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cDNA and genomic cloning of sugar beet V-type H+-ATPase subunit A and c isoforms: evidence for coordinate expression during plant development and coordinate induction in response to high salinity.

The plant V-type H+-ATPase (V-ATPase) does not only serve basic housekeeping functions but is also involved in stress-induced NaCl sequestration during salinity stress. To address the question whether the same isoforms conferring housekeeping functions are equally involved in the response to high salinity, we have isolated cDNA clones for subunits A and c, as representing the peripheral V1 complex and the membrane-integral V0 complex, respectively, from the halotolerant sugar beet (Beta vulgaris L., diploid variety). RNA blot analysis with gene-specific probes revealed a coordinate expression of the cloned subunit A and c isoforms during plant development and in response to high salinity. Also, in rapidly dividing suspension-cultured cells with 10-fold increased transcript amounts as compared to young leaf tissue, the ratio of transcripts for both genes was similar to the ratio found for transcripts in leaves of different age. We have then isolated partial genomic clones (BVA/70 for Beta V-ATPase 70 kDa subunit; BVA/16-1 for Beta V-ATPase 16 kDa subunit), including the promoter regions. Transcription start mapping revealed long 5'-UTR leader sequences (230 and 172 bases, respectively) for both genes. Both promoters contain putative G-box motifs in similar distance to the TATA boxes. For a quantitative comparison of relative promoter strength, the BVA/70 and BVA/16-1 promoters linked to the luciferase reporter gene (LUC) were delivered to sugar beet suspension-cultured cells by particle bombardment. The BVA/16-1 promoter showed a 1.7-fold higher activity as compared with the BVA/70 promoter. Salt treatment induced an increase of BVA/70 (+70%) and BVA/16-1 (+57%) promoter activities, concomitant with increased transcript amounts. The following sequences have been deposited at the EMBL database X98767: Beta vulgaris V-ATPase subunit A, cDNA clone; X98851, B. vulgaris V-ATPase subunit c isoform 1, cDNA clone; Y11038, B. vulgaris V-ATPase subunit A, partial genomic clone; Y11037, B. vulgaris V-ATPase subunit c isoform 1, partial genomic clone.

Amino Acid Sequence↗

The plastid clpP1 protease gene is essential for plant development.

Plastids of higher plants are semi-autonomous cellular organelles that have their own genome and transcription-translation machinery. Examples of plastid functions are photosynthesis and biosynthesis of starch, amino acids, lipids and pigments. Plastid functions are encoded in approximately 120 plastid genes and approximately 3,000 nuclear genes. Although many embryo and seedling lethal nuclear genes are required for chloroplast biogenesis, until now deletion of plastid genes either had no phenotypic consequence (8 genes), or caused a mutant phenotype but did not affect viability (13 genes). Here we identify an essential plastid gene. By using the CRE-lox site-specific recombination system we have deleted clpP1 (caseinolytic protease P1), one of the three genes (clpP1, ycf1 and ycf2) whose disruption had previously only been possible in a fraction of the 1,000-10,000 plastid genome copies in a cell. Loss of the clpP1 gene product, the ClpP1 protease subunit, results in ablation of the shoot system of tobacco plants, suggesting that ClpP1-mediated protein degradation is essential for shoot development.

Endopeptidases↗

Expression of streptavidin in tomato resulted in abnormal plant development that could be restored by biotin application.

Biotin is an essential cofactor for a variety of carboxylase and decarboxylase reactions and is involved in diverse metabolic pathways of all organisms. In the present study we tested the hypothesis that controlling biotin availability by the expression of Streptomyces avidinii streptavidin, would impede plant development. Transient expression of streptavidin fused to plant signal peptide, bacterial signal peptide or both, in tomato (Lycopersicon esculentum cv. VF36) plants resulted in various levels of tissue impairment, exhibited as lesion development on 1-week-old tomato seedlings. The least toxic construct was introduced to tomato (stable transformation) under the constitutive CaMV 35S promoter, and lesions appeared on stems, flower morphologies were modified and numbers and sizes of fruits were altered. Furthermore, tissue-specific expression of the streptavidin, by means of the beta-phaseolin or TobRB7 promoters, resulted in localised effects, i.e., impaired seed formation or seedless fruits, respectively, with no alteration in the morphology of the other plant organs. External application of biotin on streptavidin-expressing tomato plants prevented the degeneration symptoms and facilitated normal plant development. It can be concluded that expression of streptavidin in the plant cell can lead to local and temporal deficiencies in biotin availability, impairing developmental processes while biotin application restores plant growth cycle.

Biotin↗

Expression profiling of plant development.

Microarrays have been used to study the response of plants to many signals, including light, hormones and transcription factors. The results in each case can give an overall view of the global response to the signal or identify direct targets of the signal, and can reveal new links between different signaling pathways.

Gene Expression Profiling↗

Inducible phytoalexins in juvenile soybean genotypes predict soybean looper resistance in the fully developed plants.

The hypocotyl of different soybean genotypes was tested for its inducible phytoalexin (i.e. glyceollin or coumestrol) accumulation and its inducible soybean looper resistance in response to chemical elicitation. A very highly insect-resistant soybean genotype (PI 227687) produced significantly more phytoalexins than a relatively insect-susceptible one (Davis) in response to the same chemical elicitation. The resultant standardized hypocotyl assay allowed quick categorization of unknown soybean genotypes regarding the level of insect resistance in the fully developed plants. Glyceollin was a better indicator of inducible resistance than coumestrol. Elicitor concentration influenced the amount of glyceollin and coumestrol accumulated. Younger seedlings (4-5 d old) responded stronger to chemical elicitation than did older ones (7-10 d old). The elicited accumulation of glyceollin showed a temporal pattern that peaked at 72 h. Accumulation of coumestrol showed a gradual increase. Elicitation of phytoalexins in juvenile soybean plants by sulfhydryl-binding reagents was found to be useful for the prediction of genotypic differences in the level of insect resistance in the fully developed plants.

Journal Article↗

Balance between cell division and differentiation during plant development.

The processes which make possible that a cell gives rise to two daughter cells define the cell division cycle. In individual cells, this is strictly controlled both in time and space. In multicellular organisms extra layers of regulation impinge on the balance between cell proliferation and cell differentiation within particular ontogenic programs. In contrast to animals, organogenesis in plants is a post-embryonic process that requires developmentally programmed reversion of sets of cells from different differentiated states to a pluripotent state followed by regulated proliferation and progression through distinct differentiation patterns. This implies a fine coupling of cell division control, cell cycle arrest and reactivation, endoreplication and differentiation. The emerging view is that cell cycle regulators, in addition to controlling cell division, also function as targets for maintaining cell homeostasis during development. The mechanisms and cross talk among different cell cycle regulatory pathways are discussed here in the context of a developing plant.

Arabidopsis↗

Tall tales from sly dwarves: novel functions of gibberellins in plant development.

Gibberellins (GAs) are endogenous hormones controlling numerous aspects of plant growth and development. Our present understanding of GA physiology is based largely on genetic analysis in model plants such as Arabidopsis. In spite of the success of this approach, the discovery of additional physiological roles for GAs in seed development, pollen tube growth and meristem development indicates that the existing collection of GA-related mutants (identified partially or entirely on the basis of vegetative phenotypes) has failed to uncover all aspects of plant development that are controlled by GAs. The continued use of ever improving forward and reverse genetic techniques is expected to lead to the discovery of further novel roles for GAs in plant development.

Arabidopsis↗

E2F factors rate controls the dual role of CDE/E2F composite element: a model of E2F-regulated gene expression in plant development.

The promoters of several E2F-regulated genes identified in plants contain a variety of E2F motifs, notably a composite element consisting of a "CDE-like element" C/GGCGG on one strand, described as repressor in animals, associated with an E2F element on the complementary strand. This detailed study throughout plant development using ribonucleotide reductase promoters, allows us to propose a model, where E2F and composite elements play a dual role. Such regulation is mainly conditioned by the availability of E2F factors in tissues and during the cell cycle in tobacco.

Animals↗

Diverse functions of Polycomb group proteins during plant development.

Polycomb group (PcG) proteins play essential roles in animal and plant life cycles by controlling the expression of important developmental regulators. These structurally heterogeneous proteins form multimeric protein complexes that control higher order chromatin structure and, thereby, the expression state of their target genes. Once established, PcG proteins maintain silent gene expression states over many cell divisions providing a molecular basis for a cellular 'memory.' PcG proteins are best known for their role in the control of homeotic genes in Drosophila and mammals. In addition, they play important roles in the control of cell proliferation in vertebrate and invertebrate systems. Recent studies in plants have shown that PcG proteins regulate diverse developmental processes and, as in animals, they affect both homeotic gene expression and cell proliferation. Thus, the function of PcG proteins has been widely conserved between the plant and animal kingdoms.

Gene Expression Regulation, Developmental↗

A geminivirus replication protein interacts with a protein kinase and a motor protein that display different expression patterns during plant development and infection.

The geminivirus protein AL1 initiates viral DNA replication, regulates its own expression, and induces plant gene transcription. To better understand how AL1 interacts with host proteins during these processes, we used yeast two-hybrid library screening and a baculovirus protein interaction system to identify plant proteins that interact with AL1. These studies identified a Ser/Thr kinase, a kinesin, and histone H3 as AL1 partners. The kinase is autophosphorylated and can phosphorylate common kinase substrates in vitro. The kinesin is phosphorylated in insect cells by a cyclin-dependent kinase. Immunostaining of Nicotiana benthamiana and Arabidopsis showed that kinase protein levels and subcellular location are regulated during plant development and geminivirus infection. By contrast, the kinesin is ubiquitous even though it is associated with the spindle apparatus in mitotic cells. Together, our results establish that AL1 interacts with host proteins involved in plant cell division and development. Possible functions of these host factors in healthy and geminivirus-infected plants are discussed.

Animals↗

Gene regulatory network models for plant development.

Accumulated genetic data are stimulating the use of mathematical and computational tools for studying the concerted action of genes during cell differentiation and morphogenetic processes. At the same time, network theory has flourished, enabling analyses of complex systems that have multiple elements and interactions. Reverse engineering methods that use genomic data or detailed experiments on gene interactions have been used to propose gene network architectures. Experiments on gene interactions incorporate enough detail for relatively small developmental modules and thus allow dynamical analyses that have direct functional interpretations. Generalities are beginning to emerge. For example, biological genetic networks are robust to environmental and genetic perturbations. Such dynamical studies also enable novel predictions that can lead to further experimental tests, which might then feedback to the theoretical analyses. This interplay is proving productive for understanding plant development. Finally, both experiments on gene interactions and theoretical analyses allow the identification of frequent or fixed evolutionary solutions to developmental problems, and thus are contributing to an understanding of the genetic basis of the evolution of development and body plan.

Biological Evolution↗

Plant development is regulated by a family of auxin receptor F box proteins.

The plant hormone auxin has been implicated in virtually every aspect of plant growth and development. Auxin acts by promoting the degradation of transcriptional regulators called Aux/IAA proteins. Aux/IAA degradation requires TIR1, an F box protein that has been shown to function as an auxin receptor. However, loss of TIR1 has a modest effect on auxin response and plant development. Here we show that three additional F box proteins, called AFB1, 2, and 3, also regulate auxin response. Like TIR1, these proteins interact with the Aux/IAA proteins in an auxin-dependent manner. Plants that are deficient in all four proteins are auxin insensitive and exhibit a severe embryonic phenotype similar to the mp/arf5 and bdl/iaa12 mutants. Correspondingly, all TIR1/AFB proteins interact with BDL, and BDL is stabilized in triple mutant plants. Our results indicate that TIR1 and the AFB proteins collectively mediate auxin responses throughout plant development.

Arabidopsis↗

CHORIPETALA and DESPENTEADO: general regulators during plant development and potential floral targets of FIMBRIATA-mediated degradation.

Two Antirrhinum majus mutants, choripetala (cho) and despenteado (desp), exhibit identical highly pleiotropic phenotypes including petaloid transformation of first whorl floral organs, narrowing of both vegetative and floral organs, reduction in carpel size and fertility and delayed germination. The petaloid first whorl results from ectopic expression of the class B genes DEFICIENS and GLOBOSA and is correlated with the ectopic expression of the proposed class B/C gene regulator FIMBRIATA (FIM). Ectopic class B gene expression is apparent from the earliest point at which class B gene transcription can be detected in the wild type, indicating that the pre-patterning of the class B domain has been disrupted in these mutants. Single and double mutant analyses indicate that CHO and DESP also play a role in regulation of the class C domain. Interestingly, the cho and desp mutations partially suppress the phenotype of fim null mutants, suggesting that the F-box protein FIM may target a member of the CHO/DESP pathway for degradation. We propose that CHO and DESP are members of a 'basal regulatory function' influencing many processes throughout plant development and in particular are directly or indirectly required for the repression of class B and C genes during early stages of flower development.

Gene Expression Regulation, Developmental↗

Emerging regulatory roles of small metabolites in plant development and adaptation.

Once viewed mainly as metabolic intermediates, small metabolites are increasingly recognized as spatially and temporally regulated signals that coordinate plant development and adaptation. Understanding these metabolite-based regulatory processes could reveal new strategies to improve crop resilience, productivity, and sustainability under changing environmental conditions.

MSI↗

Plant development: local control, global patterning.

Several different lines of inquiry have converged on the conclusion that the number and plane of cell divisions is under tight control in plant development. At the same time, there are new data which show that the pattern of cell division is less important in the formation of organ-scale plant patterns than was expected previously. This apparent paradox can be resolved by recognizing the role of cell-cell communication in the control of plant cell-division patterns.

Cell Division↗

Mutation in the Arabidopsis PASTICCINO1 gene, which encodes a new FK506-binding protein-like protein, has a dramatic effect on plant development.

The pasticcino (pas) mutants of Arabidopsis thaliana are a new class of plant developmental mutants; members of this class show ectopic cell proliferation in cotyledons, extra layers of cells in the hypocotyl, and an abnormal apical meristem. This phenotype is correlated with both cell division and cell elongation defects. There are three complementation groups of pas mutants (pas1, pas2, and pas3, with, respectively 2, 1, and 4 alleles). Here we describe in more detail the pas1-1 allele, which was obtained by insertional mutagenesis. The PAS1 gene has been cloned and characterized; it encodes an immunophilin-like protein similar to the p59 FK506-binding protein (FKBP52). PAS1 is characterized by an FKBP-like domain and three tetratricopeptide repeat units. Although the presence of immunophilins in plants has already been demonstrated, the pas1-1 mutant represents the first inactivation of an FKBP-like gene in plants. PAS1 expression is altered in pas1 mutants and in the pas2 and pas3 mutants. The expression of the PAS1 gene is increased in the presence of cytokinins, a class of phytohormones originally discovered because of their ability to stimulate cell division. These results are of particular relevance as they show for the first time that an FKBP-like protein plays an important role in the control of plant development.

Amino Acid Sequence↗