Soybean(Glycine max) is an economically important oilbearing crop that is widely planted worldwide(Babu et al.,2017). Many diseases seriously limit soybean yield. Soybean root rot disease, one of the most destructive ...Soybean(Glycine max) is an economically important oilbearing crop that is widely planted worldwide(Babu et al.,2017). Many diseases seriously limit soybean yield. Soybean root rot disease, one of the most destructive soybean diseases, occurs throughout the growth period, resulting in considerable yield losses(Kamoun et al., 2015).展开更多
The creation of new soybean varieties has been limited by genomic duplication and redundancy.Efficient multiplex gene editing and large chromosomal segment deletion through clustered regularly interspaced palindromic ...The creation of new soybean varieties has been limited by genomic duplication and redundancy.Efficient multiplex gene editing and large chromosomal segment deletion through clustered regularly interspaced palindromic repeats(CRISPR)/CRISPR-associated protein(Cas)systems are promising strategies for overcoming these obstacles.CRISPR/Cpf1 is a robust tool for multiplex gene editing.However,large chromosomal excision mediated by CRISPR/Cpf1 has been reported in only a few non-plant species.Here,we report on CRISPR/LbCpf1-induced large chromosomal segment deletions in soybean using multiplex gene targeting.The CRISPR/LbCpf1 system was optimized for direct repeat and guide RNA lengths in crispr RNA(crRNA)array.The editing efficiency was evaluated using LbCpf1 driven by the CaMV35S and soybean ubiquitin promoter.The optimized system exhibited editing efficiencies of up to 91.7%.Our results showed eight gene targets could be edited simultaneously in one step when a single eight-gRNA-target crRNA array was employed,with an efficiency of up to 17.1%.We successfully employed CRISPR/LbCpf1 to produce small fragments(<1 Kb)and large chromosomal segment deletions(10 Kb-1 Mb)involving four different gene clusters in soybean.Together,these data demonstrate the power of the CRISPR/LbCpf1 platform for multiplex gene editing and chromosomal segment deletion in soybean,supporting the use of this technology in both basic research and agricultural applications.展开更多
Dear Editor, ManybacterialproteinsareinvolvedinAgrobacterium- mediated plant transformation. By contrast, relatively little is known about plant proteins that play key roles in transformation. Some of these host prot...Dear Editor, ManybacterialproteinsareinvolvedinAgrobacterium- mediated plant transformation. By contrast, relatively little is known about plant proteins that play key roles in transformation. Some of these host proteins interact with Virulence effector proteins, including VirE2, that are transferred from Agrobacterium to plants （Gelvin, 2010; Pitzschke and Hirt, 2010）. A recent study indicated that the plant protein SUPPRESSOR OF G2 ALLELE OF SKP1 （SGT1）, a co-chaperone of heat shock protein 90 （HSP90）, is required for Agrobacterium-mediated transformation （Anand et al., 2012）. These studies suggested the involvement of HSPg0 in Agrobacterium-mediated transformation.展开更多
基金supported by grants from the National Science Foundation of China (NSFC32172499, 31901957)+2 种基金China National Funds for Innovative Research Groups (31721004)the Chinese Modern Agricultural Industry Technology System (CARS004-PS14)Fundamental Research Funds for the Central Universities (JCQY201903)。
文摘Soybean(Glycine max) is an economically important oilbearing crop that is widely planted worldwide(Babu et al.,2017). Many diseases seriously limit soybean yield. Soybean root rot disease, one of the most destructive soybean diseases, occurs throughout the growth period, resulting in considerable yield losses(Kamoun et al., 2015).
基金supported by grants from National Science Foundation of China(NSFC31901957)Fundamental Research Funds for the Central Universities(JCQY201903)。
文摘The creation of new soybean varieties has been limited by genomic duplication and redundancy.Efficient multiplex gene editing and large chromosomal segment deletion through clustered regularly interspaced palindromic repeats(CRISPR)/CRISPR-associated protein(Cas)systems are promising strategies for overcoming these obstacles.CRISPR/Cpf1 is a robust tool for multiplex gene editing.However,large chromosomal excision mediated by CRISPR/Cpf1 has been reported in only a few non-plant species.Here,we report on CRISPR/LbCpf1-induced large chromosomal segment deletions in soybean using multiplex gene targeting.The CRISPR/LbCpf1 system was optimized for direct repeat and guide RNA lengths in crispr RNA(crRNA)array.The editing efficiency was evaluated using LbCpf1 driven by the CaMV35S and soybean ubiquitin promoter.The optimized system exhibited editing efficiencies of up to 91.7%.Our results showed eight gene targets could be edited simultaneously in one step when a single eight-gRNA-target crRNA array was employed,with an efficiency of up to 17.1%.We successfully employed CRISPR/LbCpf1 to produce small fragments(<1 Kb)and large chromosomal segment deletions(10 Kb-1 Mb)involving four different gene clusters in soybean.Together,these data demonstrate the power of the CRISPR/LbCpf1 platform for multiplex gene editing and chromosomal segment deletion in soybean,supporting the use of this technology in both basic research and agricultural applications.
文摘Dear Editor, ManybacterialproteinsareinvolvedinAgrobacterium- mediated plant transformation. By contrast, relatively little is known about plant proteins that play key roles in transformation. Some of these host proteins interact with Virulence effector proteins, including VirE2, that are transferred from Agrobacterium to plants （Gelvin, 2010; Pitzschke and Hirt, 2010）. A recent study indicated that the plant protein SUPPRESSOR OF G2 ALLELE OF SKP1 （SGT1）, a co-chaperone of heat shock protein 90 （HSP90）, is required for Agrobacterium-mediated transformation （Anand et al., 2012）. These studies suggested the involvement of HSPg0 in Agrobacterium-mediated transformation.
