A Remarkable New Leaf Type With Unusual Photosynthetic Tissue in a Central Asiatic Genus of Chenopodiaceae

Abstract: From the hygrohalophyte Borszczowia aralocaspica Bunge (Chenopodiaceae), a new leaf type with 1-layered chlorenchyma is described as "borszczovoid" and compared with other leaf types in subfamily Salsoloideae. The chlorenchyma is suspected to represent a unique C₄ type. Evidence is cited from anatomical studies and documented by micrographs and Carbon isotope determinations (ä¹³C values). The 1-layered photosynthetic tissue combines all essential anatomical characters of a 2-layered chlorenchyma of regular C₄ plants and is in intimate contact with concentrically arranged peripheral bundles. The ä¹³C values are − 13.03 ‰ from young stems and − 13.78 ‰ from leaves. The results are discussed in the anatomical, physiological and taxonomic framework. In addition, from distantly-related Suaeda species of section Conosperma the conospermoid leaf type is re-described. It is characterized by typical palisade and Kranz layers and differs from the C₄ suaedoid type by an external water-storaging hypodermis and an arrangement of Kranz cells reminiscent of the atriplicoid type from subfamily Chenopodioideae. From eight other species of Chenopodiaceae ä¹³C values are given for the first time.     

Structural, biochemical, and physiological characterization of C4 photosynthesis in species having two vastly different types of kranz anatomy in genus Suaeda (Chenopodiaceae)

C (4) species of family Chenopodiaceae, subfamily Suaedoideae have two types of Kranz anatomy in genus Suaeda, sections Salsina and Schoberia, both of which have an outer (palisade mesophyll) and an inner (Kranz) layer of chlorenchyma cells in usually semi-terete leaves. Features of Salsina (S. AEGYPTIACA, S. arcuata, S. taxifolia) and Schoberia type (S. acuminata, S. Eltonica, S. cochlearifoliA) were compared to C (3) type S. Heterophylla. In Salsina type, two layers of chlorenchyma at the leaf periphery surround water-storage tissue in which the vascular bundles are embedded. In leaves of the Schoberia type, enlarged water-storage hypodermal cells surround two layers of chlorenchyma tissue, with the latter surrounding the vascular bundles. The chloroplasts in Kranz cells are located in the centripetal position in Salsina type and in the centrifugal position in the Schoberia type. Western blots on C (4) acid decarboxylases show that both Kranz forms are NAD-malic enzyme (NAD-ME) type C (4) species. Transmission electron microscopy shows that mesophyll cells have chloroplasts with reduced grana, while Kranz cells have chloroplasts with well-developed grana and large, specialized mitochondria, characteristic of NAD-ME type C (4) chenopods. In both C (4) types, phosphoenolpyruvate carboxylase is localized in the palisade mesophyll, and Rubisco and mitochondrial NAD-ME are localized in Kranz cells, where starch is mainly stored. The C (3) species S. heterophylla has Brezia type isolateral leaf structure, with several layers of Rubisco-containing chlorenchyma. Photosynthetic response curves to varying CO (2) and light in the Schoberia Type and Salsina type species were similar, and typical of C (4) plants. The results indicate that two structural forms of Kranz anatomy evolved in parallel in species of subfamily Suaedoideae having NAD-ME type C (4) photosynthesis.     

Occurrence of C3 and C4 photosynthesis in cotyledons and leaves of Salsola species (Chenopodiaceae)

Most species of the genus Salsola (Chenopodiaceae) that have been examined exhibit C₄ photosynthesis in leaves. Four Salsola species from Central Asia were investigated in this study to determine the structural and functional relationships in photosynthesis of cotyledons compared to leaves, using anatomical (Kranz versus non-Kranz anatomy, chloroplast ultrastructure) and biochemical (activities of photosynthetic enzymes of the C₃ and C₄ pathways, ¹⁴C labeling of primary photosynthesis products and ¹³C/¹²C carbon isotope fractionation) criteria. The species included S. paulsenii from section Salsola, S. richteri from section Coccosalsola, S. laricina from section Caroxylon, and S. gemmascens from section Malpigipila. The results show that all four species have a C₄ type of photosynthesis in leaves with a Salsoloid type Kranz anatomy, whereas both C₃ and C₄ types of photosynthesis were found in cotyledons. S. paulsenii and S. richteri have NADP- (NADP-ME) C₄ type biochemistry with Salsoloid Kranz anatomy in both leaves and cotyledons. In S. laricina, both cotyledons and leaves have NAD-malic enzyme (NAD-ME) C₄ type photosynthesis; however, while the leaves have Salsoloid type Kranz anatomy, cotyledons have Atriplicoid type Kranz anatomy. In S. gemmascens, cotyledons exhibit C₃ type photosynthesis, while leaves perform NAD-ME type photosynthesis. Since the four species studied belong to different Salsola sections, this suggests that differences in photosynthetic types of leaves and cotyledons may be used as a basis or studies of the origin and evolution of C₄ photosynthesis in the family Chenopodiaceae.This revised version was published online in October 2005 with corrections to the Cover Date.     

A comparative anatomical and biochemical analysis in salsola (Chenopodiaceae) species with and without a Kranz type leaf anatomy: a possible reversion of C4 to C3 photosynthesis

Leaf anatomy was studied by light and electron microscopy and the leaf activities of RUBP carboxylase, PEP carboxylase, and malic enzyme were assayed in: Salsola australis and S. oreophila grown on the West Pamirs at 1800 m altitude; in S. australis grown on the East Pamirs at 3860 m; and in S. arbusculiformis grown in the Kisil-Kum desert in Middle Asia near 500 m. Carbon isotope fractionation ratio values also were measured on whole leaf tissue for 18 Salsola species field collected in these and other regions of the former USSR. S. australis leaves are cylindrical and in cross section exhibit a peripheral ring of mesophyll and then an inner ring of bundle sheath type cells; and its biochemical characteristics and deltaC values are typical of a C4 species of the NADP-malic enzyme malate-forming group. These traits were expressed independent of the plant growth altitude up to 4000 m. C4 type deltaC values were obtained in 14 of the Salsola species. Anatomical, structural, and biochemical features typical of the C4 syndrome were absent in S. oreophila and S. arbusculiformis. Four Salsola species, including these two, had C3-type deltaC values. Their cylindrical leaves in cross section exhibited two to three peripheral rings as layers of palisade parenchyma. Although their vascular bundles were surrounded by green bundle sheath cells, their organelle numbers were comparable to those in mesophyll cells. Neither bundle sheath cell wall thickenings nor dimorphic chloroplasts in two leaf cell types were observed. In S. oreophila, there was a high activity of RuBP carboxylase, but a low activity of C4 cycle enzymes. Interpretation of these data lends evidence to the hypothesis that a small group of C3 Salsola species, including S. oreophila, S. arbusculiformis, S. montana, and S. pachyphylla, arose as the result of a reversion of a C4 to a C3 type of photosynthetic CO2 fixation in the cooler climates of Middle Asia.     

Proof of C4 photosynthesis without Kranz anatomy in Bienertia cycloptera (Chenopodiaceae)

Kranz anatomy, with its separation of elements of the C4 pathway between two cells, has been an accepted criterion for function of C4 photosynthesis in terrestrial plants. However, Bienertia cycloptera (Chenopodiaceae), which grows in salty depressions of Central Asian semi-deserts, has unusual chlorenchyma, lacks Kranz anatomy, but has photosynthetic features of C4 plants. Its photosynthetic response to varying CO2 and O2 is typical of C4 plants having Kranz anatomy. Lack of night-time CO2 fixation indicates it is not acquiring carbon by Crassulacean acid metabolism. This species exhibits an independent, novel solution to function of the C4 mechanism through spatial compartmentation of dimorphic chloroplasts, other organelles and photosynthetic enzymes in distinct positions within a single chlorenchyma cell. The chlorenchyma cells have a large, spherical central cytoplasmic compartment interconnected by cytoplasmic channels through the vacuole to the peripheral cytoplasm. This compartment is filled with mitochondria and granal chloroplasts, while the peripheral cytoplasm apparently lacks mitochondria and has grana-deficient chloroplasts. Immunolocalization studies show enzymes compartmentalized selectively in the CC compartment, including Rubisco in chloroplasts, and NAD-malic enzyme and glycine decarboxylase in mitochondria, whereas pyruvate, Pi dikinase of the C4 cycle is localized selectively in peripheral chloroplasts. Phosphoenolpyruvate carboxylase, a cytosolic C4 cycle enzyme, is enriched in the peripheral cytoplasm. Our results show Bienertia utilizes strict compartmentation of organelles and enzymes within a single cell to effectively mimic the spatial separation of Kranz anatomy, allowing it to function as a C4 plant having suppressed photorespiration; this raises interesting questions about evolution of C4 mechanisms.     

Origin and evolution of C4 photosynthesis in the tribe Salsoleae (Chenopodiaceae) based on anatomical and biochemical types in leaves and cotyledons

 The Chenopodiaceae genus Salsola contains a large number of species with C₄ photosynthesis. Along with derivative genera they have a prominent position among the desert vegetation of Asia and Africa. About 130 species from Asia and Africa were investigated to determine the occurrence of C₃ versus C₄ syndrome in leaves and cotyledons, and to study specific anatomical and biochemical features of photosynthesis in both photosynthetic organs. The species studied belong to all six previously identified sections of the tribe Salsoleae based on morphological characters. Types of photosynthesis were identified using carbon ¹³C/¹²C isotope fractionation. The representatives of all systematic groups were investigated for mesophyll anatomy and biochemical subtypes by determination of enzyme activity (RUBPC, PEPC, NAD- and NADP-ME and AAT) and primary photosynthetic products. Two photosynthetic types (C₃ and C₄) and two biochemical subtypes (NAD- and NADP-ME) were identified in both leaves and cotyledons. Both Kranz and non-Kranz type anatomy were found in leaves and cotyledons, but cotyledons had more diversity in anatomical structure. Strong relationships between anatomical types and biochemical subtypes in leaves and cotyledons were shown. We found convincing evidence for a similar pattern of structural and biochemical features of photosynthesis in leaves and cotyledons within systematic groups, and evaluated their relevance at the evolutionary level. We identified six groups in tribe Salsoleae with respect to photosynthetic types and mesophyll structure in leaves and cotyledons. Two separate lineages of biochemical and anatomical evolution within Salsoleae were demonstrated based on studies of leaves and cotyledons. The sections Caroxylon, Malpighipila, Cardiandra and Belanthera have no C₃ species and only the NAD-ME C₄ subtype has been found in leaves. We suggest the C₄ species in the NADP-ME lineage evolved in Coccosalsola and Salsola sections, and originated in the subsection Arbuscula. Coccosalsola contains many species with C₃ and/or C₃-C₄ intermediate photosynthesis. Within these main evolutionary lineages, species of different taxonomic groups (sections and subsections) had differences in anatomical or/and biochemical features in leaves and cotyledons. We conclude that structural and biochemical changes in the photosynthetic apparatus in species of the tribe Salsoleae were a key factor in their evolution and broad distribution in extreme desert environments. Received January 25, 2001 Accepted July 17, 2001     

植物叶片解剖结构的量化及其在C4植物高粱中的应用

测量切片内所有细胞或细胞器的总周长,并乘以切片厚度和校正因子是一种量化植物叶片解剖结构的常用方法。前人在使用该方法时因忽视了细胞大小、切片厚度等影响估算精确度的因素,从而导致计数样本偏少并产生较大误差。通过分析该方法误差产生的原因及其与精确度的关系,明确了该方法在实际应用中不同精确度所要求对应的切片数量。在此基础上,将该方法用于C4植物高粱(Sorghum bicolor)叶片结构的量化,提出叶肉细胞与维管束鞘细胞接触面积(CABM)等参数,发展并完善了植物叶片解剖结构的量化方法。     

Leaf anatomy of the subtribe Hyptidinae (Labiatae)

The leaf anatomy of the subtribe Hyptidinae (Labiatae), which consists of four small genera and the largr genus Hyplis , is dercribed. The leaves may be dorsiventral or isobilateral. Variable characters of the lainina include: frequency and forms of trichomes, cuticular markings, presence of adaxial stoinata, thickness of leaf, thickness of adaxial epidermis, presence of a hypodermis, occurrence of sclcrified tissues (especially sclerified bundle sheath extensions, phloem and xylem fibres), mesophyll structure and venation pattern. Petiole vasculature varies from simple to complex, sometimes with medullary traces present. Most of the variable characters are related to xeromorphy and are tax-onomically useful within the framework of the present classification.     

Recent developments in mesophyll conductance in C3, C4, and crassulacean acid metabolism plants

The conductance of carbon dioxide (CO₂) from the substomatal cavities to the initial sites of CO₂ fixation (g_m) can significantly reduce the availability of CO₂ for photosynthesis. There have been many recent reviews on: (i) the importance of g_m for accurately modelling net rates of CO₂ assimilation, (ii) on how leaf biochemical and anatomical factors influence g_m, (iii) the technical limitation of estimating g_m, which cannot be directly measured, and (iv) how g_m responds to long‐ and short‐term changes in growth and measurement environmental conditions. Therefore, this review will highlight these previous publications but will attempt not to repeat what has already been published. We will instead initially focus on the recent developments on the two‐resistance model of g_m that describe the potential of photorespiratory and respiratory CO₂ released within the mitochondria to diffuse directly into both the chloroplast and the cytosol. Subsequently, we summarize recent developments in the three‐dimensional (3‐D) reaction‐diffusion models and 3‐D image analysis that are providing new insights into how the complex structure and organization of the leaf influences g_m. Finally, because most of the reviews and literature on g_m have traditionally focused on C₃ plants we review in the final sections some of the recent developments, current understanding and measurement techniques of g_m in C₄ and crassulacean acid metabolism (CAM) plants. These plants have both specialized leaf anatomy and either a spatially or temporally separated CO₂ concentrating mechanisms (C₄ and CAM, respectively) that influence how we interpret and estimate g_m compared with a C₃ plants.     

Leaf morphology and anatomy of Camellia section Camellia (Theaceae)

The delimitations of species in Camellia section Camellia have been disputed for many years, resulting from uncertain relationships among species. Leaf morphological and anatomical characters for 54 species and three varieties in this section were investigated to reveal the relationships. Principal component analysis and cluster analysis were conducted using the transformed data for quantitative and qualitative characters from leaf morphology and anatomy. Combining the results of statistical analysis with comparative leaf characters of morphology and anatomy, we discussed the taxonomic treatment of section Camellia by Chang compared with that of Ming and we conclude that section Camellia consists of c. 50 species. © 2009 The Linnean Society of London, Botanical Journal of the Linnean Society, 2009, 159 , 456–476.     

Leaf anatomy of Camellia sect. Oleifera and sect. Paracamellia (Theaceae) with reference to their taxonomic significance

Leaf anatomy of 21 species representing sect. Oleifera H. T. Chang and sect. Paracamellia Sealy in the genus Camellia was investigated using light microscopy. All the shapes of leaf epidermal cells were polygo-nal, and the patterns of anticlinal walls were sinuolate or sinuous, which were important to the classification of these two sections. The stomata were cyclocytic type and only present on the abaxial epidermis in both sampled sections. Seventeen leaf anatomical and cytologic characters were selected and used for the cluster analysis and principle components analyses performed by PAST program. Congruent results were obtained, with sect. Oleifera and sect. Paracamellia consistently grouping in two different clades, and some related species nesting together. This study provided leaf anatomical evidence for the merge of C. lanceoleosa H. T. Chang & J. S. Chiu and C. fluviatilis Hand.-Mazz.; of C. brevistyla Coh. St., C. obtusifolia H. T. Chang, and C. puniceiflora H. T. Chang; and of C. grijsii Hance and C. shensiensis H. T. Chang. In addition, it was also proved that C. tenii Sealy should be segregated from sect. Paracamellia and placed in the sect. Corallina Sealy. For further interest, we suggested that C. phaeoclada H. T. Chang be recognized as a subspecies of C. microphylla (Merr.) Chien. Overall, the results presented provided new insights into the relationships within a number of Camellia plants and sug-gested directions for future studies.     

C4 anatomy can evolve via a single developmental change

C₄ photosynthesis is a complex trait that boosts productivity in warm environments. Paradoxically, it evolved independently in numerous plant lineages, despite requiring specialised leaf anatomy. The anatomical modifications underlying C₄ evolution have previously been evaluated through interspecific comparisons, which capture numerous changes besides those needed for C₄ functionality. Here, we quantify the anatomical changes accompanying the transition between non‐C₄ and C₄ phenotypes by sampling widely across the continuum of leaf anatomical traits in the grass Alloteropsis semialata. Within this species, the only trait that is shared among and specific to C₄ individuals is an increase in vein density, driven specifically by minor vein development that yields multiple secondary effects facilitating C₄ function. For species with the necessary anatomical preconditions, developmental proliferation of veins can therefore be sufficient to produce a functional C₄ leaf anatomy, creating an evolutionary entry point to complex C₄ syndromes that can become more specialised.     

Mesophyll cell properties for some C3 and C4 species with high photosynthetic rates

Leaves of twelve C₃ species and six C₄ species were examined to understand better the relationship between mesophyll cell properties and the generally high photosynthetic rates of these plants. The CO₂ diffusion conductance expressed per unit mesophyll cell surface area (g_CO2^cell) cell was determined using measurements of the net rate of CO₂ uptake, water vapor conductance, and the ratio of mesophyll cell surface area to leaf surface area (A^mes/A). A^mes/A averaged 31 for the C₃ species and 16 for the C₄ species. For the C₃ species g_CO2^cell ranged from 0.12 to 0.32 mm s^-1, and for the C₄ species it ranged from 0.55 to 1.5 mm s^-1, exceeding a previously predicted maximum of 0.5 mm s^-1. Although the C₃ species Cammissonia claviformis did not have the highest g_CO2^cell, the combination of the highest A^mes and highest stomatal conductance resulted in this species having the greatest maximum rate of CO₂ uptake in low oxygen, 93 μmol m^-2 s^-1 (147 mg dm^-2 h^-1). The high g_CO2^cell of the C₄ species Amaranthus retroflexus (1.5 mm s^-1) was in part attributable to its thin cell wall (72 nm thick).     

Variations in the dorso-ventral organization of leaf structure and Kranz anatomy coordinate the control of photosynthesis and associated signalling at the whole leaf level in monocotyledonous species

Photosynthesis and associated signalling are influenced by the dorso-ventral properties of leaves. The degree of adaxial/abaxial symmetry in stomatal numbers, photosynthetic regulation with respect to light orientation and the total section areas of the bundle sheath (BS) cells and the surrounding mesophyll (M) cells on the adaxial and abaxial sides of the vascular bundles were compared in two C₄[ Zea mays (maize) and Paspalum dilatatum ] and one C₃[ Triticum turgidum (Durum wheat)] monocotyledonous species. The C₃ leaves had a higher degree of dorso-ventral symmetry than the C₄ leaves. Photosynthetic regulation was the same on each side of the wheat leaves, as were stomatal numbers and the section area of the BS relative to that of the M cells (BS/M section area ratio). In contrast, photosynthetic regulation in maize and P. dilatatum leaves showed a marked surface-specific response to light orientation. Compared to the adaxial sides of the C₄ monocotyledonous leaves, the abaxial surfaces had more stomata and the BS/M section area ratio was significantly higher. Differences in dorso-ventral structure, particularly in Kranz anatomy, serve not only to maximize photosynthetic capacity with respect light orientation in C₄ monocotyledonous leaves but also allow adaxial and abaxial-specific signalling from the respective M cells.     

Photosynthesis in Salsola Species (Chenopodiaceae) from Southern Africa Relative to their C4 Syndrome Origin and their African-Asian Arid Zone Migration Pathways

Abstract: Over 60 Salsola species of Chenopodiaceae from South Africa were studied for their photosynthesis type, using δ¹³C analysis and light microscopy of leaf anatomy. These species cover about 70 % of the total list of Southern African Salsola species and grow naturally in South and Southwest African desert regions. All species are shrubby forms and belong to the single subsection Caroxylon. Only C₄ photosynthesis was found in the Salsola species determined with ¹³C/¹²C carbon isotope discrimination values that ranged from - 11.04 to - 14.03 % (PDB), plus the presence of a Kranz type assimilation tissue anatomy. The apparent absence of C₃ in Salsola in South and Southwest Africa and the known presence of C₃ and C₃ - C₄ intermediate photosynthesis in Caroxylon, Salsola species in Asia strongly indicate that the genus Salsola originated in Asia and later migrated to South Africa.     

Leaf anatomy is not correlated to CAM function in a C3+CAM hybrid species,Yucca gloriosa

Background and AimsCrassulacean acid metabolism (CAM) is often considered to be a complex trait, requiring orchestration of leaf anatomy and physiology for optimal performance. However, the observation of trait correlations is based largely on comparisons between C₃ and strong CAM species, resulting in a lack of understanding as to how such traits evolve and the level of intraspecific variability for CAM and associated traits.MethodsTo understand intraspecific variation for traits underlying CAM and how these traits might assemble over evolutionary time, we conducted detailed time course physiological screens and measured aspects of leaf anatomy in 24 genotypes of a C₃+CAM hybrid species, Yucca gloriosa (Asparagaceae). Comparisons were made to Y. gloriosa’s progenitor species, Y. filamentosa (C₃) and Y. aloifolia (CAM).Key ResultsBased on gas exchange and measurement of leaf acids, Y. gloriosa appears to use both C₃ and CAM, and varies across genotypes in the degree to which CAM can be upregulated under drought stress. While correlations between leaf anatomy and physiology exist when testing across all three Yucca species, such correlations break down at the species level in Y. gloriosa.ConclusionsThe variation in CAM upregulation in Y. gloriosa is a result of its relatively recent hybrid origin. The lack of trait correlations between anatomy and physiology within Y. gloriosa indicate that the evolution of CAM, at least initially, can proceed through a wide combination of anatomical traits, and more favourable combinations are eventually selected for in strong CAM plants.     

C3 photosynthesis in Aristida longifolia: Implication for photosynthetic diversification in Aristidoideae (Poaceae)

Only a small percentage of plant species undergo C(4) photosynthesis. Despite its rarity, the C(4) pathway has evolved numerous times from C(3) ancestors, with as many as 18 independent origins in grasses alone. We report non-Kranz (C(3)) anatomy in Aristida longifolia, a species in a genus of ca. 300 species previously thought to possess only Kranz (C(4)) anatomy. Leaf blade transections of A. longifolia show widely spaced vascular bundles, nonradiate chlorenchyma, and few or no chloroplasts in cells of the sheaths surrounding the vascular bundle, all features indicative of C(3) photosynthesis. Carbon isotope ratios range from -27.68 to -29.71%, likewise indicative of C(3) photosynthesis. We also reconstruct the phylogeny of Aristidoideae, comprising Aristida, Sartidia (C(3)), and Stipagrostis (C(4)), using a sample of 11 species, including A. longifolia, and DNA sequences of the nuclear ribosomal internal transcribed spacer region and the chloroplast rpl16 intron and trnL-trnF region. Sartidia and Stipagrostis resolve as sisters, and sister to this clade is Aristida. Aristida longifolia resolves as sister to the remaining species in the genus. C(3) photosynthesis is hypothesized to be ancestral in Aristidoideae, which means the C(4) pathway evolved twice in the subfamily-in Stipagrostis and early in the diversification of the Aristida clade.     

Remarkable leaf anatomical variations in Neurachne and its allies (Poaceae) in relation to G3 and G4 photosynthesis

The leaf blade anatomy of all species of Neurachne, Paraneurachne and Thyridolepis has been examined, with special reference to features reliably indicative of photosynthetic pathway. Neurachne alopecuroidea, N. queenslandica and N. tenuifolia exhibit C₃ anatomy, as do all three Thyridolepis species. Paraneurachne muelleri, N. munroi and N. minor all have C₄ anatomy, of the rare ' Alloteropsis-type'. However, all the species exhibit unusual anatomical features, and N. lanigera cannot be confidently assigned as C₃ or C, on anatomical grounds alone. Taxonomic considerations confirm that the three genera are closely related to one another in the context of the eu-panicoid assemblage as a whole, and that the genus Neurachne itself, although it exhibits variation in photosynthetic pathway, is not amenable to further taxonomic subdivision.     

Morphology, anatomy and histochemistry of Salicornioideae (Chenopodiaceae) fruits and seeds

BACKGROUND AND AIMS: The subfamily Salicornioideae (Chenopodiaceae) are a taxonomically difficult group largely due to the lack of diagnostic characters available to delineate tribal- and generic-level boundaries; a consequence of their reduced floral and vegetative features. This study examined the variation in fruits and seeds across both tribes of the Salicornioideae to assess if characters support traditional taxonomic sections. METHODS: Light microscopy, environmental scanning electron microscopy and anatomical ultra-thin sectioning were employed to examine variation in fruits and seeds. Sixty-eight representatives across 14 of the 15 genera currently recognized within the tribes Halopeplideae and Salicornieae were examined to determine whether characters support current taxonomic groups. KEY RESULTS: Characters such as seed coat structure, embryo shape, seed orientation, the forms of seed storage proteins and carbohydrates show variation within the Salicornioideae and may be phylogenetically useful. The campylotropous ovule typical of the Chenopodiaceae generally results in a curved embryo; however, many Halosarcia and Sclerostegia species have straight embryos and in Salicornia and Sarcocornia the large peripheral embryo appears bent rather than curved. Seed coat ornamentation of Microcnemum and Arthrocnemum is distinct from other Salicornioideae as the elongated epidermal cells of the exotesta have convex walls. Histochemical stains of anatomical sections of cotyledon cells showed protein bodies were variable in shape, and starch grains were present in some species, namely Salicornia bigelovii, S. europaea and Allenrolfea occidentalis. CONCLUSIONS: While fruits and seeds were found to be variable within the subfamily, no synapomorphic characters support the tribe Halopeplideae as these genera have crustaceous seed coats, curved embryos and abundant perisperm; features characteristic of many of the tribe Salicornieae. The endemic Australian genera are closely related and few seed and fruit characters are diagnostic at the generic level. Nineteen characters identified as being potentially informative will be included in future phylogenetic analyses of the subfamily.