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Fixation of hybrid vigor in rice:synthetic apomixis generated by genome editing

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aBIOTECH 2020年第1期1卷 15-20页

作者： kejian wang State Key Laboratory of Rice Biology China National Rice Research InstituteChinese Academy of Agricultural SciencesHangzhou 310006China

Apomixis is an asexual reproduction process in which clonal seeds are formed without meiosis and *** of its potential in permanently preserving hybrid vigor,apomixis has attracted a great deal of interests from plant biologists and the seed ***,despite of decades of effort,introgression of apomixis traits from wild relatives into major crops has remained ***,synthetic apomixis has been proposed as an alternative to fix hybrid *** this article,I present the development of the MiMe(Mitosis instead of Meiosis),which turns meiosis into mitosis and leads to the production of clonal ***-like clonal seeds are generated when MiMe plants are crossed to special genome elimination lines,which contain an altered centromere-specific histone 3(CENH3).Furthermore,induction of haploid plants from egg cells can be achieved by either egg cell-specific expression of BABY BOOM1(BBM1),or disruption of MATRILINEAL(MTL)using CRISPR/Cas9 gene-editing *** apomixis is established and clonal seeds are produced by simultaneous engineering MiMe with altering BBM1 expression or MTL ***,I discuss how to further improve the apomixis strategy and its applications in crop breeding.

关键词： Apomixis Heterosis Hybrid vigor Genome editing

Development and Application of CRISPR/Cas System in Rice

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Rice science 2019年第2期26卷 69-76,I0004-I0006页

作者： REN Jun HU Xixun wang kejian wang Chun State Key Laboratory of Rice Biology China National Rice Research Institute Chinese Academy of Agricultural Sciences

In the past several years, the CRISPR(clustered regularly interspaced short palindromic repeats)/Cas(CRISPR-associated protein) system has been harnessed as an efficient and powerful tool for targeted genome editing in different prokaryotic and eukaryotic species. Here, we review the development and application of CRISPR/Cas system in rice, emphasizing different varieties of CRISPR/Cas systems have been applied, strategies for multiplex editing, methods for precise gene insertion and replacement, and efficient systems for base editing and site-specific transcriptional regulation. In addition, the biosecurity of CRISPR/Cas system is also discussed, including transgene-free methods and off-target effects of CRISPR/Cas system. Thus, the development and application of CRISPR/Cas system will have significant impact on functional genomic research and variety improvement in rice.

关键词： rice CRISPR Cas genome editing protospacer adjacent motif

A simple and efficient method for CRISPR/Cas9-induced mutant screening

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Journal of Genetics and Genomics 2017年第4期44卷 207-213页

作者： Yufeng Hua Chun wang Jian Huang kejian wang State Key Laboratory of Rice Biology China National Rice Research InstituteChinese Academy of Agricultural Sciences School of Biology & Basic Medical Sciences Soochow University

The clustered regularly interspaced short palindromic repeats（CRISPR）/CRISPR-associated protein 9（Cas9） system provides a technological breakthrough in mutant generation. Several methods such as the polymerase chain reaction（PCR）/restriction enzyme（RE） assay, T7 endonuclease I（T7EI） assay, Surveyor nuclease assay, PAGE-based genotyping assay, and high-resolution melting（HRM） analysis-based assay have been developed for screening CRISPR/Cas9-induced mutants. However, these methods are timeand labour-intensive and may also be sequence-limited or require very expensive equipment. Here, we described a cost-effective and sensitive screening technique based on conventional PCR, annealing at critical temperature PCR（ACT-PCR）, for identifying mutants. ACT-PCR requires only a single PCR step followed by agarose gel electrophoresis. We demonstrated that ACT-PCR accurately distinguished CRISPR/Cas9-induced mutants from wild type in both rice and zebrafish. Moreover, the method can be adapted for accurately determining mutation frequency in cultured cells. The simplicity of ACT-PCR makes it particularly suitable for rapid, large-scale screening of CRISPR/Cas9-induced mutants in both plants and animals.

关键词： ACT-PCR CRISPR/Cas9 Genome editing Mutant screening

CRISPR/Cas systems: The link between functional genes and genetic improvement

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The Crop Journal 2021年第3期9卷 678-687页

作者： Yong Huang Huirong Dong Meiqi Shang kejian wang State Key Laboratory of Rice Biology China National Rice Research InstituteChinese Academy of Agricultural SciencesHangzhou 310006ZhejiangChina

With the ever-increasing human population and deteriorating environmental conditions,there is an urgent need to breed environmentally friendly and resource-conserving rice cultivars to achieve sustainable agricultural development and food ***,conventional rice improvement strategies,such as hybrid breeding,are time-consuming and laborious processes and may not be able to keep pace with increasing food demand in the *** genome-editing technologies,especially clustered regularly interspaced short palindromic repeat(CRISPR)/CRISPR-associated protein(CRISPR/Cas),permit efficient targeted genome modification and offer great promise for the creation of desired plants with higher yield,improved grain quality,and resistance to herbicides,diseases,and insect *** is also great potential for tapping heterosis using the CRISPR/Cas *** this review,we focus on the most essential applications of CRISPR/Cas genome editing to rice genetic improvement,considering traits such as yield,quality,and herbicide,disease and insect-pest *** discuss applications of CRISPR/Cas to the exploitation of ***,we outline perspectives for future rice breeding using genome-editing technologies.

关键词： CRISPR/Cas Genome editing Rice Application Heterosis

Generation of marker-free transgenic rice using CRISPR/Cas9 system controlled by ?oral speci?c promoters

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Journal of Genetics and Genomics 2019年第1期46卷 61-64页

作者： Junjie wang Chun wang kejian wang State Key Laboratory of Rice Biology China National Rice Research Institute Chinese Academy of Agricultural Sciences

Recently, the CRISPR/Cas9 system has been used as a powerful tool for genome editing in many species (Jinek et al., 2012;Cong et al., 2013;Wright et al., 2016;Li et al., 2017;Deng et al., 2018). The CR1SPR/Cas9 system can not only be used as a useful technology to disrupt endogenous genes but also expand numerous other applications, such as precise base editing (Komor et al., 2016;Zong et al., 2017), regulation of gene expression (Gilbert et al., 2013), and gene replacement or insertion (wang et al., 2017).

关键词： the CRISPR/Cas9 system expand numerous precise base

Biodegradation and Mechanical Property of Polylactic Acid/Thermoplastic Starch Blends with Poly (ethylene glycol)

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Journal of Wuhan University of Technology(Materials Science) 2013年第1期28卷 157-162页

作者：薛平 wang kejian 贾明印 YANG Meijuan Institute of Plastics Machinery and Engineering Beijing University of Chemical Technology

The effects of adding poly （ethylene glycol）（PEG） into polylactic acid/thermoplastic starch blends （PLA/TPS） on the properties were investigated by DSC, SEM and mechanical property-testing. The blends of PLA/TPS blended with increasing content PEG exhibited lower temperature of glass transition （T） and lower temperature of melting （T） as well as higher melt flow index （MFI）, which indicates the plasticization and proeessability of the composites were dramatically improved. The tensile strength, flexural strength and izod impact strength of PLA/TPS （80/20） increased at first and then decreased with increasing content of PEG due to stronger interfacial adhesion. The optimized mechanical property can be obtained for the blend with 3 wt % PEG. The samples containing PEG after soil burial for 5 months showed quicker degradation being accompanied with large weight loss and mechanical properties loss.

关键词： biodegradation mechanical property polylactic acid starch

Regulation of metallothionein gene expression in response to benzo[a]pyrene exposure and bacterial challenge in marine cultured black porgy(Acanthopagrus schlegelii)

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Chinese Journal Of Geochemistry 2014年第4期33卷 404-410页

作者： YANG Ming PAN Chenyuan BO Jun wang kejian School of Environmental and Chemical Engineering Shanghai University State Key Laboratory of Marine Environmental Science Xiamen University

Metallothioneins(MTs) are normally considered to be sensitive indicators of heavy metal pollution, but it is less clear whether the MT gene can actively respond to other environmental stresses. In this study, an MT m RNA molecular sequence of 471 bp(full length) was identified in marine cultured black porgy(Acanthopagrus schlegelii), encoding 60 amino acids containing 20 cysteine residues. The MT sequence was highly homologous to that of other fish belonging to the MT superfamily type 1 family. The three dimensional structure of the deduced MT peptide was composed of two metal-binding domains capable of ligating divalent heavy metals. The MT m RNA transcripts were detected in the 11 tested tissues and the highest quantity was present in the liver. Stresses by two factors, benzo[a]pyrene(B[a]p) exposure and bacterial challenge, were evaluated on MT gene expression. The level of MT gene transcripts in the liver significantly declined 24 h post B[a]p exposure and the quantity was significantly correlated with the exposure time during a 24 h period. In contrast, MT gene expression in the liver was significantly increased 48 h post bacterial infection and the quantity was significantly correlated with the infection time during this period of 48 h. Our results indicated that MT gene expression in black porgy liver was sensitive to environmental stresses other than just the heavy metal pollution reported, suggesting that the development of a reliable biomarker for heavy metal pollution will be more complex than expected.

关键词：金属硫蛋白基因苯并[a]芘细菌感染海洋养殖曝光量黑鲷重金属污染 mRNA转录

The rice codebook: From reading to editing

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Molecular Plant 2022年第4期15卷 569-572页

作者： kejian wang Huanbin Zhou Qian Qian State Key Laboratory of Rice Biology China National Rice Research InstituteChinese Academy of Agricultural SciencesHangzhou 310006China State Key Laboratory for Biology of Plant Diseases and Insect Pests Institute of Plant ProtectionChinese Academy of Agricultural SciencesBeijing 100193China

Rice(Oryza sativa)is the main staple food crop that feeds more than half of the world's *** its relatively small genome size,abundant genetic resources,and highly efficient transformation technology,rice also remains as an excellent model system for monocot *** 2002,the International Rice Genome Sequencing Project(IRGSP)released the draft genome sequence of rice variety Nipponbare,the first crop genetic codebook deciphered right after the human genome(Yu et al.,2002).Since then,rice genome annotation,large-scale genome resequencing of diverse rice germplasm accessions,a comprehensive map of rice genome variations,and high-quality reference genome sequence,especially the gap-free reference genome,were completed successively in the next two decades along with the development of high-throughput DNA sequencing techniques(wang et al.,2018;Li et al.,2021;Song et al.,2021).Consequently,rice functional genomics research has boomed rapidly,greatly reshaping our knowledge of plant molecular biology and designed crop breeding.

关键词： sativa germplasm successively

Development and Application of Prime Editing in Plants

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Rice science 2023年第6期30卷 509-522页

作者： LIU Tingting ZOU Jinpeng YANG Xi wang kejian RAO Yuchun wang Chun State Key Laboratory of Rice Biology and Breeding China National Rice Research InstituteHangzhou 310006China College of Life Science and Technology Huazhong Agricultural UniversityWuhan 430070China College of Life Sciences Zhejiang Normal UniversityJinhua 321004China

Clustered regularly interspaced palindromic repeats(CRISPR)/CRISPR-associated protein(Cas)-mediated genome editing has greatly accelerated progress in plant genetic research and agricultural breeding by enabling targeted genomic ***,the prime editing system,derived from the CRISPR/Cas system,has opened the door for even more precise genome *** editing has the capability to facilitate all 12 types of base-to-base conversions,as well as desired insertions or deletions of fragments,without inducing double-strand breaks and requiring donor DNA *** a short time,prime editing has been rapidly verified as functional in various plants,and can be used in plant genome functional analysis as well as precision breeding of *** this review,we summarize the emergence and development of prime editing,highlight recent advances in improving its efficiency in plants,introduce the current applications of prime editing in plants,and look forward to future prospects for utilizing prime editing in genetic improvement and precision molecular breeding.

关键词： prime editing CRISPR/Cas precision genome editing crop breeding