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| 1 | Systematic identification of endogenous RNA polymeraseⅢpromoters for efficient RNA guidebased genome editing technologies in maize显示文摘Single-guide RNA(sg RNA) is one of the two core components of the CRISPR(clustered regularly interspaced short palindromic repeat)/Cas(CRISPR-associated) genome-editing technology. We established an in vitro Traffic Light Reporter(TLR) system, which is designated as the same colors as traffic lights such as green, red and yellow were produced in cells. The TLR can be readily used in maize mesophyll protoplast for a quick test of promoter activity. The TLR assay indicates the variation in transcription activities of the seven Pol III promoters, from 3.4%(U6-1) to over 21.0%(U6-6). The U6-2 promoter, which was constructed to drive sg RNA expression targeting the Zm Wx1 gene, yielded mutation efficiencies ranging from 48.5% to 97.1%. Based on the reported and unpublished data, the in vitro TLR assay results were confirmed to be a readily system and may be extended to other plant species amenable to efficient genome editing via CRISPR/Cas. Our efforts provide an efficient method of identifying native Pol III-recognized promoters for RNA guide-based genome-editing systems in maize. | Xiantao Qi Le Dong Changlin Liu Long Mao Fang Liu Xin Zhang Beijiu Cheng Chuanxiao Xie | 2018 | The Crop Journal2018，6，3： | 7 |
| 2 | Creation of targeted inversion mutations in plants using an RNA-guided endonuclease显示文摘Inversions are DNA rearrangements that are essential for plant gene evolution and adaptation to environmental changes. We demonstrate the creation of targeted inversions and previously reported targeted deletion mutations via delivery of a pair of RNA-guided endonucleases(RGENs) of CRISPR/Cas9. The efficiencies of the targeted inversions were2.6% and 2.2% in the Arabidopsis FLOWERING TIME(At FT) and TERMINAL FLOWER 1(At TFL1)loci, respectively. Thus, we successfully established an approach that can potentially be used to introduce targeted DNA inversions of interest for functional studies and crop improvement. | Congsheng Zhang Changlin Liu Jianfeng Weng Beijiu Cheng Fang Liu Xinhai Li Chuanxiao Xie | 2017 | The Crop Journal2017，5，1： | 6 |
| 3 | Conversion of a normal maize hybrid into a waxy version using in vivo CRISPR/Cas9 targeted mutation activity显示文摘Waxy maize is a specialty maize that produces mainly amylopectin starch with special food or industrial values. The objective of this study was to overcome the limitations of wx mutant allele acquisition and breeding efficiency by conversion of parental lines from normal to waxy maize. The intended mutation activity was achieved by in vivo CRISPR/Cas9 machinery involving desired-target mutation of the Wx locus in the ZC01 background,abbreviated as ZC01-DTM^(wx). Triple selection was applied to segregants to obtain high genome background recovery with transgene-free wx mutations. The targeted mutation was identified, yielding six types of mutations among progeny crossed with ZC01-DTM^(wx).The amylopectin contents of the endosperm starch in mutant lines and hybrids averaged94.9%, while those of the wild-type controls were significantly(P < 0.01) lower, with an average of 76.9%. Double selection in transgene-free lines was applied using the Bar strip test and Cas9 PCR screening. The genome background recovery ratios of the lines were determined using genome-wide SNP data. That of lines used as male parents was as high as98.19% and that of lines used as female parents was as high as 86.78%. Conversion hybrids and both parental lines showed agronomic performance similar to that of their wild-type counterparts. This study provides a practical example of the efficient extension of CRISPR/Cas9 targeted mutation to industrial hybrids for transformation of a recalcitrant species. | Xiantao Qi Hao Wu Haiyang Jiang Jinjie Zhu Changling Huang Xin Zhang Changlin Liu Beijiu Cheng | 2020 | The Crop Journal2020，8，3： | 4 |
| 4 | Establishment of an efficient seed fluorescence reporter-assisted CRISPR/Cas9 gene editing in maize显示文摘Genome editing by clustered regularly interspaced short palindromic sequences(CRISPR)/CRISPRassociated protein 9(Cas9)has revolutionized functional gene analysis and genetic improvement.While reporter-assisted CRISPR/Cas systems can greatly facilitate the selection of genome-edited plants produced via stable transformation,this approach has not been well established in seed crops.Here,we established the seed fluorescence reporter(SFR)-assisted CRISPR/Cas9 systems in maize(Zea mays L.),using the red fluorescent Ds RED protein expressed in the endosperm(En-SFR/Cas9),embryos(Em-SFR/Cas9),or both tissues(Em/En-SFR/Cas9).All three SFRs showed distinct fluorescent patterns in the seed endosperm and embryo that allowed the selection of seeds carrying the transgene of having segregated the transgene out.We describe several case studies of the implementation of En-SFR/Cas9,Em-SFR/Cas9,and Em/En-SFR/Cas9 to identify plants not harboring the genomeediting cassette but carrying the desired mutations at target genes in single genes or in small-scale mutant libraries,and report on the successful generation of single-target mutants and/or mutant libraries with En-SFR/Cas9,Em-SFR/Cas9,and Em/En-SFR/Cas9.SFR-assisted genome editing may have particular value for application scenarios with a low transformation frequency and may be extended to other important monocot seed crops. | Yuanyuan Yan Jinjie Zhu Xiantao Qi Beijiu Cheng Changlin Liu Chuanxiao Xie | 2021 | Journal of Integrative Plant Biology2021，63，9： | 3 |
| 5 | Anaerobic biodegradation,physical and structural properties of normal and high-amylose maize starch films显示文摘Biodegradable plastics have attracted considerable attention in recent years due to their biodegradability,biocompatibility and non-toxicity.In this study,normal maize starch(containing 25%amylose)and high-amylose maize starch(containing 80%amylose)were served as model materials to prepare starch/polyvinyl alcohol(PVA)blends.To comprehensively study the effects of amylose contents on the film performances,the mechanical properties,water resistance and anaerobic biodegradability of the two films were examined.Moreover,the processes of anaerobic degradation were investigated by evolutions of biogas production,pH in reactors and the changes of film structures and compositions.The results indicated that amylose content played an important role in the microstructures of starch film as well as mechanical properties and water resistance,whereas it had no significant influence on anaerobic biodegradability of the films.Nonetheless,the structure of high-amylose maize starch/PVA film was more suitable and beneficial to the anaerobic biodegradation than that of the normal maize starch/PVA film,because it could effectively avoid accumulation of volatile fatty acids,which contributed to the stable biogas production,short fermentation period and non-souring in the reactor. | Liu Weiwei Xue Juan Cheng Beijiu Zhu Suwen Ma Qing Ma Huan | 2016 | International Journal of Agricultural and Biological Engineering2016，9，5： | 3 |
| 6 | Genome-Wide Analysis of BURP Domain-Containing Genes in Populus trichocarpa显示文摘BURP domain-containing proteins have a conserved structure and are found extensively in plants.The functions of the proteins in this family are diverse,but remain unknown in Populus trichocarpa.In the present study,a complete genome of P.trichocarpa was analyzed bioinformatically.A total of 18 BURP family genes,named PtBURPs,were identified and characterized according to their physical positions on the P.trichocarpa chromosomes.A phylogenetic tree was generated from alignments of PtBURP protein sequences,while phylogenetic relationships were also examined between PtBURPs and BURP family genes in other plants,including rice,soybean,maize and sorghum.BURP genes in P.trichocarpa were classified into five classes,namely PG1β-like,BNM2-like,USP-like,RD22-like and BURP V.The multiple expectation maximization for motif elicitation(MEME) and multiple protein sequence alignments of PtBURPs were also performed.Results from the transcript level analyses of 10 PtBURP genes under different stress conditions revealed the expression patterns in poplar and led to a discussion on genome duplication and evolution,expression profiles and function of PtBURP genes. | Yuanhua Shao Guo Wei Ling Wang Qing Dong Yang Zhao Beijiu Chen Yan Xiang | 2011 | Journal of Integrative Plant Biology2011，53，9： | 2 |
| 7 | Genome-wide identification and characterization of maize expansin genes expressed in endosperm显示文摘 | Wei Zhang Hanwei Yan Weijun Chen Jinyang Liu Cuiping Jiang Haiyang Jiang Suwen Zhu Beijiu Cheng | 2014 | Molecular Genetics and Genomics2014，，6： | 1 |
| 8 | Genome-wide survey and characterization of the WRKY gene family in Populus trichocarpa显示文摘 | Hongsheng He Qing Dong Yuanhua Shao Haiyang Jiang Suwen Zhu Beijiu Cheng Yan Xiang | 2012 | Plant Cell Reports2012，，7： | 1 |
| 9 | Bacterially expressed dsRNA protects maize against SCMV infection显示文摘 | Defang Gan Jiao Zhang Haibo Jiang Tong Jiang Suwen Zhu Beijiu Cheng | 2010 | Plant Cell Reports2010，，11： | 1 |
| 10 | SDG721 and SDG705 are required for rice growth显示文摘H3K4me3 plays important roles in development, transcription, and environmental responses. Here,we report that SDG721(SET-domain group protein 721)and SDG705 are involved in regulating rice development.SDG721 and SDG705 encode TRITHORAX-like proteins,which appear to modulate H3K4methylation levels. Loss of SDG721 and SDG705 function resulted in GA-deficient phenotypes, including semi-dwarfism, reduced cell length, and reduced panicle branching. The transcripts levels and H3K4me3 levels of GA biosynthesis genes and GA signaling pathway genes were downregulated in the sdg721 sdg705 plants. Together, these results suggest that SDG721 and SDG705 regulate H3K4methylation,which is crucial for plant development in rice. | Pengfei Jiang Shiliang Wang Aziz Ul Ikram Zuntao Xu Haiyang Jiang Beijiu Cheng Yong Ding | 2018 | Journal of Integrative Plant Biology2018，60，7： | 1 |
| 11 | Overexpression of a maize WRKY58 gene enhances drought and salt tolerance in transgenic rice显示文摘 | Ronghao Cai Yang Zhao Yufu Wang Yongxiang Lin Xiaojian Peng Qian Li Yuwei Chang Haiyang Jiang Yan Xiang Beijiu Cheng | 2014 | Plant Cell, Tissue and Organ Culture (PCTOC)2014，，3： | 1 |
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