ABSTRACT
How do genes modify cellular growth to create morphological diversity? We study this problem in two related plants with differently shaped leaves: Arabidopsis thaliana (simple leaf shape) and Cardamine hirsuta (complex shape with leaflets). We use live imaging, modeling, and genetics to deconstruct these organ-level differences into their cell-level constituents: growth amount, direction, and differentiation. We show that leaf shape depends on the interplay of two growth modes: a conserved organ-wide growth mode that reflects differentiation; and a local, directional mode that involves the patterning of growth foci along the leaf edge. Shape diversity results from the distinct effects of two homeobox genes on these growth modes: SHOOTMERISTEMLESS broadens organ-wide growth relative to edge-patterning, enabling leaflet emergence, while REDUCED COMPLEXITY inhibits growth locally around emerging leaflets, accentuating shape differences created by patterning. We demonstrate the predictivity of our findings by reconstructing key features of C. hirsuta leaf morphology in A. thaliana. VIDEO ABSTRACT.
Subject(s)
Arabidopsis/growth & development , Cardamine/growth & development , Plant Leaves/growth & development , Arabidopsis/genetics , Cardamine/genetics , Cell Lineage/genetics , Computational Biology/methods , Gene Expression Regulation, Plant/genetics , Plant Leaves/genetics , Plant Proteins/metabolismABSTRACT
How tissue-level information encoded by fields of regulatory gene activity is translated into the patterns of cell polarity and growth that generate the diverse shapes of different species remains poorly understood. Here, we investigate this problem in the case of leaf shape differences between Arabidopsis thaliana, which has simple leaves, and its relative Cardamine hirsuta that has complex leaves divided into leaflets. We show that patterned expression of the transcription factor CUP-SHAPED COTYLEDON1 in C. hirsuta (ChCUC1) is a key determinant of leaf shape differences between the two species. Through inducible genetic perturbations, time-lapse imaging of growth, and computational modeling, we find that ChCUC1 provides instructive input into auxin-based leaf margin patterning. This input arises via transcriptional regulation of multiple auxin homeostasis components, including direct activation of WAG kinases that are known to regulate the polarity of PIN-FORMED auxin transporters. Thus, we have uncovered a mechanism that bridges biological scales by linking spatially distributed and species-specific transcription factor expression to cell-level polarity and growth, to shape diverse leaf forms.
Subject(s)
Arabidopsis Proteins , Arabidopsis , Cell Polarity , Gene Expression Regulation, Plant , Indoleacetic Acids , Plant Leaves , Indoleacetic Acids/metabolism , Plant Leaves/growth & development , Plant Leaves/genetics , Plant Leaves/metabolism , Cell Polarity/genetics , Arabidopsis/genetics , Arabidopsis/growth & development , Arabidopsis/metabolism , Arabidopsis Proteins/metabolism , Arabidopsis Proteins/genetics , Cardamine/genetics , Cardamine/metabolism , Cardamine/growth & development , Transcription Factors/metabolism , Transcription Factors/geneticsABSTRACT
Two interrelated problems in biology are understanding the regulatory logic and predictability of morphological evolution. Here, we studied these problems by comparing Arabidopsis thaliana, which has simple leaves, and its relative, Cardamine hirsuta, which has dissected leaves comprising leaflets. By transferring genes between the two species, we provide evidence for an inverse relationship between the pleiotropy of SHOOTMERISTEMLESS (STM) and BREVIPEDICELLUS (BP) homeobox genes and their ability to modify leaf form. We further show that cis-regulatory divergence of BP results in two alternative configurations of the genetic networks controlling leaf development. In C. hirsuta, ChBP is repressed by the microRNA164A (MIR164A)/ChCUP-SHAPED COTYLEDON (ChCUC) module and ChASYMMETRIC LEAVES1 (ChAS1), thus creating cross-talk between MIR164A/CUC and AS1 that does not occur in A. thaliana. These different genetic architectures lead to divergent interactions of network components and growth regulation in each species. We suggest that certain regulatory genes with low pleiotropy are predisposed to readily integrate into or disengage from conserved genetic networks influencing organ geometry, thus rapidly altering their properties and contributing to morphological divergence.
Subject(s)
Arabidopsis/growth & development , Arabidopsis/genetics , Cardamine/growth & development , Cardamine/genetics , Gene Regulatory Networks/genetics , Homeodomain Proteins/genetics , Plant Leaves , Plant Proteins/genetics , Arabidopsis/anatomy & histology , Arabidopsis Proteins/genetics , Arabidopsis Proteins/metabolism , Cardamine/anatomy & histology , Gene Expression Regulation, Plant , Homeodomain Proteins/metabolism , MicroRNAs/genetics , MicroRNAs/metabolism , Plant Leaves/anatomy & histology , Plant Leaves/genetics , Plant Leaves/growth & development , Plant Proteins/metabolismABSTRACT
Cardamine violifolia (Brassicaceae) is a novel selenium(Se) hyperaccumulation plant with rich nutrients, and serves as a good source of special vegetables in Enshi, China. The present study aimed to investigate the effects of the application of selenate, selenite, and Se yeast (50-800 mg/L) on the growth, Se accumulation, nutrient uptake, and antioxidant response of C. violifolia. The results showed that the Se accumulation efficiency was selenate > selenite > Se yeast, the maximum Se concentration could achieve over 7000 mg/kg, and about 90% was organic Se. The major Se speciation found was mainly SeCys2 and the proportion of various Se species were affected by the Se forms and concentrations. Besides, the plant growth, nutrition quality indexes, element uptakes, and antioxidant responses indicated that 200 mg/L selenate was optimum for C. violifolia to accumulate Se without much impacts, while to obtain more proportion of organic Se, 200 mg/L selenite might be a better choice.
Subject(s)
Antioxidants/metabolism , Cardamine/physiology , Selenic Acid/metabolism , Selenious Acid/metabolism , Selenium/metabolism , Yeast, Dried/chemistry , Bioaccumulation , Cardamine/chemistry , Cardamine/enzymology , Cardamine/growth & development , China , Dose-Response Relationship, Drug , Elements , Nutritional Physiological Phenomena , Nutritive ValueABSTRACT
A key question in biology is to understand how interspecies morphological diversities originate. Plant roots present a huge interspecific phenotypical variability, mostly because roots largely contribute to adaptation to different kinds of soils. One example is the interspecific cortex layer number variability, spanning from one to several. Here, we review the latest advances in the understanding of the mechanisms expanding and/or restricting cortical layer number in Arabidopsis thaliana and their involvement in cortex pattern variability among multi-cortical layered species such as Cardamine hirsuta or Oryza sativa.
Subject(s)
Arabidopsis/growth & development , Cardamine/growth & development , Oryza/growth & development , Plant Roots/growth & development , Arabidopsis/anatomy & histology , Cardamine/anatomy & histology , Oryza/anatomy & histology , Plant Roots/anatomy & histologyABSTRACT
Neofunctionalization following gene duplication is thought to be one of the key drivers in generating evolutionary novelty. A gene duplication in a common ancestor of land plants produced two classes of KNOTTED-like TALE homeobox genes, class I (KNOX1) and class II (KNOX2). KNOX1 genes are linked to tissue proliferation and maintenance of meristematic potentials of flowering plant and moss sporophytes, and modulation of KNOX1 activity is implicated in contributing to leaf shape diversity of flowering plants. While KNOX2 function has been shown to repress the gametophytic (haploid) developmental program during moss sporophyte (diploid) development, little is known about KNOX2 function in flowering plants, hindering syntheses regarding the relationship between two classes of KNOX genes in the context of land plant evolution. Arabidopsis plants harboring loss-of-function KNOX2 alleles exhibit impaired differentiation of all aerial organs and have highly complex leaves, phenocopying gain-of-function KNOX1 alleles. Conversely, gain-of-function KNOX2 alleles in conjunction with a presumptive heterodimeric BELL TALE homeobox partner suppressed SAM activity in Arabidopsis and reduced leaf complexity in the Arabidopsis relative Cardamine hirsuta, reminiscent of loss-of-function KNOX1 alleles. Little evidence was found indicative of epistasis or mutual repression between KNOX1 and KNOX2 genes. KNOX proteins heterodimerize with BELL TALE homeobox proteins to form functional complexes, and contrary to earlier reports based on in vitro and heterologous expression, we find high selectivity between KNOX and BELL partners in vivo. Thus, KNOX2 genes confer opposing activities rather than redundant roles with KNOX1 genes, and together they act to direct the development of all above-ground organs of the Arabidopsis sporophyte. We infer that following the KNOX1/KNOX2 gene duplication in an ancestor of land plants, neofunctionalization led to evolution of antagonistic biochemical activity thereby facilitating the evolution of more complex sporophyte transcriptional networks, providing plasticity for the morphological evolution of land plant body plans.
Subject(s)
Arabidopsis/genetics , Evolution, Molecular , Gene Duplication , Homeodomain Proteins/genetics , Life Cycle Stages/genetics , Plant Proteins/genetics , Arabidopsis/growth & development , Cardamine/genetics , Cardamine/growth & development , Diploidy , Gene Expression Regulation, Plant , Haploidy , Phenotype , Plant Leaves/genetics , Plant Leaves/growth & developmentABSTRACT
The developmental basis for the generation of divergent leaf forms is largely unknown. Here we investigate this problem by studying processes that distinguish development of two related species: Arabidopsis thaliana, which has simple leaves, and Cardamine hirsuta, which has dissected leaves with individual leaflets. Using genetics, expression studies and cell lineage tracing, we show that lateral leaflet formation in C. hirsuta requires the establishment of growth foci that form after leaf initiation. These growth foci are recruited at the leaf margin in response to activity maxima of auxin, a hormone that polarizes growth in diverse developmental contexts. Class I KNOTTED1-like homeobox (KNOX) proteins also promote leaflet initiation in C. hirsuta, and here we provide evidence that this action of KNOX proteins is contingent on the ability to organize auxin maxima via the PINFORMED1 (PIN1) auxin efflux transporter. Thus, differential deployment of a fundamental mechanism polarizing cellular growth contributed to the diversification of leaf form during evolution.
Subject(s)
Arabidopsis/genetics , Cardamine/genetics , Plant Leaves/growth & development , Arabidopsis/growth & development , Arabidopsis/metabolism , Arabidopsis Proteins/metabolism , Biological Evolution , Cardamine/growth & development , Cardamine/metabolism , Cell Cycle , Cell Lineage , Gene Expression Regulation, Developmental , Homeodomain Proteins/metabolism , Indoleacetic Acids/metabolism , Membrane Transport Proteins/metabolism , Mutation , Plant Leaves/genetics , Plant Proteins/metabolismABSTRACT
The evolution of plant reproductive strategies has led to a remarkable diversity of structures, especially within the flower, a structure characteristic of the angiosperms. In flowering plants, sexual reproduction depends notably on the development of the gynoecium that produces and protects the ovules. In Arabidopsis thaliana, ovule initiation is promoted by the concerted action of auxin with CUC1 (CUP-SHAPED COTYLEDON1) and CUC2, two genes that encode transcription factors of the NAC family (NAM/ATAF1,2/CUC). Here we highlight an additional role for CUC2 and CUC3 in Arabidopsis thaliana ovule separation. While CUC1 and CUC2 are broadly expressed in the medial tissue of the gynoecium, CUC2 and CUC3 are expressed in the placental tissue between developing ovules. Consistent with the partial overlap between CUC1, CUC2 and CUC3 expression patterns, we show that CUC proteins can physically interact, both in yeast cells and in planta. We found that the cuc2;cuc3 double mutant specifically harbours defects in ovule separation, producing fused seeds that share the seed coat, and suggesting that CUC2 and CUC3 promote ovule separation in a partially redundant manner. Functional analyses show that CUC transcription factors are also involved in ovule development in Cardamine hirsuta. Additionally we show a conserved expression pattern of CUC orthologues between ovule primordia in other phylogenetically distant species with different gynoecium architectures. Taken together these results suggest an ancient role for CUC transcription factors in ovule separation, and shed light on the conservation of mechanisms involved in the development of innovative structures.
Subject(s)
Ovule/growth & development , Ovule/metabolism , Arabidopsis/growth & development , Arabidopsis/metabolism , Arabidopsis Proteins/genetics , Arabidopsis Proteins/metabolism , Cardamine/growth & development , Cardamine/metabolism , Gene Expression Regulation, Plant , Magnoliopsida/growth & development , Magnoliopsida/metabolism , Plant Proteins/genetics , Plant Proteins/metabolism , Plants, Genetically Modified/growth & development , Plants, Genetically Modified/metabolism , Transcription Factors/genetics , Transcription Factors/metabolismABSTRACT
A key question in biology is how differences in gene function or regulation produce new morphologies during evolution. Here we investigate the genetic basis for differences in leaf form between two closely related plant species, Arabidopsis thaliana and Cardamine hirsuta. We report that in C. hirsuta, class I KNOTTED1-like homeobox (KNOX) proteins are required in the leaf to delay cellular differentiation and produce a dissected leaf form, in contrast to A. thaliana, in which KNOX exclusion from leaves results in a simple leaf form. These differences in KNOX expression arise through changes in the activity of upstream gene regulatory sequences. The function of ASYMMETRIC LEAVES1/ROUGHSHEATH2/PHANTASTICA (ARP) proteins to repress KNOX expression is conserved between the two species, but in C. hirsuta the ARP-KNOX regulatory module controls new developmental processes in the leaf. Thus, evolutionary tinkering with KNOX regulation, constrained by ARP function, may have produced diverse leaf forms by modulating growth and differentiation patterns in developing leaf primordia.
Subject(s)
Arabidopsis/growth & development , Arabidopsis/genetics , Cardamine/growth & development , Cardamine/genetics , Plant Leaves/growth & development , Plant Leaves/genetics , Arabidopsis Proteins/genetics , Biological Evolution , Gene Expression Regulation, Developmental , Gene Expression Regulation, Plant , Genes, Homeobox , Genes, Plant , Homeodomain Proteins/genetics , Molecular Sequence Data , Plant Proteins/genetics , Plants, Genetically Modified , Promoter Regions, Genetic , RNA, Messenger/genetics , RNA, Plant/genetics , Species Specificity , Transcription Factors/geneticsABSTRACT
The mutually exclusive relationship between ARP and KNOX1 genes in the shoot apical meristem and leaf primordia in simple leaved plants such as Arabidopsis has been well characterized. Overlapping expression domains of these genes in leaf primordia have been described for many compound leaved plants such as Solanum lycopersicum and Cardamine hirsuta and are regarded as a characteristic of compound leaved plants. Here, we present several datasets illustrating the co-expression of ARP and KNOX1 genes in the shoot apical meristem, leaf primordia, and developing leaves in plants with simple leaves and simple primordia. Streptocarpus plants produce unequal cotyledons due to the continued activity of a basal meristem and produce foliar leaves termed "phyllomorphs" from the groove meristem in the acaulescent species Streptocarpus rexii and leaves from a shoot apical meristem in the caulescent Streptocarpus glandulosissimus. We demonstrate that the simple leaves in both species possess a greatly extended basal meristematic activity that persists over most of the leaf's growth. The area of basal meristem activity coincides with the co-expression domain of ARP and KNOX1 genes. We suggest that the co-expression of ARP and KNOX1 genes is not exclusive to compound leaved plants but is associated with foci of meristematic activity in leaves.
Subject(s)
Gene Expression Regulation, Plant , Homeodomain Proteins/genetics , Meristem/genetics , Plant Leaves/genetics , Plant Proteins/genetics , Amino Acid Sequence , Cardamine/genetics , Cardamine/growth & development , Genes, Plant , Meristem/growth & development , Meristem/metabolism , Molecular Sequence Data , Plant Leaves/growth & development , Plant Leaves/metabolismABSTRACT
Cardamine hirsuta, a small crucifer closely related to the model organism Arabidopsis thaliana, offers high genetic tractability and has emerged as a powerful system for studying the genetic basis for diversification of plant form. Contrary to A. thaliana, which has simple leaves, C. hirsuta produces dissected leaves divided into individual units called leaflets. Leaflet formation requires activity of Class I KNOTTED1-like homeodomain (KNOX) proteins, which also promote function of the shoot apical meristem (SAM). In C. hirsuta, KNOX genes are expressed in the leaves whereas in A. thaliana their expression is confined to the SAM, and differences in expression arise through cis-regulatory divergence of KNOX regulation. KNOX activity in C. hirsuta leaves delays the transition from proliferative growth to differentiation thus facilitating the generation of lateral growth axes that give rise to leaflets. These axes reflect the sequential generation of cell division foci across the leaf proximodistal axis in response to auxin activity maxima, which are generated by the PINFORMED1 (PIN1) auxin efflux carriers in a process that resembles organogenesis at the SAM. Delimitation of C. hirsuta leaflets also requires the activity of CUP SHAPED COTYLEDON (CUC) genes, which direct formation of organ boundaries at the SAM. These observations show how species-specific deployment of fundamental shoot development networks may have sculpted simple versus dissected leaf forms. These studies also illustrate how extending developmental genetic studies to morphologically divergent relatives of model organisms can greatly help elucidate the mechanisms underlying the evolution of form.
Subject(s)
Cardamine/growth & development , Plant Leaves/growth & development , Cardamine/anatomy & histology , Genes, Homeobox/physiology , Genes, Plant , Homeodomain Proteins/genetics , Homeodomain Proteins/physiology , Insulator Elements/physiology , Meristem/anatomy & histology , Meristem/growth & development , Plant Growth Regulators/physiology , Plant Leaves/anatomy & histologySubject(s)
Plant Leaves/growth & development , Plant Leaves/genetics , Arabidopsis/genetics , Arabidopsis/growth & development , Cardamine/genetics , Cardamine/growth & development , Gene Expression Regulation, Developmental , Gene Expression Regulation, Plant , Genes, Homeobox , Genes, Plant , Plants, Genetically Modified , Promoter Regions, Genetic , Species SpecificityABSTRACT
Current advances in developmental genetics are increasingly underpinned by comparative approaches as more powerful experimental tools become available in non-model organisms. Cardamine hirsuta is related to the model plant Arabidopsis thaliana and comparisons between these two experimentally tractable species have advanced our understanding of development and diversity. The power of forward genetics to uncover new biology was evident in the isolation of REDUCED COMPLEXITY, a gene which is present in C. hirsuta but lost in A. thaliana, and shapes crucifer leaf diversity. Transferring two Knotted1-like homeobox genes between C. hirsuta and A. thaliana revealed a constraint imposed by pleiotropy on the evolutionary potential of cis regulatory change to modify leaf shape. FLOWERING LOCUS C was identified as a heterochronic gene that underlies natural leaf shape variation in C. hirsuta.
Subject(s)
Cardamine/genetics , Evolution, Molecular , Plant Development/genetics , Plant Proteins/genetics , Arabidopsis/genetics , Arabidopsis/growth & development , Cardamine/growth & development , Gene Expression Regulation, Plant , Plant Leaves/anatomy & histology , Plant Leaves/genetics , Plant Leaves/growth & developmentABSTRACT
Plants flower in response to many varied cues, such as temperature, photoperiod, and age. The floral transition of Cardamine flexuosa, a herbaceous biennial-to-perennial plant, requires exposure to cold temperature, a treatment known as vernalization. C. flexuosa younger than 5 weeks old are not fully responsive to cold treatment. We demonstrate that the levels of two age-regulated microRNAs, miR156 and miR172, regulate the timing of sensitivity in response to vernalization. Age and vernalization pathways coordinately regulate flowering through modulating the expression of CfSOC1, a flower-promoting MADS-box gene. The related annual Arabidopsis thaliana, which has both vernalization and age pathways, does not possess an age-dependent vernalization response. Thus, the recruitment of age cue in response to environmental signals contributes to the evolution of life cycle in plants.
Subject(s)
Cardamine/growth & development , Cold Temperature , Flowers/growth & development , Gene Expression Regulation, Plant , MADS Domain Proteins/genetics , Plant Proteins/genetics , Cardamine/genetics , Flowers/genetics , MicroRNAs/metabolism , Time FactorsABSTRACT
Diversity in leaf shape is produced by alterations of the margin: for example, deep dissection leads to leaflet formation and less-pronounced incision results in serrations or lobes. By combining gene silencing and mutant analyses in four distantly related eudicot species, we show that reducing the function of NAM/CUC boundary genes (NO APICAL MERISTEM and CUP-SHAPED COTYLEDON) leads to a suppression of all marginal outgrowths and to fewer and fused leaflets. We propose that NAM/CUC genes promote formation of a boundary domain that delimits leaflets. This domain has a dual role promoting leaflet separation locally and leaflet formation at distance. In this manner, boundaries of compound leaves resemble boundaries functioning during animal development.
Subject(s)
Gene Expression Regulation, Plant , Genes, Plant , Plant Leaves/growth & development , Transcription Factors/genetics , Amino Acid Sequence , Aquilegia/genetics , Aquilegia/growth & development , Aquilegia/metabolism , Cardamine/genetics , Cardamine/growth & development , Cardamine/metabolism , Gene Expression Profiling , Gene Silencing , Solanum lycopersicum/genetics , Solanum lycopersicum/growth & development , Solanum lycopersicum/metabolism , Molecular Sequence Data , Pisum sativum/genetics , Pisum sativum/growth & development , Phylogeny , Plant Leaves/genetics , Plant Leaves/metabolism , Plant Proteins/genetics , Plant Proteins/metabolism , Plants, Genetically Modified , Solanum tuberosum/genetics , Solanum tuberosum/growth & development , Solanum tuberosum/metabolism , Transcription Factors/chemistry , Transcription Factors/metabolismABSTRACT
BACKGROUND AND AIMS: Hybridization is an important evolutionary phenomenon, and therefore a detailed understanding of the dynamics of interspecific gene flow and resulting morphological and genetic patterns is of widespread interest. Here hybridization between the polyploids Cardamine pratensis and C. raphanifolia at four localities is explored. Using different types of data, the aim is to provide simultaneous and direct comparisons between genotype and phenotype variation patterns in the studied hybrid populations. METHODS: Evidence of hybridization has been gathered from morphology, molecular markers (amplified fragment length polymorphism and chloroplast DNA sequences), pollen viability, karyology and nuclear DNA content. KEY RESULTS: All data support extensive gene flow occurring in the hybrid populations. A wide range of morphological and genetic variation is observed, which includes both parental and intermediate types. Unbalanced pollen fertility and several ploidy levels are recorded. CONCLUSIONS: Incongruence reported between genotype and phenotype suggests that parental phenotypes are affected by introgression, and intermediate hybrid phenotypes can be genetically closer to one of the parents. Thus, it is evident that morphology, when used alone, can be misleading for interpreting hybridization, and critical evaluation of other data is needed.