Plants have mechanisms to recognize and reject pollen from other species. Although widespread, these mechanisms are less well understood than the self‐incompatibility (
The collaborative non‐self‐recognition model for S‐
- NSF-PAR ID:
- 10246763
- Publisher / Repository:
- Wiley-Blackwell
- Date Published:
- Journal Name:
- The Plant Journal
- Volume:
- 87
- Issue:
- 6
- ISSN:
- 0960-7412
- Page Range / eLocation ID:
- p. 606-616
- Format(s):
- Medium: X
- Sponsoring Org:
- National Science Foundation
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Summary SI ) mechanisms plants use to reject pollen from close relatives. Previous studies have shown that some interspecific reproductive barriers (IRB s) are related toSI in the Solanaceae. For example, the pistilSI proteins S‐RN ase andHT protein function in a pistil‐sideIRB that causes rejection of pollen from self‐compatible (SC ) red/orange‐fruited species in the tomato clade. However, S‐RN ase‐independentIRB s also clearly contribute to rejecting pollen from these species. We investigated S‐RN ase‐independent rejection ofSolanum lycopersicum pollen bySC Solanum pennellii LA 0716,SC .Solanum habrochaites LA 0407, andSC Solanum arcanum LA 2157, which lack functional S‐RN ase expression. We found that all three accessions expressHT proteins, which previously had been known to function only in conjunction with S‐RN ase, and then usedRNA i to test whether they also function in S‐RN ase‐independent pollen rejection. SuppressingHT SC S. pennellii LA 0716 allowsS. lycopersicum pollen tubes to penetrate farther into the pistil inHT HT SC .Solanum habrochaites LA 0407 and inSC S. arcanum LA 2157 allowsS. lycopersicum pollen tubes to penetrate to the ovary and produce hybrids that, otherwise, would be difficult to obtain. Thus,HT proteins are implicated in both S‐RN ase‐dependent and S‐RN ase‐independent pollen rejection. The results support the view that overall compatibility results from multiple pollen–pistil interactions with additive effects. -
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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 proteins
At SLAH 3,At ALMT 12,At TMEM 16 andAt CCC as the putative anion transporters responsible for these currents.At CCC ‐GFP was observed at the shank andAt SLAH 3‐GFP at the tip and shank of thePT plasma membrane. Both are likely to carry the majority of anion current at negative potentials, as extracellular anionic fluxes measured at the tip ofPT s with an anion vibrating probe were significantly lower inslah3 −/− andccc −/− mutants, but unaffected inalmt12 −/− andtmem16 −/− . We further characterised the effect ofpH andGABA by patch clamp. Strong regulation by extracellularpH was observed in the wild‐type, but not intmem16 −/− . Our results are compatible withAt TMEM 16 functioning as an anion/H+cotransporter and therefore, as a putativepH sensor.GABA presence: (1) inhibited the overall currents, an effect that is abrogated in thealmt12 −/− and (2) reduced the current inAt ALMT 12 transfectedCOS ‐7 cells, strongly suggesting the direct interaction ofGABA withAt ALMT12.Our data show that
At SLAH 3 andAt CCC activity is sufficient to explain the major component of extracellular anion fluxes, and unveils a possible regulatory system linkingPT growth modulation bypH ,GABA , and [Ca2+]cytthrough anionic transporters. -
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Summary The mitochondrial and chloroplast
mRNA s of the majority of land plants are modified through cytidine to uridine (C‐to‐U)RNA editing. Previously, forward and reverse genetic screens demonstrated a requirement for pentatricopeptide repeat (PPR ) proteins forRNA editing. Moreover, chloroplast editing factorsOZ 1,RIP 2,RIP 9 andORRM 1 were identified in co‐immunoprecipitation (co‐IP) experiments, albeit the minimal complex sufficient for editing activity was never deduced. The current study focuses on isolated, intact complexes that are capable of editing distinct sites. Peak editing activity for four sites was discovered in size‐exclusion chromatography (SEC) fractions ≥ 670 kDa, while fractions estimated to be approximately 413 kDa exhibited the greatest ability to convert a substrate containing the editing siterps14 C80.RNA content peaked in the ≥ 670 kDa fraction. Treatment of active chloroplast extracts withRN ase A abolished the relationship of editing activity with high‐MW fractions, suggesting a structuralRNA component in native complexes. By immunoblotting,RIP 9,OTP 86,OZ 1 andORRM 1 were shown to be present in active gel filtration fractions, thoughOZ 1 andORRM 1 were mainly found in low‐MW inactive fractions. Active editing factor complexes were affinity‐purified using anti‐RIP 9 antibodies, and orthologs to putativeArabidopsis thaliana RNA editing factorPPR proteins,RIP 2,RIP 9,RIP 1,OZ 1,ORRM 1 andISE 2 were identified via mass spectrometry. Western blots from co‐IP studies revealed the mutual association ofOTP 86 andOZ 1 with nativeRIP 9 complexes. Thus,RIP 9 complexes were discovered to be highly associated with C‐to‐URNA editing activity and other editing factors indicative of their critical role in vascular plant editosomes. -
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Premise Herbarium specimens have been used to detect climate‐induced shifts in flowering time by using the day of year of collection (
DOY ) as a proxy for first or peak flowering date. Variation among herbarium sheets in their phenological status, however, undermines the assumption thatDOY accurately represents any particular phenophase. Ignoring this variation can reduce the explanatory power of pheno‐climatic models (PCM s) designed to predict the effects of climate on flowering date.Methods Here we present a protocol for the phenological scoring of imaged herbarium specimens using an ImageJ plugin, and we introduce a quantitative metric of a specimen's phenological status, the phenological index (
PI ), which we use inPCM s to control for phenological variation among specimens ofStreptanthus tortuosus (Brassicaceeae) when testing for the effects of climate onDOY . We demonstrate that includingPI as an independent variable improves model fit.Results Including
PI inPCM s increased the modelR 2relative toPCM s that excludedPI ; regression coefficients for climatic parameters, however, remained constant.Discussion Our protocol provides a simple, quantitative phenological metric for any observed plant. Including
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