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Creating a novel herbicide-tolerance OsALS allele using CRISPR/Cas9-mediated gene editing

The Crop Journal 2021年第2期9卷 305-312页

作者： Fangquan Wang Yang Xu Wenqi Li Zhihui Chen Jun Wang Fangjun Fan yajun tao Yanjie Jiang Qian-Hao Zhu Jie Yang Institute of Food Crops Jiangsu Academy of Agricultural Sciences/Nanjing Branch of Chinese National Center for Rice ImprovementNanjing 210014JiangsuChina Jiangsu Co-Innovation Center for Modern Production Technology of Grain Crops Yangzhou UniversityYangzhou 225009JiangsuChina Provincial Key Laboratory of Agrobiology Jiangsu Academy of Agricultural SciencesNanjing 210014JiangsuChina CSIRO Agriculture and Food GPO Box 1700CanberraACT 2601Australia Institute of Life Science Jiangsu UniversityZhenjiang 212013JiangsuChina

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.

关键词： Rice CRISPR/Cas9 ALS Herbicide-tolerance Crop breeding

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Intron-targeted gene insertion in rice using CRISPR/Cas9: A case study of the Pi-ta gene

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The Crop Journal 2020年第3期8卷 424-431页

作者： Yang Xu Fangquan Wang Zhihui Chen Jun Wang Wen-Qi Li Fangjun Fan yajun tao Ling Zhao Weigong Zhong Qian-Hao Zhu Jie Yang Institute of Food Crops Jiangsu Academy of Agricultural Sciences/Nanjing Branch of Chinese National Center for Rice ImprovementNanjing 210014JiangsuChina Jiangsu Co-Innovation Center for Modern Production Technology of Grain Crops Yangzhou UniversityYangzhou 225009JiangsuChina Provincial Key Laboratory of Agrobiology Jiangsu Academy of Agricultural SciencesNanjing 210014JiangsuChina CSIRO Agriculture and Food GPO Box 1700Canberra ACT 2601Australia Institute of Life Science Jiangsu UniversityZhenjiang 212013JiangsuChina

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.

关键词： Rice CRISPR/Cas9 Pi-ta Genome editing Molecular breeding

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Regulating surface electron structure of PtNi nanoalloy via boron doping for high‐current‐density Li‐O2 batteries with low overpotential and long‐life cyclability

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SmartMat 2024年第1期5卷 110-120页

作者： yajun Ding Yuanchao Huang Yuejiao Li tao Zhang Zhong‐Shuai Wu State Key Laboratory of Catalysis Dalian Institute of Chemical PhysicsChinese Academy of SciencesDalianChina State Key Laboratory of Silicon Materials and School of Materials Science and Engineering Zhejiang UniversityHangzhouChina Institute of Advanced Semiconductors Hangzhou Innovation CenterZhejiang UniversityHangzhouChina University of Chinese Academy of Sciences BeijingChina State Key Laboratory of High Performance Ceramics and Superfine Microstructure Shanghai Institute of CeramicsChinese Academy of SciencesShanghaiChina Dalian National Laboratory for Clean Energy Chinese Academy of SciencesDalianChina

The realization of high‐efficiency,reversible,stable,and safe Li‐O2 batteries is severely hindered by the large overpotential and side reactions,especially at high rate conditions.Therefore,rational design of cathode catalysts with high activity and stability is crucial to overcome the terrible issues at high current density.Herein,we report a surface engineering strategy to adjust the surface electron structure of boron(B)‐doped PtNi nanoalloy on carbon nanotubes(PtNiB@CNTs)as an efficient bifunctional cathodic catalyst for high‐rate and long‐life Li‐O2 batteries.Notably,the Li‐O2 batteries assembled with as‐prepared PtNiB@CNT catalyst exhibit ultrahigh discharge capacity of 20510 mA·h/g and extremely low overpotential of 0.48 V at a high current density of 1000 mA/g,both of which outperform the most reported Pt‐based catalysts recently.Meanwhile,our Li‐O2 batteries offer excellent rate capability and ultra‐long cycling life of up to 210 cycles at 1000 mA/g under a fixed capacity of 1000 mA·h/g,which is two times longer than those of Pt@CNTs and PtNi@CNTs.Furthermore,it is revealed that surface engineering of PtNi nanoalloy via B doping can efficiently tailor the electron structure of nanoalloy and optimize the adsorption of oxygen species,consequently delivering excellent Li‐O2 battery performance.Therefore,this strategy of regulating the nanoalloy by doping nonmetallic elements will pave an avenue for the design of high‐performance catalysts for metal‐oxygen batteries.

关键词： B doping bifunctional catalyst Li‐O2 battery low charge overpotential PtNi nanoalloy

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基于HRM技术开发水稻糊化温度基因ALK功能标记

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中国水稻科学 2024年第1期38卷 106-110页

作者：王军周晶陶亚军李文奇朱建平范方军王芳权许扬陈智慧蒋彦婕李霞杨杰江苏省农业科学院粮食作物研究所/农业农村部淮河下游种质创制重点实验室南京210014 扬州大学生物科学与技术学院江苏扬州225009 扬州大学/江苏省粮食作物现代产业技术协同创新中心江苏扬州225009

【目的】糊化温度是影响稻米蒸煮食味品质的重要指标,ALK是控制水稻糊化温度的主效基因,开发可快速高通量鉴定ALK基因型的功能标记有助于提高水稻品质改良的效率。【方法】根据ALK基因4198位、4329位和4330位存在的单碱基差异,设计基于... 详细信息

【目的】糊化温度是影响稻米蒸煮食味品质的重要指标,ALK是控制水稻糊化温度的主效基因,开发可快速高通量鉴定ALK基因型的功能标记有助于提高水稻品质改良的效率。【方法】根据ALK基因4198位、4329位和4330位存在的单碱基差异,设计基于HRM技术检测的基因功能标记。【结果】通过PCR检测结合测序分析,筛选了ALK基因2个功能区域的功能标记ALKH4、ALKH5。利用ALKH4、ALKH5对81份籼稻品种、279份粳稻品种进行了ALK基因型检测,结果发现,16份粳稻和19份籼稻为G-GC基因型;51份粳稻为A-GC基因型;212份粳稻和62份籼稻为G-TT基因型。【结论】基于HRM技术开发的基因功能标记ALKH4、ALKH5可以快速高通量鉴定控制糊化温度ALK不同基因型,具有重要的应用价值。

关键词：水稻糊化温度 ALK 功能标记 HRM技术

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Rational Design of Grain Size to Improve Rice Yield and Quality

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Rice science 2023年第1期30卷 1-5页

作者： tao yajun WANG Jun XU Yang WANG Fangquan LI Wenqi JIANG Yanjie CHEN Zhihui FAN Fangjun ZHU Jianping LI Xia YANG Jie Institute of Food Crops Jiangsu Academy of Agricultural Sciences/Nanjing Branch of Chinese National Center for Rice ImprovementNanjing 210014China Jiangsu Co-Innovation Center for Modern Production Technology of Grain Crops Yangzhou UniversityYangzhou 225009China

Grain size, determined by grain length, grain width and grain thickness, is associated with grain yield and quality. Many genes controlling grain size were cloned and their related regulatory mechanisms were clearly clarified. However, whether these genes can be directly introduced into japonica rice for grain size improvement is unknown.

关键词： Yield Rational size

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Duplicated chalcone synthase(CHS)genes modulate flavonoid production in tea plants in response to light stress

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Journal of Integrative Agriculture 2024年第6期23卷 1940-1955页

作者： Mingzhuo Li Wenzhao Wang Yeru Wang Lili Guo yajun Liu Xiaolan Jiang Liping Gao tao Xia State Key Laboratory of Tea Plant Biochemistry and Utilization Anhui Agricultural UniversityHefei 230036China School of Life Sciences Anhui Agricultural UniversityHefei 230036China College of Horticulture Northwest A&F UniversityYangling 712100China Department of Plant and Microbial Biology North Carolina State UniversityRaleigh 27606NCUSA

In tea plants,the abundant flavonoid compounds are responsible for the health benefits for the human body and define the astringent flavor profile.While the downstream mechanisms of flavonoid biosynthesis have been extensively studied,the role of chalcone synthase(CHS)in this secondary metabolic process in tea plants remains less clear.In this study,we compared the evolutionary profile of the flavonoid metabolism pathway and discovered that gene duplication of CHS occurred in tea plants.We identified three CsCHS genes,along with a CsCHS-like gene,as potential candidates for further functional investigation.Unlike the CsCHS-like gene,the CsCHS genes effectively restored flavonoid production in Arabidopsis chs-mutants.Additionally,CsCHS transgenic tobacco plants exhibited higher flavonoid compound accumulation compared to their wild-type counterparts.Most notably,our examination of promoter and gene expression levels for the selected CHS genes revealed distinct responses to UV-B stress in tea plants.Our findings suggest that environmental factors such as UV-B exposure could have been the key drivers behind the gene duplication events in CHS.

关键词： tea flavonoids biosynthesis CHS gene duplication UV-B stress

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Effect of VEGF/GREDVY Modified Surface on Vascular Cells Behavior

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Journal of Wuhan University of Technology(Materials Science) 2024年第1期39卷 244-254页

作者：魏来 TAN Jianying LI Li WANG Huanran LIU Sainan ZENG Zheng LIU tao WANG Jian 陈俊英 WENG yajun School of Materials Science and Engineering Southwest Jiaotong UniversityChengdu 610031 China

We synthesized B-He/B-GREDVY and immobilized them on avidin-coated surfaces.To examine the immobilization of molecules in the material, the following experiments were performed:fluorescein isothiocyanate(FITC) fluorescence staining, water contact angle and atomic force microscopy(AFM) measurements. Besides, the biological evaluation experiments were also performed, such as platelets adhesion and activation, the culturing of smooth muscle cells(SMC) and endothelial cells(EC). These experimental results show that the modified surfaces could prevent the hyperproliferation of SMC, and promote the proliferation and migration of EC and EPC. Furthermore, the adding of VEGF improved the EC adhesion in a dynamic environment. Generally, it is expected that the modified surfaces could be used to accelerate the formation of the newly endothelial layer for the construction of platforms for coronary artery stent therapy.

关键词： biotin-GREDVY VEGF anticoagulation endothelial cells endothelial progenitor cells

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Suppression of OsSPDS2 improves grain size and yield production in rice(Oryza sativa L.)

Suppression of OsSPDS2 improves grain size and yield product...

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2017中国长三角遗传学大会

作者： yajun tao Jun Wang Yong Zhou Guohua Liang Jiangsu Key Laboratory of Crop Genetics and Physiology/Co-lnnovation Center for Modern Production Technology of Grain Crops Key Laboratory of Plant Functional Genomics of the Ministry of EducationYangzhou University Institute of Food Crops Jiangsu Academy of Agricultural Sciences

Polyamines are a class of low-molecular-weight aliphatic cations with strong biological activities,and they are involved in all stages of growth and development in plants.However,the mechanism of spermidine synthase regulating seed size has been little examined so far.We cloned spermidine synthase 2(OsSPDS2) and constructed over-expression,RNA interference(RNAi) vectors.From seeding stage,we observed that the plants with OsSPDS2 suppressed showed stronger growing vigor compared with OsSPDS2-OE lines.When tillering,the difference becomes more pronounced.At maturity,we measured several agronomic traits of Nipponbare and transgenic lines.The suppressed lines(RNAil and RNAi2) with down-regulated 0 s5 PD52 expression showed higher plant height,bigger seed size and higher grain yield,displaying stronger growing vigor.Oppositely,the up-regulated OsSPDS2 resulted smaller plant architecture compared to Nipponbare.Unexpectedly,we found OsSPDS2 also affect seed dormancy.Our results suggest that OsSPDS2 is involved into polyaminesynthetic pathway and affect the synthesis of ethylene to regulate seed size and seed dormancy in rice.

关键词： Rice OsSPDS2 Grain size Yield production

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Efficient Improvement of Nutritional Content in Rice Grains by Precise Base Editing of OsROS1

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Rice science 2023年第6期30卷 499-503页

作者： XU Yang WANG Fangquan LI Wenqi WANG Jun tao yajun FAN Fangjun CHEN Zhihui JIANG Yanjie ZHU Jianping LI Xia ZHU Qianhao YANG Jie Key Laboratory of Germplasm Innovation in Downstream of Huaihe River Ministry of Agriculture and Rural Affairs/Institute of Food CropsJiangsu Academy of Agricultural SciencesNanjing 210014China Jiangsu Co-Innovation Center for Modern Production Technology of Grain Crops Yangzhou UniversityYangzhou 225009China Institute of Life Science Jiangsu UniversityZhenjiang 212013China Commonwealth Scientific and Industrial Research Organisation Agriculture and Food GPO Box 1700CanberraACT 2601Australia

Aleurone forms the outermost layer of the rice endosperm and plays a critical role in apoplastic nutrient uptake during endosperm development.Thickening the aleurone layer has been proposed to significantly increase the nutrient content of rice grains.In this study,we used a CRISPR/Cas9-mediated precise base editing method to target OsROS1,a gene associated with aleurone thickness,in the background of the japonica glutinous rice cultivar Zhennuo 19。

关键词： CRISPR/Cas9 cultivar precise

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Camellia sinensis CsMYB4a participates in regulation of stamen growth by interaction with auxin signaling transduction repressor CsAUX/IAA4

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The Crop Journal 2024年第1期12卷 188-201页

作者： Guoliang Ma Mingzhuo Li Yingling Wu Changjuan Jiang Yifan Chen Dawei Xing Yue Zhao yajun Liu Xiaolan Jiang tao Xia Liping Gao School of Life Science Anhui Agricultural UniversityHefei 230036AnhuiChina Plant for Human Health Institute Department of Plant and Microbial BiologyNorth Carolina State UniversityKannapolisNC 28081USA State Key Laboratory of Tea Plant Biology and Utilization/Key Laboratory of Tea Biology and Tea Processing of Ministry of Agriculture/Anhui Provincial Laboratory of Tea Plant Biology and Utilization Anhui Agricultural UniversityHefei 230036AnhuiChina

Subgroup 4(Sg4)members of the R2R3-MYB are generally known as negative regulators of the phenylpropanoid pathway in plants.Our previous research showed that a R2R3-MYB Sg4 member from Camellia sinensis(CsMYB4a)inhibits expression of some genes in the phenylpropanoid pathway,but its physiological function in the tea plant remained unknown.Here,CsMYB4a was found to be highly expressed in anther and filaments,and participated in regulating filament growth.Transcriptome analysis and exogenous auxin treatment showed that the target of CsMYB4a might be the auxin signal pathway.Auxin/indole-3-acetic acid 4(AUX/IAA4),a repressor in auxin signal transduction,was detected from a yeast two-hybrid screen using CsMYB4a as bait.Gene silencing assays showed that both CsIAA4 and CsMYB4a regulate filament growth.Tobacco plants overexpressing CsIAA4 were insensitive to exogenous a-NAA,consistent with overexpression of CsMYB4a.Protein-protein interaction experiments revealed that CsMYB4a interacts with N-terminal of CsIAA4 to prevent CsIAA4 degradation.Knock out of the endogenous NtIAA4 gene,a CsIAA4 homolog,in tobacco alleviated filament growth inhibition and a-NAA insensitivity in plants overexpressing CsMYB4a.All results strongly suggest that CsMYB4a works synergistically with CsIAA4 and participates in regulation of the auxin pathway in stamen.

关键词： AUX/IAA4 Auxin signaling CsMYB4a Subgroup 4 R2R3-MYB

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