Drought-responsive genes expressed predominantly in root tissues are enriched with homotypic cis-regulatory clusters in promoters of major cereal crops

The root appears to be the most relevant organ for breeding drought stress tolerance.However, our knowledge about temporal and spatial regulation of drought-associated genes in the root remains fragmented, especially in crop plants. We performed a meta-analysis of expression divergence of essential drought-inducible genes and analyzed their association with cis-elements in model crops and major cereal crops. Our analysis of42 selected drought-inducible genes revealed that these are expressed primarily in roots,followed by shoot, leaf, and inflorescence tissues, especially in wheat. Quantitative real-time RT-PCR analysis confirmed higher expression of TaDREB2 and TaAQP7 in roots,correlated with extensive rooting and drought-stress tolerance in wheat. A promoter scan up to 2 kb upstream of the translation start site using phylogenetic footprinting revealed708 transcription factor binding sites, including drought response elements(DREs), auxin response elements(Aux REs), MYCREs/MYBREs, ABAREs, and ERD1 in 19 selected genes.Interestingly, these elements were organized into clusters of overlapping transcription factor binding sites known as homotypic clusters(HCTs), which modulate drought physiology in plants. Taken together, these results revealed the expression preeminence of major drought-inducible genes in the root, suggesting its crucial role in drought adaptation. The occurrence of HCTs in drought-inducible genes highlights the putative evolutionary modifications of crop plants in developing drought adaptation. We propose that these DNA motifs can be used as molecular markers for breeding drought-resilient cultivars, particularly in the cereal crops.

Combination of 6-Benzylaminopurine and Thidiazuron Promotes Highly Efficient Shoot Regeneration from Cotyledonary Node of Mature Peanut (Arachis hypogaea L.) Cultivars

Efficient in vitro plantlet regeneration is an important step to successfully transform genes for the improvement of agronomic traits.A combination of 6-benzylaminopurine(BAP)and thidiazuron(TDZ)plant growth regulators was applied to evaluate shoot regeneration capacity whereasα-naphthalene acetic acid(NAA)combination with 6-benzylaminopurine(BAP),and 2,4-dichlorophenoxyacetic acid(2,4-D)with 6-benzylaminopurine were tested to optimize root induction for two peanut cultivars.The result showed combination(BAP with TDZ)was found to be effective in promoting shoot.The highest shoot regeneration frequency(93%)was obtained on a medium supplemented with 4 mg/L BAP and 0.5 mg/L TDZ while an average regeneration frequency(87%)was achieved in a medium containing combinations of 2 mg/L BAP with 1 mg/L TDZ.The shooting rate increased for both cultivars as the concentrations of BAP increased and TDZ decreased.The highest rooting rate(93%)was obtained on a medium supplemented with 3.5 mg/L NAA with 2.5 mg/L BAP for both cultivars.The rooting rate increased as the concentration of auxin to cytokinin ratio increased.The maximum rooting rate(83%)was obtained on MS medium supplemented with 0.3 mg/L 2,4-D with 0.2 mg/L BAP for the cultivar N3.The result indicated that BAP with NAA was much better than BAP with 2,4-D in rooting rate.Thus,the protocol developed was genotype independent and effective for peanut tissue culture.

Exploration of Genetic Pattern of Phenological Traits in Wheat (Triticum aestivum L.) under Drought Stress

Drought is the major detrimental environmental factor for wheat(Triticum aestivum L.)production.The exploration of genetic patterns underlying drought tolerance is of great significance.Here we report the gene actions controlling the phenological traits using the line×tester model studying 27 crosses and 12 parents under normal irrigation and drought conditions.The results interpreted via multiple analysis(mean performance,correlations,principal component,genetic analysis,heterotic and heterobeltiotic potential)disclosed highly significant differences among germplasm.The phenological waxiness traits(glume,boom,and sheath)were strongly interlinked.Flag leaf area exhibits a positive association with peduncle and spike length under drought.The growing degree days(heat-units)greatly influence spikelets and grains per spike,however,the grain yield/plant was significantly reduced(17.44 g to 13.25 g)under drought.The principal components based on eigenvalue indicated significant PCs(first-seven)accounted for 79.9%and 73.9%of total variability under normal irrigation and drought,respectively.The investigated yield traits showed complex genetic behaviour.The genetic advance confronted a moderate to high heritability for spikelets/spike and grain yield/plant.The traits conditioned by dominant genetic effects in normal irrigation were inversely controlled by additive genetic effects under drought and vice versa.The magnitude of dominance effects for phenological and yield traits,i.e.,leaf twist,auricle hairiness,grain yield/plant,spikelets,and grains/spike suggests that selection by the pedigree method is appropriate for improving these traits under normal irrigation conditions and could serve as an indirect selection index for improving yield-oriented traits in wheat populations for drought tolerance.However,the phenotypic selection could be more than effective for traits conditioned by additive genetic effects under drought.We suggest five significant cross combinations based on heterotic and heterobeltiotic potential of wheat genotypes for improved yield and enhanced biological production of wheat in advanced generations under drought.

种子表型平台的改进及其在芥菜种子活力研究中的应用

为了研究芥菜萌发期的耐盐性,本研究通过增加喷雾装置等方面的改进,构建了新一代的高通量、低成本种子表型平台SeedGerm V 2.0,可实现无人值守运行。基于该种子表型平台,分析了地方特色芥菜品种水东甜芥菜在不同浓度NaCl胁迫下的种子萌发动力学特征。结果表明,种子平台能够自动采集芥菜种子萌发数据并进行分析,可以用于盐胁迫研究。较低NaCl溶液对芥菜种子萌发率有一定促进作用,但随着盐浓度的升高,芥菜种子的萌发率迅速下降,浓度达到0.300 mol/L时发芽基本停止,表明其具有较强的耐盐性。估算了芥菜种子萌发的积盐模型(Halothermal time,HaloTT)参数,其中积盐常数θ_(halo)为2.26 mol/(L·h),中位值NaCl_(b(50))为0.22 mol/L,标准方差δ_(NaCl b)为0.067 mol/L。利用上述参数得到水东甜芥菜种子萌发预测方程:Probit(g)=(NaCl+2.26/t g-0.22)/0.067,可以准确预测不同盐浓度下芥菜种子的发芽时间和发芽率,对于芥菜的育种和生产具有重要参考价值。

百脉根BIO和豌豆突变位点ELE2的比较基因组定位(英文)

豆科两侧对称花的花瓣具有背腹(DV)的分化以及可变的器官内部(IN)非对称性,在大小与形状上显示出不同的发育特征;因而花瓣的发育为克隆决定植物器官的形状与大小的关键基因提供了很好的实验系统。本研究对百脉根中BIO基因进行研究。百脉根bio突变体具有多效性,既影响花器官内部的对称性也影响器官的大小和育性,豌豆ele突变体的表型与bio相似。定位结果表明BIO和ELE2位于豆科基因组的共线性区段,提示BIO和ELE2可能是同源基因突变所致。本研究利用比较基因组定位方法,将BIO和ELE2候选基因锚定在豆科模式植物百脉根和蒺藜苜蓿基因组含有11个同源基因的BAC重叠群上。BIO和ELE2基因的克隆将有助于揭示豆科花瓣形态和大小调控的分子机理,进而为豆科作物遗传改良提供分子理论基础。

A Genomic Variation Map Provides Insights into the Genetic Basis of Spring Chinese Cabbage (Brassica rapa ssp.pekinensis)Selection

Chinese cabbage is the most consumed leafy crop in East Asian countries.However,premature bolting induced by continuous low temperatures severely decreases the yield and quality of the Chinese cabbage, and therefore restricts its planting season and geographic distribution.In the past 40years,spring Chinese cabbage with strong winterness has been selected to meet the market demand.Here,we report a genome variation map of Chinese cabbage generated from the resequencing data of 194 geographically diverse accessions of three ecotypes.In-depth analyses of the selection sweeps and genome-wide patterns revealed that spring Chinese cabbage was selected from a specific population of autumn Chinese cabbage around the area of Shandong peninsula in northern China.We identified 23 genomic loci that underwent intensive selection,and further demonstrated by gene expression and haplotype analyses that the incorporation of elite alleles of VERNALISATION INSENTIVE 3.1(BrVIN3.1)and FLOWER LOCUS C 1(BrFLC1)is a determinant genetic source of variation during selection.Moreover,we showed that the quantitative response of BrVIN3.1 to cold due to the sequence variations in the cis elements of the BrVlN3.1 promoter significantly contributes to bolting-time variation in Chinese cabbage.Collectively, our study provides valuable insights into the genetic basis of spring Chinese cabbage selection and will facilitate the breeding of bolting-resistant Varieties by molecular-marker-assisted selection,transgenic or gene editingapproaches.

Transcriptional and Post-transcriptional Modulation of SQU and KEW Activities in the Control of Dorsal-Ventral Asymmetric Flower Development in Lotus japonicus

In Papilionoideae legume, Lotusjaponicus, the development of dorsal-ventral （DV） asymmetric flowers is mainly controlled by two TB1/CYCLOIDEA/PCF （TCP） genes, SQUARED STANDARD （SQU） and KEELED WINGS IN LOTUS （KEW）, which determine dorsal and lateral identities, respectively. However, the molecular basis of how these two highly homologous genes orchestrate their diverse functions remains unclear. Here, we analyzed their expression levels, and investigated the transcriptional activities of SQUand KEW. We demonstrated that SQU possesses both activation and repression activities, while KEW acts only as an activator. They form homo- and heterodimers, and then collaboraUvely regulate their expression at the transcription level. Furthermore, we identified two types of post-transcriptional modifications, phosphor- ylation and ATP/GTP binding, both of which could affect their transcriptional activities. Mutations in ATP/ GTP binding motifs of SQU and KEW lead to failure of phosphorylation, and transgenic plants bearing the mutant proteins display defective DV asymmetric flower development, indicating that the two conjugate modifications are essential for their diverse functions. Altogether, SQU and KEW activities are precisely modulated at both transcription and post-transcription levels, which might link DV asymmetric flower development to different physiological status and/or signaling pathways.

Genetic Analysis of ele Mutants and Comparative Mapping of ele1 Locus in the Control of Organ Internal Asymmetry in Garden Pea

Previous study has shown that during zygomorphic development in garden pea （Pisum sativum L.）, the organ internal （IN） asymmetry of lateral and ventral petals was regulated by a genetic locus, SYMMETRIC PETAL 1 （SYP1）, while the dorsoventral （DV） asymmetry was determined by two CYC-like TCP genes or the PsCYC genes, KEELED WINGS （K） and LOBED STANDARD 1 （LST1）. In this study, two novel loci, ELEPHANT EAR-LIKE LEAF 1 （ELE1） and ELE2 were characterized. These mutants exhibit a similar defect of IN asymmetry as syp1 in lateral and ventral petals, but also display pleiotropic effects of enlarged organ size. Genetic analysis showed that ELE1 and ELE2 were involved in same genetic pathway and the enlarged size of petals but not compound leaves in e/e2 was suppressed by introducing k and Ist1, indicating that the enlargement of dorsal petal in e/e2 requires the activities of K and LST1. An experimental framework of comparative genomic mapping approach was set up to map and clone LjELE1 locus in Lotus japonicus. Cloning the ELE1 gene will shed light on the underlying molecular mechanism during zygomorphic development and further provide the molecular basis for genetic improvement on legume crops.

LATHYROIDES, Encoding a WUSCHEL-Related Homeoboxl Transcription Factor, Controls Organ Lateral Growth, and Regulates Tendril and Dorsal Petal Identities in Garden Pea （Pisum sativum L.）

During organ development, many key regulators have been identified in plant genomes, which play a conserved role among plant species to control the organ identities and/or determine the organ size and shape. It is intriguing whether these key regulators can acquire diverse function and be integrated into different molecular pathways among different species, giving rise to the immense diversity of organ forms in nature. In this study, we have characterized and cloned LATHYROIDES （LATH）, a classical locus in pea, whose mutation displays pleiotropic alteration of lateral growth of organs and predominant effects on tendril and dorsal petal development. LATH encodes a WUSCHEL-related home- oboxl （WOX1） transcription factor, which has a conserved function in determining organ lateral growth among different plant species. Furthermore, we showed that LATH regulated the expression level of TENDRIL-LESS （TL）, a key factor in the control of tendril development in compound leaf, and LATH genetically interacted with LOBED STANDARD （LST）, a floral dorsal factor, to affect the dorsal petal identity. Thus, LATH plays multiple roles during organ development in pea： it maintains a conserved function controlling organ lateral outgrowth, and modulates organ identities in compound leaf and zygomorphic flower development, respectively. Our data indicated that a key regulator can play important roles in different aspects of organ development and dedicate to the complexity of the molecular mechanism in the control of organ development so as to create distinct organ forms in different species.

Genetic pathways controlling inflorescence architecture and development in wheat and barley

Modifications of inflorescence architecture have been crucial for the successful domestication of wheat and barley, which are central members of the Triticeae tribe that provide essential grains for the human diet. Investigation of the genes and alleles that underpin domestication-related traits has provided valuable insights into the molecular regulation of inflorescence development of the Triticeae, and further investigation of modified forms of architecture are proving to be equally fruitful.The identified genes are involved in diverse biological processes, including transcriptional regulation, hormone biosynthesis and metabolism, post-transcriptional and post-translational regulation, which alter inflorescence architecture by modifying the development and fertility of lateral organs, called spikelets and florets. Recent advances in sequencing capabilities and the generation of mutant populations are accelerating the identification of genes that influence inflorescence development, which is important given that genetic variation for this trait promises to be a valuable resource for optimizing grain production. This review assesses recent advances in our understanding of the genes controlling inflorescence development in wheat and barley, with the aim of highlighting the importance of improvements in developmental biology for optimizing the agronomic performance of staple crop plants.

Bacterial Vector-Borne Plant Diseases:Unanswered Questions and Future Directions

Vector-borne plant diseases have significant ecological and economic impacts,affecting farm profitability and forest composition throughout the world.Bacterial vector-borne pathogens have evolved sophisticated strategies to interact with their hemipteran insect vectors and plant hosts.These pathogens reside in plant vascular tissue,and their study represents an excellent opportunity to uncover novel biological mechanisms regulating intracellular pathogenesis and to contribute to the control of some of the world's most invasive emerging diseases.In this perspective,we highlight recent advances and major unanswered questions in the realm of bacterial vector-borne disease,focusing on liberibacters,phytoplasmas,spiroplasmas,and Xylella fastidiosa.