This is an addendum to our recent paper published in The Plant Journal (52:352–61). The major findings were: (1) trichomes on the leaves of gl3-sst sim double mutants developed as large multi-cellular clusters whereas wild type trichomes are composed of single cells; (2) ectopic CYCD3;1 expression in gl3-sst trichomes also resulted in trichome cluster formation; and (3) that GL1 expression is prolonged in the gl3-sst sim trichome clusters. This addendum shows that ectopic CYCD3;1 expression in gl3-sst also enhanced GL1 expression. An analysis of the GL1 promoter found two overlapping potential E2F binding sites in a region of the promoter known to be essential for GL1 function. This finding indicates that GL1 may be directly regulated by the activity of a CYCD3/CDKA complex that phosphorylates E2F-RB bound to the GL1 promoter.
plant cell cycle; endoreduplication; glabra1; plant development
Two orientation devices are described which are currently in use at the J. Hillis Miller Health Center Library.
One is a taped tour which requires a portable recorder with earphones attached. Tapes are now available for nursing students, physical therapy students, and Health Center staff. The tour includes location information and description of the card catalog and certain basic index and abstract services.
The second orientation device is a short instruction tape on the use of Index Medicus which is attached to the Index Medicus table. This is heard through a telephone apparatus.
It is anticipated that the tape technique will be expanded to apply to other students and other library tools. It is also believed that this technique may be used in other libraries.
Information about the supplies and equipment used is given as an addendum.
A network of shared intermediates/components and/or common molecular outputs in biotic and abiotic stress signaling has long been known, but the possibility of effective influence between differently triggered stresses (co-protection) is less studied. Recent observations show that wounding induces transient protection in tomato (Solanum lycopersicum L.) to four pathogens with a range of lifestyles, locally and systemically. The contribution of ethylene (ET) in basal but also in wound-induced resistance to each pathogen, although dispensable, is demonstrated to be positive (Botrytis cinerea, Phytophthora capsici) or negative (Fusarium oxysporum, Pseudomonas syringae pv. tomato). Furthermore, the expression of several defense markers is influenced locally and/or systemically by wounding and ET, and might be part of that core of conserved molecular responses whereby an abiotic stress such as wounding imparts co-resistance to biotic stress. In this addendum, we speculate on some of the physiological responses to wounding that might contribute to the modulation of resistance in a more pathogen-specific manner.
tomato; phytophthora; fusarium; wounding; ethylene; defense mechanisms; electric fields; zoospores; tylosis
Antisense oligodeoxynucleotide (ODN) inhibition emerges as an effective means for probing gene function in plant cells. Employing this method we have established the importance of the SUSIBA2 transcription factor for regulation of starch synthesis in barley endosperm, and arrived at a model for the role of the SUSIBAs in sugar signaling and source-sink commutation during cereal endosperm development. In this addendum we provide additional data demonstrating the suitability of the antisense ODN technology in studies on starch branching enzyme activities in barley leaves. We also comment on the mechanism for ODN uptake in plant cells.
antisense ODN; barley; SBE; SUSIBA2
Glucose functions in plants both as a metabolic resource as well as a hormone that regulates expression of many genes. Arabidopsis hexokinase1 (HXK1) is the best understood plant glucose sensor/transducer, yet we are only now appreciating the cellular complexity of its signaling functions. We have recently shown that one of the earliest detectable responses to plant glucose treatments are extensive alterations of cellular F-actin. Interestingly, AtHXK1 is predominantly located on mitochondria, yet also can interact with actin. A normal functioning actin cytoskeleton is required for HXK1 to act as an effector in glucose signaling assays. We have suggested that HXK1 might alter F-actin dynamics and thereby influence the formation and/or stabilization of cytoskeleton-bound polysomes. In this Addendum, we have extended our initial observations on the subcellular targeting of HXK1 and its interaction with F-actin. We then further consider the cellular context in which HXK1 might regulate gene expression.
Arabidopsis; F-actin; glucose signaling; hexokinase; hTalin; mitochondria; polysomes; protoplasts; transient expression assay; fluorescence microscopy
In animal cells, nitric oxide and NO-derived molecules have been shown to mediate post-translational modifications such as S-nitrosylation and protein tyrosine nitration which are associated with cell signalling and pathological processes, respectively. In plant cells, knowledge of the function of these post-translational modifications under physiological and stress conditions is still very rudimentary. In this addendum, we briefly examine how reactive nitrogen species (RNS) can exert important effects on proteins that could mediate signalling processes in plants.
nitrosative stress; nitric oxide synthase; S-nitrosoglutathione; nitro-tyrosine; post-translational modifications; S-nitrosylation
The formation of neuronal synapses is a finely organized process that involves the presynaptic assembly of the machinery responsible for neurotransmitter release and the postsynaptic recruitment of neurotransmitter receptors and scaffold proteins to the postsynaptic density (PSD). The molecular cues guiding the establishment of synaptic connections are now beginning to be identified. Recent studies indicate that cell adhesion molecules (CAMs) participate prominently in the key steps of synapse formation, inducing trans-synaptic adhesion and promoting a precise alignment of pre- and postsynaptic terminals. This addendum describes a new mechanism of cell-cell interaction that combines features of both diffusible and membrane-bound synaptogenic factors. It particularly points out the key role played by GDNF triggering trans-homophilic binding between GFRα1 molecules and cell adhesion between GFRα1-expressing cells. In this model GFRα1 functions as a ligand-induced cell adhesion molecule (LICAM) to establish precise synaptic contacts and promote the assembly of presynaptic terminals. In this overview, I summarize the current concepts of synapse formation in the limelight of this new mechanism of ligand-induced cell adhesion.
GDNF; GFRα1; LICAM; cell adhesion; synapse formation
Remodeling of joints is a key feature of inflammatory and degenerative joint disease. Bone erosion, cartilage degeneration and growth of bony spurs termed osteophytes are key features of structural joint pathology in the course of arthritis, which lead to impairment of joint function. Understanding their molecular mechanisms is essential to tailor targeted therapeutic approaches to protect joint architecture from inflammatory and mechanical stress. This addendum summarizes the new insights in the molecular regulation of bone formation in the joint and its relation to bone resorption. It describes how inflammatory cytokines impair bone formation and block the repair response of joints towards inflammatory stimuli. It particularly points out the key role of Dickkopf-1 protein, a regulator of the Wingless signaling and inhibitor of bone formation. This new link between inflammation and bone formation is also crucial for explaining the generation of osteophytes, bony spurs along joints, which are characterized by new bone and cartilage formation. This mechanism is largely dependent on an activation of wingless protein signaling and can lead to complete joint fusion. This addendum summarized the current concepts of joint remodeling in the limelight of these new findings.
joint remodeling; arthritis; bone formation; bone erosion; osteoblasts; osteoclasts; Dickkopf; wingless proteins
For a long time it has been believed that lignification has an important role in host defense against pathogen invasion. Recently, by using an RNAi gene-silencing assay we showed that monolignol biosynthesis plays a critical role in cell wall apposition (CWA)-mediated defense against powdery mildew fungus penetration into diploid wheat. Silencing monolignol genes led to super-susceptibility of wheat leaf tissues to an appropriate pathogen, Blumeria graminis f. sp. tritici (Bgt), and compromised penetration resistance to a non-appropriate pathogen, B. graminis f. sp. hordei. Autofluorescence of CWA regions was reduced significantly at the fungal penetration sites in silenced cells. Our work indicates an important role for monolignol biosynthetic genes in effective CWA formation against pathogen penetration. In this addendum, we show that silencing of monolignol genes also compromised penetration resistant to Bgt in a resistant wheat line. In addition, we discuss possible insights into how lignin biosynthesis contributes to host defense.
monolignol; papilla autofluorescence; methylated lignin; defense lignin; cereal
Hormonal regulation of root development is a long known phenomenon. In the past decades, the molecular mechanisms of individual hormonal pathways and their impact on root development have been studied. Recent genetic and molecular studies suggest importance of interactions of the individual hormonal pathways and their components. In our paper1 we show impact of endogenous cytokinin on the root architecture and its interaction with auxin in Arabidopsis thaliana. In this addendum we discuss our results in the light of significant recent papers that deal with cytokinin-auxin interactions and we point out spatiotemporal specificity of these interactions in the root development.
root meristem; hormonal regulation; cytokinin-auxin interaction; spatiotemporal specificity; Arabidopsis thaliana
Regulation of mRNA decay rates appears to be an important control point in determining the abundance of gene transcripts. Rapid change in decay rates of mRNAs could provide prompt responses of the plants to environmental fluctuations. SOS1 is a plasma-membrane Na+/H+ antiporter crucial for salt tolerance in Arabidopsis. In our recent paper in The Plant Journal, we have shown that SOS1 mRNA is inherently instable at normal growth conditions, but its stability is substantially elevated by salt and oxidative stress treatments. Salt stress-induced SOS1 mRNA stability is mediated by reactive oxygen species (ROS) produced, at least in part, through NADPH oxidases. We proposed a hypothetical model for the signaling pathway controlling SOS1 mRNA stability. In this addendum, we discuss the possible involvement of other components in conferring inherent instability and stress-induced stability of SOS1 mRNA.
mRNA stability; salt stress; signaling transduction; SOS1
After germination, seedlings follow one of two developmental programs, photo- and skotomorphogenesis, and the choice is determined by the interplay between environmental signals (light) and endogenous cues (the plant hormones gibberellins among others). In the December issue of Plant Journal we describe a molecular mechanism that allows the integration of light and gibberellin signaling to tightly control the switch between skoto- and photomorphogenesis. On one hand, the stability of HY5, a transcription factor required by light to promote photomorphogenesis, is enhanced in the light and in situations with compromised GA biosynthesis. And, on the other hand, the promotion of growth during etiolation is exerted by the PIF family of transcription factors, whose abundance is enhanced by the absence of light, and whose activity is regulated by functional interaction with gibberellin signaling. In this addendum we propose that the control of the activity of light-dependent transcription factors by gibberellins is a common theme in other developmental processes, such as shade avoidance and photoperiodic regulation of cell expansion.
gibberellins; photomorphogenesis; shade avoidance; cell expansion
Plant growth and development are tightly regulated by both plant growth substances and environmental factors such as temperature. Taking into account the above, it was reasonable to point out that indole-3-acetic acid (IAA), the most abundant type of auxin in plants, could be involved in temperature- dependent growth of plant cells. We have recently shown that growth of maize coleoptile segments in the presence of auxin (IAA) and fusicoccin (FC) shows the maximum value in the range 30–35°C and 35–40°C, respectively. Furthermore, simultaneous measurements of growth and external medium pH indicated that FC at stressful temperatures was not only much more active in the stimulation of growth, but was also more effective in acidifying the external medium than IAA. The aim of this addendum is to determine interrelations between the action of IAA and FC (applied together with IAA) on growth and medium pH of maize coleoptile segments incubated at high temperature (40°C), which was optimal for FC but not for IAA.
auxin; fusicoccin; coleoptile segments; elongation growth; medium pH
Brain derived neurotrophic factor (BDNF) is crucial for the formation of appropriate synaptic connections during development and for learning and memory in adults. Secretion of this neurotrophin is under activity-dependent control. Understanding which patterns of physiological activity regulate BDNF secretion is therefore an important step in the comprehension of its role. We have recently shown that back propagation of action potentials (bAPs) is the principal triggering mechanism of dendritic BDNF secretion occurring during ongoing neuronal activity in neuronal cultures. In the present addendum we discuss possible implications of bAPs-induced BDNF secretion on the construction and reorganization of neuronal networks.
BDNF; peptide secretion; back propagating action potentials; synaptic plasticity; neuromodulation; acetylcholine
Using a live-cell imaging approach to study individual micro-tubules, we have compared microtubule behavior between net-like and aligned cortical arrays. In contrast to previous studies, a steep angled collision between the growing end of a microtubule and a preexisting microtubule was found to favor crossover. Frequencies of microtubule crossovers, bundling and catastrophes are similar regardless of whether the cell exhibited a net-like or aligned microtubule array. In the predominantly aligned array of petiole cells, severing occurs at the sites of microtubule crossovers and serves to remove unaligned microtubules and to increase microtubule density. Severing was observed to be rare in net-like arrays. Microtubule severing is carried out by the katanin enzyme. In this addendum, we present new insights into the possible mechanism of crossing over and preliminary data looking at organization of the array in a katanin mutant.
cortical array; collision; catastrophe; severing; katanin; petiole; pavement; cotyledons; YFP
Recent large scale phosphoproteomics studies have helped identify many phosphorylation sites of both membrane and soluble proteins. In most cases the relevance of specific sites has yet to be established whereas in a small number of cases their potency in modulating protein activity is evident. With the increasing amount of data it is becoming clear that phosphosites are often conserved within protein families, pointing to generic regulatory mechanisms. In addition, such mechanisms may be conserved across species. In this addendum evidence is presented for these phenomena occurring in rice and Arabidopsis.
Arabidopsis; kinase; phosphoproteomics; rice
It is well known for a long time, that nitric oxide (NO) functions in variable physiological and developmental processes in plants, however the source of this signaling molecule in the diverse plant responses is very obscure.1 Although existance of nitric oxide sythase (NOS) in plants is still questionable, LNMMA (NG-monomethyl-L-arginine)-sensitive NO generation was observed in different plant species.2,3 In addition, nitrate reductase (NR) is confirmed to have a major role as source of NO.4,5 This multifaced molecule acts also in auxin-induced lateral root (LR) formation, since exogenous auxin enhanced NO levels in regions of Arabidopsis LR initiatives. Our results pointed out the involvement of nitrate reductase enzyme in auxin-induced NO formation. In this addendum, we speculate on auxin-induced NO production in lateral root primordial formation.
atnoa1; indole-3-butyric acid; nia1; nia2 double mutant; nitric oxide
Polarity is a fundamental cell property essential for differentiation, proliferation and morphogenesis in unicellular and multicellular organisms. We have recently demonstrated that phosphatidylinositol 3-kinase (PI3K) activity is required for the establishment of anterior-posterior axis, leading to asymmetrical localization of F-actin in migrating monospores of the red alga Porphyra yezoensis. We also showed that the formation of the apical-basal axis via adhesion of monospores to the substratum after the cessation of migration requires newly synthesized proteins and does not depend on PI3K activity. However, little is known about the mechanism and regulation of axis conversion during development of monospores. In this addendum, we report our investigation as to the role of the cell wall in axis conversion. Our results indicate that inhibition of cell wall synthesis prevented the development of germlings. Also, defects in the cell wall disrupted the asymmetrical distribution of F-actin and inhibited the adhesion to the substratum that is required for establishment of apical-basal axis. Hence, we conclude that the cell wall is critical for the maintenance of cell polarity in migrating cells, which is indirectly involved in axis conversion via enabling monospores to adhere to the substratum.
BFA; cell polarity; cell wall; F-actin; monospores; PI3K; Porphyra yezoensis
Oxidative stress, arising from an imbalance in the generation and removal of reactive oxygen species (ROS), is a challenge faced by all aerobic organisms. In plants, different pathways sense ROS from extracellular sources or organelles such as mitochondria, chloroplast or peroxisome. In our recent paper on Plant Molecular Biology1 we have studied the Arabidopsis thaliana early response to the generation of superoxide anion in chloroplasts during active photosynthesis. Transcript profile analysis revealed that the expression level of various genes encoding heat shock proteins (Hsps), increased after a short term of oxidative stress treatment. Furthermore, there was an induction of heat shock transcription factors HsfA2 and HsfA4A that were reported to be regulators of genes involved in stress response of Arabidopsis.1,2
In this addendum, we complement the expression analysis of two Hsp genes encoding Hsp70 and a 17.6 kDa class I small heat-shock protein (sHsp), and discuss their plausible role during oxidative stress, considering our data and other recently published papers.
heat shock factor; heat shock protein; chloroplast; oxidative stress; signalling
Accumulating evidence indicates that plant growth promoting rhizobacteria (PGPR) influence plant growth and development by the production of phytohormones such as auxins, gibberellins, and cytokinins. Little is known on the genetic basis and signal transduction components that mediate the beneficial effects of PGPRs in plants. We recently reported the identification of a Bacillus megaterium strain that promoted growth of A. thaliana and P. vulgaris seedlings. In this addendum, the role of cytokinin signaling in mediating the plant responses to bacterial inoculation was investigated using A. thaliana mutants lacking one, two or three of the putative cytokinin receptors CRE1, AHK2 and AHK3, and RPN12 a gene involved in cytokinin signaling. We show that plant growth promotion by B. megaterium is reduced in AHK2-2 single and double mutant combinations and in RPN12. Furthermore, the triple cytokinin-receptor CRE1-12/AHK2-2/AHK3-3 knockout was insensitive to inoculation in terms of growth promotion and root developmental responses. Our results indicate that cytokinin receptors play a complimentary role in plant growth promotion by B. megaterium.
Arabidopsis; plant growth stimulation; root development; rhizobacteria
Copper amine oxidases (CuAO) and flavin-containing amine oxidases (PAO) are hydrogen peroxide (H2O2)-producing enzymes responsible for the oxidative de-amination of polyamines. Currently, a key role has been ascribed to apoplastic amine oxidases in plants, i.e., to behave as H2O2-delivering systems in the cell wall during cell growth and differentiation as well as in the context of host-pathogen interactions. Indeed, H2O2 is the co-substrate for the peroxidase-driven reactions during cell-wall maturation and a key signalling molecule in defence mechanisms. We recently demonstrated the involvement of an apoplastic PAO in the wound-healing process of the Zea mays mesocotyl. Experimental evidence indicated a similar role for an apoplastic PAO in Nicotiana tabacum. In this addendum we suggest that a CuAO activity is also involved in this healing event.
polyamine oxidase; copper amine oxidase; hydrogen peroxide; wound healing
In parallel to evoking the accumulation of stress-related transcripts, exposure to low level 900 MHz EMF affected the levels of ATP, the main energy molecule of the cell. Its concentration dropped rapidly (27% after 30 min) in response to EMF exposure, along with a 18% decrease in the adenylate energy charge (AEC), a good marker of cell energy status. One could interpret this decrease in ATP and AEC in a classical way, i.e., as the result of an increase in cellular energy usage, but recent work brings exciting new insights in pointing out a signalling function for ATP, especially in the stress physiology context where it could trigger both reactive oxygen species and calcium movement (this latter being involved in plant responses to EMF exposure). In this addendum, we discuss our results within this new perspective for ATP function.
EMF; ATP; eATP; tomato; stress
In eukaryotes, most genetic material resides in a separate membrane-bound compartment known as the nucleus. Transport of cargo, such as RNA and protein, across this barrier is facilitated by the nuclear pore complex (NPC). In the July issue of Plant Physiology, we showed that a component of the NPC, Arabidopsis thaliana TPR (AtTPR), is required for normal development. Two striking phenotypes of attpr mutants are that they are early flowering and show an accumulation of polyadenylated RNA in the nucleus. In addition, the expression of several microRNAs (miRNAs) is reduced in attpr mutants. In this addendum, we have examined the effect of AtTPR on the expression of miRNA targets. Our results show that miRNA targets are more likely to be upregulated than other transcripts in attpr mutants. For example, when comparing the nuclear RNA pool between wild-type and attpr plants, we found that 75% of the miRNA targets showing a significant change in transcript level are upregulated in attpr mutants. Although the targets of some miRNAs were upregulated, other miRNA targets were relatively unaffected by attpr mutations. Thus it appears that AtTPR may be required for the proper expression or localization of a subset of miRNAs.
nuclear export; flowering time; FLC; AtTPR
Sensing chemical signal secreted from host root and find the best site for penetration are crucial for initiating infection of Phytophthora zoospore. G protein α subunit of P. sojae participates in not only the chemotaxis to soybean isoflavone, but also finding penetrating site. Furthermore, although calcium signal pathways are influenced by Gα, other signal pathways also influenced by G protein remain to be discovered. In this addendum, we describe an RGS protein, PsRGS6, is expressed downregulated in zoospores of Gα silenced mutant. This result indicates that the expression of Gα and RGS protein may be influenced by each other. Some differences between Gα mutants of P. infestans and P. sojae may be due to the different developmental procedures.
G protein α subunit; penetrating site; regulator of G protein signaling (RGS); phytophthora
Reactive oxygen species (ROS) are commonly found in plants as natural by-products of the metabolism but their production is greatly enhanced under abiotic stresses. Particular metabolites and enzymes belonging to the ascorbate-glutathione cycle are able to scavenge these deleterious molecules and modulate the cellular redox-status. In the March issue of Journal of Plant Physiology, we have shown that drought stress induces a raise in glutathione reductase (GR) activity and gene expression that could be related to the intensity of the drought treatment and the drought susceptibility of the bean cultivar (cowpea and/or common bean). In the present addendum we show new data on GR specific activity during progressive drought stress and recovery of the drought-susceptible bean cultivar which can be related to the previously found dual-targeted GR gene expression. Furthermore, since in leguminous plants homoglutathione (hGSH) is generally the most abundant low molecular weight thiol form, we discuss on the occurrence of a (homo)glutathione reductase activity in beans.
common bean; cowpea; drought stress; (homo)glutathione; (homo)glutathione reductase; legumes; Phaseolus vulgaris; recovery; Vigna unguiculata