Precise editing of <i><scp>CLAVATA</scp></i> genes in <i>Brassica napus</i> L. regulates multilocular silique development

Yang, Yang; Zhu, Kaiyu; Li, Huailin; Han, Shaoqing; Meng, Qingwei; Khan, Shahid Ullah; Fan, Chuchuan; Xie, Kabin; Zhou, Yongming

doi:10.1111/pbi.12872

Cited by 140 publications

(137 citation statements)

References 48 publications

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“…The binary pYLCRIPSR/Cas9 multiplex genome targeting vector system was utilized for gene editing in this investigation (Ma et al , ). The selection of sequence‐specific sgRNAs in the target gene, CRISPR/Cas9 construct assembly and Agrobacterium tumefaciens ‐mediated hypocotyl transformation in B. napus were conducted as previously described (Yang et al , ). The oligos employed in constructing the sgRNA vectors are listed in Table S1.…”

Section: Methodsmentioning

confidence: 99%

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

Zhai

Cai

et al. 2019

Plant Biotechnology Journal

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159

138

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Yellow seed is a desirable trait with great potential for improving seed quality in Brassica crops. Unfortunately, no natural or induced yellow seed germplasms have been found in Brassica napus, an important oil crop, which likely reflects its genome complexity and the difficulty of the simultaneous random mutagenesis of multiple gene copies with functional redundancy. Here, we demonstrate the first application of CRISPR/Cas9 for creating yellow‐seeded mutants in rapeseed. The targeted mutations of the BnTT8 gene were stably transmitted to successive generations, and a range of homozygous mutants with loss‐of‐function alleles of the target genes were obtained for phenotyping. The yellow‐seeded phenotype could be recovered only in targeted mutants of both BnTT8 functional copies, indicating that the redundant roles of BnA09.TT8 and BnC09.TT8b are vital for seed colour. The BnTT8 double mutants produced seeds with elevated seed oil and protein content and altered fatty acid (FA) composition without any serious defects in the yield‐related traits, making it a valuable resource for rapeseed breeding programmes. Chemical staining and histological analysis showed that the targeted mutations of BnTT8 completely blocked the proanthocyanidin (PA)‐specific deposition in the seed coat. Further, transcriptomic profiling revealed that the targeted mutations of BnTT8 resulted in the broad suppression of phenylpropanoid/flavonoid biosynthesis genes, which indicated a much more complex molecular mechanism underlying seed colour formation in rapeseed than in Arabidopsis and other Brassica species. In addition, gene expression analysis revealed the possible mechanism through which BnTT8 altered the oil content and fatty acid composition in seeds.

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Section: Methodsmentioning

confidence: 99%

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

Zhai

Cai

et al. 2019

Plant Biotechnology Journal

Self Cite

159

138

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“…In recent years, the CLV ‐ WUS pathway has become an attractive target of genome editing for crop improvement (Rodriguez‐Leal et al , ), which has produced a series of exciting achievements. The genome editing of CLV3 orthologues can produce multilocular siliques in several dry fruit crops, including Brassica rapa and B. napus (Fan et al , ; Yang et al , ). In addition, the genome editing of CLV1/2/3 also produced multiple locular fruits with increased size in fleshy fruit crops, such as tomato and groundcherry (Lemmon et al , ; Li et al , ; Rodrı´guez‐Leal et al , ; Xu et al , ; Zsögön et al , ).…”

Section: Receptor Kinase Signallingmentioning

confidence: 99%

“…Interestingly, many of these genes have a conserved function in regulating fruit development in both dry and fleshy fruit types, suggesting broad applicability of common genome edits. For instance, CYP78A9 and CLV‐WUS pathway genes can regulate both silique size in Arabidopsis /rapeseed (Fan et al , ; Shi et al , ; Xiao et al , ; Xu et al , ; Yang et al , ) and fruit size in cherry/tomato (Lemmon et al , ; Li et al , ; Qi et al , ; Rodriguez‐Leal et al ., ; Xu et al , ). Besides, many of these genes have similar functions in regulating fruit size in Arabidopsis and other crops.…”

Section: Conclusion and Future Prospectsmentioning

confidence: 99%

“…Besides, many of these genes have similar functions in regulating fruit size in Arabidopsis and other crops. For example, ARF18 , ARP1 , BoMF2 , DRM1 , CYP78A9 , CLV1 , CLV3 and SMG7 can regulate silique length in Arabidopsis and rapeseed (Kang et al , ; Lee et al , ; Li et al , ; Liu et al , ; Shi et al , ; Xiao et al , ; Yang et al , ). More importantly, many of the fruit size regulatory genes have expression activity and additional effects on other organs, such as roots, stems, leaves, flowers and seeds, indicating a cooperative/synergistic regulation.…”

Section: Conclusion and Future Prospectsmentioning

confidence: 99%

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Genetic and signalling pathways of dry fruit size: targets for genome editing‐based crop improvement

Hussain

Shi

Scheben

et al. 2020

Plant Biotechnology Journal

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Summary Fruit is seed‐bearing structures specific to angiosperm that form from the gynoecium after flowering. Fruit size is an important fitness character for plant evolution and an agronomical trait for crop domestication/improvement. Despite the functional and economic importance of fruit size, the underlying genes and mechanisms are poorly understood, especially for dry fruit types. Improving our understanding of the genomic basis for fruit size opens the potential to apply gene‐editing technology such as CRISPR/Cas to modulate fruit size in a range of species. This review examines the genes involved in the regulation of fruit size and identifies their genetic/signalling pathways, including the phytohormones, transcription and elongation factors, ubiquitin‐proteasome and microRNA pathways, G‐protein and receptor kinases signalling, arabinogalactan and RNA‐binding proteins. Interestingly, different plant taxa have conserved functions for various fruit size regulators, suggesting that common genome edits across species may have similar outcomes. Many fruit size regulators identified to date are pleiotropic and affect other organs such as seeds, flowers and leaves, indicating a coordinated regulation. The relationships between fruit size and fruit number/seed number per fruit/seed size, as well as future research questions, are also discussed.

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“…Alternatively, grain weight was enhanced via knockout of GW2 , GW5 and TGW6 in rice (Xu et al ), and of GASR7 in wheat (Zhang et al ). Increased locule and seed number per silique were obtained in rapeseed via knockout of both homeoalleles of the CLV3 gene (Yang et al ). Taking an even more sophisticated approach in tomato, Rodríguez‐Leal et al () demonstrated that a whole range of variability in locule number and fruit size can be obtained by producing an array of CLV3 promoter variants with different fragment deletions.…”

Section: Applications Of Targeted Genome Modification By Nhejmentioning

confidence: 99%

The CRISPR/Cas revolution continues: From efficient gene editing for crop breeding to plant synthetic biology

Kumlehn

Pietralla

Hensel

et al. 2018

JIPB

100

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Since the discovery that nucleases of the bacterial CRISPR (clustered regularly interspaced palindromic repeat)‐associated (Cas) system can be used as easily programmable tools for genome engineering, their application massively transformed different areas of plant biology. In this review, we assess the current state of their use for crop breeding to incorporate attractive new agronomical traits into specific cultivars of various crop plants. This can be achieved by the use of Cas9/12 nucleases for double‐strand break induction, resulting in mutations by non‐homologous recombination. Strategies for performing such experiments − from the design of guide RNA to the use of different transformation technologies − are evaluated. Furthermore, we sum up recent developments regarding the use of nuclease‐deficient Cas9/12 proteins, as DNA‐binding moieties for targeting different kinds of enzyme activities to specific sites within the genome. Progress in base deamination, transcriptional induction and transcriptional repression, as well as in imaging in plants, is also discussed. As different Cas9/12 enzymes are at hand, the simultaneous application of various enzyme activities, to multiple genomic sites, is now in reach to redirect plant metabolism in a multifunctional manner and pave the way for a new level of plant synthetic biology.

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Precise editing of CLAVATA genes in Brassica napus L. regulates multilocular silique development

Cited by 140 publications

References 48 publications

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

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

Genetic and signalling pathways of dry fruit size: targets for genome editing‐based crop improvement

The CRISPR/Cas revolution continues: From efficient gene editing for crop breeding to plant synthetic biology

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