Targeted mutagenesis of <i>BnTT8</i> homologs controls yellow seed coat development for effective oil production in <i>Brassica napus</i> L.

Zhai, Yungu; Yu, Kaidi; Cai, Shaoxin; Hu, Limin; Amoo, Olalekan; Lei, Xu; Yang, Yang; Ma, Boyuan; Jiao, Yangmiao; Zhang, Chaofeng; Khan, Muhammad Hafeez Ullah; Khan, Shahid Ullah; Fan, Chuchuan; Zhou, Yongming

doi:10.1111/pbi.13281

Cited by 143 publications

(122 citation statements)

References 57 publications

(126 reference statements)

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“…Though two TT18 genes (Bra.A01TT18 and Bra.A03TT18) in B. rapa were not involved in the Bra.TT8-containing primary network, a positive regulatory relationship between TT8 and TT18 could be observed due to their similar weak expression in the yellow-seeded B. rapa accession. A similar network has also been identified in B. napus [40,42,43], Thus, we infer that the regulatory network of the TT8-involved complex was evolutionarily conserved between Arabidopsis and Brassica crop species. Considering the divergence of the two genera, the conserved TT8 genes might be optimal target loci for manipulation in the breeding of yellow-seeded varieties of Brassica and related crops.…”

Section: The Conserved Dream Complex May Control Seed Size As An Upstsupporting

confidence: 76%

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Deciphering the transcriptional regulatory networks that control size, color, and oil content in Brassica rapa seeds

Niu

Wu²,

et al. 2020

Preprint

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Background Brassica rapa is an important oilseed and vegetable crop species and is the A subgenome donor of two important oilseed Brassica crops, Brassica napus and Brassica juncea. Although seed size (SZ), seed color (SC), and oil content (OC) substantially affect seed yield and quality, the mechanisms regulating these traits in Brassica crops remain unclear. Results We collected seeds from a pair of B. rapa accessions with significantly different SZ, SC, and OC at seven seed developmental stages (every 7 days from 7 to 49 days after pollination), and identified 24,835 differentially expressed genes (DEGs) from seven pairwise comparisons between accessions at each developmental stage. K-means clustering identified a group of cell cycle-related genes closely connected to variation in SZ of B. rapa. A weighted correlation analysis using the WGCNA package in R revealed two important co-expression modules comprising genes whose expression was positively correlated with SZ increase and negatively correlated with seed yellowness, respectively. Upregulated expression of cell cycle-related genes in one module was important for the G2/M cell cycle transition, and the transcription factor Bra.A05TSO1 seemed to positively stimulate the expression of two CYCB1;2 genes to promote seed development. In the second module, a conserved complex regulated by the transcription factor TT8 appear to determine SC through downregulation of TT8 and its target genes TT3, TT18, and ANR. Further, upregulation of genes involved in triacylglycerol biosynthesis and storage in the seed oil body may increase OC. We further validated the accuracy of the transcriptome data by quantitative real-time PCR of 15 DEGs. Finally, we used our results to construct detailed models to clarify the regulatory mechanisms underlying variations in SZ, SC, and OC in B. rapa. Conclusions This study based on transcriptome comparison provides insight into the regulatory mechanisms underlying the variations of SZ, SC, and OC in plants. The findings hold great promise for improving seed yield, quality and OC through genetic engineering of critical genes in future molecular breeding.

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Section: The Conserved Dream Complex May Control Seed Size As An Upstsupporting

confidence: 76%

“…yellow sarson [38]. Recent studies have also revealed that simultaneous natural mutations of two homologous TT8 genes in allotetraploid B. juncea resulted in the yellow-seeded trait [39], and a similar mutant also could be created in B. napus using CRISPR/Cas9 technology [40].…”

Section: The Conserved Dream Complex May Control Seed Size As An Upstmentioning

confidence: 99%

Deciphering the transcriptional regulatory networks that control size, color, and oil content in Brassica rapa seeds

Niu

Wu²,

et al. 2020

Preprint

View full text Add to dashboard Cite

show abstract

“…Though two TT18 genes (Bra.A01TT18 and Bra.A03TT18) in B. rapa were not involved in the Bra.TT8-containing primary network, a positive regulatory relationship between TT8 and TT18 could be observed due to their similar weak expression in the yellow-seeded B. rapa accession. A similar network has also been identified in B. napus [42,44,45], Thus, we infer that the regulatory network of the TT8-involved complex was evolutionarily conserved between Arabidopsis and Brassica crop species. Considering the divergence of the two genera, the conserved TT8 genes might be optimal target loci for manipulation in the breeding of yellow-seeded varieties of Brassica and related crops.…”

Section: Tt8 Might Be a Conserved Key Target For Creating Yellow-seedsupporting

confidence: 76%

“…yellow sarson [40]. Recent studies have also revealed that simultaneous natural mutations of two homologous TT8 genes in allotetraploid B. juncea resulted in the yellowseeded trait [41], and a similar mutant also could be created in B. napus using CRISPR/Cas9 technology [42].…”

Section: Tt8 Might Be a Conserved Key Target For Creating Yellow-seedmentioning

confidence: 99%

Deciphering the transcriptional regulatory networks that control size, color, and oil content in Brassica rapa seeds

Niu

Wu²,

et al. 2020

Preprint

View full text Add to dashboard Cite

Background: Brassica rapa is an important oilseed and vegetable crop species and is the A subgenome donor of two important oilseed Brassica crops, Brassica napus and Brassica juncea. Although seed size (SZ), seed color (SC), and oil content (OC) substantially affect seed yield and quality, the mechanisms regulating these traits in Brassica crops remain unclear.Results: We collected seeds from a pair of B. rapa accessions with significantly different SZ, SC, and OC at seven seed developmental stages (every 7 days from 7 to 49 days after pollination), and identified 28,954 differentially expressed genes (DEGs) from seven pairwise comparisons between accessions at each developmental stage. K-means clustering identified a group of cell cycle-related genes closely connected to variation in SZ of B. rapa. A weighted correlation analysis using the WGCNA package in R revealed two important co-expression modules comprising genes whose expression was positively correlated with SZ increase and negatively correlated with seed yellowness, respectively. Upregulated expression of cell cycle-related genes in one module was important for the G2/M cell cycle transition, and the transcription factor Bra.A05TSO1 seemed to positively stimulate the expression of two CYCB1;2 genes to promote seed development. In the second module, a conserved complex regulated by the transcription factor TT8 appear to determine SC through downregulation of TT8 and its target genes TT3, TT18, and ANR. In the third module, WRI1 and FUS3 were conserved to increase the seed OC, and Bra.A03GRF5 was revealed as a key transcription factor on lipid biosynthesis. Further, upregulation of genes involved in triacylglycerol biosynthesis and storage in the seed oil body may increase OC. We further validated the accuracy of the transcriptome data by quantitative real-time PCR of 15 DEGs. Finally, we used our results to construct detailed models to clarify the regulatory mechanisms underlying variations in SZ, SC, and OC in B. rapa.Conclusions: This study provides insight into the regulatory mechanisms underlying the variations of SZ, SC, and OC in plants based on transcriptome comparison. The findings hold great promise for improving seed yield, quality and OC through genetic engineering of critical genes in future molecular breeding.

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“…Seed coat development competes for sucrose with reserve component synthesis in embryo and endosperm. Recently, studies had demonstrated that the amount of PAs in the seed coat is negatively correlated with the amount of lipid content in the embryo in Arabidopsis and rapeseed [11,12]. Thus, strategies of manipulating the PAs biosynthesis pathway in seed coat could be used to increase the seed lipid content.…”

Section: Introductionmentioning

confidence: 99%

CRISPR-Cas9 Mediated Knockout of NtAn1 to Enhance the Lipid Accumulation in Tobacco Seed for Biodiesel Production

Tian¹,

Liu²,

Li³

et al. 2020

Preprint

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Background: Tobacco seed lipid is a promising non-edible feedstock for biodiesel production. In order to meet the increasing demand, achieving high seed lipid content is one of the major goals in tobacco seed production. The TT8 gene and its homologs negatively regulate seed lipid accumulation in Arabidopsis and Brassica species. We speculated that manipulating the homolog genes of TT8 in tobacco could enhance the accumulation of seed lipid.Results: In this present study, we found that the TT8 homolog genes in tobacco, NtAn1a and NtAn1b, were highly expressed in developing seed. Targeted mutagenesis of NtAn1 genes were created by the CRISPR-Cas9 based gene editing technology. Due to the defect of PAs biosynthesis, mutant seeds showed a phenotype of yellow seed coat. Seed lipid accumulation was enhanced by about 18% and 15% in two targeted mutant lines, respectively. Protein content was also significantly increased in mutant seeds. In addition, the seed yield related traits were not affected by the targeted mutagenesis of NtAn1 genes. Thus, the overall lipid productivity of the NtAn1 knockout mutants were dramatically enhanced. Conclusion: Tobacco NtAn1 genes regulate both PAs and lipid accumulation in the process of seed development. Targeted mutagenesis of NtAn1 genes could generate a yellow-seeded tobacco variety with high lipid and protein content. Furthermore, the present results revealed that CRISPR-Cas9 system could be employed in tobacco seed de novo domestication for biodiesel feedstock production.

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Targeted mutagenesis of BnTT8 homologs controls yellow seed coat development for effective oil production in Brassica napus L.

Cited by 143 publications

References 57 publications

Deciphering the transcriptional regulatory networks that control size, color, and oil content in Brassica rapa seeds

Deciphering the transcriptional regulatory networks that control size, color, and oil content in Brassica rapa seeds

Deciphering the transcriptional regulatory networks that control size, color, and oil content in Brassica rapa seeds

CRISPR-Cas9 Mediated Knockout of NtAn1 to Enhance the Lipid Accumulation in Tobacco Seed for Biodiesel Production

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