Drought is a major abiotic stress factor limiting maize production, and elucidating the genetic control of root system architecture and plasticity to water-deficit stress is a crucial problemto improve drought adaptability. In this study, 13 root and shoot traits and genetic plasticity were evaluated in a recombinant inbred line (RIL) population under well-watered (WW) and water stress (WS) conditions. Significant phenotypic variation was observed for all observed traits both under WW and WS conditions. Most of the measured traits showed significant genotype-environment interaction (GEI) in both environments. Strong correlations were observed among traits in the same class. Multi-environment (ME) and multi-trait (MT) QTL analyses were conducted for all observed traits. A total of 48 QTLs were identified by ME, including 15 QTLs associated with 9 traits showing significant QTL-by-Environment interactions (QEI). QTLs associated with crown root angle (CRA2) and crown root length (CRL1) were identified as having antagonistic pleiotropic effects, while 13 other QTLs showed signs of conditional neutrality (CN), including 9 and 4 QTLs detected under WW and WS conditions, respectively. MT analysis identified 14 pleiotropic QTLs for 13 traits, SNP20 (1@79.2 cM) was associated with the length of crown root (CR), primary root (PR), and seminal root (SR) and might contribute to increases in root length under WS condition. Taken together, these findings contribute to our understanding of the phenotypic and genotypic patterns of root plasticity in response to water deficiency, which will be useful to improve drought tolerance in maize.

Maize requires more water than most other crops; therefore, the water use efficiency of this crop must be improved for maize production under undesirable land and changing environmental conditions.To elucidate the genetic control of drought in maize, we evaluated approximately 5000 inbred lines from 30 linkage-association joint mapping populations under two contrasting water regimes for seven drought-related traits, including yield and anthesis-silking interval (ASI). The joint linkage analysis was conducted to identify 220 quantitative trait loci (QTLs) under well-watered conditions and 169 QTLs under water-stressed conditions. The genome-wide association analysis identified 365 single nucleotide polymorphisms (SNPs) associated with drought-related traits, and these SNPs were located in 354 candidate genes. Fifty-two of these genes showed significant differential expression in the inbred line B73 under the well-watered and water-stressed conditions. In addition, genomic predictions suggested that the moderate-density SNPs obtained through genotyping-by-sequencing were able to make accurate predictions in the nested association mapping population for drought-related traits with moderate-to-high heritability under the water-stressed conditions.The results of the present study provide important information that can be used to understand the genetic basis of drought stress responses and facilitate the use of beneficial alleles for the improvement of drought tolerance in maize.

The diversity of metabolites found in plants is by far greater than in most other organisms. Metabolic profiling techniques, which measure many of these compounds simultaneously, enabled investigating the regulation of metabolic networks and proved to be useful for predicting important agronomic traits. However, little is known about the genetic basis of metabolites in crops such as maize. Here, a set of 289 diverse maize inbred lines was genotyped with 56,110 SNPs and assayed for 118 biochemical compounds in the leaves of young plants, as well as for agronomic traits of mature plants in field trials. Metabolite concentrations had on average a repeatability of 0.73 and showed a correlation pattern that largely reflected their functional grouping. Genome-wide association mapping with correction for population structure and cryptic relatedness identified for 26 distinct metabolites strong associations with SNPs, explaining up to 32.0% of the observed genetic variance. On nine chromosomes, we detected 15 distinct SNP-metabolite associations, each of which explained more then 15% of the genetic variance. For lignin precursors, including p-coumaric acid and caffeic acid, we found strong associations (P values to ) with a region on chromosome 9 harboring cinnamoyl-CoA reductase, a key enzyme in monolignol synthesis and a target for improving the quality of lignocellulosic biomass by genetic engineering approaches. Moreover, lignin precursors correlated significantly with lignin content, plant height, and dry matter yield, suggesting that metabolites represent promising connecting links for narrowing the genotype-phenotype gap of complex agronomic traits.

Maize is a globally valuable commodity and one of the most extensively studied genetic model organisms. However, we know surprisingly little about the extent and potential utility of the genetic variation found in wild relatives of maize. Here, we characterize a high-density genomic variation map from 744 genomes encompassing maize and all wild taxa of the genus Zea, identifying over 70 million single-nucleotide polymorphisms. The variation map reveals evidence of selection within taxa displaying novel adaptations. We focus on adaptive alleles in highland teosinte and temperate maize, highlighting the key role of flowering-time-related pathways in their adaptation. To show the utility of variants in these data, we generate mutant alleles for two flowering-time candidate genes. This work provides an extensive sampling of the genetic diversity of Zea, resolving questions on evolution and identifying adaptive variants for direct use in modern breeding.© 2022. The Author(s), under exclusive licence to Springer Nature America, Inc.

密植是提高作物单位面积产量、促进粮食增产的重要途径之一。叶夹角是影响玉米(Zea mays)密植的关键因子。中国农业大学田丰课题组最近克隆了2个调控玉米叶夹角的数量性状位点(QTL)——UPA1和UPA2, 揭示了这2个位点的功能基因(brd1和ZmRAVL1)通过油菜素内酯(BR)信号通路调控叶夹角。UPA2位于ZmRAVL1上游9.5 kb, 可与DRL1蛋白结合。另一个影响玉米叶夹角的蛋白LG1可以激活ZmRAVL1的表达; DRL1蛋白与LG1蛋白直接互作抑制LG1对ZmRAVL1的激活表达。玉米祖先种大刍草(teosinte)的UPA2位点序列与DRL1蛋白结合能力更强, 导致大刍草ZmRAVL1的表达受到更强的抑制, 下调表达的ZmRAVL1进一步使下游基因brd1的表达下调, 进而降低叶环区的内源BR水平, 导致叶夹角变小。将大刍草的UPA2等位基因导入到玉米中或对玉米中ZmRAVL1进行基因编辑, 在密植条件下均可显著提高玉米产量。上述发现为高产玉米品种的分子育种改良提供了重要理论基础和基因资源。

Drought is one of the environmental factors that severely restrict plant distribution and crop production. Recently, we reported that the high-affinity potassium transporter OsHAK1 plays important roles in K acquisition and translocation in rice over low and high K concentration ranges, however, knowledge on the regulatory roles of OsHAK1 in osmotic/drought stress is limited. Here, transcript levels of OsHAK1 were found transiently elevated by water deficit in roots and shoots, consistent with the enhanced GUS activity in transgenic plants under stress. Under drought conditions, OsHAK1 knockout mutants (KO) presented lower tolerance to the stress and displayed stunted growth at both the vegetative and reproductive stages. Phenotypic analysis of OsHAK1 overexpression seedlings (Ox) demonstrated that they present better tolerance to drought stress than wild-type (WT). Compared to WT seedlings, OsHAK1 overexpressors had lower level of lipid peroxidation, higher activities of antioxidant enzymes (POX and CAT) and higher proline accumulation. Furthermore, qPCR analysis revealed that OsHAK1 act as a positive regulator of the expression of stress-responsive genes as well as of two well-known rice channel genes (OsTPKb and OsAKT1) involved in K homeostasis and stress responses in transgenic plants under dehydration. Most important, OsHAK1-Ox plants displayed enhanced drought tolerance at the reproductive stage, resulting in 35% more grain yield than WT under drought conditions, and without exhibiting significant differences under normal growth conditions. Consequently, OsHAK1 can be considered to be used in molecular breeding for improvement of drought tolerance in rice.

Drought is a major environmental disaster that causes crop yield loss worldwide. Metabolites are involved in various environmental stress responses of plants. However, the genetic control of metabolomes underlying crop environmental stress adaptation remains elusive.Here, we perform non-targeted metabolic profiling of leaves for 385 maize natural inbred lines grown under well-watered as well as drought-stressed conditions. A total of 3890 metabolites are identified and 1035 of these are differentially produced between well-watered and drought-stressed conditions, representing effective indicators of maize drought response and tolerance. Genetic dissections reveal the associations between these metabolites and thousands of single-nucleotide polymorphisms (SNPs), which represented 3415 metabolite quantitative trait loci (mQTLs) and 2589 candidate genes. 78.6% of mQTLs (2684/3415) are novel drought-responsive QTLs. The regulatory variants that control the expression of the candidate genes are revealed by expression QTL (eQTL) analysis of the transcriptomes of leaves from 197 maize natural inbred lines. Integrated metabolic and transcriptomic assays identify dozens of environment-specific hub genes and their gene-metabolite regulatory networks. Comprehensive genetic and molecular studies reveal the roles and mechanisms of two hub genes, Bx12 and ZmGLK44, in regulating maize metabolite biosynthesis and drought tolerance.Our studies reveal the first population-level metabolomes in crop drought response and uncover the natural variations and genetic control of these metabolomes underlying crop drought adaptation, demonstrating that multi-omics is a powerful strategy to dissect the genetic mechanisms of crop complex traits.© 2021. The Author(s).

We find that the DREB subfamily transcription factor, CmERF053, has a novel function to regulate the development of shoot branching and lateral root in addition to affecting abiotic stress. Dehydration-responsive element binding proteins (DREBs) are important plant transcription factors that regulate various abiotic stresses. Here, we isolated an APETALA2/ethylene-responsive factor (AP2/ERF) transcription factor from chrysanthemum (Chrysanthemum morifolium 'Jinba'), CmERF053, the expression of which was rapidly up-regulated by main stem decapitation. Phylogenetic analysis indicated that it belongs to the A-6 group of the DREB subfamily, and the subcellular localization assay confirmed that CmERF053 was a nuclear protein. Overexpression of CmERF053 in Arabidopsis exhibited positive effects of plant lateral organs, which had more shoot branching and lateral roots than did the wild type. We also found that the expression of CmERF053 in axillary buds was induced by exogenous cytokinins. These results suggested that CmERF053 may be involved in cytokinins-related shoot branching pathway. In this study, an altered auxin distribution was observed during root elongation in the seedlings of the overexpression plants. Furthermore, overexpress CmERF053 gene could enhance drought tolerance. Together, these findings indicated that CmERF053 plays crucial roles in regulating shoot branching, lateral root, and drought stress in plant. Moreover, our study provides potential application value for improving plant productivity, ornamental traits, and drought tolerance.

钙调磷酸酶B类互作蛋白激酶CIPK（CBL interacting protein kinases）是植物钙离子信号通路中响应非生物逆境胁迫的重要蛋白激酶之一。本研究以拟南芥和水稻中CIPK家族基因序列信息为基础，利用玉米参考基因组B73和生物信息学分析方法，全基因组范围内鉴定玉米CIPK基因家族成员，分析CIPK家族基因的进化关系、基因结构、基因表达模式和对干旱胁迫的响应。本研究共鉴定出44个玉米CIPK家族基因，并将其分为5个亚家族，每个亚家族有不同的外显子-内含子和UTR的结构特征；基于基因差异表达分析，筛选出5个与抗旱性相关的候选基因ZmCIPK3、ZmCIPK7、ZmCIPK44、ZmCIPK25和ZmCIPK28；进一步的遗传数据表明，干旱胁迫下ZmCIPK3拟南芥转基因株系的存活率明显高于野生型，提高了拟南芥的抗旱性；同时，干旱胁迫下ZmCIPK3拟南芥转基因株系中抗旱性相关生化指标过氧化物酶（POD）、超氧化物歧化酶（SOD）活性显著高于野生型，而丙二醛（MDA）和脯氨酸（Pro）的含量显著低于野生型。本研究在玉米全基因组水平上鉴定了CIPK基因家族成员，分析了其在不同抗旱性材料、不同水分处理下的基因表达模式，明确了ZmCIPK3是一个抗旱性候选基因。

Increased planting densities have boosted maize yields. Upright plant architecture facilitates dense planting. Here, we cloned () and, two quantitative trait loci conferring upright plant architecture. is controlled by a two-base sequence polymorphism regulating the expression of a B3-domain transcription factor () located 9.5 kilobases downstream. exhibits differential binding by DRL1 (DROOPING LEAF1), and DRL1 physically interacts with LG1 (LIGULELESS1) and represses LG1 activation of regulates (), which underlies, altering endogenous brassinosteroid content and leaf angle. The allele that reduces leaf angle originated from teosinte, the wild ancestor of maize, and has been lost during maize domestication. Introgressing the wild allele into modern hybrids and editing enhance high-density maize yields.Copyright © 2019 The Authors, some rights reserved; exclusive licensee American Association for the Advancement of Science. No claim to original U.S. Government Works.

AP2/ERF (APETALA2/ethylene-responsive factor)转录因子是植物中最大的转录因子家族之一, 在调控植物生长发育和响应逆境胁迫等方面起重要作用。探究玉米(Zea mays L.) AP2/ERF家族基因功能将为玉米新种质创制提供重要的基因资源。本研究克隆获得了ZmEREB211 (Gene ID: 103647485)基因, 利用生物信息学分析、实时荧光定量PCR等技术对该基因的基本特性、组织表达特性及响应逆境胁迫表达模式等进行了分析; 对转基因拟南芥株系进行了相应逆境胁迫处理和表型鉴定。结果显示: 该基因只包含1个外显子, cDNA全长为792 bp, 编码263个氨基酸; ZmEREB211蛋白分子量为27.9 kD, 理论等电点为6.01, 具有AP2家族所特有的保守结构域; ZmEREB211基因在玉米根系中的表达量最高, 且在幼根中的表达量高于成熟根中的表达量; 同时该基因在脱水、高盐、干旱和低温等处理条件下均有不同程度的诱导表达。在分别含有不同浓度梯度的NaCl、甘露醇(mannitol)和茉莉酸(jasmonic acid, JA)的1/2 MS培养基上, 转ZmEREB211基因拟南芥株系的根长显著长于野生型。在干旱和高盐处理下, 盆栽转基因拟南芥株系较野生型株系表现出更强的耐受性, 且苗期的绿叶数显著多于野生型, 过氧化物酶(POD)活性和叶绿素含量均显著高于野生型。研究表明ZmEREB211可能参与调控玉米根系生长发育, 对高盐、干旱、渗透等逆境胁迫及JA激素处理均能起到正向的调控作用。本研究为进一步解析ZmEREB211在玉米中的生物学功能提供了重要的参考依据。

玉米生长发育过程中会遭受干旱、高温、高盐及营养元素匮乏等多种非生物胁迫, 导致其产量和品质下降, 造成严重的农业减产。MYB类转录因子在植物中广泛分布, 参与植物生长发育和环境响应, 筛选及鉴定出具有抗逆功能的MYB类转录因子能为玉米抗逆遗传改良提供理论依据。本研究从玉米干旱处理的材料中克隆了一个R2R3-MYB家族转录因子基因ZmMYB12, 其响应自然干旱、ABA及PEG处理, 基因表达被诱导上调。病毒诱导沉默ZmMYB12后的玉米植株对干旱更加敏感, 活性氧积累更多, 根系更小; 复水后ZmMYB12沉默植株存活率更低, 表明ZmMYB12是干旱正调控因子。进一步构建ZmMYB12稳定过表达拟南芥, 干旱处理后ZmMYB12过表达株系活性氧积累更少, 侧根更多, 抗旱能力增强。同时, 发现ZmMYB12过表达拟南芥在低磷胁迫下侧根数量更多, 根系酸化程度增加, 叶绿素及花青素含量更多, 体内无机磷含量高于野生型, 表明ZmMYB12参与到低磷胁迫过程中, 并且提高了植株对磷元素的吸收及利用率。研究表明ZmMYB12调控干旱抗性并响应低磷胁迫, 为作物抗旱及耐低磷育种提供了一个良好的基因资源。

Drought stress severely limits the growth, development, and productivity of crops, and therefore understanding the mechanisms by which plants respond to drought is crucial. In this study, we cloned a maize NAC transcription factor, ZmNAC49, and identified its function in response to drought stress. We found that ZmNAC49 is localized in the nucleus and has transcriptional activation activity. ZmNAC49 expression is rapidly and strongly induced by drought stress, and overexpression enhances stress tolerance in maize. Overexpression also significant decreases the transpiration rate, stomatal conductance, and stomatal density in maize. Detailed study showed that ZmNAC49 overexpression affects the expression of genes related to stomatal development, namely ZmTMM, ZmSDD1, ZmMUTE, and ZmFAMA. In addition, we found that ZmNAC49 can directly bind to the promoter of ZmMUTE and suppress its expression. Taken together, our results show that the transcription factor ZmNAC49 represses ZmMUTE expression, reduces stomatal density, and thereby enhances drought tolerance in maize.© The Author(s) 2020. Published by Oxford University Press on behalf of the Society for Experimental Biology. All rights reserved. For permissions, please email: journals.permissions@oup.com.

TIFY, previously known as ZIM, comprises a plant-specific family annotated as transcription factors that might play important roles in stress response. Despite TIFY proteins have been reported in Arabidopsis and rice, a comprehensive and systematic survey of ZmTIFY genes has not yet been conducted. To investigate the functions of ZmTIFY genes in this family, we isolated and characterized 30 ZmTIFY (1 TIFY, 3 ZML, and 26 JAZ) genes in an analysis of the maize (Zea mays L.) genome in this study. The 30 ZmTIFY genes were distributed over eight chromosomes. Multiple alignment and motif display results indicated that all ZmTIFY proteins share two conserved TIFY and Jas domains. Phylogenetic analysis revealed that the ZmTIFY family could be divided into two groups. Putative cis-elements, involved in abiotic stress response, phytohormones, pollen grain, and seed development, were detected in the promoters of maize TIFY genes. Microarray data showed that the ZmTIFY genes had tissue-specific expression patterns in various maize developmental stages and in response to biotic and abiotic stresses. The results indicated that ZmTIFY4, 5, 8, 26, and 28 were induced, while ZmTIFY16, 13, 24, 27, 18, and 30 were suppressed, by drought stress in the maize inbred lines Han21 and Ye478. ZmTIFY1, 19, and 28 were upregulated after infection by three pathogens, whereas ZmTIFY4, 13, 21, 23, 24, and 26 were suppressed. These results indicate that the ZmTIFY family may have vital roles in response to abiotic and biotic stresses. The data presented in this work provide vital clues for further investigating the functions of the genes in the ZmTIFY family.

The TIFY family is a novel plant-specific gene family involved in the regulation of diverse plant-specific biologic processes, such as development and responses to phytohormones, in Arabidopsis. However, there is limited information about this family in monocot species. This report identifies 20 TIFY genes in rice, the model monocot species. Sequence analysis indicated that rice TIFY proteins have conserved motifs beyond the TIFY domain as was previously shown in Arabidopsis. On the basis of their protein structures, members of the TIFY family can be divided into two groups. Transcript level analysis of OsTIFY genes in tissues and organs revealed different tempo-spatial expression patterns, suggesting that expression and function vary by stage of plant growth and development. Most of the OsTIFY genes were predominantly expressed in leaf. Nine OsTIFY genes were responsive to jasmonic acid and wounding treatments. Interestingly, almost all the OsTIFY genes were responsive to one or more abiotic stresses including drought, salinity, and low temperature. Over-expression of OsTIFY11a, one of the stress-inducible genes, resulted in significantly increased tolerance to salt and dehydration stresses. These results suggest that the OsTIFY family may have important roles in response to abiotic stresses. The data presented in this report provide important clues for further elucidating the functions of the genes in the OsTIFY family.