skip to main content


Title: The long and short of the S ‐locus in Turnera (Passifloraceae)
Summary

Distyly is an intriguing floral adaptation that increases pollen transfer precision and restricts inbreeding. It has been a model system in evolutionary biology since Darwin. Although theS‐locus determines the long‐ and short‐styled morphs, the genes were unknown inTurnera. We have now identified these genes.

We used deletion mapping to identify, and then sequence,BACclones and genome scaffolds to constructS/shaplotypes. We investigated candidate gene expression, hemizygosity, and used mutants, to explore gene function.

Thes‐haplotype possessed 21 genes collinear with a region of chromosome 7 of grape. TheS‐haplotype possessed three additional genes and two inversions.TsSPH1was expressed in filaments and anthers,TsYUC6in anthers andTsBAHDin pistils. Long‐homostyle mutants did not possessTsBAHDand a short‐homostyle mutant did not expressTsSPH1.

Three hemizygous genes appear to determine S‐morph characteristics inT. subulata. Hemizygosity is common to all distylous species investigated, yet the genes differ. The pistil candidate gene,TsBAHD, differs from that ofPrimula, but both may inactivate brassinosteroids causing short styles.TsYUC6is involved in auxin synthesis and likely determines pollen characteristics.TsSPH1is likely involved in filament elongation. We propose an incompatibility mechanism involvingTsYUC6andTsBAHD.

 
more » « less
Award ID(s):
1759906
NSF-PAR ID:
10459731
Author(s) / Creator(s):
 ;  ;  ;  ;  ;  
Publisher / Repository:
Wiley-Blackwell
Date Published:
Journal Name:
New Phytologist
Volume:
224
Issue:
3
ISSN:
0028-646X
Page Range / eLocation ID:
p. 1316-1329
Format(s):
Medium: X
Sponsoring Org:
National Science Foundation
More Like this
  1. Summary

    Symbiotic nitrogen fixation in legumes is mediated by an interplay of signaling processes between plant hosts and rhizobial symbionts. In legumes, several secreted protein families have undergone expansions and play key roles in nodulation. Thus, identifying lineage‐specific expansions (LSEs) of nodulation‐associated genes can be a strategy to discover candidate gene families.

    Using bioinformatic tools, we identified 13LSEs of nodulation‐related secreted protein families, each unique to eitherGlycine,ArachisorMedicagolineages. In theMedicagolineage, nodule‐specific Polycystin‐1, Lipoxygenase, Alpha Toxin (PLAT) domain proteins (NPDs) expanded to five members. We examinedNPDfunction usingCRISPR/Cas9 multiplex genome editing to createMedicago truncatulaNPDknockout lines, targeting one to fiveNPDgenes.

    Mutant lines with differing combinations ofNPDgene inactivations had progressively smaller nodules, earlier onset of nodule senescence, or ineffective nodules compared to the wild‐type control. Double‐ and triple‐knockout lines showed dissimilar nodulation phenotypes but coincided in upregulation of aDHHC‐type zinc finger and an aspartyl protease gene, possible candidates for the observed disturbance of proper nodule function.

    By postulating that gene family expansions can be used to detect candidate genes, we identified a family of nodule‐specificPLATdomain proteins and confirmed that they play a role in successful nodule formation.

     
    more » « less
  2. Summary

    We investigated the molecular basis and physiological implications of anion transport during pollen tube (PT) growth inArabidopsis thaliana(Col‐0).

    Patch‐clamp whole‐cell configuration analysis of pollen grain protoplasts revealed three subpopulations of anionic currents differentially regulated by cytoplasmic calcium ([Ca2+]cyt). We investigated the pollen‐expressed proteinsAtSLAH3,AtALMT12,AtTMEM16 andAtCCCas the putative anion transporters responsible for these currents.

    AtCCCGFPwas observed at the shank andAtSLAH3‐GFPat the tip and shank of thePTplasma membrane. Both are likely to carry the majority of anion current at negative potentials, as extracellular anionic fluxes measured at the tip ofPTs with an anion vibrating probe were significantly lower inslah3−/−andccc−/−mutants, but unaffected inalmt12−/−andtmem16−/−. We further characterised the effect ofpHandGABAby patch clamp. Strong regulation by extracellularpHwas observed in the wild‐type, but not intmem16−/−. Our results are compatible withAtTMEM16 functioning as an anion/H+cotransporter and therefore, as a putativepHsensor.GABApresence: (1) inhibited the overall currents, an effect that is abrogated in thealmt12−/−and (2) reduced the current inAtALMT12 transfectedCOS‐7 cells, strongly suggesting the direct interaction ofGABAwithAtALMT12.

    Our data show thatAtSLAH3 andAtCCCactivity is sufficient to explain the major component of extracellular anion fluxes, and unveils a possible regulatory system linkingPTgrowth modulation bypH,GABA, and [Ca2+]cytthrough anionic transporters.

     
    more » « less
  3. Summary

    The collaborative non‐self‐recognition model for S‐RNase‐based self‐incompatibility predicts that multiple S‐locus F‐box proteins (SLFs) produced by pollen of a givenS‐haplotype collectively mediate ubiquitination and degradation of all non‐self S‐RNases, but not self S‐RNases, in the pollen tube, thereby resulting in cross‐compatible pollination but self‐incompatible pollination. We had previously used pollen extracts containingGFP‐fused S2SLF1 (SLF1 with anS2‐haplotype) ofPetunia inflatafor co‐immunoprecipitation (Co‐IP) and mass spectrometry (MS), and identified PiCUL1‐P (a pollen‐specific Cullin1), PiSSK1 (a pollen‐specific Skp1‐like protein) and PiRBX1 (a conventional Rbx1) as components of theSCFS2–SLF1complex. Using pollen extracts containing PiSSK1:FLAG:GFPfor Co‐IP/MS, we identified two additionalSLFs (SLF4 andSLF13) that were assembled intoSCFSLFcomplexes. As 17SLFgenes (SLF1toSLF17) have been identified inS2andS3pollen, here we examined whether all 17SLFs are assembled into similar complexes and, if so, whether these complexes are unique toSLFs. We modified the previous Co‐IP/MSprocedure, including the addition of style extracts from four differentS‐genotypes to pollen extracts containing PiSSK1:FLAG:GFP, to perform four separate experiments. The results taken together show that all 17SLFs and anSLF‐like protein,SLFLike1 (encoded by anS‐locus‐linked gene), co‐immunoprecipitated with PiSSK1:FLAG:GFP. Moreover, of the 179 other F‐box proteins predicted byS2andS3pollen transcriptomes, only a pair with 94.9% identity and another pair with 99.7% identity co‐immunoprecipitated with PiSSK1:FLAG:GFP. These results suggest thatSCFSLFcomplexes have evolved specifically to function in self‐incompatibility.

     
    more » « less
  4. Summary

    Plants mount coordinated immune responses to defend themselves against pathogens. However, the cellular components required for plant immunity are not fully understood. The jasmonate‐mimicking coronatine (COR) toxin produced byPseudomonas syringaepv.tomato(Pst)DC3000 functions to overcome plant immunity. We previously isolated eight Arabidopsis (scord) mutants that exhibit increased susceptibility to aCOR‐deficient mutant ofPstDC3000. Among them, thescord6mutant exhibits defects both in stomatal closure response and in restricting bacterial multiplication inside the apoplast. However, the identity ofSCORD6remained elusive.

    In this study, we aim to identify theSCORD6gene.

    We identifiedSCORD6via next‐generation sequencing and found it to beMURUS1(MUR1), which is involved in the biosynthesis ofGDPl‐fucose.

    Discovery ofSCORD6asMUR1led to a series of experiments that revealed a multi‐faceted role ofl‐fucose biosynthesis in stomatal and apoplastic defenses as well as in pattern‐triggered immunity and effector‐triggered immunity, including glycosylation of pattern‐recognition receptors. Furthermore, compromised stomatal and/or apoplastic defenses were observed in mutants of several fucosyltransferases with specific substrates (e.g.O‐glycan,N‐glycan or theDELLAtranscriptional repressors). Collectively, these results uncover a novel and broad role ofl‐fucose and protein fucosylation in plant immunity.

     
    more » « less
  5. Summary

    Plant smallRNAs (sRNAs) modulate key physiological mechanisms through post‐transcriptional and transcriptional silencing of gene expression. SmallRNAs fall into two major categories: those are reliant onRNA‐dependentRNApolymerases (RDRs) for biogenesis and those that are not. KnownRDR1/2/6‐dependentsRNAs include phased and repeat‐associated short interferingRNAs, while knownRDR1/2/6‐independentsRNAs are primarily microRNAs (miRNA) and other hairpin‐derivedsRNAs. In this study we produced and analyzedsRNA‐seq libraries fromrdr1/rdr2/rdr6triple mutant plants. We found 58 previously annotated miRNAloci that were reliant onRDR1, ‐2, or ‐6function, casting doubt on their classification. We also found 38RDR1/2/6‐independentsRNAloci that are notMIRNAs or otherwise hairpin‐derived, and did not fit into other known paradigms forsRNAbiogenesis. These 38sRNA‐producing loci have as‐yet‐undescribed biogenesis mechanisms, and are frequently located in the vicinity of protein‐coding genes. Altogether, our analysis suggests that these 38 loci represent one or more undescribed types ofsRNAinArabidopsis thaliana.

     
    more » « less