Dehiscence Zone Research Articles

A comprehensive transcriptome analysis of silique development and dehiscence in Arabidopsis and Brassica integrating genotypic, interspecies and developmental comparisons

Asynchronous flowering of Brassica napus (canola) leads to seeds and siliques at varying stages of maturity as harvest approaches. This range of maturation can result in premature silique dehiscence (pod shattering), resulting in yield losses, which may be worsened by environmental stresses. Therefore, a goal for canola crop improvement is to reduce shattering in order to maximize yield. We performed a comprehensive transcriptome analysis on the dehiscence zone (DZ) and valve of Arabidopsis and Brassica siliques in shatter resistant and sensitive genotypes at several developmental stages. Among known Arabidopsis dehiscence genes, we confirmed that homologs of SHP1/2, FUL, ADPG1, NST1/3 and IND were associated with shattering in B. juncea and B. napus. We noted a correlation between reduced pectin degradation genes and shatter-resistance. Tension between lignified and non-lignified cells in the silique DZ plays a major role in dehiscence. Light microscopy revealed a smaller non-lignified separation layer in relatively shatter-resistant B. juncea relative to B. napus and this corresponded to increased expression of peroxidases involved in monolignol polymerization. Sustained repression of auxin biosynthesis, transport and signaling in B. juncea relative to B. napus may cause differences in dehiscence zone structure and cell wall constituents. Tension on the dehiscence zone is a consequence of shrinkage and loss of flexibility in the valves, which is caused by senescence and desiccation. Reduced shattering was generally associated with upregulation of ABA signaling and down-regulation of ethylene and jasmonate signaling, corresponding to more pronounced stress responses and reduced senescence and photosynthesis. Overall, we identified 124 cell wall related genes and 103 transcription factors potentially involved in silique dehiscence.

Evolution of fruit development genes in flowering plants.

The genetic mechanisms regulating dry fruit development and opercular dehiscence have been identified in Arabidopsis thaliana. In the bicarpellate silique, valve elongation and differentiation is controlled by FRUITFULL (FUL) that antagonizes SHATTERPROOF1-2 (SHP1/SHP2) and INDEHISCENT (IND) at the dehiscence zone where they control normal lignification. SHP1/2 are also repressed by REPLUMLESS (RPL), responsible for replum formation. Similarly, FUL indirectly controls two other factors ALCATRAZ (ALC) and SPATULA (SPT) that function in the proper formation of the separation layer. FUL and SHP1/2 belong to the MADS-box family, IND and ALC belong to the bHLH family and RPL belongs to the homeodomain family, all of which are large transcription factor families. These families have undergone numerous duplications and losses in plants, likely accompanied by functional changes. Functional analyses of homologous genes suggest that this network is fairly conserved in Brassicaceae and less conserved in other core eudicots. Only the MADS box genes have been functionally characterized in basal eudicots and suggest partial conservation of the functions recorded for Brassicaceae. Here we do a comprehensive search of SHP, IND, ALC, SPT, and RPL homologs across core-eudicots, basal eudicots, monocots and basal angiosperms. Based on gene-tree analyses we hypothesize what parts of the network for fruit development in Brassicaceae, in particular regarding direct and indirect targets of FUL, might be conserved across angiosperms.

Development of the pericarp, septa and zone of dehiscence in the fruit of Cassia tora L.

The pod of <em>Cassia tora</em> is the product of a single carpel consisting of 20-30 seeds enclosed one in each compartment formed by the ingrowth of the endocarpic tissue. There are three distinct histological zones of the pericarp Epicarp (a), Mesocarp (b) and Endocarp (c). The paracytic stomata and uniseriate multicellular trichomes are found only in the outermost layer of the epicarp. The epicarp is single layered. The outer mesocarp in later stages of its development has many collenchyma, particularly at dorsal and ventral margins. The mesocarp develops from the homogenous mesodermal tissue of the ovary wall. The middle mesocarpic tissue becomes sclerenchymatous above the :main bundles providing rigidity at the two sutures where the zones of dehiscence develop inbetween them. The endocarp develops from the inner epidermis and 2-3 hypodermal layers. Due to frequent periclinal and anticlinal divisions and enlargement of the cells in the inner hypodermal layers, the septa are formed which never fuse; thus the so called compartment of the seed is not completely closed and the seeds slide off on separation of the valves. The inner hypodermal layers form the band of sclerenchyma above the septal cells. The inner margins without septa and sclerenchyma provide easy opening of the two valves from the chamber side.

Role of the FUL–SHP network in the evolution of fruit morphology and function

Arabidopsis research in the last decade has started to unravel the genetic networks directing gynoecium and fruit patterning in this model species. Only recently, the work from several groups has also started to address the conservation of these networks in a wide number of species with very different fruit morphologies, and we are now beginning to understand how they might have evolved. This review summarizes recent advances in this field, focusing mainly on MADS-box genes with a well-known role in dehiscence zone development, while also discussing how these studies may contribute to expand our views on fruit evolution.

Over expression of the wheat BEL1-like gene TaqSH1 affects floral organ abscission in Arabidopsis thaliana

Seed shattering is one of the major domestication traits of crops. In wheat, except for the Q gene whose mutation renders free threshing and changing of rachis fragility, not much is known about the molecular mechanism for this process. We report here the cloning and characterization of TaqSH1, the ortholog of the rice seed shattering gene qSH1. TaqSH1 encodes a BEL1-like protein that is conserved between monocots and eudicots. TaqSH1 was located on the homoeologous group 3, a potential new genetic locus for seed threshability in wheat. Over expression of TaqSH1 in Arabidopsis resulted in dwarfed plants. The inflorescences of transgenic plants were more compact with larger pedicel angles. Scanning Electron Microscopy (SEM) showed that the transgenic siliques had a narrower replum where the dehiscence zone was altered. In addition, abscission of petals was significantly delayed due to delayed abscission zone development. Real-time PCR assays showed that over expression of TaqSH1 down regulated known Arabidopsis abscission related genes IDA, HAESA, KNAT1/6 and SHP1/2 in the transgenic plants. Taken together, our data suggest that TaqSH1 may represent another example of conserved mechanisms across monocots and eudicots for fruit/grain abscission and should have potential application in genetic manipulation of wheat seed shattering.

Evidence that an evolutionary transition from dehiscent to indehiscent fruits in Lepidium (Brassicaceae) was caused by a change in the control of valve margin identity genes

In the Brassicaceae, indehiscent fruits evolved from dehiscent fruits several times independently. Here we use closely related wild species of the genus Lepidium as a model system to analyse the underlying developmental genetic mechanisms in a candidate gene approach. ALCATRAZ (ALC), INDEHISCENT (IND), SHATTERPROOF1 (SHP1) and SHATTERPROOF2 (SHP2) are known fruit developmental genes of Arabidopsis thaliana that are expressed in the fruit valve margin governing dehiscence zone formation. Comparative expression analysis by quantitative RT-PCR, Northern blot and in situ hybridization show that their orthologues from Lepidium campestre (dehiscent fruits) are similarly expressed at valve margins. In sharp contrast, expression of the respective orthologues is abolished in the corresponding tissue of indehiscent Lepidium appelianum fruits, indicating that changes in the genetic pathway identified in A.thaliana caused the transition from dehiscent to indehiscent fruits in the investigated species. As parallel mutations in different genes are quite unlikely, we conclude that the changes in gene expression patterns are probably caused by changes in upstream regulators of ALC, IND and SHP1/2, possible candidates from A.thaliana being FRUITFULL (FUL), REPLUMLESS (RPL) and APETALA2 (AP2). However, neither expression analyses nor functional tests in transgenic plants provided any evidence that the FUL or RPL orthologues of Lepidium were involved in evolution of fruit indehiscence in Lepidium. In contrast, stronger expression of AP2 in indehiscent compared to dehiscent fruits identifies AP2 as a candidate gene that deserves further investigation.

Citrus sinensis L. Osbeck orthologs of FRUITFULL and SHATTERPROOF are differentially expressed during fruit development

Several regulatory steps and genes involved in fruit development were identified and characterized in Arabidopsis thaliana. FRUITFULL (FUL) and SHATTERPROOF (SHP), which belong to the MADS-box family of transcription factors, act together to promote the differentiation of the dehiscence zone and thus control the process of pod shattering in Arabidopsis. Homologs to these genes have been described in fleshy fruit species, but the specific nature of the regulatory hierarchy and interactions among these key regulators remains elusive in most plant species. Here, Citrus sinensis putative orthologs to FUL and SHP, named CsFUL and CsSHP respectively, were characterized. Phylogenetic comparisons indicated that CsFUL belongs to FRUITFUL sub-clade within the AP1/SQUA major clade while CsSHP falls into PLENA sub-clade from the AG/PLE clade. CsFUL and CsSHP protein sequences possess all of the characteristic conserved domains commonly found in A- and C-lineages of MIKC MADS-box proteins, respectively. Semi-quantitative RT-PCR showed preferential expression of both genes in developing fruits. In situ hybridization and a detailed analysis of Citrus fruit development using scanning electron microscopy allowed further characterization of these genes during C. sinensis fruit development. CsFUL and CsSHP are differentially expressed in exocarp, mesocarp and endocarp tissues in early stages of fruit development but their expression diminishes with fruit maturation. Moreover, the co-localization of CsFUL and CsSHP mRNA during oil glands and juice vesicle formation suggests a potential role in the development of such structures. Altogether, there results might contribute to a better understanding of the molecular mechanisms involved in Citrus fruit development.

A novel approach to dissect the abscission process in Arabidopsis.

Abscission is the consequence of a specialized layer of cells undergoing a complex series of molecular and biochemical events. Analysis of the specific molecular changes associated with abscission is hampered by contamination from neighboring nonseparating tissues. Moreover, studies of abscission frequently involve the examination of events that take place in isolated segments of tissue exposed to nonphysiological concentrations of ethylene or indole-3-acetic acid for protracted periods (more than 24 h) of time. To resolve these problems, we have adopted the use of a transgenic line of Arabidopsis (Arabidopsis thaliana) where the promoter of an abscission-specific polygalacturonase gene (At2g41850/ARABIDOPSIS DEHISCENCE ZONE POLYGALACTURONASE2) has been fused to a green fluorescent protein reporter. RNA was extracted from green fluorescent protein-tagged cells, released from abscising floral organs, and used to generate a complementary DNA library. This library was used to probe a microarray, and a population of abscission-related transcripts was studied in detail. Seven novel abscission-related genes were identified, four of which encode proteins of unknown function. Reverse transcription-polymerase chain reaction analyses and promoter fusions to the β-glucuronidase reporter gene confirmed the expression of these genes in the abscission zone and revealed other places of expression during seedling development. Three of these genes were studied further by crossing reporter lines to the abscission mutants inflorescence deficient in abscission (ida) and blade-on-petiole1 (bop1)/bop2 and an IDA-overexpressing line. Phenotypic analysis of an At3g14380 transfer DNA insertion line indicates that this gene plays a functional role in floral organ shedding. This strategy has enabled us to uncover new genes involved in abscission, and their possible contribution to the process is discussed.

Invited Commentary

Invited Commentary

Effect of Structural Differentiation of Fruit Dehiscence Zone on Pod Dehis-cence in Oilseed Rape

Effect of Structural Differentiation of Fruit Dehiscence Zone on Pod Dehis-cence in Oilseed Rape

SPATULA and ALCATRAZ, are partially redundant, functionally diverging bHLH genes required for Arabidopsis gynoecium and fruit development

The Arabidopsis gynoecium is a complex organ that facilitates fertilization, later developing into a dehiscent silique that protects seeds until their dispersal. Identifying genes important for development is often hampered by functional redundancy. We report unequal redundancy between two closely related genes, SPATULA (SPT) and ALCATRAZ (ALC), revealing previously unknown developmental roles for each. SPT is known to support septum, style and stigma development in the flower, whereas ALC is involved in dehiscence zone development in the fruit. ALC diverged from a SPT-like ancestor following gene duplication coinciding with the At-β polyploidy event. Here we show that ALC is also involved in early gynoecium development, and SPT in later valve margin generation in the silique. Evidence includes the increased severity of early gynoecium disruption, and of later valve margin defects, in spt-alc double mutants. In addition, a repressive version of SPT (35S:SPT-SRDX) disrupts both structures. Consistent with redundancy, ALC and SPT expression patterns overlap in these tissues, and the ALC promoter carries two atypical E-box elements identical to one in SPT required for valve margin expression. Further, SPT can heterodimerize with ALC, and 35S:SPT can fully complement dehiscence defects in alc mutants, although 35S:ALC can only partly complement spt gynoecium disruptions, perhaps associated with its sequence simplification. Interactions with FRUITFULL and SHATTERPROOF genes differ somewhat between SPT and ALC, reflecting their different specializations. These two genes are apparently undergoing subfunctionalization, with SPT essential for earlier carpel margin tissues, and ALC specializing in later dehiscence zone development.

Exploring a new model of cell wall regulation: identification and expression of two putative SHINEs transcription factors in Eucalyptus

Background Eucalyptus forests are a competitive and efficient alternative to convert carbon from the atmosphere in cellulose, an important source for paper manufacture and bioenergy production. To obtain transgenic Eucalyptus with important traits improved it is necessary to make modifications in genes that affect the final phenotype. One interesting gene that follows this requisite was recently found: this is the AtSHN2 gene (Arabidopsis thaliana SHINE 2). AtSHN2 codifies to a Transcription Factor known as “Arabidopsis SHINE/WAX INDUCER”. Instead of inducing drought tolerance in transgenic rice (Oryza sativa), AtSHN2 overexpression causes: i) 34% increase in the cellulose content; ii) 45% reduction in lignin content and iii) increase in wood digestibly (elevated S:G ratio) with no compromise in plant strength and performance [1]. The discovery of AtSHN2 function in plant cell wall formation, led Ambavaram and collaborators [1] to perform other studies and ultimately to propose the following model: AtSHN2 regulates positively MYB transcription factors (TF) related to cellulose synthesis and it downregulates MYBTF’s related to lignin formation. At the same time, SHINE can repress NAC TFthat controls MYB expression[1]. As a consequence of the interesting phenotype achieved through AtSHN2 overexpression in rice, this work focused on the identification and analyses of AtSHN orthologues in Eucalyptus. Bioinformatics tools were used to search for AtSHN similar genes in Eucalyptus. Moreover, the expression profile of the corresponding genes in Eucalyptus was evaluated to prove their role as AtSHN. To carry it on, the expression experiments were done with flower, leaf and xylem. If the Eucalyptus putativeSHINE’s has the same function of the AtSHN’s,, gene expression in flower tissues will be the highest [2]. This is because it is known that AtSHN’s genes are preferentially expressed in abscission and dehiscence zones, a phenomenon that usually occurs in lots of flower tissues.

The mechanism for explosive seed dispersal in Cardamine hirsuta (Brassicaceae)

Although many highly successful weed species use a ballistic seed dispersal mechanism, little is known about the mechanics of this process. Bittercress (Cardamine hirsuta) siliques are morphologically similar to Arabidopsis siliques, but they can project their seeds up to 5 m, while Arabidopsis seeds are dispersed by gravity. Comparison of these species should enable us to determine which structures might be responsible for ballistic seed dispersal. Sections of Arabidopsis and bittercress siliques were immunolabeled with antibodies raised against a variety of polysaccharide epitopes. In bittercress, the second endocarp layer (enB) of the valve had strongly asymmetrical cell wall thickenings, whereas the analogous cells in Arabidopsis were reinforced symmetrically and to a lesser extent. Additionally, an accumulation of mucilaginous pectins was found between the first and second endocarp (enA and enB) layers in the bittercress valve that was not present in Arabidopsis. However, in both species, highly de-esterified homogalacturonan was lost in the dehiscence zone (at the carpel/replum interface) as the siliques matured, thus allowing for separation of the valve at maturity. Ballistic seed dispersal in bittercress may involve the contraction of the outer pericarp tissue against the highly asymmetrically thickened enB cells, which are hypothesized to bend in one direction preferentially. The stress generated by the differential drying of the inner and outer layers of the valve is released suddenly as the adhesion between the cells of the dehiscence zone is lost, leading to a rapid coiling of the valve and dispersal of the seeds.

Identification and genetic characterization of an ethylene-dependent polygalacturonase from apricot fruit

During fruit ripening a loss of firmness occurs, which is a key factor limiting postharvest life. In apricot, Prunus armeniaca L., a wide range of fruit firmness at commercial maturity has been observed in different cultivars. Endopolygalacturonase (endoPG) activity has been reported to be associated with differences in firmness in many fruit species, but never in apricot. In this paper, we reported the identification of an apricot cDNA ( PaPG) coding for an endoPG-like protein with 393 amino acids. Protein sequence comparison with known polygalacturonases (PGs) revealed that multiple features as conserved domains and functional residues and a predicted signal peptide were present in PaPG. Moreover, a phylogenetic analysis of this and other plant PGs placed PaPG into a clade containing endoPGs expressed in fruit, abscission and dehiscence zones without a propeptide sequence, very close to PRF5 from peach ( Prunus persica L. Batsch). PaPG gene expression increased during postharvest storage of the fruit, correlating with fruit softening and ethylene release, and it responded to exogenous ethylene treatments. We localized the PaPG gene in apricot linkage group 4 after genetic mapping based on SNP analysis, in a position apparently syntenic to the PRF5 locus from peach. Results obtained offer genetic evidence supporting the hypothesis that PaPG and PRF5 are orthologous genes, and consequently position PaPG as a gene of interest for studies on fruit softening in apricot, and contribute to the development of molecular tools for breeding apricots with longer shelf life.

Ethephon and Aventrol as tools to enhance spring rape productivity

Abstract Ethephon and Aventrol were used as tools to provoke the processes taking part in the formation of rape seed yield and quality. Investigations on spring rape (Brassica napus L.) cultivars ‘Terra’ and ‘Landmark’ were carried out from 2008–2010. Ethephon (10 mM) and Aventrol (1 l/ha — pinolene 960 g/l) were used on different plant growth stages: BBCH-62–64 and BBCH-72–74, BBCH-80–82, respectively. Impact of ethephon manifested itself as activation of ethylene evocation by siliqua and a slight activation of growth of siliqua dehiscence zone. Siliqua cell plasmalemma and tonoplast H+-ATPases activation under the influence of ethephon occurred but did not lead to the destruction of transmembrane electrochemical potential. Extra seed yield and crude fat yield increased; tendency towards a lowering of the saturated/unsaturated fatty acid ratio was observed. Under the influence of Aventrol the dehiscence zone of siliqua was more closed when compared to the control and the ethephon treated variants, seed loss was significantly lowered and transmembrane cation transport was not damaged. The seed yield increased, and this was due to the accumulation of extra crude fat. Aventrol did not change the fatty acid content in rape seed oil. The positive impacts of ethephon and Aventrol for spring rape seed yield formation and possible mechanisms are discussed.

Cadmium affects the expression of ELF4, a circadian clock gene in Arabidopsis

The transcript abundance of clock-associated genes in Arabidopsis thaliana was monitored in a time point experiment, in plants treated with cadmium (Cd) for 24 h and upon two weeks Cd exposure and maintained in 12-h light/12-h dark cycles. The mRNA abundance of several molecular components associated with the circadian clock was modulated by Cd in plants treated for 24 h. In particular the CIRCADIAN CLOCK ASSOCIATED1 (CCA1), LATE ELONGATED HYPOCOTYL (LHY), TIMING OF CAB EXPRESSION1 (TOC1), EARLY FLOWERING3 (ELF3) and EARLY FLOWERING4 (ELF4) exhibited an amplified transcription peak and GIGANTEA (GI) showed a postponed and increased mRNA peak. Short-term Cd treatment did not affect the transcription level of CONSTANS (CO) or ZEITLUPE (ZTL). Long-term Cd exposure did not alter rhythmicity and expression amplitude with the exception of ELF4 that showed higher RNA abundance in plants Cd treated. In addition, measurement of flowering time in wild-type and ELF4-overexpressing plants exposed to Cd indicated that it is not altered by the environmental stress posed by Cd toxicity. The ELF4 promoter study indicated that this gene is expressed in leaves as well as in stamen, abscission and dehiscence zones. Long-term Cd exposure significantly modified ELF4 expression in these organs. The presence of this metal in polluted sites could therefore impact plant responsiveness to photoperiod and temperature and hence developmental decisions.

A fertilization-independent developmental program triggers partial fruit development and senescence processes in pistils of Arabidopsis.

The pistil is the specialized plant organ that enables appropriate pollination and ovule fertilization, after which it undergoes growth and differentiation to become a fruit. However, in most species, if ovules are not fertilized around anthesis the pistil irreversibly loses its growth capacity. We used physiological, molecular, and transcriptomic tools to characterize the post-anthesis development of the unfertilized Arabidopsis (Arabidopsis thaliana) pistil. Surprisingly, developmental processes that have been previously described in developing Arabidopsis fruits, such as the collapse of the adaxial epidermis, differentiation of a sclerenchyma layer in the adaxial subepidermis and the dehiscence zone, and valve dehiscence, were also observed in the unfertilized pistil. We determined that senescence is first established in the transmitting tract, stigma, and ovules immediately after anthesis, and that the timing of senescence in the stigma and ovules correlates with the loss of fruit-set responsiveness of the pistil to pollen and the hormone gibberellin (GA), respectively. Moreover, we showed that mutants with altered ovule development have impaired fruit-set response to the GA gibberellic acid, which further indicates that the presence of viable ovules is required for fruit-set responsiveness to GAs in the unfertilized pistil. Our data suggest that a fertilization-independent developmental program controls many of the processes during post-anthesis development, both in unfertilized pistils and seeded fruits, and point to a key role of the ovule in the capacity of pistils to undergo fruit set in response to GA.

Isolation of a Polygalacturonase Gene from Citrus sinensis Fruit and Its Expression Relative to Fruit Mastication Trait, Fruit Development, and Calcium or Boron Treatments

Polygalacturonase (PG) plays a significant role in pectin solubilization and depolymerization in vivo and is involved in numerous plant developmental processes. In this study, a polygalacturonase gene (CitPG, EF185420) was isolated from Citrus sinensis. It is 1,686 bp with a putative open reading frame of 1,338 bp encoding 445 amino acids. Phylogenetic analysis showed that CitPG belongs to clade B which includes PGs expressed in fruits or dehiscence zones. CitPG expression profiles during fruit ripening were compared between two pairs of varieties which differ in their mastication traits: C. sinensis cv. Fengjiewancheng (FJWC) vs. C. sinensis cv. Fengjie 72-1(FJ72-1) and Citrus reticulata cv. Kinokuni (NM) vs. C. reticulata cv. Miguang (MG), in which the mastication trait of “FJWC” and “NM” are inferior to their respective partner cultivars. Results indicated that the CitPG transcript level was significantly lower in the inferior mastication variety at the late ripening stage or at all ripening stages. Calcium or boron treatment resulted in a significant reduction in CitPG transcription level in fruit pulp at harvest time (235 days after anthesis). This was accompanied by an increase in protopectin content. In addition, the mRNA level of CitPG in the fruit peel and pulp increased during the fruit's rapid cell enlargement stage and was inversely correlated with protopectin levels. These results suggest that CitPG is associated with the favorable enhancement of the fruit mastication trait and is involved in dissolving the protopectin during cell enlargement.

Regulation of tissue-specific expression of SPATULA, a bHLH gene involved in carpel development, seedling germination, and lateral organ growth in Arabidopsis

SPATULA is a bHLH transcription factor that promotes growth of tissues arising from the carpel margins, including the septum and transmitting tract. It is also involved in repressing germination of newly harvested seeds, and in inhibiting cotyledon, leaf, and petal expansion. Using a reporter gene construct, its expression profile was fully defined. Consistent with its known functions, SPT was expressed in developing carpel margin tissues, and in the hypocotyls and cotyledons of germinating seedlings, and in developing leaves and petals. It was also strongly expressed in tissues where no functions have been identified to date, including the dehiscence zone of fruits, developing anthers, embryos, and in the epidermal initials and new stele of root tips. The promoter region of SPT was dissected by truncation and deletion, and two main regions occupied by tissue-specific enhancers were identified. These were correlated with eight regions conserved between promoter regions of Arabidopsis, Brassica oleracea, and Brassica rapa. When transformed into Arabidopsis, the B. oleracea promoter drove expression in reproductive tissues mostly comparable to the equivalent Arabidopsis promoter. There is genetic evidence that SPT function in the gynoecium is associated with the perception of auxin. However, site-directed mutagenesis of three putative auxin-response elements had no detectable effect on SPT expression patterns. Even so, disruption of a putative E-box variant adjacent to one of these resulted in a loss of valve dehiscence zone expression. This expression was also specifically lost in mutants of another bHLH gene INDEHISCENT, indicating that IND may directly regulate SPT expression through this variant E-box.

Analysis of promoter activity of members of the PECTATE LYASE-LIKE (PLL) gene family in cell separation in Arabidopsis

BackgroundPectate lyases depolymerize pectins by catalyzing the eliminative cleavage of α-1,4-linked galacturonic acid. Pectate lyase-like (PLL) genes make up among the largest and most complex families in plants, but their cellular and organismal roles have not been well characterized, and the activity of these genes has been assessed only at the level of entire organs or plant parts, potentially obscuring important sub-organ or cell-type-specific activities. As a first step to understand the potential functional diversity of PLL genes in plants and specificity of individual genes, we utilized a reporter gene approach to document the spatial and temporal promoter activity for 23 of the 26 members of the Arabidopsis thaliana (Arabidopsis) PLL gene family throughout development, focusing on processes involving cell separation.ResultsNumerous PLL promoters directed activity in localized domains programmed for cell separation, such as the abscission zones of the sepal, petal, stamen, and seed, as well as the fruit dehiscence zone. Several drove activity in cell types expected to facilitate separation, including the style and root endodermal and cortical layers during lateral root emergence. However, PLL promoters were active in domains not obviously programmed for separation, including the stipule, hydathode and root axis. Nearly all PLL promoters showed extensive overlap of activity in most of the regions analyzed.ConclusionsOur results document potential for involvement of PLL genes in numerous aspects of growth and development both dependent and independent of cell separation. Although the complexity of the PLL gene family allows for enormous potential for gene specialization through spatial or temporal regulation, the high degree of overlap of activity among the PLL promoters suggests extensive redundancy. Alternatively, functional specialization might be determined at the post-transcriptional or protein level.