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CRISPR/Cas9 gene editing and natural variation analysis demonstrate the potential for HvARE1 in improvement of nitrogen use efficiency in barley

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Journal of Integrative Plant Biology 2022年第3期64卷 756-770页

作者： Sakura D.Karunarathne Yong Han Xiao-Qi Zhang Chengdao Li Western Crop Genetics Alliance College of ScienceHealthEngineering and EducationMurdoch UniversityPerthWA6150 Australia Western Australian State Agricultural Biotechnology Centre Murdoch UniversityPerthWA6150 Australia Department of Primary Industries and Regional Development PerthWA6151 Australia

Nitrogen is a major determinant of grain yield and quality.As excessive use of nitrogen fertilizer leads to environmental pollution and high production costs,improving nitrogen use efficiency(NUE)is fundamental for a sustainable agriculture.Here,we dissected the role of the barley abnormal cytokinin response1 repressor 1(Hv ARE1)gene,a candidate for involvement in NUE previously identified in a genome-wide association study,through natural variation analysis and clustered regularly interspacedshort palindromic repeats(CRISPR)/CRISPRassociated protein 9(Cas9)-mediated gene editing.Hv ARE1 was predominantly expressed in leaves and shoots,with very low expression in roots under low nitrogen conditions.Agrobacterium-mediated genetic transformation of immature embryos(cv.Golden Promise)with single guide RNAs targeting Hv ARE1 generated 22 T0 plants,from which four T1 lines harbored missense and/or frameshift mutations based on genotyping.Mutant are1 lines exhibited an increase in plant height,tiller number,grain protein content,and yield.Moreover,we observed a 1.5-to2.8-fold increase in total chlorophyll content in the flag leaf at the grain filling stage.Delayed senescence by 10–14 d was also observed in mutant lines.Barley are1 mutants had high nitrogen content in shoots under low nitrogen conditions.These findings demonstrate the potential of ARE1 in NUE improvement in barley.

关键词： ARE1 barley CRISPR/Cas9 gene editing leaf senescence NUE

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The alleviation of ammonium toxicity in plants

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Journal of Integrative Plant Biology 2023年第6期65卷 1362-1368页

作者： Chengbin Xiao Yuan Fang Suomin Wang Kai He Ministry of Education Key Laboratory of Cell Activities and Stress Adaptations School of Life SciencesLanzhou UniversityLanzhou 730000China State Key Laboratory of Grassland Agro‐Ecosystems College of Pastoral Agriculture Science and TechnologyLanzhou UniversityLanzhou 730000China

Nitrogen(N)is an essential macronutrient for plants and profoundly affects crop yields and qualities.Ammonium(NH4+)and nitrate(NO3-)are major inorganic N forms absorbed by plants from the surrounding environments.Intriguingly,NH4+is usually toxic to plants when it serves as the sole or dominant N source.It is thus important for plants to coordinate the utilization of NH4+and the alleviation of NH4+toxicity.To fully decipher the molecular mechanisms underlying how plants minimize NH4+toxicity may broadly benefit agricultural practice.In the current minireview,we attempt to discuss recent discoveries in the strategies for mitigating NH4+toxicity in plants,which may provide potential solutions for improving the nitrogen use efficiency(NUE)and stress adaptions in crops.

关键词： alleviation ammonium toxicity channel nitrogen NUE transporter

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Balanced nitrogen–iron sufficiency boosts grain yield and nitrogen use efficiency by promoting tillering

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Molecular Plant 2023年第12期16卷 1661-1677页

作者： Ying Song Guang-Yu Wan Jing-Xian Wang Zi-Sheng Zhang Jin-Qiu Xia Liang-Qi Sun Jie Lu Chuan-Xi Ma Lin-Hui Yu Cheng-Bin Xiang Jie Wu Division of Life Sciences and Medicine Division of Molecular&Cell BiophysicsHefei National Science Center for Interdisciplinary Sciences at the MicroscaleMOE Key Laboratory for Membraneless Organelles and Cellular DynamicsUniversity of Science and Technology of ChinaThe Innovation Academy of Seed DesignChinese Academy of SciencesHefeiAnhui Province 230027China School of Agronomy Anhui Agricultural UniversityHefeiAnhui Province 230036China State Key Laboratory of Crop Stress Biology for Arid Areas and Institute of Future Agriculture Northwest A&F UniversityYanglingShaanxi 712100China

Crop yield plays a critical role in global food security.For optimal plant growth and maximal crop yields,nutrients must be balanced.However,the potential significance of balanced nitrogen-iron(N-Fe)for improving crop yield and nitrogen use efficiency(NUE)has not previously been addressed.Here,we show that balanced N-Fe sufficiency significantly increases tiller number and boosts yield and NUE in rice and wheat.NIN-like protein 4(OsNLP4)plays a pivotal role in maintaining the N-Fe balance by coordinately regulating the expression of multiple genes involved in N and Fe metabolism and signaling.OsNLP4 also suppresses OsD3 expression and strigolactone(SL)signaling,thereby promoting tillering.Balanced N-Fe sufficiency promotes the nuclear localization of OsNLP4 by reducing H_(2)O_(2) levels,reinforcing the functions of OsNLP4.Interestingly,we found that OsNLP4 upregulates the expression of a set of H2O2-scavenging genes to promote its own accumulation in the nucleus.Furthermore,we demonstrated that foliar spraying of balanced N-Fe fertilizer at the tillering stage can effectively increase tiller number,yield,and NUE of both rice and wheat in the field.Collectively,these findings reveal the previously unrecognized effects of N-Fe balance on grain yield and NUE as well as the molecular mechanism by which the OsNLP4-OsD3 module integrates N-Fe nutrient signals to downregulate SL signaling and thereby promote rice tillering.Our study sheds light on how N-Fe nutrient signals modulate rice tillering and provide potential innovative approaches that improve crop yield with reduced N fertilizer input for benefitting sustainable agriculture worldwide.

关键词： nitrogen-iron balance tller NUE grain yield OsNLP4 rice

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Genome-wide association study and transcriptome analysis reveal new QTL and candidate genes for nitrogen-deficiency tolerance in rice

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The Crop Journal 2022年第4期10卷 942-951页

作者： Qing Li Xueli Lu Changjian Wang Lan Shen Liping Dai Jinli He Long Yang Peiyuan Li Yifeng Hong Qiang Zhang Guojun Dong Jiang Hu Guangheng Zhang Deyong Ren Zhenyu Gao Longbiao Guo Qian Qian Li Zhu Dali Zeng State Key Laboratory of Rice Biology China National Rice Research InstituteChinese Academy of Agricultural SciencesHangzhou 311401ZhejiangChina

The development of rice cultivars with improved nitrogen use efficiency(NUE)is desirable for sustainable agriculture.Achieving this goal depends in part on understanding how rice responds to low soil nitrogen(N)and identifying causative genes underlying this trait.To identify quantitative trait loci(QTL)or genes associated with low N response,we conducted a genome-wide association study(GWAS)using a diverse panel of 230 rice accessions and performed a transcriptomic investigation of rice accessions with differential responses to low N stress at two N levels.We detected 411 GWAS-associated genes in 5 QTL and 2722 differentially expressed genes in response to low N,of which 24 were identified by both methods and ranked according to gene annotations,literature queries,gene expression,and genetic diversity analysis.The large-scale datasets obtained from this study reveal low N-responsive characteristics and provide insights towards understanding the regulatory mechanisms of N-deficiency tolerance in rice,and the candidate genes or QTL would be valuable resources for increasing rice NUE via molecular biotechnology.

关键词： Nitrogen NUE Rice GWAS RNA-seq

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A molecular framework underlying low-nitrogeninduced early leaf senescence in Arabidopsis thaliana

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Molecular Plant 2023年第4期16卷 756-774页

作者： Hongmei Fan Shuxuan Quan Qing Ye Lei Zhang Wei Liu Ning Zhu Xiaoqi Zhang Wenyuan Ruan Keke Yi Nigel M.Crawford Yong Wang State Key Laboratory of Crop Biology College of Life SciencesShandong Agricultural UniversityTai'anShandong 271018China Key Laboratory of Plant Nutrition and Fertilizer Ministry of AgricultureInstitute of Agricultural Resources and Regional PlanningChinese Academy of Agricultural SciencesBeijing 10081China Section of Cell and Developmental Biology Division of Biological ScienceUniversity of California at San DiegoLa JollaCA 92093-0116USA

Nitrogen(N)deficiency causes early leaf senescence,resulting in accelerated whole-plant maturation and severely reduced crop yield.However,the molecular mechanisms underlying N-deficiency-induced early leaf senescence remain unclear,even in the model species Arabidopsis thaliana.In this study,we identified Growth,Development and Splicing 1(GDS1),a previously reported transcription factor,as a new regulator of nitrate(NO3)signaling by a yeast-one-hybrid screen using a NO3enhancer fragment from the promoter of NRT2.1.We showed that GDS1 promotes NO3 signaling,absorption and assimilation by affecting the expression of multiple NO3 regulatory genes,including Nitrate Regulatory Gene2(NRG2).Interestingly,we observedthat gds1mutants show early leaf senescence as well as reduced NO3-contentand Nuptake under N-deficient conditions.Further analyses indicated that GDS1 binds to the promoters of several senescence-related genes,including Phytochrome-lnteracting Transcription Factors 4 and 5(PIF4 and PIF5)and represses their expression.Interestingly,we found that N deficiency decreases GDS1 protein accumulation,and GDS1 could interact with Anaphase Promoting Complex Subunit 10(APC10).Genetic and biochemical experiments demonstrated that Anaphase Promoting Complex or Cyclosome(APC/C)promotes the ubiquitination and degradation of GDS1 under N deficiency,resulting in loss of PIF4 and PiF5 repression and consequent early leaf senescence.Furthermore,we discovered that overexpression of GDS1 could delay leaf senescence and improve seed yield and N-use efficiency(NUE)in Arabidopsis.In summary,our study uncovers a molecular framework illustrating a new mechanism underlying low-N-induced early leaf senescence and provides potential targets for genetic improvement of crop varieties with increased yield and NUE.

关键词： nitrate signaling GDS1 nitrogen-deficiency-induced leaf senescence PIF4/PIF5 APC/C ubiquitina-tion NUE

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Hole fertilization in the root zone facilitates maize yield and nitrogen utilization by mitigating potential N loss and improving mineral N accumulation

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Journal of Integrative Agriculture 2023年第4期22卷 1184-1198页

作者： SHI Wen-xuan ZHANG Qian LI Lan-tao TAN Jin-fang XIE Ruo-han WANG Yi-lun College of Resources and Environment Henan Agricultural UniversityZhengzhou 450002P.R.China School of Agriculture Shenzhen Campus of Sun Yat-sen UniversityShenzhen 518107P.R.China

Reducing environmental impacts and improving N utilization are critical to ensuring food security in China.Although root-zone fertilization has been considered an effective strategy to improve nitrogen use efficiency (NUE),the effect of controlled-release urea (CRU) applied in conjunction with normal urea in this mode is unclear.Therefore,a 3-year field experiment was conducted using a no-N-added as a control and two fertilization modes (FF,furrow fertilization by manual trenching,i.e.,farmer fertilizer practice;HF:root-zone hole fertilization by point broadcast manually) at 210 kg N ha^(–1) (controlled-release:normal fertilizer=5:5),along with a 1-year in-situ microplot experiment.Maize yield,NUE and N loss were investigated under different fertilization modes.The results showed that compared with FF,HF improved the average yield and N recovery efficiency by 8.5 and 22.3%over three years,respectively.HF had a greater potential for application than FF treatment,which led to increases in dry matter accumulation,total N uptake,SPAD value and LAI.In addition,HF remarkably enhanced the accumulation of ^(15)N derived from fertilizer by 17.2%compared with FF,which in turn reduced the potential loss of^(15)N by 43.8%.HF increased the accumulation of N in the tillage layer of soils at harvest for potential use in the subsequent season relative to FF.Hence,HF could match the N requirement of summer maize,sustain yield,improve NUE and reduce environmental N loss simultaneously.Overall,root-zone hole fertilization with blended CRU and normal urea can represent an effective and promising practice to achieve environmental integrity and food security on the North China Plain,which deserves further application and investigation.

关键词： maize yield hole fertilization NUE ^(15)N-labeled blended urea ^(15)N loss

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OPPORTUNITIES AND APPROACHES FOR MANIPULATING SOIL-PLANT MICROBIOMES FOR EFFECTIVE CROP NITROGEN USE IN AGROECOSYSTEMS

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Frontiers of Agricultural Science and Engineering 2022年第3期9卷 333-343页

作者： Jingjing PENG Olatunde OLADELE Xiaotong SONG Xiaotang JU Zhongjun JIA Hangwei HU Xuejun LIU Shuikuan BEI Anhui GE Limei ZHANG Zhenling CUI College of Resources and Environmental Sciences National Academy of Agriculture Green DevelopmentKey Laboratory of Plant-Soil InteractionsMinistry of EducationChina Agricultural UniversityBeijing 100193China State Key Laboratory of Urban and Regional Ecology Research Center for Eco-Environmental SciencesChinese Academy of SciencesBeijing 100085China College of Tropical Crops Hainan UniversityHaikou 570228China Key Labortatory of Mollisols Agroecology Northeast Institute of Geography and AgroecologyChinese Academy of SciencesChangchun 130102China Faculty of Veterinary and Agricultural Sciences The University of MelbourneParkvilleVIC.3010Australia University of Chinese Academy of Sciences Beijing 100049China.

Soil microbiomes drive the biogeochemical cycling of nitrogen and regulate soil N supply and loss,thus,pivotal nitrogen use efficiency(NUE).Meanwhile,there is an increasing awareness that plant associated microbiomes and soil food web interactions is vital for modulating crop productivity and N uptake.The rapid advances in modern omics-based techniques and biotechnologies make it possible to manipulate soil-plant microbiomes for improving NUE and reducing N environmental impacts.This paper summarizes current progress in research on regulating soil microbial N cycle processes for NUE improvement,plant-microbe interactions benefiting plant N uptake,and the importance of soil microbiomes in promoting soil health and crop productivity.We also proposes a potential holistic(rhizosphere-root-phyllosphere)microbe-based approach to improve NUE and reduce dependence on mineral N fertilizer in agroecosystems,toward nature-based solution for nutrient management in intensive cropping systems.

关键词： nitrogen microbiome NUE rhizosphere phyllosphere soil food web

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Mitigating growth-stress tradeoffs via elevated TOR signaling in rice

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Molecular Plant 2024年第2期17卷 240-257页

作者： Wei Li Jiaqi Liu Zeqi Li Ruiqiang Ye Wenzhen Chen Yuqing Huang Yue Yuan Yi Zhang Huayi Hu Peng Zheng Zhongming Fang Zeng Tao Shiyong Song Ronghui Pan Jian Zhang Jumim Tu Jen Sheen Hao Du State Key Laboratory of Rice Biology College of Agriculture and BiotechnologyZhejiang UniversityYu-Hang-Tang Road No.866Hangzhou 310058China ZJU-Hangzhou Global Scientific and Technological Innovation Center Zhejiang UniversityHangzhou 311215China Hainan Institute of Zhejiang University Sanya 572025China National Key Laboratory of Plant Molecular Genetics CASCenter for Excellence in Molecular Plant SciencesShanghai lnstitute of Plant Physiology and EcologyChineseAcademy of SciencesShanghai 200032China Key Laboratory of Plant Resource Conservation and Germplasm Innovation in Mountainous Region(Ministry of Education) College of Agricultural SciencesGuizhouUniversityGuiyang 550025China State Key Laboratory of Rice Biology and Breeding China National Rice Research InstituteHangzhou 311400China Department of Molecular Biology and Center for Computational and Integrative Biology Massachusetts General Hospitaland Department of GeneticsHarvard Medical SchoolBostonMA02114USA

Rice production accounts for approximately half of the freshwater resources utilized in agriculture,result-ing in greenhouse gas emissions such as methane(CH4)from flooded paddy fields.To address this chal-lenge,environmentally friendly and cost-effective water-saving techniques have become widely adopted in rice cultivation.However,the implementation of water-saving treatments(WsTs)in paddy-field rice has been associated with a substantial yield loss of up to 50%as well as a reduction in nitrogen use efficiency(NUE).In this study,we discovered that the target of rapamycin(TOR)signaling pathway is compromised in rice under WsT.Polysome profiling-coupled transcriptome sequencing(polysome-seq)analysis unveiled a substantial reduction in global translation in response to WST associated with the downregulation of TOR activity.Molecular,biochemical,and genetic analyses revealed new insights into the impact of the positive TOR-S6K-RPS6 and negative TOR-MAF1 modules on translation repression under WST.Intriguingly,ammonium exhibited a greater ability to alleviate growth constraints under WsT by enhancing TOR signaling,which simultaneously promoted uptake and utilization of ammonium and nitrogen allocation.We further demonstrated that TOR modulates the ammonium transporter AMT1;1 as well as the amino acid permease APP1 and dipeptide transporter NPF7.3 at the translational level through the 5'untranslated region.Collectively,these findings reveal that enhancing TOR signaling could mitigate rice yield penalty due to WST by regulating the processes involved in protein synthesis and NUE.Our study will contribute to the breeding of new rice varieties with increased water and fertilizer utilization efficiency.

关键词： target of rapamycin TOR water-saving rice low-carbon agriculture drought nitrogen use efficiency NUE

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Physiological evaluation of nitrogen use efficiency of different apple cultivars under various nitrogen and water supply conditions

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Journal of Integrative Agriculture 2020年第3期19卷 709-720页

作者： WANG Qian LIU Chang-hai HUANG Dong DONG Qing-long LI Peng-min Steve van NOCKER MA Feng-wang State Key Laboratory of Crop Stress Biology for Arid Areas/Shaanxi Key Laboratory of Apple College of HorticultureNorthwest A&F UniversityYangling 712100P.R.China Department of Horticulture Michigan State UniversityEast LansingMI 48824USA

Nitrogen(N) deficiency is a common problem for apple(Malus×domestica) production in arid regions of China.However,N utilization efficiency(NUE) of different apple cultivars grown under low N conditions in arid regions has not been evaluated.In this study,NUE was assessed for one-year-old seedlings of six apple cultivars,Golden Delicious,Qinguan,Jonagold,Honeycrisp,Fuji and Pink Lady,grafted onto Malus hupehensis Rehd.rootstocks.Four treatments were used,including control water with control N(CWCN),limited water with control N(LWCN),control water with low N(CWLN) and limited water with low N(LWLN).Our results showed that growth indices such as biomass,plant height and stem diameter,and photosynthetic rate of all cultivars decreased in the order CWCN>CWLN>LWCN>LWLN.When subjected to LWLN treatment,Qinguan showed better growth and photosynthetic characters than other tested cultivars.Additionally,Qinguan and Pink Lady had higher NUE,while Honeycrisp and Jonagold had lower NUE,based on the determination of biomass,photosynthetic parameters,chlorophyll content,the maximal photochemical efficiency of PSII(Fv/Fm),15 N and N contents.

关键词： apple drought N deficiency physiological indices 15N-labeling NUE

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有机、无机肥配施对关中冬小麦氮素利用和土壤碳氮动态的影响

有机、无机肥配施对关中冬小麦氮素利用和土壤碳氮动态的影响

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作者：彭凯西北农林科技大学

学位级别：硕士

为追求持续高产,满足日益增长的口粮需求,农业生产中过度施用氮肥现象日益严重,作物高产与农田生态环境间的矛盾已不容忽视。有机无机肥配施被认为是实现农业可持续发展的有效施肥方式,然而,对有机无机肥配施的最佳比例不同研究往往有... 详细信息

为追求持续高产,满足日益增长的口粮需求,农业生产中过度施用氮肥现象日益严重,作物高产与农田生态环境间的矛盾已不容忽视。有机无机肥配施被认为是实现农业可持续发展的有效施肥方式,然而,对有机无机肥配施的最佳比例不同研究往往有不同的衡量标准,这些标准通常只考虑了产量、土壤养分、温室气体等单一因素,综合考量多种衡量因素进而筛选出最佳比例还有待进一步研究。因此,本研究在等氮条件下设置5个施肥处理和一个不施肥对照,施肥处理中有机肥氮替代无机肥氮比例分别为0、15%、30%、60%、100%(不施肥、100U、15S+85U、30S+70U、60S+40U、100S),探究不同配施比例下冬小麦产量和氮素吸收利用、土壤固碳效应、土壤氮素时空变化规律、农田温室气体排放特征。通过以上研究内容,本研究旨兼顾产量和环境效益,从而筛选出关中地区最佳有机无机肥配施比例,为当地农业可持续发展提供一定依据。本研究主要结果以下:(1)小麦成熟期地上部各处理氮素积累量存在明显差异,不施肥处理氮素积累量最低;随着有机肥配施比例增加,茎、叶、穗氮素积累量表现出先增加后降低趋势,30S+70U处理穗氮素积累量最高,与单施无机肥相比,两年平均增加42.11%。穗中N积累量高于茎叶,并且穗N积累量随有机肥配施比例增加而降低。低量有机肥配施比例(15-30%)下作物产量相比不施肥明显增加,第一年15S+85U处理产量较高,相比不施肥增加1736.22 kg/hm,增幅30.31%;第二年30S+70U处理产量较高,相比不施肥增加1455.61 kg/hm,增幅24.54%;全量有机肥配施下作物产量与不施肥无明显差异。(2)有机无机肥配施能显著增加土壤颗粒有机碳(Particle Organic Carban POC)含量,POC含量随有机肥用量增加而增加,而不同处理矿物结合有机碳(Mineral binding Organic Carbon MOC)差异不明显;POC含量的增加进而导致土壤有机碳(Soil Organic Carban SOC)的增加,各处理中以100S下有机碳储量最高,第二个小麦季100S处理有机碳储量相比不施肥增加15.39 Mg/hm,增幅56.23%;各处理对有机碳储量的促进作用表现为:高量有机肥配施>低量有机肥配施。有机无机肥配施能有效增加0-20cm土壤全氮含量;低量有机肥配施无机肥为整个小麦生育期提供了较高水平的NH-N和NO-N含量,60S+40U处理能有效降低深层土壤NH-N淋溶损失;30S+70U处理土壤中尿素N百分比和残留量较高,表明有机无机肥配施有利于无机肥氮保留在土壤中。(3)三种温室气体中以CO累计排放量最高,保持在1234.70-2121.01 kg·hm·yr;有机无机肥配施增加了NO和CO累计排放量,第二个小麦季60S+40U处理NO和CO累计排放量相比100U分别增加0.90、89.21 kg·hm·yr;总体上三种温室气体累计排放处理间量差异不明显。综上所述,等氮条件下低量有机无机肥配施能有效促进小麦成熟期氮素向籽粒转运,进而显著提高小麦氮素利用率,最终增加作物产量;同时,有机无机肥配能进一步减少速效氮向深层土淋溶,并且有机无机肥配施与单施无机肥温室气体累计排放量整体上无明显差异,因此,在施氮量不变的前提下,关中地区15-30%有机肥替代无机肥能够减少无机氮肥施用量,兼顾产量与生态效益。

关键词：有机无机肥配施 NUE 温室气体有机碳储量 15N

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