Edyta M. Gola

The lack of mitochondrial AtFtsH4 protease alters Arabidopsis leaf morphology at the late stage of rosette development under short-day photoperiod.

AtFtsH4 is one of four inner membrane-bound mitochondrial ATP-dependent metalloproteases in Arabidopsis thaliana, called AAA proteases, whose catalytic site is exposed to the intermembrane space. In the present study, we used a reverse-genetic approach to investigate the physiological role of the AtFtsH4 protease. We found that loss of AtFtsH4 did not significantly affect Arabidopsis growth under optimal conditions (long days); however, severe morphological and developmental abnormalities in late rosette development occurred under short-day conditions. The asymmetric shape and irregular serration of expanding leaf blades were the most striking features of the ftsh4 mutant phenotype. The severe abnormal morphology of the leaf blades was accompanied by ultrastructural changes in mitochondria and chloroplasts. These abnormalities correlated with elevated levels of reactive oxygen species and carbonylated mitochondrial proteins. We found that two classes of molecular chaperones, Hsp70 and prohibitins, were over-expressed in ftsh4 mutants during late vegetative growth under both short- and long-day conditions. Taken together, our data indicate that lack of AtFtsH4 results in impairment of organelle development and Arabidopsis leaf morphology under short-day conditions.

Mitochondrial protease AtFtsH4 protects ageing Arabidopsis rosettes against oxidative damage under short-day photoperiod.

Mitochondrial AtFtsH4 protease, whose catalytic site is exposed to the intermembrane space, is one of four inner membrane-bound FtsH proteases in Arabidopsis. We found that the loss of AtFtsH4 altered Arabidopsis leaf morphology at the late stage of rosette growth under short-day photoperiod, while such changes were not observed in ftsh4 mutants grown under long days. These morphological changes were correlated with elevated levels of both reactive oxygen species (ROS) and carbonylated proteins, which strongly suggested that ageing ftsh4 plants experienced oxidative stress. This view was supported by the accumulation of electron-dense material, presumably containing aggregated oxidized proteins, in mitochondria of ftsh4 plants with the most strongly malformed leaf blades. Taken together, our data published in the May issue of Plant J 1 suggest a link between the lack of AtFtsH4 protease, oxidative stress and altered leaf morphology at the late rosette stage under short days. Here, we present evidence that the onset of altered leaf morphology in ftsh4 correlates with an increase in the abundance of AtFtsH4 transcript observed in wild-type Arabidopsis growing under the same conditions. We also discuss how the lack of AtFtsH4 may cause oxidative stress towards the end of the vegetative growth in short days.

Slime cells on the surface of Eragrostis seeds maintain a level of moisture around the grain to enhance germination

Eragrostis is a cosmopolitan genus of the family Poaceae. Several wild species, including E. pilosa (L.) Beauv., are harvested for food, but the only cultivated crop-species is tef [E. tef (Zucc.) Trotter]. Despite its importance as a staple food and its plasticity to diverse environmental conditions, little is known about the structural and physiological strategies that adapt tef seeds to endure diverse and variable moisture regimes. Here, we report the presence of slime cells, a type of modified epidermal cell, covering the fruit of tef and its wild relative, E. pilosa. The slime produced by Eragrostis belongs to the ‘true’slime type, since it is exclusively composed of pectins. Pectin forms uniform layers on the cell wall inner surface, which are confined by a thin cellulose layer to prevent release into the cell lumen. In the presence of water, pectins quickly hydrate, causing swelling of the slime cells. This is followed by their detachment, which may be controlled by a thin cuticle layer on the fruit surface. The ability of slime to absorb and maintain moisture around the grain is thought to be an adaptive feature for Eragrostis growing in dry habitats. This retention of water by slime may create conditions that are suitable for rapid germination.

The mitochondrial protease AtFTSH4 safeguards Arabidopsis shoot apical meristem function.

The shoot apical meristem (SAM) ensures continuous plant growth and organogenesis. In LD 30 °C, plants lacking AtFTSH4, an ATP-dependent mitochondrial protease that counteracts accumulation of internal oxidative stress, exhibit a puzzling phenotype of premature SAM termination. We aimed to elucidate the underlying cellular and molecular processes that link AtFTSH4 with SAM arrest. We studied AtFTSH4 expression, internal oxidative stress accumulation, and SAM morphology. Directly in the SAM we analysed H2O2 accumulation, mitochondria behaviour, and identity of stem cells using WUS/CLV3 expression. AtFTSH4 was expressed in proliferating tissues, particularly during the reproductive phase. In the mutant, SAM, in which internal oxidative stress accumulates predominantly at 30 °C, lost its meristematic fate. This process was progressive and stage-specific. Premature meristem termination was associated with an expansion in SAM area, where mitochondria lost their functionality. All these effects destabilised the identity of the stem cells. SAM termination in ftsh4 mutants is caused both by internal oxidative stress accumulation with time/age and by the tissue-specific role of AtFTSH4 around the flowering transition. Maintaining mitochondria functionality within the SAM, dependent on AtFTSH4, is vital to preserving stem cell activity throughout development.

Dichotomous branching: the plant form and integrity upon the apical meristem bifurcation

The division of the apical meristem into two independently functioning axes is defined as dichotomous branching. This type of branching typically occurs in non-vascular and non-seed vascular plants, whereas in seed plants it presents a primary growth form only in several taxa. Dichotomy is a complex process, which requires a re-organization of the meristem structure and causes changes in the apex geometry and activity. However, the mechanisms governing the repetitive apex divisions are hardly known. Here, an overview of dichotomous branching is presented, occurring in structurally different apices of phylogenetically distant plants, and in various organs (e.g., shoots, roots, rhizophores). Additionally, morphogenetic effects of dichotomy are reviewed, including its impact on organogenesis and mechanical constraints. At the end, the hormonal and genetic regulation of the dichotomous branching is discussed.

Impermanency of Initial Cells in Huperzia lucidula (Huperziaceae) Shoot Apices

Structural changes in apical meristems accompany morphological specialization and adaptation of vascular-plant sporophytes to changing environments. Analysis of cell clones at the shoot apical meristem surface of a representative of the basal vascular plants made it possible to trace the dynamics of initial cells, which are recorded in cell wall arrangements. The cellular pattern at the summit of the Huperzia lucidula (Michx.) Trevisan shoot apical meristem is usually composed of four initial cells, forming a tetrad at the apex center, and their derivative clones. Our data show that the apical initial cells present at a given time in a tetrad often are transient and replaced by new initials in the ontogeny of a given Huperzia apex. Specifically, the process of dichotomous branching leads to extensive cell division, resulting in a pool of potential initial cells from which the new initial centers of the twin axes are selected. The ability to specify newly arisen cells as initials during shoot dichotomy indicates that a mechanism for selection and replacement of initial cells may exist in lycopods. Such a cellular mechanism, which could play an important role in protecting the genetic identity and integrity of meristematic cells, would be expected to emerge early in the phylogeny of vascular plants.

Validation of leaf undulation traits in the taxonomy of Epipactis muelleri Godfery, 1921 (Orchidaceae, Neottieae)

Leaf undulation is used as a diagnostic feature in the taxonomy of different taxa of the Epipactis (Orchidaceae) genus. It is considered a specifically important trait, apart from the column (gynostemium) structure, in the identification of E. muelleri. However, leaf undulation is a common developmental phenomenon in various plants, including orchids, thus the main goal of the research presented was to validate the usefulness of this feature in Epipactis taxonomy. In the course of this study, the leaf structure of six Epipactis species was analyzed, i.e., E. albensis, E. atrorubens, E. helleborine, E. muelleri, E. palustris and E. purpurata. The leaf margin outline and the cellular pattern of the leaf surface, as well as the presence of papillae, were examined in detail. The results obtained showed that in all taxa analyzed, representing different groups of the Epipactis genus, leaf undulation was present. The phenomenon was related to leaf blade development and the local differential growth of leaf sectors rather than to the taxonomic position of the species. Therefore, leaf undulation studied in Helleborines does not have a diagnostic value as a non-programmed intrinsic feature and should not be applied to taxa identification. The valid trait in Epipactis classification (including E. muelleri) is the column structure.

Meristems of Seedless Vascular Plants: The State of the Art

Over the past several decades, plant meristems have been a focus of developmental research. These unique structures, containing stem cells, have the ability to self-maintain and provide cells for growth and development, supporting the longevity of a plant as well as plants’ architectural complexity and diversity of forms. In addition, meristems are structurally diversified across the plant kingdom. Therefore, unraveling the key processes and mechanisms governing meristems functioning in a wide variety of plants is crucial from a developmental perspective. In ferns and lycophytes, which are basal groups of seedless vascular plants, meristem regulation is still not fully understood at the cellular and developmental levels, for example, relating to self-maintenance and protection of the undamaged genetic information. Furthermore, the research extending our knowledge on the accompanying molecular mechanism is in the infancy. In this review, we have gathered scattered information concerning not only apical but also lateral and intercalary meristems in seedless vascular plants. We hope that provided here the state of the art will guide the readers to their future experimental ideas to improve the functional understanding of meristems in these so interesting plants.

Impairment of Meristem Proliferation in Plants Lacking the Mitochondrial Protease AtFTSH4

Shoot and root apical meristems (SAM and RAM, respectively) are crucial to provide cells for growth and organogenesis and therefore need to be maintained throughout the life of a plant. However, plants lacking the mitochondrial protease AtFTSH4 exhibit an intriguing phenotype of precocious cessation of growth at both the shoot and root apices when grown at elevated temperatures. This is due to the accumulation of internal oxidative stress and progressive mitochondria dysfunction. To explore the impacts of the internal oxidative stress on SAM and RAM functioning, we study the expression of selected meristem-specific (STM, CLV3, WOX5) and cell cycle-related (e.g., CYCB1, CYCD3;1) genes at the level of the promoter activity and/or transcript abundance in wild-type and loss-of-function ftsh4-1 mutant plants grown at 30 °C. In addition, we monitor cell cycle progression directly in apical meristems and analyze the responsiveness of SAM and RAM to plant hormones. We show that growth arrest in the ftsh4-1 mutant is caused by cell cycle dysregulation in addition to the loss of stem cell identity. Both the SAM and RAM gradually lose their proliferative activity, but with different timing relative to CYCB1 transcriptional activity (a marker of G2-M transition), which cannot be compensated by exogenous hormones.

Leaf arrangements are invalid in the taxonomy of orchid species

The selection and validation of proper distinguishing characters are of crucial importance in taxonomic revisions. The modern classifications of orchids utilize the molecular tools, but still the selection and identification of the material used in these studies is for the most part related to general species morphology. One of the vegetative characters quoted in orchid manuals is leaf arrangement. However, phyllotactic diversity and ontogenetic changeability have not been analysed in detail in reference to particular taxonomic groups. Therefore, we evaluated the usefulness of leaf arrangements in the taxonomy of the genus Epipactis Zinn, 1757. Typical leaf arrangements in shoots of this genus are described as distichous or spiral. However, in the course of field research and screening of herbarium materials, we indisputably disproved the presence of distichous phyllotaxis in the species Epipactis purpurata Sm. and confirmed the spiral Fibonacci pattern as the dominant leaf arrangement. In addition, detailed analyses revealed the presence of atypical decussate phyllotaxis in this species, as well as demonstrated the ontogenetic formation of pseudowhorls. These findings confirm ontogenetic variability and plasticity in E. purpurata. Our results are discussed in the context of their significance in delimitations of complex taxa within the genus Epipactis.