napus (AACC). Three different interspecific crossing approaches were undertaken. Approaches 1 and 2 were designed directly to develop yellow-seeded B. napus while approach 3 was designed to produce a yellow-seeded CC genome species. Approaches 1 and 2 differed in the steps taken after trigenomic interspecific hybrids (ABC) were generated from B. carinata × B. rapa crosses. The aim of approach 1 was to transfer the yellow seed colour genes from the A to the C genome as an intermediate step in developing yellow-seeded B. napus . For this purpose, the ABC hybrids were crossed with black-seeded B. napus and the three-way interspecific hybrids were self-pollinated for a number of generations. The F7 generation resulted in the yellowish-brown-seeded B. napus line, No. 06. Crossing this line with the B. napus line No. 01, resynthesized from a black-seeded B. alboglabra × B. rapa var. `yellow sarson' cross (containing the yellow seed colour genes in its AA genome), yielded yellow-seeded B. napus . This result indicated that the yellow seed colour genes were transferred from the A to the C genome in the yellowish-brown seed colour line No. 06. In approach 2, trigenomic diploids (AABBCC) were generated from the above-mentioned trigenomic haploids (ABC). The seed colour of the trigenomic diploid was brown, in contrast to the yellow seed colour of the parental species. Trigenomic diploids were crossed with the resynthesized B. napus line No. 01 to eliminate the B genome chromosomes, and to develop yellow-seeded B. napus with the AA genome of `yellow sarson' and the CC genome of B. carinata with yellow seed colour genes. This interspecific cross failed to generate any yellow-seeded B. napus . Approach 3 was to develop... [ABSTRACT FROM AUTHOR]

Yellow seeded Brassica napus was developed through interspecific crosses with the two mustard species, B. juncea and B. carinata. The objective of these two interspecific crosses was the introgression of genes for yellow seed colour from the A genome of B. juncea and C genome of B. carinata into the A and C genomes of B. napus, respectively. The inter- specific F[Sub1] generations were backcrossed to B. napus in an attempt to eliminate B genome chromosomes and to improve fertility. Backcross F[Sub2] plants of the (B. napus × B. juncea) × B. napus cross were then crossed with backcross F[Sub2] plants of the (B. napus × B. carinata) × B. napus cross. The objective of this intercrossing was to combine the A and C genome yellow seeded characteristics of the two backcross populations into one genotype. The F[Sub2] generation of the backcross F2 intercrosses was grown in the field, plants were individually harvested and visually rated for seed colour. Ninety-one yellow seeded plants were identified among the 4858 plants inspected. This result indicated that the interspecific crossing scheme was successful in developing yellow seeded B. napus. [ABSTRACT FROM AUTHOR]

Yellow seed coat color is a desirable characteristic in rapeseed (Brassica napus), as it is associated with higher oil content and higher quality of meal. Alternative splicing (AS) is a vital post-transcriptional regulatory process contributing to plant cell differentiation and organ development. To identify novel transcripts and differences at the isoform level that are associated with seed color in B. napus, we compared 31 RNA-seq libraries of yellow- and black-seeded B. napus at five different developmental stages. AS events in the different samples were highly similar, and intron retention accounted for a large proportion of the observed AS pattern. AS mainly occurred in the early and middle stage of seed development. Weighted gene co-expression network analysis (WGCNA) identified 23 co-expression modules composed of differentially spliced genes, and we picked out two of the modules whose functions were highly associated with seed color. In the two modules, we found candidate DAS (differentially alternative splicing) genes related to the flavonoid pathway, such as TT8 (BnaC09g24870D), TT5 (BnaA09g34840D and BnaC08g26020D), TT12 (BnaC06g17050D and BnaA07g18120D), AHA10 (BnaA08g23220D and BnaC08g17280D), CHI (BnaC09g50050D), BAN (BnaA03g60670D) and DFR (BnaC09g17150D). Gene BnaC03g23650D, encoding RNA-binding family protein, was also identified. The splicing of the candidate genes identified in this study might be used to develop stable, yellow-seeded B. napus. This study provides insight into the formation of seed coat color in B. napus. [ABSTRACT FROM AUTHOR]

The oil yields, fatty acid compositions, and total protein contents of three varieties of Turkish poppy were investigated. Solvent extraction of yellow seed gave the highest oil yield (49.2%), while white seed (36.8%) and blue seed (33.6) showed considerably lower yields. Yellow seed was also subjected to mechanical extraction, yielding 32.3% oil. Solvent extraction of the oilcake from the press gave 17.6% additional oil. Fatty acid compositions of the oils were determined by GGIMS and major components were found to be linoleic (56.4-69.2%), oleic (16.1-19.4%), and palmitic (10.6-16.3%,) depending on the color of the seeds. The values of proximate analysis, nutritional elements and heavy metals content were also reported for the most widely cultivated yellow seed. [ABSTRACT FROM AUTHOR]

Brassica napus is one of the important oil crops grown worldwide, and oil quality improvement is a major goal in rapeseed breeding. Yellow seed is an excellent trait, which has great potential in improving seed quality and economic value. In this study, we created stable yellow seed mutants using a CRISPR/Cas9 system and obtained the yellow seed phenotype only when the four alleles of two BnTT2 homologues were knocked out, indicating that the two BnTT2 homologues had conserved but redundant functions in regulating seed color. Histochemical staining and flavonoid metabolic analysis proved that the  BnTT2 mutation hindered the synthesis and accumulation of proanthocyanidins. Transcriptome analysis also showed that the BnTT2 mutation inhibited the expression of genes in the phenylpropanoid and flavonoid biosynthetic pathway, which might be regulated by the complex of BnTT2, BnTT8 and BnTTG1. In addition, the homozygous mutants of BnTT2 homologues increased oil content and improved fatty acid composition with higher linoleic acid (C18:2) and linolenic acid (C18:3), which could be used for the genetic improvement of rapeseed. Overall, this research showed that the BnTT2 mutation can be used for yellow seed breeding and oil improvement, which is of great significance in improving the economic value of rapeseeds.

A mutant with yellow seedcoat colour was isolated by NAYAR (1968) in the mustard variety ‘Rai-5’. This mutant was crossed to the national check cultivar ‘Varuna’ in order to develop improved strains with yellow seedcoat. Four such strains with yellow seeds were evaluated for their seed yield, yield components and percent oil. Two strains TM-9 and TM-17 were more productive than ‘Varuna’ in seed yield. All the yellow seeded strains showed higher oil percentage as compared to ‘Varuna’. The seedcoat in the yellow seeded strains accounts for 14–15 % of the seed weight as compared to 18 % in the black seeded ‘Varuna’. The higher proportion of the cotyledons and embryo accounts for the increased oil percentage in the yellow seeded types. [ABSTRACT FROM AUTHOR]

Brassica napus L. is rich in phenolic components and it has natural antioxidant characteristics which are important to human health. In the present study, the total phenolic and flavonoid contents of developing seeds of yellow- and black-seeded B. napus were compared. Both phenolic and flavonoid contents were significantly higher at 5 weeks after flowering (WAF) in black seeds (6.44 ± 0.97 mg EE/g phenolics and 3.78 ± 0.05 mg EE/g flavonoids) than yellow seeds (2.80 ± 0.13 mg/g phenolics and 0.83 ± 0.01 mg/g flavonoids). HPLC–DAD–ESI/MS analysis revealed different content of 56 phenolic components between yellow and black-seeded B. napus, including kaempferol-3-O-glucoside, isorhamnetin-3-O-glucoside, quercetin-3-O-sophoroside, procyanidin B2 ([DP 2]), which were significantly reduced in yellow seeds compared with black seeds. Applying the 2,2-diphenyl-1-picrylhydrazyl (DPPH) and 2,2′-azinobis-(3-ethylbenzothiazoline-6-sulfonic acid) (ABTS) radical assay, we found maximum clearance of DPPH and ABTS in the late developmental stages of yellow and black seeds. Additionally, the ferric reducing antioxidant power (FRAP) value maximized at 5 WAF in black seeds (432.52 ± 69.98 μmol Fe (II)/g DW) and 6 WAF in yellow seeds (274.08 ± 2.40 μmol Fe (II)/g DW). Generally, antioxidant ability was significantly reduced in yellow-seeded B. napus compared to black rapeseed, and positive correlations between antioxidation and flavonoid content were found in both yellow- and black-seeded B. napus. [ABSTRACT FROM AUTHOR]

Lines of Brassica juncea with tetralocular (four-valved) pods were produced by interspecific hybridization between dilocular (tow-valved) B. juncea var. Pusa Barani and tetralocular yellow-seed turnip rape (Brassica rapa ssp. trilocularis) var. YID-1. Some selected lines were significantly superior to their parents for some agronomic traits, Including seed yield and harvest index. Line 11 was the best among these and, recorded 70% yield advantage in plot yield. [ABSTRACT FROM AUTHOR]