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CrisprStitch:Fast evaluation of the efficiency of CRISPR editing systems

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plant Communications 2024年第3期5卷 13-16页

作者： Yangshuo Han Guanqing Liu Yuechao Wu Yu Bao Yong Zhang Tao Zhang Jiangsu key laboratory of Crop Genomics and Molecular Breeding/Zhongshan Biological Breeding laboratory/key laboratory of plant Functional Genomics of the Ministry of Education Agricultural College of Yangzhou UniversityYangzhou 225009China Jiangsu Co-Innovation Center for Modern Production Technology of Grain Crops/Jiangsu key laboratory of Crop Genetics and physiology Yangzhou UniversityYangzhou 225009China Chongqing key laboratory of plant Resource Conservation and Germplasm Innovation Integrative Science Center of Germplasm Creation in Western China(Chongqing)Science CitySchool of Life SciencesSouthwest UniversityChongqing 400715China

Dear Editor,Targeted genome-editing technology using designed nucleases has been rapidly advancing and catalyzing significant breakthroughs in the life ***-designed CRISPR editing experiments can be efficiently evaluated through nextgeneration sequencing(NGS).Nevertheless,NGS data analysis currently lacks a user-friendly pipeline capable of automatically calculating mutations and evaluating editing efficiency in bulk,as well as providing a more comprehensive visualization of results.

关键词： breakthrough editing efficiency

Rice canopy temperature is affected by nitrogen fertilizer

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Journal of Integrative Agriculture 2024年第3期23卷 824-835页

作者： Min Jiang Zhang Chen Yuan Li Xiaomin Huang Lifen Huang Zhongyang Huo Jiangsu key laboratory of Crop Genetics and physiology/Jiangsu key laboratory of Crop Cultivation and physiology/Agricultural College Yangzhou UniversityYangzhou 225009China Jiangsu Co-lnnovation Center for Modern Production Technology of Grain Crops Yangzhou UniversityYangzhou 225009China

Canopy temperature strongly influences crop yield formation and is closely related to plant physiological ***, the effects of nitrogen treatment on canopy temperature and rice growth have yet to be comprehensively examined. We conducted a two-year field experiment with three rice varieties(HD-5, NJ-9108, and YJ-805) and three nitrogen treatments(zero-N control(CK), 200 kg ha~(–1)(MN), and 300 kg ha~(–1)(HN)). We measured canopy temperature using a drone equipped with a high-precision camera at the six stages of the growth period. Generally,canopy temperature was significantly higher for CK than for MN and HN during the tillering, jointing, booting, and heading stages. The temperature was not significantly different among the nitrogen treatments between the milky and waxy stages. The canopy temperature of different rice varieties was found to follow the order: HD-5>NJ-9108>YJ-805, but the difference was not significant. The canopy temperature of rice was mainly related to plant traits, such as shoot fresh weight(correlation coefficient r=–0.895), plant water content(–0.912), net photosynthesis(–0.84), stomatal conductance(–0.91), transpiration rate(–0.90), and leaf stomatal area(–0.83). A structural equation model(SEM) showed that nitrogen fertilizer was an important factor affecting the rice canopy *** study revealed:(1) A suite of plant traits was associated with the nitrogen effects on canopy temperature,(2) the heading stage was the best time to observe rice canopy temperature, and(3) at that stage, canopy temperature was negatively correlated with rice yield, panicle number, and grain number per panicle. This study suggests that canopy temperature can be a convenient and accurate indicator of rice growth and yield prediction.

关键词： canopy temperature rice physiological and biochemical characteristics yield

Biosynthetic pathway of prescription cucurbitacin IIa and high-level production of key triterpenoid intermediates in engineered yeast and tobacco

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plant Communications 2024年第6期5卷 154-172页

作者： Geng Chen Zhaokuan Guo Yanyu Shu Yan Zhao Lei Qiu Shaofeng Duan Yuan Lin Simei He Xiaobo Li Xiaolin Feng Guisheng Xiang Bo Nian Yina Wang Zhiyuan Li Chongkang Yang Yang Shi Yingchun Lu Guanze Liu Shengchao Yang Guanghui Zhang Bing Hao state key laboratory of Conservation and Utilization of Bio-resources in Yunnan The Key Laboratory of Medicinal Plant Biology of Yunnan ProvinceNational&Local Joint Engineering Research Center on Germplasms Innovation&Utilization of Chinese Medicinal Materials in Southwest ChinaYunnan Agricultural UniversityKunming 650201China Yunnan Characteristic plant Extraction laboratory KunmingYunnan 650106China

Cucurbitacin IIa is a triterpenoid isolated exclusively from Hemsleya plants and a non-steroidal anti-inflammatory drug that functions as the main ingredient of prescription Hemslecin capsules and tablets in *** biology provides new strategies for production of such valuable cucurbitacins at a large scale;however,the biosynthetic pathway of cucurbitacin IIa has been unknown,and the heterologous production of cucurbitacins in galactose medium has been expensive and low *** this study,we characterized the functions of genes encoding two squalene epoxidases(HcSE1-2),six oxidosqualene cyclases(HcOSC1-6),two CYP450s(HcCYP87D20 and HcCYP81Q59),and an acyltransferase(HcAT1)in cucurbitacin IIa biosynthesis by heterologous expression in Saccharomyces cerevisiae and Nicotiana *** achieved high-level production of the key cucurbitacin precursor 11-carbonyl-20b-hydroxy-Cuol from glucose in yeast via modular engineering of the mevalonate pathway and optimization of P450 expression *** resulting yields of 46.41 mg/l 11-carbonyl-20b-hydroxy-Cuol and 126.47 mg/l total cucurbitacin triterpenoids in shake flasks are the highest yields yet reported from engineered ***,production of 11-carbonyl-20b-hydroxy-Cuol by transient gene expression in tobacco resulted in yields of 1.28 mg/g dry weight in *** work reveals the key genes involved in biosynthesis of prescription cucurbitacin IIa and demonstrates that engineered yeast cultivated with glucose can produce high yields of key triterpenoid *** describe a low-cost and highly efficient platform for rapid screening of candidate genes and high-yield production of pharmacological triterpenoids.

关键词： cucurbitacin IIa oxidosqualene cyclase OSC 11-carbonyl-20b-hydroxy-Cuol Saccharomyces cerevisiae Nicotiana benthamiana Hemsleya chinensis

Effect of mutations on acetohydroxyacid synthase(AHAS)function in Cyperus difformis L.

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Journal of Integrative Agriculture 2024年第1期23卷 177-186页

作者： Xiaotong Guo Xiangju Li Zheng Li Licun Peng Jingchao Chen Haiyan Yu Hailan Cui state key laboratory for Biology of plant Diseases and Insect Pests Institute of Plant ProtectionChinese Academy of Agricultural SciencesBeijing 100193China College of Resources and Environmental Sciences China Agricultural UniversityBeijing 100193China

Cyperus difformis *** a troublesome weed in paddy fields and has attracted attention due to its resistance to acetohydroxyacid synthase(AHAS)*** was found that the amino acid mutation in AHAS was the primary cause for the resistance of Cyperus ***,the effect of different mutations on AHAS function is not clear in Cyperus *** confirm the effect of mutations on AHAS function,six biotypes were collected,including Pro197Arg,Pro197Ser,Pro197Leu,Asp376Glu,Trp574Leu and wild type,from Hunan,Anhui,Jiangxi and Jiangsu provinces,China and the function of AHAS was *** AHAS in vitro inhibition assay results indicated that the mutations decreased the sensitivity of AHAS to pyrazosulfuron-ethyl,in which the I_(50)(the half maximal inhibitory concentration)of wild type AHAS was 0.04μmol L^(-1)and Asp376Glu,Pro197Leu,Pro197Arg,Pro197Ser and Trp574Leu mutations were 3.98,11.50,40.38,38.19 and 311.43μmol L^(-1),*** the determination of enzyme kinetics parameters,the Km and the maximum reaction velocity(Vmax)of the wild type were 5.18 mmol L^(-1)and 0.12 nmol mg^(-1)min^(-1),respectively,and the Km values of AHAS with Asp376Glu,Trp574Leu,Pro197Leu and Pro197Ser mutations were 0.38-0.93 times of the wild *** Km value of the Pro197Arg mutation was 1.14times of the wild type,and the Vmax values of the five mutations were 1.17-3.33-fold compared to the wild *** was found that the mutations increased the affinity of AHAS to the substrate,except for the Pro197Arg *** a concentration of 0.0032-100 mmol L^(-1)branched-chain amino acids(BCAAs),the sensitivity of the other four mutant AHAS biotypes to feedback inhibition decreased,except for the Pro197Arg *** study elucidated the effect of different mutations on AHAS function in Cyperus difformis and provided ideas for further study of resistance development.

关键词： acetohydroxyacid synthase(AHAS) mutation enzyme function Cyperus difformis

Integrative regulatory mechanisms of stomatal movements under changing climate

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Journal of Integrative plant Biology 2024年第3期66卷 368-393页

作者： Jingbo Zhang Xuexue Chen Yajing Song Zhizhong Gong state key laboratory of Nutrient Use and Management College of Resources and Environmental SciencesNational Academy of Agriculture Green DevelopmentChina Agricultural UniversityBeijing 100193China state key laboratory of plant Cell and Chromosome Engineering Institute of Genetics and Developmental BiologyChinese Academy of SciencesBeijing 100101China state key laboratory of plant Environmental Resilience College of Biological SciencesChina Agricultural UniversityBeijing 100094China Institute of Life Science and Green Development School of Life SciencesHebei UniversityBaoding 071001China

Global climate change-caused drought stress,high temperatures and other extreme weather profoundly impact plant growth and development,restricting sustainable crop *** cope with various environmental stimuli,plants can optimize the opening and closing of stomata to balance CO_(2)uptake for photosynthesis and water loss from *** cells perceive and integrate various signals to adjust stomatal pores through turgor pressure *** mechanisms and signaling networks underlying the stomatal movements in response to environmental stresses have been extensively studied and *** review focuses on the molecular mechanisms of stomatal movements mediated by abscisic acid,light,CO_(2),reactive oxygen species,pathogens,temperature,and other *** discussed the significance of elucidating the integrative mechanisms that regulate stomatal movements in helping design smart crops with enhanced water use efficiency and resilience in a climate-changing world.

关键词： environmental stress guard cell plant regulatory mechanisms stomatal movements

Genetic variation in a heat shock transcription factor modulates cold tolerance in maize

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Molecular plant 2024年第9期17卷 1423-1438页

作者： Lei Gao Lingling Pan Yiting Shi Rong Zeng Minze Li Zhuoyang Li Xuan Zhang Xiaoming Zhao Xinru Gong Wei Huang Xiaohong Yang Jinsheng Lai Jianru Zuo Zhizhong Gong Xiqing Wang Weiwei Jin Zhaobin Dong Shuhua Yang state key laboratory of plant Environmental Resilience Frontiers Science Center for Molecular Design BreedingCollege of Biological SciencesCenter for Crop Functional Genomics and Molecular BreedingChina Agricultural UniversityBeijing 100193China state key laboratory of Maize Bio-breeding National Maize Improvement CenterFrontiers Science Center for Molecular Design BreedingDepartment of Plant Geneticsand BreedingChinaAgricultural UniversityBeijingChina state key laboratory of plant Genomics and National plant Gene Research Center Institute of Genetics and Developmental BiologyChinese Academy of SciencesBeijing100101China

Understanding how maize(Zea mays)responds to cold stress is crucial for facilitating breeding programs of cold-tolerant *** extensive utilization of the genome-wide association study(GWAs)approach for exploring favorable natural alleles associated with maize cold tolerance,few studies have successfully identified candidate genes that contribute to maize cold *** this study,we used a diverse panel of inbred maize lines collected from different germplasm sources to perform a GWAS on var-iations in the relative injured area of maize true leaves during cold stress-a trait very closely correlated with maize cold *** identified HsF21,which encodes a B-class heat shock transcription factor(HSF)that positively regulates cold tolerance at both the seedling and germination *** varia-tions in the promoter of the cold-tolerant HSF21Hap1 allele led to increased HSF21 expression under cold stress by inhibiting binding of the basic leucine zipper bziP68 transcription factor,a negative regulator of cold *** integrating transcriptome deep sequencing,DNA affinity purification sequencing,and targeted lipidomic analysis,we revealed the function of HsF21 in regulating lipid metabolism homeo-stasis to modulate cold tolerance in *** addition,we found that HsF21 confers maize cold tolerance without incurring yield ***,this study establishes HsF21 as a key regulator that en-hances cold tolerance in maize,providing valuable genetic resources for breeding of cold-tolerant maize varieties.

关键词： heat shock factor HSF21 cold tolerance natural variation bZiP68 lipid metabolism maize

Potassium alleviated high nitrogen-induced apple growth inhibition by regulating photosynthetic nitrogen allocation and enhancing nitrogen utilization capacity

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Horticultural plant Journal 2024年第1期10卷 1-14页

作者： Xinxiang Xu Guangyuan Liu Jingquan Liu Mengxue Lyu Fen Wang Yue Xing Hao Meng Min Li Yu Jiang Ge Tian Zhanling Zhu Yuanmao Jiang Shunfeng Ge state key laboratory of Crop Biology College of Horticulture Science and EngineeringShandong Agricultural UniversityTai'anShandong 271018China key laboratory of biochemistry and Molecular Biology in Universities of Shandong College of Biological and Agricultural EngineeringWeifang UniversityWeifangShandong 261061China

There is a close relationship between potassium(K)and nitrogen(N).However,the roles of K under high N conditions remain *** a hydroponics approach,we monitored the morphological,physiological,and molecular changes in M9T337 apple(Malus domestica)rootstocks under different nitrate(10 and 30 mmol·L^(-1)NO_(3)^(-))and K supply(0.5,6,10,and 20 mmol·L_(-1)K^(+))*** revealed that high nitrate inhibited the root growth of M9T337 rootstocks,downregulated the expressions of K transporter genes(MdPT5,MdHKT1,and MdATK1),and reduced the net NO3-and K+influx at the surface of roots,thereby resulting in an N/K imbalance in *** investigation showed that 10 mmol·L^(-1)K increased the activity of N metabolic enzymes(NR,GS,NiR,and GOGAT),upregulated the expressions of genes related to nitrate uptake and transport(MdNRT1.1,MdNRT1.2,MdNRT1.5,and MdNRT2.4),promoted15N transport from the roots to the shoots,optimized leaf N distribution,and improved photosynthetic N utilization efficiency under high nitrate *** results suggest that the negative effects of high nitrate may be related to the N/K imbalance and that reducing N/K in plants by increasing K supply level can effectively alleviate the inhibition of N assimilation by high nitrate stress.

关键词： Apple rootstock K level High nitrate stress N metabolism ^(15)N

A novel secreted protein FgHrip1 from Fusarium graminearum triggers immune responses in plants

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Journal of Integrative Agriculture 2024年第11期23卷 3774-3787页

作者： Zhenchao Fu Huiqian Zhuang Vincent Ninkuu Jianpei Yan Guangyue Li Xiufen Yang Hongmei Zeng state key laboratory for Biology of plant Diseases and Insect Pests Institute of Plant Protection Chinese Academy of Agricultural Sciences

Fusarium graminearum, the primary pathogenic fungus responsible for Fusarium head blight(FHB) in wheat,secretes abundant chemical compounds that interact with host plants. In this study, a secreted protein FgHrip1,isolated from the culture filtrate of F. graminearum, was found to induce typical cell death in tobacco. The FgHrip1 gene was then cloned and expressed in Escherichia coli. Further bioassay analysis showed that the recombinant FgHrip1 induced early defense induction events, such as reactive oxygen species(ROS) production,callose deposition, and up-regulation of defense-related genes in tobacco. Furthermore, FgHrip1 significantly enhanced immunity in tobacco seedlings against Pseudomonas syringae pv. tabaci 6605(Pst. 6605) and tobacco mosaic virus(TMV). FgHrip1-treated wheat spikes also exhibited defense-related transcript accumulation and developed immunity against FHB infection. Whereas the expression of FgHrip1 was induced during the infection process, the deletion of the gene impaired the virulence of F. graminearum. Our results suggest that FgHrip1triggers immunity and induces disease resistance in tobacco and wheat, thereby providing new insight into strategy for biocontrol of FHB.

关键词： FgHrip1 secreted protein plant immunity Fusarium graminearum wheat

Eubacterium coprostanoligenes alleviates chemotherapy-induced intestinal mucositis by enhancing intestinal mucus barrier

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Acta Pharmaceutica Sinica B 2024年第4期14卷 1677-1692页

作者： Dongsheng Bai Jiawei Zhao Runde Wang Jiaying Du Chen Zhou Chunyang Gu Yuxiang Wang Lulu Zhang Yue Zhao Na Lu state key laboratory of Natural Medicines Jiangsu Key Laboratory of Carcinogenesis and InterventionDepartment of PhysiologySchool of Basic Medicine and Clinical PharmacyChina Pharmaceutical UniversityNanjing 210009China Department of Clinical Pharmacology School of PharmacyNanjing Medical UniversityNanjing 211166China

Chemotherapy-induced mucositis represents a severe adverse outcome of cancer treatment,significantly curtailing the efficacy of these treatments and,in some cases,resulting in fatal *** identifying intestinal epithelial cell damage as a key factor in chemotherapy-induced mucositis,the paucity of effective treatments for such damage is *** our study,we discovered that Eubacterium coprostanoligenes promotes mucin secretion by goblet cells,thereby fortifying the integrity of the intestinal mucus *** enhanced barrier function serves to resist microbial invasion and sub-sequently reduces the inflammatory ***,this effect remains unobtrusive to the anti-tumor efficacy of chemotherapy ***,*** up-regulates the expression of AUF1,leading to the stabilization of Muc2 mRNA and an increase in mucin synthesis in goblet *** espe-cially significant finding is that *** activates the AhR pathway,thereby promoting the expression of *** summary,our results strongly indicate that *** enhances the intestinal mucus barrier,effec-tively alleviating chemotherapy-induced intestinal mucositis by activating the AhR/AUFl pathway,consequently enhancing Muc2 mRNA stability.

关键词： Chemotherapy Mucositis Eubacterium coprostanoligenes MUC2 AUF1