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Strable, J.

Publications and source records attributed to Strable, J..

9 recordsLinked to original sources

The teosinte mexicana chromosomal inversion Inv4m modulates maize flowering time, plant height, and growth regulation gene networks

Chromosomal inversions facilitate local adaptation by maintaining co-adapted allele complexes as a single inherited unit, but the genes driving their phenotypic effects are rarely identified. Inv4m is an inversion from the highland teosinte Zea mays ssp. mexicana that is nearly fixed at 2500 masl in Mexican traditional varieties but absent in temperate maize. While association studies link Inv4m to flowering time, its key adaptive trait, the mechanisms connecting it to this phenotype remain unknown. Here, we generate near isogenic lines by introgressing the highland Michoacan 21 (Mi21) Inv4m haplotype into temperate B73 through eight backcross generations. We assemble the Inv4m-Mi21 NIL genome and, aligning to three Zea reference genomes, provide the first precise delimitation of Inv4m, a 15.2 Mb region with breakpoints overlapping knob repeat arrays. In field trials across Pennsylvania and North Carolina, Inv4m consistently accelerated flowering, whereas its plant height effect reversed sign between regions, a genotype by environment interaction consistent with environment-dependent fitness. Using RNA-seq, we identify 465 differentially expressed genes, the strongest from a cluster of JUMONJI histone demethylases (JMJ). Comparing five Zea genomes, the B73 reference carries a lineage-specific tandem expansion of five JMJ paralogs while highland genotypes carrying Inv4m retain a single ancestral copy, a difference that accounts for most of the clusters differential expression. We then show Inv4m disrupts growth related coexpression modules, with the JMJ cluster losing connectivity, and traced a trans regulatory network linking it to cell proliferation genes including pcna2 and sec6. In summary, we identify candidate genes and networks underlying Inv4ms effects and propose that part of this inversions adaptive value may reside in a dosage-sensitive regulator whose action propagates through a downstream network of genes that are involved in growth and flowering and are important for highland adaptation. Recombination suppression may thus protect a co-adapted regulatory architecture rather than independent alleles alone. Graphical Abstract O_FIG O_LINKSMALLFIG WIDTH=194 HEIGHT=200 SRC="FIGDIR/small/732082v1_ufig1.gif" ALT="Figure 1"> View larger version (106K): org.highwire.dtl.DTLVardef@85400org.highwire.dtl.DTLVardef@c2f820org.highwire.dtl.DTLVardef@301c0aorg.highwire.dtl.DTLVardef@80e715_HPS_FORMAT_FIGEXP M_FIG C_FIG

Plant Biology↗

Transit amplifying cells balance growth and differentiation in above-ground meristems

In both animals and plants, stem cell niches balance between cell-renewal and the generation of progeny cells that differentiate into specialized tissues. The multipotent and highly proliferative transit amplifying cells (TACs) integrate signals from stem cells and their differentiating progeny cells. Here we used spatial transcriptomics mapped to individual cells to illustrate the localization of TACs in maize meristems based on transcriptional gradients. Through genetic interactions and fluctuations in the transcriptional gradients we show that the multiplicative cell divisions are independently controlled from the TAC cell divisions. The dynamic nature of transcriptional variation in response to cell state or environment, together with the potential to improve yield by their modulation highlights the importance of finetuned modulation of networks rather than constitutive perturbations for crop improvement.

plant biology↗

Regulatory variation controlling architectural pleiotropy in maize

An early event in plant organogenesis is establishment of a boundary between the meristem and differentiating lateral organ. In maize (Zea mays), evidence suggests a common gene network functions at boundaries of distinct organs and contributes to pleiotropy between leaf angle and tassel branch number, two agronomic traits. To uncover regulatory variation at the nexus of these two traits, we used regulatory network topologies derived from specific developmental contexts to guide multivariate genome-wide association analyses. In addition to defining network plasticity around core pleiotropic loci, we identified new transcription factors that contribute to phenotypic variation in canopy architecture, and structural variation that contributes to cis-regulatory control of pleiotropy between tassel branching and leaf angle across maize diversity. Results demonstrate the power of informing statistical genetics with context-specific developmental networks to pinpoint pleiotropic loci and their cis-regulatory components, which can be used to fine-tune plant architecture for crop improvement.

plant biology↗

Cytoplasmic Male Sterility and Abortive Seed Traits Generated through Mitochondrial Genome Editing Coupled with Allotopic Expression of atp1 in Tobacco

Allotopic expression is the term given for the deliberate relocation of gene function from an organellar genome to the nuclear genome. We hypothesized that the allotopic expression of an essential mitochondrial gene using a promoter that expressed efficiently in all cell types except those responsible for male reproduction would yield a cytoplasmic male sterility (CMS) phenotype once the endogenous mitochondrial gene was inactivated via genome editing. To test this, we repurposed the mitochondrially encoded atp1 gene of tobacco to function in the nucleus under the transcriptional control of a CaMV 35S promoter (construct 35S:nATP1), a promoter that has been shown to be minimally expressed in early stages of anther development. The endogenous atp1 gene was eliminated ({Delta}atp1) from 35S:nATP1 tobacco plants using custom-designed meganucleases directed to the mitochondria. Vegetative growth of most 35S:nATP1/{Delta}atp1 plants appeared normal, but upon flowering produced malformed anthers that failed to shed pollen. When 35S:nATP1/{Delta}atp1 plants were cross-pollinated, ovary/capsule development appeared normal, but the vast majority of the resultant seeds were small, largely hollow and failed to germinate, a phenotype akin to the seedless trait known as stenospermocarpy. Characterization of the mitochondrial genomes from three independent {Delta}atp1 events suggested that spontaneous recombination over regions of microhomology and substoichiometric shifting were the mechanisms responsible for atp1 elimination and genome rearrangement in response to exposure to the atp1-targeting meganucleases. Should the results reported here in tobacco prove to be translatable to other crop species, then multiple applications of allotopic expression of an essential mitochondrial gene followed by its elimination through genome editing can be envisaged. Depending on the promoter(s) used to drive the allotopic gene, this technology may have potential application in the areas of: (1) CMS trait development for use in hybrid seed production; (2) seedless fruit production; and (3) transgene containment.

plant biology↗

Interspecies transfer of syntenic RAMOSA1 orthologs and promoter cis sequences impacts maize inflorescence architecture

Grass inflorescences support floral structures that each bear a single grain, where variation in branch architecture directly impacts yield. The maize RAMOSA1 (ZmRA1) transcription factor acts as a key regulator of inflorescence development by imposing branch meristem determinacy. Here, we show RA1 transcripts accumulate in boundary domains adjacent to spikelet meristems in Sorghum bicolor (Sb) and Setaria viridis (Sv) inflorescences similar as in the developing maize tassel and ear. To evaluate functional conservation of syntenic RA1 orthologs and promoter cis sequences in maize, sorghum and setaria, we utilized interspecies gene transfer and assayed genetic complementation in a common inbred background by quantifying recovery of normal branching in highly ramified ra1-R mutants. A ZmRA1 transgene that includes endogenous upstream and downstream flanking sequences recovered normal tassel and ear branching in ra1-R. Interspecies expression of two transgene variants of the SbRA1 locus, modeled as the entire endogenous tandem duplication or just the non-frameshifted downstream copy, complemented ra1-R branching defects and induced novel fasciation and branch patterns. The SvRA1 locus lacks conserved, upstream noncoding cis sequences found in maize and sorghum; interspecies expression of an SvRA1 transgene did not or only partially recovered normal inflorescence forms. Driving expression of the SvRA1 coding region by the ZmRA1 upstream region, however, recovered normal inflorescence morphology in ra1-R. These data leveraging interspecies gene transfer suggest that cis-encoded temporal regulation of RA1 expression is a key factor in modulating branch meristem determinacy that ultimately impacts grass inflorescence architecture.

plant biology↗

Plant stem cell organization and differentiation at single-cell resolution

Plants maintain populations of pluripotent stem cells in shoot apical meristems (SAMs), which continuously produce new aboveground organs. We used single-cell RNA sequencing to achieve an unbiased characterization of the transcriptional landscape of the maize shoot stem-cell niche and its differentiating cellular descendants. Stem cells housed in the SAM tip are engaged in genome integrity maintenance and exhibit a low rate of cell division, consistent with their contributions to germline and somatic cell fates. Surprisingly, we find no evidence for a canonical stem cell organizing center subtending these cells. In addition, we use trajectory inference to trace the gene expression changes that accompany cell differentiation. These data provide a valuable scaffold on which to better dissect the genetic control of plant shoot morphogenesis.

plant biology↗

The FUSED LEAVES1/ADHERENT1 Regulatory Module Is Required For Maize Cuticle Development And Organ Separation

In land plants all aerial epidermal cells are covered by the cuticle, an extracellular hydrophobic layer. The cuticle represents a primary barrier between cells and the external environment, provides protection against abiotic and biotic stresses, and prevents organ fusion during development. Here we report the cloning and characterization of a classic mutant of maize called adherent1 (ad1), first described a century ago, and we show that AD1 encodes a 3-KETOACYL-CoA SYNTHASE involved in the deposition of cuticular wax on the epidermis of leaves and inflorescences. ad1 mutants show decreased amounts of various wax components as well as a range of organ fusion defects during vegetative and reproductive development. Accordingly, we find that AD1 is strongly expressed in the epidermis of various developing organs where it is directly regulated by the MYB transcription factor FUSED LEAVES1 (FDL1), which in turn controls a series of additional genes involved in cuticle formation. Altogether, our results identify a major pathway of cuticle biosynthesis essential for the development of maize plants, and a key regulatory module that is conserved across monocot and eudicot species. One sentence summaryThe classic maize mutant adherent1, first isolated a century ago, is affected in an enzyme responsible for cuticle formation that is regulated by the MYB transcription factor FUSED LEAVES1.

plant biology↗

Maize nested introgression library provides evidence for the involvement of liguleless1 in resistance to northern leaf blight

Plant disease resistance is largely governed by complex genetic architecture. In maize, few disease resistance loci have been characterized. Near-isogenic lines (NILs) are a powerful genetic tool to dissect quantitative trait loci (QTL). We analyzed an introgression library of maize near-isogenic lines (NILs), termed a nested NIL (nNIL) library for resistance to northern leaf blight (NLB) caused by the fungal pathogen Setosphaeria turcica. The nNIL library was comprised of 412 BC5F4 NILs that originated from 18 diverse donor parents and a common recurrent parent, B73. Single nucleotide polymorphisms identified through genotyping by sequencing (GBS) were used to define introgressions and for association analysis. NILs that conferred resistance and susceptibility to NLB were comprised of introgressions that overlapped known NLB QTL. Genome-wide association analysis and stepwise regression further resolved five QTL regions, and implicated several candidate genes, including Liguleless1 (Lg1), a key determinant of leaf architecture in cereals. Two independently-derived mutant alleles of lg1 inoculated with S. turcica showed enhanced susceptibility to NLB. In the maize nested association mapping population, leaf angle was positively correlated with NLB in five recombinant inbred line (RIL) populations, and negatively correlated with NLB in four RIL populations. This study demonstrates the power of a nNIL library combined with high density SNP coverage to resolve QTLs. Furthermore, the role of lg1 in leaf architecture and in resistance to NLB has important applications in crop improvement.\n\nSignificance Statement (120 words)Understanding the genetic basis of disease resistance is important for crop improvement. We analyzed response to northern leaf blight (NLB) in a maize population consisting of 412 near-isogenic lines (NILs) derived from 18 diverse donor parents backcrossed to a recurrent parent, B73. NILs were genotyped by sequencing to detect introgressed segments. We identified NILs with greater resistance or susceptibility to NLB than B73. Genome-wide association analysis, coupled with stepwise regression, identified 5 candidate loci for NLB resistance, including the liguleless1 gene. The LIGULELESS1 transcription factor is critical in development of the leaf ligular region and influences leaf angle. We found that liguleless1 mutants are significantly more susceptible to NLB, uncovering a pleiotropic role for liguleless1 in development and disease resistance.

plant biology↗

Maize YABBY drooping leaf genes regulate floret development and floral meristem determinacy

Floret units in cereals produce grain, directly impacting yield. Here we report mutations in the maize CRABS CLAW (CRC) co-orthologs drooping leaf1 (drl1) and drl2 alter the development of ear and tassel florets. Pistillate florets of drl1 ears appear sterile and display ectopic unfused carpels that fail to enclose an expanded nucellus. Staminate florets of drl1 tassels have extra stamens and retain fertile anthers. Natural variation and transposon alleles of drl2 enhance drl1 floret phenotypes by reducing floral meristem (FM) determinacy. The drl paralogs are co-expressed in lateral floral organ primordia, but not within the FM. Together, the expression patterns and indeterminate mutant FMs suggest that the drl genes regulate FM activity and impose meristem determinacy by a non-cell autonomous signal. Genetic interaction analyses of drl mutants with maize floral mutants indicate that the drl genes are required throughout floret development, illustrating their importance for proper floret patterning in maize.

plant biology↗