Weeds and weedy rice plague commercial rice fields in many countries. Developingherbicide-tolerance rice is the most efficient strategy to control weed proliferation. CRISPR/Cas9-mediated gene editing, which generates...Weeds and weedy rice plague commercial rice fields in many countries. Developingherbicide-tolerance rice is the most efficient strategy to control weed proliferation. CRISPR/Cas9-mediated gene editing, which generates small InDels and nucleotide substitutions atand around target sites using error-prone non-homologous end joining DNA repairing, hasbeen widely adopted for generation of novel crop germplasm with a wide range of geneticvariation in important agronomic traits. We created a novel herbicide-tolerance allele inrice by targeting the acetolactate synthase (OsALS) gene using CRISPR/Cas9-mediated geneediting. The novel allele (G628W) arose from a G-to-T transversion at position 1882 of OsALSand conferred a high level of herbicide tolerance. Transgene-free progeny carryinghomozygous G628W allele were identified and showed agronomic performance similar tothat of wild-type plants, suggesting that the G628W allele is a valuable resource fordeveloping elite rice varieties with strong herbicide tolerance. To promote use of the G628Wallele and to accelerate introgression and/or pyramiding of the G628W allele with other elitealleles, we developed a DNA marker for the G628W allele that accurately and robustlydistinguished homozygous from heterozygous segregants. Our result further demonstratesthe feasibility of CRISPR/Cas9-mediated gene editing in creating novel genetic variation forcrop breeding.展开更多
Intron-targeted gene insertion strategy using CRISPR/Cas9(clustered regularly interspaced short palindromic repeats/CRISPR-associated Cas9) has been shown to be a potential tool for crop genetic improvement by targete...Intron-targeted gene insertion strategy using CRISPR/Cas9(clustered regularly interspaced short palindromic repeats/CRISPR-associated Cas9) has been shown to be a potential tool for crop genetic improvement by targeted mutagenesis or gene replacement of an elite allele into widely cultivated rice varieties. The rice blast resistant protein Pi-ta, differs from its susceptible counterpart, pi-ta, by a single amino acid in exon 2. To create new materials resistant to the rice blast disease, we inserted a genomic fragment containing the exon 2 and 3′ untranslated region(3′ UTR) of Pi-ta into intron 1 of pi-ta in rice materials susceptible to rice blast using the intron-targeted insertion strategy. The gene insertion frequency was3.8%. Several novel transgene-free progeny with stably inherited homozygous insert were identified in the T_1 generation, which have been crossed to rice germplasm bearing other resistance gene(R gene) for pyramiding of R genes. This work verified the feasibility of using the genome editing technology in improvement of qualitative agronomic trait in crops.展开更多
Dear Editor,Rice(Oryza sativa L.),one of the most important staple crops,feeds more than half of the world’s population.The appli-cation of nitrogen(N)fertilizer has greatly enhanced rice production but has also incr...Dear Editor,Rice(Oryza sativa L.),one of the most important staple crops,feeds more than half of the world’s population.The appli-cation of nitrogen(N)fertilizer has greatly enhanced rice production but has also increased the cost of rice production and environmental pollution.Based on previous research,O.sativa subsp.japonica rice varieties generally have lower nitro-gen use efficiency(NUE)than O.sativa subsp.indica rice varieties(Zhang and Chu 2020).Therefore,breeding new japonica rice varieties that not only produce high yields,have superior taste and texture,and possess disease resistance but also exhibit improved NUE is increasingly essential.展开更多
基金This study was supported by the National Transgenic Science and Technology Program(2018ZX08001-02B)the Jiangsu Agricultural Science and Technology Innovation Fund(CX(19)3059)the Jiangsu Province Key Research and Development Program(Modern Agriculture,BE2017345-2).
文摘Weeds and weedy rice plague commercial rice fields in many countries. Developingherbicide-tolerance rice is the most efficient strategy to control weed proliferation. CRISPR/Cas9-mediated gene editing, which generates small InDels and nucleotide substitutions atand around target sites using error-prone non-homologous end joining DNA repairing, hasbeen widely adopted for generation of novel crop germplasm with a wide range of geneticvariation in important agronomic traits. We created a novel herbicide-tolerance allele inrice by targeting the acetolactate synthase (OsALS) gene using CRISPR/Cas9-mediated geneediting. The novel allele (G628W) arose from a G-to-T transversion at position 1882 of OsALSand conferred a high level of herbicide tolerance. Transgene-free progeny carryinghomozygous G628W allele were identified and showed agronomic performance similar tothat of wild-type plants, suggesting that the G628W allele is a valuable resource fordeveloping elite rice varieties with strong herbicide tolerance. To promote use of the G628Wallele and to accelerate introgression and/or pyramiding of the G628W allele with other elitealleles, we developed a DNA marker for the G628W allele that accurately and robustlydistinguished homozygous from heterozygous segregants. Our result further demonstratesthe feasibility of CRISPR/Cas9-mediated gene editing in creating novel genetic variation forcrop breeding.
基金supported by the National Transgenic Major Project of China(2018ZX08001-02B)the Key Research and Development Program of Jiangsu Province(Modern Agriculture,BE2018381-2)+1 种基金the National Science Foundation of Jiangsu Province(BK20170610)the Exploratory Project of the Jiangsu Academy of Agricultural Sciences(ZX(17)2014)。
文摘Intron-targeted gene insertion strategy using CRISPR/Cas9(clustered regularly interspaced short palindromic repeats/CRISPR-associated Cas9) has been shown to be a potential tool for crop genetic improvement by targeted mutagenesis or gene replacement of an elite allele into widely cultivated rice varieties. The rice blast resistant protein Pi-ta, differs from its susceptible counterpart, pi-ta, by a single amino acid in exon 2. To create new materials resistant to the rice blast disease, we inserted a genomic fragment containing the exon 2 and 3′ untranslated region(3′ UTR) of Pi-ta into intron 1 of pi-ta in rice materials susceptible to rice blast using the intron-targeted insertion strategy. The gene insertion frequency was3.8%. Several novel transgene-free progeny with stably inherited homozygous insert were identified in the T_1 generation, which have been crossed to rice germplasm bearing other resistance gene(R gene) for pyramiding of R genes. This work verified the feasibility of using the genome editing technology in improvement of qualitative agronomic trait in crops.
基金supported by the STI 2030-Major Projects(2023ZD04066)the Jiangsu Seed Industry Revitalization Project(JBGS[2021]011 and JBGS[2021]039)+1 种基金the Ministry of Agriculture and Rural Affairs of China,the Jiangsu Provincial Key Research and Development Program(BE2021374 and BE2020339)the National Natural Science Foundation of China(32201898).
文摘Dear Editor,Rice(Oryza sativa L.),one of the most important staple crops,feeds more than half of the world’s population.The appli-cation of nitrogen(N)fertilizer has greatly enhanced rice production but has also increased the cost of rice production and environmental pollution.Based on previous research,O.sativa subsp.japonica rice varieties generally have lower nitro-gen use efficiency(NUE)than O.sativa subsp.indica rice varieties(Zhang and Chu 2020).Therefore,breeding new japonica rice varieties that not only produce high yields,have superior taste and texture,and possess disease resistance but also exhibit improved NUE is increasingly essential.
