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Expansion and improvement of ChinaMu by MuT-seq and chromosome-level assembly of the Mu-starter genome

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Journal of Integrative Plant Biology 2024年第4期66卷 645-659页

作者： Lei Liang Yuancong Wang Yanbin Han Yicong Chen Mengfei Li Yibo Wu Zeyang Ma Han Zhao Rentao Song State Key Laboratory of maize Bio-breeding Frontiers Science Center for Molecular Design BreedingJoint International Research Laboratory of Crop Molecular BreedingNational Maize Improvement CenterCollege of Agronomy and BiotechnologyChina Agricultural UniversityBeijing 100091China institute of Crop Germplasm and Biotechnology Jiangsu Provincial Key Laboratory of AgrobiologyJiangsu Academy of Agricultural SciencesNanjing 210014China Sanya institute of China Agricultural University Sanya 572025China Hainan Yazhou Bay Seed Laboratory Sanya 572025China

ChinaMu is the largest sequence-indexed Mutator(Mu)transposon insertional library in maize(Zea mays).In this study,we made significant improvements to the size and quality of the ChinaMu library.We developed a new Mu-tag isolation method Mu-Tn5-seq(MuT-seq).Compared to the previous method used by ChinaMu,MuT-seq recovered 1/3 more germinal insertions,while requiring only about 1/14 of the sequencing volume and 1/5 of the experimental time.Using MuT-seq,we identified 113,879 germinal insertions from 3,168 Mu-active F1 families.We also assembled a high-quality genome for the Mu-active line Mu-starter,which harbors the initial active MuDR element and was used as the pollen donor for the mutation population.Using the Mu-starter genome,we recovered 33,662(15.6%)additional germinal insertions in 3,244(7.4%)genes in the Mu-starter line.The Mu-starter genome also improved the assignment of 117,689(54.5%)germinal insertions.The newly upgraded ChinaMu dataset currently contains 215,889 high-quality germinal insertions.These insertions cover 32,224 pan-genes in the Mu-starter and B73Ref5 genomes,including 23,006(80.4%)core genes shared by the two genomes.As a test model,we investigated Mu insertions in the pentatricopeptide repeat(PPR)superfamily,discovering insertions for 92%(449/487)of PPR genes in ChinaMu,demonstrating the usefulness of ChinaMu as a functional genomics resource for maize.

关键词： ChinaMu Mutator MuT-seq PPR gene Zea mays

Striking a growth–defense balance:Stress regulators that function in maize development

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Journal of Integrative Plant Biology 2024年第3期66卷 424-442页

作者： Shiyi Xie Hongbing Luo Wei Huang Weiwei Jin Zhaobin Dong maize Engineering and Technology Research Center of Hunan Province College of AgronomyHunan Agricultural UniversityChangsha 410128China State Key Laboratory of maize Bio-breeding National Maize Improvement CenterFrontiers Science Center for Molecular Design BreedingChina Agricultural UniversityBeijing 100193China Tianjin Key Laboratory of Intelligent breeding of Major Crops Fresh Corn Research Center of BTHCollege of Agronomy&Resources and EnvironmentTianjin Agricultural UniversityTianjin 300384China

maize(Zea mays)cultivation is strongly affected by both abiotic and biotic stress,leading to reduced growth and productivity.It has recently become clear that regulators of plant stress responses,including the phytohormones abscisic acid(ABA),ethylene(ET),and jasmonic acid(JA),together with reactive oxygen species(ROS),shape plant growth and development.Beyond their well established functions in stress responses,these molecules play crucial roles in balancing growth and defense,which must be finely tuned to achieve high yields in crops while maintaining some level of defense.In this review,we provide an in-depth analysis of recent research on the developmental functions of stress regulators,focusing specifically on maize.By unraveling the contributions of these regulators to maize development,we present new avenues for enhancing maize cultivation and growth while highlighting the potential risks associated with manipulating stress regulators to enhance grain yields in the face of environmental challenges.

关键词： stress regulators maize development ABA ET JA ROS

Sucrose-associated SnRK1a1-mediated phosphorylation of Opaque2 modulates endosperm filling in maize

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molecular Plant 2024年第5期17卷 788-806页

作者： Tao Yang Yunqin Huang Longyu Liao Shanshan Wang Haoyu Zhang Jingying Pan Yongcai Huang Xiaoling Li Di Chen Tao Liu Xiaoduo Lu Yongrui Wu State Key Laboratory of Crop Gene Exploration and Utilization in Southwest China Sichuan Agricultural UniversityChengdu 611130SichuanChina National Key Laboratory of Plant molecular Genetics CAS Center for Excellence in Molecular Plant SciencesShanghai lnstitute of Plant Physiology and EcologyChinese Academy of SciencesShanghai 200032China institute of molecular breeding for maize Qilu Normal UniversityJinanChina

During maize endosperm filling,sucrose not only serves as a source of carbon skeletons for storage-reserve synthesis but also acts as a stimulus to promote this process.However,the molecular mechanisms underlying sucrose and endosperm filling are poorly understood.In this study,we found that sucrose promotes the expression of endosperm-filling hub gene Opaque2(O2),coordinating with storage-reserve accumulation.We showed that the protein kinase SnRK1a1 can attenuate O2-mediated transactivation,but sucrose can release this suppression.Biochemical assays revealed that SnRK1a1 phosphorylates O2 at serine 41(S41),negatively affecting its protein stability and transactivation ability.We observed that mutation of SnRK1a1 results in larger seeds with increased kernel weight and storage reserves,while overexpression of SnRK1a1 causes the opposite effect.Overexpression of the native O2(O2-OE),phospho-dead(O2-SA),and phospho-mimetic(O2-SD)variants all increased 100-kernel weight.Although O2-SA seeds exhibit smaller kernel size,they have higher accumulation of starch and proteins,resulting in larger vitreous endosperm and increased test weight.O2-SD seeds display larger kernel size but unchanged levels of storage reserves and test weight.O2-OE seeds show elevated kernel dimensions and nutrient storage,like a mixture of O2-SA and O2-SD seeds.Collectively,our study discovers a novel regulatory mechanism of maize endosperm filling.Identification of S41 as a SnRK1-mediated phosphorylation site in O2 offers a potential engineering target for enhancing storage-reserve accumulation and yield in maize.

关键词： maize Opaque2 SnRK1 endosperm phosphorylation sucrose

Genome-wide association mapping and genomic prediction of stalk rot in two mid-altitude tropical maize populations

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The Crop Journal 2024年第2期12卷 558-568页

作者： Junqiao Song Angela Pacheco Amos Alakonya Andrea S.Cruz-Morales Carlos Muoz-Zavala Jingtao Qu Chunping Wang Xuecai Zhang Felix San Vicente Thanda Dhliwayo College of Agronomy Henan University of Science and TechnologyLuoyang 471000HenanChina International maize and Wheat Improvement Center(CIMMYT) El BatanMexico Anyang Academy of Agricultural Sciences Anyang 455000HenanChina CIMMYT-China Specialty maize Research Center Crop Breedingand Cultivation Research InstituteShanghai Academy of Agricultural SciencesShanghai 201403China

maize stalk rot reduces grain yield and quality.Information about the genetics of resistance to maize stalk rot could help breeders design effective breeding strategies for the trait.Genomic prediction may be a more effective breeding strategy for stalk-rot resistance than marker-assisted selection.We performed a genome-wide association study(GWAS)and genomic prediction of resistance in testcross hybrids of 677 inbred lines from the Tuxpe?o and non-Tuxpe?o heterotic pools grown in three environments and genotyped with 200,681 single-nucleotide polymorphisms(SNPs).Eighteen SNPs associated with stalk rot shared genomic regions with gene families previously associated with plant biotic and abiotic responses.More favorable SNP haplotypes traced to tropical than to temperate progenitors of the inbred lines.Incorporating genotype-by-environment(G×E)interaction increased genomic prediction accuracy.

关键词： maize stalk rot Genome-wide association mapping Haplotype analysis Genomic prediction G×E interaction

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Computational tools for plant genomics and breeding

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Science China Life Sciences 2024年第8期 1579-1590页

作者： Hai Wang Mengjiao Chen Xin Wei Rui Xia Dong Pei Xuehui Huang Bin Han State Key Laboratory of maize Bio-breeding Frontiers Science Center for Molecular Design BreedingJoint International Research Laboratory of Crop Molecular BreedingNational Maize Improvement CenterCollege of Agronomy and BiotechnologyChina Agricultural University Sanya institute of China Agricultural University Hainan Yazhou Bay Seed Laboratory State Key Laboratory of Tree Genetics and breeding Key Laboratory of Tree Breeding and Cultivation of the State Forestry and Grassland AdministrationResearch Institute of ForestryChinese Academy of Forestry Shanghai Key Laboratory of Plant molecular Sciences College of Life SciencesShanghai Normal University College of Horticulture South China Agricultural University National Center for Gene Research CAS Center for Excellence in Molecular Plant SciencesChinese Academy of Sciences

Plant genomics and crop breeding are at the intersection of biotechnology and information technology.Driven by a combination of highthroughput sequencing,molecular biology and data science,great advances have been made in omics technologies at every step along the central dogma,especially in genome assembling,genome annotation,epigenomic profiling,and transcriptome profiling.These advances further revolutionized three directions of development.One is genetic dissection of complex traits in crops,along with genomic prediction and selection.The second is comparative genomics and evolution,which open up new opportunities to depict the evolutionary constraints of biological sequences for deleterious variant discovery.The third direction is the development of deep learning approaches for the rational design of biological sequences,especially proteins,for synthetic biology.All three directions of development serve as the foundation for a new era of crop breeding where agronomic traits are enhanced by genome design.

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Genetic and lipidomic analyses reveal the key role of lipid metabolism for cold tolerance in maize

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Journal of Genetics and Genomics 2024年第3期51卷 326-337页

作者： Lei Gao Haifang Jiang Minze Li Danfeng Wang Hongtao Xiang Rong Zeng Limei Chen Xiaoyan Zhang Jianru Zuo Shuhua Yang Yiting Shi State Key Laboratory of Plant Environmental Resilience College of Biological SciencesFrontiers Science Center for Molecular Design BreedingCenter for Crop Functional Genomics and Molecular BreedingChina Agricultural UniversityBeijing 100193China State Key Laboratory of Wheat&maize Crop Science College of Life SciencesHenan Agricultural UniversityZhengzhouHenan 450002China State Key Laboratory of Plant Genomics Institute of Genetics and Developmental BiologyChinese Academy of SciencesBeijing 100101China University of Chinese Academy of Sciences Beijing 100049China Suihua Branch of Heilongjiang Academy of Agricultural Machinery Sciences SuihuaHeilongjiang 152052China

Lipid remodeling is crucial for cold tolerance in plants.However,the precise alternations of lipidomics during cold responses remain elusive,especially in maize(Zea mays L.).In addition,the key genes responsible for cold tolerance in maize lipid metabolism have not been identified.Here,we integrate lipidomic,transcriptomic,and genetic analysis to determine the profile of lipid remodeling caused by cold stress.We find that the homeostasis of cellular lipid metabolism is essential for maintaining cold tolerance of maize.Also,we detect 210 lipid species belonging to 13 major classes,covering phospholipids,glycerides,glycolipids,and free fatty acids.Various lipid metabolites undergo specific and selective alterations in response to cold stress,especially mono-/di-unsaturated lysophosphatidic acid,lysophosphatidylcholine,phosphatidylcholine,and phosphatidylinositol,as well as polyunsaturated phosphatidic acid,monogalactosyldiacylglycerol,diacylglycerol,and triacylglycerol.In addition,we identify a subset of key enzymes,including ketoacyl-acyl-carrier protein synthase II(KAS II),acyl-carrier protein 2(ACP2),male sterility33(Ms33),and stearoyl-acyl-carrier protein desaturase 2(SAD2)involved in glycerolipid biosynthetic pathways are positive regulators of maize cold tolerance.These results reveal a comprehensive lipidomic profile during the cold response of maize and provide genetic resources for enhancing cold tolerance in crops.

关键词： maize Cold stress Lipid metabolism Transcriptome Lipidomic

IbNIEL-mediated degradation of IbNAC087 regulates jasmonic acid-dependent salt and drought tolerance in sweet potato

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Journal of Integrative Plant Biology 2024年第2期66卷 176-195页

作者： Xu Li Zhen Wang Sifan Sun Zhuoru Dai Jun Zhang Wenbin Wang Kui Peng Wenhao Geng Shuanghong Xia Qingchang Liu Hong Zhai Shaopei Gao Ning Zhao Feng Tian Huan Zhang Shaozhen He Key Laboratory of Sweet Potato Biology and Biotechnology Ministry of Agriculture and Rural Affairs/Beijing Key Laboratory of Crop Genetic Improvement/Laboratory of Crop Heterosis&Utilization and Joint Laboratory for International Cooperation in Crop Molecular BreedingMinistry of EducationCollege of Agronomy&BiotechnologyChina Agricultural UniversityBeijing 100193China Sanya institute of China Agricultural University Sanya 572025China State Key Laboratory of Plant Physiology and Biochemistry National Maize Improvement CenterKey Laboratory of Biology and Genetic Improvement of MaizeBeijing Key Laboratory of Crop Genetic ImprovementChina Agricultural UniversityBeijing 100193China

Sweet potato(Ipomoea batatas[L.]Lam.)is a crucial staple and bioenergy crop.Its abiotic stress tolerance holds significant importance in fully utilizing marginal lands.Transcriptional processes regulate abiotic stress responses,yet the molecular regulatory mechanisms in sweet potato remain unclear.In this study,a NAC(NAM,ATAF1/2,and CUC2)transcription factor,IbNAC087,was identified,which is commonly upregulated in salt-and drought-tolerant germplasms.Overexpression of IbNAC087 increased salt and drought tolerance by increasing jasmonic acid(JA)accumulation and activating reactive oxygen species(ROS)scavenging,whereas silencing this gene resulted in opposite phenotypes.JA-rich IbNAC087-OE(overexpression)plants exhibited more stomatal closure than wild-type(WT)and IbNAC087-Ri plants under NaCl,polyethylene glycol,and methyl jasmonate treatments.IbNAC087 functions as a nuclear transcriptional activator and directly activates the expression of the key JA biosynthesisrelated genes lipoxygenase(IbLOX)and allene oxide synthase(IbAOS).Moreover,IbNAC087 physically interacted with a RING-type E3 ubiquitin ligase NAC087-INTERACTING E3 LIGASE(IbNIEL),negatively regulating salt and drought tolerance in sweet potato.IbNIEL ubiquitinated IbNAC087 to promote 26S proteasome degradation,which weakened its activation on IbLOX and IbAOS.The findings provide insights into the mechanism underlying the IbNIEL-IbNAC087 module regulation of JA-dependent salt and drought response in sweet potato and provide candidate genes for improving abiotic stress tolerance in crops.

关键词： abiotic tolerance IbNAC087 IbNIEL JA sweet potato

Orchestrating seed storage protein and starch accumulation toward overcoming yield–quality trade-off in cereal crops

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Journal of Integrative Plant Biology 2024年第3期66卷 468-483页

作者： Shuanghe Cao Bingyan Liu Daowen Wang Awais Rasheed Lina Xie Xianchun Xia Zhonghu He State Key Laboratory of Crop Gene Resources and breeding/National Wheat Improvement Center Institute of Crop SciencesBeijing 100081China College of Agronomy Henan Agricultural UniversityZhengzhou 450002China International maize and Wheat Improvement Center(CIMMYT)China Office Chinese Academy of Agricultural SciencesBeijing 100081China

Achieving high yield and good quality in crops is essential for human food security and health.However,there is usually disharmony between yield and quality.Seed storage protein(SSP)and starch,the predominant components in cereal grains,determine yield and quality,and their coupled synthesis causes a yield–quality trade-off.Therefore,dissection of the underlying regulatory mechanism facilitates simultaneous improvement of yield and quality.Here,we summarize current findings about the synergistic molecular machinery underpinning SSP and starch synthesis in the leading staple cereal crops,including maize,rice and wheat.We further evaluate the functional conservation and differentiation of key regulators and specify feasible research approaches to identify additional regulators and expand insights.We also present major strategies to leverage resultant information for simultaneous improvement of yield and quality by molecular breeding.Finally,future perspectives on major challenges are proposed.

关键词： cereal crops grain yield and quality seed storage protein starch synergistic regulation

Two H3K36 methyltransferases differentially associate with transcriptional activity and enrichment of facultative heterochromatin in rice blast fungus

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aBIOTECH 2024年第1期5卷 1-16页

作者： Mengting Xu Ziyue Sun Huanbin Shi Jiangnan Yue Xiaohui Xiong Zhongling Wu Yanjun Kou Zeng Tao Ministry of Agriculture Key Laboratory of molecular Biology of Crop Pathogens and Insects Institute of BiotechnologyZhejiang UniversityHangzhou 310058China State Key Lab of Rice Biology and breeding China National Rice Research InstituteHangzhou 310021China

Di-and tri-methylation of lysine 36 on histone H3(H3K36me2/3)is catalysed by histone methyltransferase Set2,which plays an essential role in transcriptional regulation.Although there is a single H3K36 methyltransferase in yeast and higher eukaryotes,two H3K36 methyltransferases,Ash1 and Set2,were present in many filamentous fungi.However,their roles in H3K36 methylation and transcriptional regulation remained unclear.Combined with methods of RNA-seq and ChIP-seq,we revealed that both Ash1 and Set2 are redundantly required for the full H3K36me2/3 activity in Magnaporthe oryzae,which causes the devastating worldwide rice blast disease.Ash1 and Set2 distinguish genomic H3K36me2/3-marked regions and are differentially associated with repressed and activated transcription,respectively.Furthermore,Ash1-catalysed H3K36me2 was co-localized with H3K27me3 at the chromatin,and Ash1 was required for the enrichment and transcriptional silencing of H3K27me3-occupied genes.With the different roles of Ash1 and Set2,in H3K36me2/3 enrichment and transcriptional regulation on the stress-responsive genes,they differentially respond to various stresses in M.oryzae.Overall,we reveal a novel mechanism by which two H3K36 methyltransferases catalyze H3K36me2/3 that differentially associate with transcriptional activities and contribute to enrichment of facultative heterochromatin in eukaryotes.

关键词： Ash1 Facultative heterochromatin H3K36me2/3 Magnaporthe oryzae Set2 Transcriptional regulation