Plant Hormone Homeostasis Research Articles

Genome-Wide Identification, Phylogeny, Evolution and Expression Patterns of AP2/ERF Genes and Cytokinin Response Factors in Brassica rapa ssp. pekinensis

The AP2/ERF transcription factor family is one of the largest families involved in growth and development, hormone responses, and biotic or abiotic stress responses in plants. In this study, 281 AP2/ERF transcription factor unigenes were identified in Chinese cabbage. These superfamily members were classified into three families (AP2, ERF, and RAV). The ERF family was subdivided into the DREB subfamily and the ERF subfamily with 13 groups (I– XI) based on sequence similarity. Duplication, evolution and divergence of the AP2/ERF genes in B. rapa and Arabidopsis thaliana were investigated and estimated. Cytokinin response factors (CRFs), as a subclade of the AP2/ERF family, are important transcription factors that define a branch point in the cytokinin two-component signal (TCS) transduction pathway. Up to 21 CRFs with a conserved CRF domain were retrieved and designated as BrCRFs. The amino acid sequences, conserved regions and motifs, phylogenetic relationships, and promoter regions of the 21 BrCRFs were analyzed in detail. The BrCRFs broadly expressed in various tissues and organs. The transcripts of BrCRFs were regulated by factors such as drought, high salinity, and exogenous 6-BA, NAA, and ABA, suggesting their involvement in abiotic stress conditions and regulatory mechanisms of plant hormone homeostasis. These results provide new insight into the divergence, variation, and evolution of AP2/ERF genes at the genome-level in Chinese cabbage.

Identification, expression, and comparative genomic analysis of the IPT and CKX gene families in Chinese cabbage (Brassica rapa ssp. pekinensis)

BackgroundCytokinins (CKs) have significant roles in various aspects of plant growth and development, and they are also involved in plant stress adaptations. The fine-tuning of the controlled CK levels in individual tissues, cells, and organelles is properly maintained by isopentenyl transferases (IPTs) and cytokinin oxidase/dehydrogenases (CKXs). Chinese cabbage is one of the most economically important vegetable crops worldwide. The whole genome sequencing of Brassica rapa enables us to perform the genome-wide identification and functional analysis of the IPT and CKX gene families.ResultsIn this study, a total of 13 BrIPT genes and 12 BrCKX genes were identified. The gene structures, conserved domains and phylogenetic relationships were analyzed. The isoelectric point, subcellular localization and glycosylation sites of the proteins were predicted. Segmental duplicates were found in both BrIPT and BrCKX gene families. We also analyzed evolutionary patterns and divergence of the IPT and CKX genes in the Cruciferae family. The transcription levels of BrIPT and BrCKX genes were analyzed to obtain an initial picture of the functions of these genes. Abiotic stress elements related to adverse environmental stimuli were found in the promoter regions of BrIPT and BrCKX genes and they were confirmed to respond to drought and high salinity conditions. The effects of 6-BA and ABA on the expressions of BrIPT and BrCKX genes were also investigated.ConclusionsThe expansion of BrIPT and BrCKX genes after speciation from Arabidopsis thaliana is mainly attributed to segmental duplication events during the whole genome triplication (WGT) and substantial duplicated genes are lost during the long evolutionary history. Genes produced by segmental duplication events have changed their expression patterns or may adopted new functions and thus are obtained. BrIPT and BrCKX genes respond well to drought and high salinity stresses, and their transcripts are affected by exogenous hormones, such as 6-BA and ABA, suggesting their potential roles in abiotic stress conditions and regulatory mechanisms of plant hormone homeostasis. The appropriate modulation of endogenous CKs levels by IPT and CKX genes is a promising approach for developing economically important high-yielding and high-quality stress-tolerant crops in agriculture.

Characterization and expression patterns of microRNAs involved in rice grain filling.

MicroRNAs (miRNAs) are upstream gene regulators of plant development and hormone homeostasis through their directed cleavage or translational repression of the target mRNAs, which may play crucial roles in rice grain filling and determining the final grain weight and yield. In this study, high-throughput sequencing was performed to survey the dynamic expressions of miRNAs and their corresponding target genes at five distinct developmental stages of grain filling. In total, 445 known miRNAs and 45 novel miRNAs were detected with most of them expressed in a developmental stage dependent manner, and the majority of known miRNAs, which increased gradually with rice grain filling, showed negatively related to the grain filling rate. Detailed expressional comparisons revealed a clear negative correlation between most miRNAs and their target genes. It was found that specific miRNA cohorts are expressed in a developmental stage dependent manner during grain filling and the known functions of these miRNAs are involved in plant hormone homeostasis and starch accumulation, indicating that the expression dynamics of these miRNAs might play key roles in regulating rice grain filling.

Inhibition of tomato (Solanum lycopersicum L.) root growth by cyanamide is due to altered cell division, phytohormone balance and expansin gene expression

Cyanamide (CA) has been reported as a natural compound produced by hairy vetch (Vicia villosa Roth.) and it was shown also to be an allelochemical, responsible for strong allelopathic potential in this species. CA phytotoxicity has been demonstrated on various plant species, but to date little is known about its mode of action at cellular level. Treatment of tomato (Solanum lycopersicum L.) roots with CA (1.2 mM) resulted in inhibition of growth accompanied by alterations in cell division, and imbalance of plant hormone (ethylene and auxin) homeostasis. Moreover, the phytotoxic effect of CA was also manifested by modifications in expansin gene expression, especially in expansins responsible for cell wall remodeling after the cytokinesis (LeEXPA9, LeEXPA18). Based on these results the phytotoxic activity of CA on growth of roots of tomato seedlings is likely due to alterations associated with cell division.Electronic supplementary materialThe online version of this article (doi:10.1007/s00425-012-1722-y) contains supplementary material, which is available to authorized users.

Opprimo ergo sum—Evasion and Suppression in the Root Endophytic Fungus Piriformospora indica

The genetically tractable endophytic fungus Piriformospora indica is able to colonize the root cortex of a great variety of different plant species with beneficial effects to its hosts, and it represents a suitable model system to study symbiotic interactions. Recent cytological studies in barley and Arabidopsis showed that, upon penetration of the root, P. indica establishes a biotrophic interaction during which fungal cells are encased by the host plasma membrane. Large-scale transcriptional analyses of fungal and plant responses have shown that perturbance of plant hormone homeostasis and secretion of fungal lectins and other small proteins (effectors) may be involved in the evasion and suppression of host defenses at these early colonization steps. At later stages, P. indica is found more often in moribund host cells where it secretes a large variety of hydrolytic enzymes that degrade proteins. This strategy of colonizing plants is reminiscent of that of hemibiotrophic fungi, although a defined shift to necrotrophy with massive host cell death is missing. Instead, the association with the plant root leads to beneficial effects for the host such as growth promotion, increased resistance to root as well as leaf pathogens, and increased tolerance to abiotic stresses. This review describes current advances in understanding the components of the P. indica endophytic lifestyle from molecular and genomic analyses.

Floral induction in longan ( Dimocarpus longan, Lour.) trees : IV. The essentiality of mature leaves for potassium chlorate induced floral induction and associated hormonal changes

This study was conducted to elucidate the function of leaves and their role in the alteration of plant hormone homeostasis during potassium chlorate (KClO 3) induced “off season” floral induction (FI) of longan. KClO 3 application had almost no effect on the concentration of gibberellins in receptive apical and lateral shoot meristems, whereas the auxin IAA initially decreased, but later remarkably increased. The most significant changes caused by KClO 3 treatments was an increase in zeatin/zeatin-riboside (Z/ZR) type cytokinins (CKs) 8 days after treatment in the shoot apical and close by lateral buds as well as in the tissue beneath these buds. The 6–7-fold increase in CK concentrations were greatly reduced in the absence of leaves (leaves removed between the apical bud and a girdle). A significant exception was a strong accumulation of isopentenyl adenine/isopentenyl adenosine (iP/iPA) type CKs in wood and bark beneath the bud of defoliated shoots that were isolated against the rest of functional leaves of the tree by a girdle. This accumulation of iP/iPA, the immediate precursors of Z/ZR, together with the strong increase of Z/ZR in the shoot apical bud (SAB) after KClO 3 application indicate that leaves may be involved in the biosynthesis of CKs. Overall, the results suggest that cytokinins play a significant role in KClO 3 induced “off season” floral induction in longan. This role may not be restricted to the biosynthesis of these hormones but also to their mobilization and accumulation in the SAB of newly flushing shoots.

PLASTOCHRON3/GOLIATH encodes a glutamate carboxypeptidase required for proper development in rice

Most aerial parts of the plant body are products of the continuous activity of the shoot apical meristem (SAM). Leaves are the major component of the aerial plant body, and their temporal and spatial distribution mainly determines shoot architecture. Here we report the identification of the rice gene PLASTOCHRON3 (PLA3)/GOLIATH (GO) that regulates various developmental processes including the rate of leaf initiation (the plastochron). PLA3/GO encodes a glutamate carboxypeptidase, which is thought to catabolize small acidic peptides and produce small signaling molecules. pla3 exhibits similar phenotypes to pla1 and pla2- a shortened plastochron, precocious leaf maturation and rachis branch-to-shoot conversion in the reproductive phase. However, in contrast to pla1 and pla2, pla3 showed pleiotropic phenotypes including enlarged embryo, seed vivipary, defects in SAM maintenance and aberrant leaf morphology. Consistent with these pleiotropic phenotypes, PLA3 is expressed in the whole plant body, and is involved in plant hormone homeostasis. Double mutant analysis revealed that PLA1, PLA2 and PLA3 are regulated independently but function redundantly. Our results suggest that PLA3 modulates various signaling pathways associated with a number of developmental processes.

What can we learn from clubroots: alterations in host roots and hormone homeostasis caused by Plasmodiophora brassicae

The clubroot disease of cruciferous crops is caused by an obligate biotrophic protist, Plasmodiophora brassicae. The disease is characterized by the development of large root galls accompanied by changes in source-sink relations and the hormonal balance within the plant. Since the disease is difficult to control, it is of high economic interest to understand the events leading to gall formation. In this review we will give an overview on the current knowledge of changes brought about in the host root by this obligate biotrophic pathogen. Emphasis will be on the regulation of changes in plant hormone homeostasis, mainly auxins and cytokinins; the possible role of secondary metabolites, especially indole glucosinolates, in gall formation and auxin homeostasis will be discussed. Also, results from mutant analysis and microarrays using the model plant Arabidopsis thaliana are presented.

Pseudomonas syringae pv. tomato hijacks the Arabidopsis abscisic acid signalling pathway to cause disease.

We have found that a major target for effectors secreted by Pseudomonas syringae is the abscisic acid (ABA) signalling pathway. Microarray data identified a prominent group of effector-induced genes that were associated with ABA biosynthesis and also responses to this plant hormone. Genes upregulated by effector delivery share a 42% overlap with ABA-responsive genes and are also components of networks induced by osmotic stress and drought. Strongly induced were NCED3, encoding a key enzyme of ABA biosynthesis, and the abscisic acid insensitive 1 (ABI1) clade of genes encoding protein phosphatases type 2C (PP2Cs) involved in the regulation of ABA signalling. Modification of PP2C expression resulting in ABA insensitivity or hypersensitivity led to restriction or enhanced multiplication of bacteria, respectively. Levels of ABA increased rapidly during bacterial colonisation. Exogenous ABA application enhanced susceptibility, whereas colonisation was reduced in an ABA biosynthetic mutant. Expression of the bacterial effector AvrPtoB in planta modified host ABA signalling. Our data suggest that a major virulence strategy is effector-mediated manipulation of plant hormone homeostasis, which leads to the suppression of defence responses.

Xanthine dehydrogenase and aldehyde oxidase impact plant hormone homeostasis and affect fruit size in 'Hass' avocado.

The contribution of xanthine dehydrogenase (XDH, EC 1.1.1.204) to fruit size was investigated using the normal and small-fruit variants of Persea americana Mill. cv. 'Hass'. Inhibition of XDH by treatment of normal fruit, in the linear phase of growth (phase II), with allopurinol (Allo) arrested fruit growth. Adenine (Ade), a less effective inhibitor of this enzyme, also arrested fruit growth when applied in phase II and slowed fruit growth when applied in phase III. A time-course study on the activity of XDH in mesocarp tissue from normal and small fruit showed that maximum activity occurred late in phase II and that the peak in activity was absent in mesocarp of the small fruit. Feeding Ade to growing fruit in phase III caused a transient decline in fruit growth (measured as change in fruit length). Thereafter, growth resumed although fruit size was irreversibly affected. Treatment of fruit with Ade and Ade-containing cytokinins altered activity of another molybdenum enzyme, aldehyde oxidase (EC 1.2.3.1). Cytokinin oxidase was induced by cytokinin and auxin. Purine catabolism via hypoxanthine/xanthine was operative in normal fruit and in mesocarp from the small-fruit variant and as expected, Allo treatment caused accumulation of xanthine and adenine. In the absence of an increase in XDH during growth of the small-fruit phenotype, low levels of Ade were interpreted as resulting from respiration-enhanced adenylate depletion. Stress and/or pathogen induction of the alternative oxidase pathway is proposed as a possible cause.

A novel short-root gene that affects specifically early root development in rice ( Oryza sativa L.)

We have isolated a novel short-root mutant, srt5, from an M 2 population of rice ( Oryza sativa L. cv. Oochikara) treated with NaN 3, and mapped its causal gene to chromosome 2. The mutant showed extreme inhibition of seminal root, crown root and lateral root elongation, and altered root hair formation at the seedling stage. Histological observation of seminal root tips indicated that the short-root phenotype was due to the reduced cell size and cell number. At later stages of plant development, roots of the mutant elongated acceleratingly and recovered normal morphology 45 days after transplantation, and adult plants of the mutant resembled the wild type except for reduced pollen and seed fertilities. Seminal root growth of the mutant was partially rescued by exogenously applied ABA, suggesting that ABA biosynthesis might be affected in srt5 plants. Moreover, the mutant also showed high resistance to 2,4-D, GA 3 and KIN based on seminal root growth assays, but the resistance to these hormones disappeared when ABA was concomitantly applied. Thus, the wild type allele of srt5 seems to be of great importance for early root development by altering plant hormone homeostasis, in which ABA plays a critical role.

A rapid increase in cytokinin levels and enhanced ethylene evolution precede Al3+-induced inhibition of root growth in bean seedlings (Phaseolus vulgaris L.)

The role of plant hormones in aluminium (Al3+)-inducedinhibition of root growth was investigated in roots of Phaseolusvulgaris L. cv. Strike. Changes in ethylene evolution and changes inthe content and composition of cytokinins (CKs), at intervals up to 150min after treatment with Al3+, were determined by gaschromatography (GC) and combined gas chromatography-mass spectrometry (GC-MS),respectively. Seedlings were cultivated in a continuously aerated nutrientsolution at pH 4.5 with and without Al3+. The growth rate ofAl3+-treated roots was significantly decreased after 360min. Ethylene evolution from excised root tips doubled after 15min of Al3+ treatment and reached a maximum 30min after treatment. Levels of CK nucleotides declined 60 to 80%after only 5 min of Al3+ treatment whereas the zeatin(Z) content increased six-fold. The increase in Z continued over the entire 150min-sampling period and reached a level 80 times higher than thatin roots not exposed to Al3+. These results show thatAl3+-induced inhibition of root growth is preceded by significantchanges in CK content and composition and enhanced ethylene evolution. Since CKcan induce ethylene production, the rapid increase in CK, particularly Z anddihydrozeatin (dZ), may contribute to inhibition of root-growth either directlyor indirectly by affecting plant hormone homeostasis.

Fruit size: Towards an understanding of the metabolic control of fruit growth using avocado as a model system

Final fruit size is the consequence of complex metabolic events that occur between fruit set and maturation. Disruption of these biochemical and molecular processes at any stage during fruit growth will impact on final fruit size. Because fruit size is a function of cell number rather than cell size, factors affecting cell division cycle activity assume importance. In this paper, we focus attention on the metabolic control of fruit growth using avocado as a model system. Three areas of current interest are highlighted, viz. the contribution by isoprenoid metabolism in the control of cell proliferation, the role played by carbohydrate content and composition in signalling changes in metabolite status and gene expression and maintenance of plant hormone homeostasis. Central to the process of fruit growth and control of final fruit size by cell division is 3‐hydroxy‐3‐methylglutaryl coenzyme A reductase (HMGR) and activity of the sucrose non‐fermenting 1‐related protein kinase (SnRK1) complex. It is argued that sugar content and composition of sink cells impact on SnRK1 (and hexokinase) to modulate expression of sugar‐metabolizing enzymes, HMGR and molybdenum cofactor (MoCo)‐containing enzymes. These changes, in turn, impact on hormone metabolism by affecting allocation of the purine‐derived MoCo to aldehyde oxidase and thus the endogenous concentration of indole‐3‐acetic acid, abscisic acid and cytokinin (CK) to alter plant hormone homeostasis. These aspects are integrated into a model to explain the metabolic control of avocado fruit growth and final fruit size.

Plant hormone homeostasis and the control of avocado fruit size

Control of plant hormone homeostasis is crucial for normal organdevelopment in plants. To elucidate the contribution of plant hormonehomeostasis to fruit growth, tissue distribution and activity of xanthinedehydrogenase (XDH), abscisic aldehyde (AB-ald)- and indole acetaldehyde(IA-ald) oxidase, and cytokinin oxidase (CKOX) were determined in seed, seedcoat and mesocarp of normal 'Hass’ avocado and its small-fruitphenotype during the linear phase of growth. Activity of these enzymes wasrelated to the tissue content of indole-3-acetic acid (IAA) and abscisic acid(ABA). IA-ald oxidase was present only in seed tissue whereas AB-ald oxidase andXDH activity was found in seed and mesocarp tissue. Seed of the small'Hass’ fruit had increased XDH and AB-ald oxidase activity and highendogenous ABA, but reduced IA-ald oxidase activity and adenine. There was nodifference in seed, seed coat and mesocarp CKOX activity between normal andsmall fruit. Inhibition of XDH activity in whole fruit by treatment withallopurinol decreased IAA and increased ABA of seed tissue. In mesocarp ofripening fruit allopurinol increased ABA and IAA but had no effect on levels ofiP. Results indicate that activity of IA-ald and AB-ald oxidases in avocadofruit contribute to maintenance of the IAA/ABA ratio in seed and mesocarp tissueand that increased AB-ald oxidase, or reduced IA-ald oxidase, may be part of thesyndrome associated with the appearance of a small-fruit phenotype.