CRISPR/Cas9‐mediated targeted mutagenesis of <i>GmFT2a</i> delays flowering time in soya bean

Cai, Yupeng; Chen, Li; Liu, Xiujie; Chen, Guo; Sun, Shi; Wu, Cunxiang; Jiang, Bingjun; Han, Tianfu; Hou, Wensheng

doi:10.1111/pbi.12758

Cited by 262 publications

(148 citation statements)

References 56 publications

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“…Accelerated maturity in rice and tomato was achieved via knockout of OsHd2 , Hd4 , Hd5 (Li et al ) and of SlSP5G (Soyk et al ), respectively. By contrast, knockout of GmFT2 entailed delayed flowering time in soybean (Cai et al ). Li et al () demonstrated that mutagenesis of the long non‐coding RNA1459 results in delayed tomato fruit ripening.…”

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

102

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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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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

102

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“…In soybean, at least 10 FT homologs have been identified (Kong et al ., ). GmFT2a and GmFT5a were found to be strictly photoperiod‐regulated, and have been shown to promote flowering in Arabidopsis and soybean (Kong et al ., ; Sun et al ., ; Cai et al ., ). The functions of the other GmFTs in soybean remain unclear, although GmFT2b , 3a , 3b , 4 , 5b , and 6 have been preliminarily studied using heterologous expression in Arabidopsis (Fan et al ., ; Zhai et al ., ; Cao et al ., ).…”

Section: Introductionmentioning

confidence: 97%

“…It is thus conceivable that some ratio of a flowering promoter and a flowering inhibitor, as specified by photoperiodic signals, may represent a sort of balance that regulates whether plants undertake vegetative growth or reproductive development In soybean, at least 10 FT homologs have been identified (Kong et al, 2010). GmFT2a and GmFT5a were found to be strictly photoperiod-regulated, and have been shown to promote flowering in Arabidopsis and soybean (Kong et al, 2010;Sun et al, 2011;Cai et al, 2017). The functions of the other GmFTs in soybean remain unclear, although GmFT2b, 3a, 3b, 4, 5b, and 6 have been preliminarily studied using heterologous expression in Arabidopsis (Fan et al, 2014;Zhai et al, 2014;Cao et al, 2016a).…”

Section: Introductionmentioning

confidence: 99%

Functional diversification of Flowering Locus T homologs in soybean: GmFT1a and GmFT2a/5a have opposite roles in controlling flowering and maturation

et al. 2017

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Summary Soybean flowering and maturation are strictly regulated by photoperiod. Photoperiod‐sensitive soybean varieties can undergo flowering reversion when switched from short‐day (SD) to long‐day (LD) conditions, suggesting the presence of a ‘floral‐inhibitor’ under LD conditions.We combined gene expression profiling with a study of transgenic plants and confirmed that GmFT1a, soybean Flowering Locus T (FT) homolog, is a floral inhibitor. GmFT1a is expressed specifically in leaves, similar to the flowering‐promoting FT homologs GmFT2a/5a. However, in Zigongdongdou (ZGDD), a model variety for studying flowering reversion, GmFT1a expression was induced by LD but inhibited by SD conditions. This was unexpected, as it is the complete opposite of the expression of flowering promoters GmFT2a/5a. Moreover, the key soybean maturity gene E1 may up‐regulate GmFT1a expression. It is also notable that GmFT1a expression was conspicuously high in late‐flowering varieties. Transgenic overexpression of GmFT1a delayed flowering and maturation in soybean, confirming that GmFT1a functions as a flowering inhibitor.This discovery highlights the complex impacts of the functional diversification of the FT gene family in soybean, and implies that antagonism between flowering‐inhibiting and flowering‐promoting FT homologs in this highly photoperiod‐sensitive plant may specify vegetative vs reproductive development.

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“…Multiple reports have described the generation of somatic mutations using CRISPR/Cas9 in soya bean (Cai et al, 2015;Du et al, 2016;Jacobs et al, 2015;Lowder et al, 2015;Michno et al, 2015;Tang et al, 2016). To date however, few studies have demonstrated whole-plant soya bean mutagenesis (Cai et al, 2017;Li et al, 2015). Here, we describe a set of plant expression vectors that can accommodate either the CRISPR/Cas9 or TALEN entry vectors to target the genes encoding the machinery proteins involved in small RNA processing in soya bean and M. truncatula Curtin et al, 2017;Zhang et al, 2013).…”

Section: Introductionmentioning

confidence: 99%

CRISPR/Cas9 and TALENs generate heritable mutations for genes involved in small RNA processing of Glycine max and Medicago truncatula

Curtin

Xiong

Michno

et al. 2017

Plant Biotechnology Journal

147

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SummaryProcessing of double‐stranded RNA precursors into small RNAs is an essential regulator of gene expression in plant development and stress response. Small RNA processing requires the combined activity of a functionally diverse group of molecular components. However, in most of the plant species, there are insufficient mutant resources to functionally characterize each encoding gene. Here, mutations in loci encoding protein machinery involved in small RNA processing in soya bean and Medicago truncatula were generated using the CRISPR/Cas9 and TAL‐effector nuclease (TALEN) mutagenesis platforms. An efficient CRISPR/Cas9 reagent was used to create a bi‐allelic double mutant for the two soya bean paralogous Double‐stranded RNA‐binding2 (GmDrb2a and GmDrb2b) genes. These mutations, along with a CRISPR/Cas9‐generated mutation of the M. truncatula Hua enhancer1 (MtHen1) gene, were determined to be germ‐line transmissible. Furthermore, TALENs were used to generate a mutation within the soya bean Dicer‐like2 gene. CRISPR/Cas9 mutagenesis of the soya bean Dicer‐like3 gene and the GmHen1a gene was observed in the T0 generation, but these mutations failed to transmit to the T1 generation. The irregular transmission of induced mutations and the corresponding transgenes was investigated by whole‐genome sequencing to reveal a spectrum of non‐germ‐line‐targeted mutations and multiple transgene insertion events. Finally, a suite of combinatorial mutant plants were generated by combining the previously reported Gmdcl1a, Gmdcl1b and Gmdcl4b mutants with the Gmdrb2ab double mutant. Altogether, this study demonstrates the synergistic use of different genome engineering platforms to generate a collection of useful mutant plant lines for future study of small RNA processing in legume crops.

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CRISPR/Cas9‐mediated targeted mutagenesis of GmFT2a delays flowering time in soya bean

Cited by 262 publications

References 56 publications

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

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

Functional diversification of Flowering Locus T homologs in soybean: GmFT1a and GmFT2a/5a have opposite roles in controlling flowering and maturation

CRISPR/Cas9 and TALENs generate heritable mutations for genes involved in small RNA processing of Glycine max and Medicago truncatula

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