Tissue-specific expression of stabilized SOLITARY-ROOT/IAA14 alters lateral root development in Arabidopsis

Auxin is important for lateral root (LR) initiation and subsequent LR primordium development. However, the roles of tissue-specific auxin signaling in these processes are poorly understood. We analyzed transgenic Arabidopsis plants expressing the stabilized mutant INDOLE-3 ACETIC ACID 14 (IAA14)/SOLITARY-ROOT (mIAA14) protein as a repressor of the auxin response factors (ARFs), under the control of tissue-specific promoters. We showed that plants expressing the mIAA14-glucocorticoid receptor (GR) fusion protein under the control of the native IAA14 promoter had the solitary-root/iaa14 mutant phenotypes, including the lack of LR formation under dexamethasone (Dex) treatment, indicating that mIAA14-GR is functional in the presence of Dex. We then demonstrated that expression of mIAA14-GR under the control of the stele-specific SHORT-ROOT promoter suppressed LR formation, and showed that mIAA14-GR expression in the protoxylem-adjacent pericycle also blocked LR formation, indicating that the normal auxin response mediated by auxin/indole-3 acetic acid (Aux/IAA) signaling in the protoxylem pericycle is necessary for LR formation. In addition, we demonstrated that expression of mIAA14-GR under either the ARF7 or the ARF19 promoter also suppressed LR formation as in the arf7 arf19 double mutants, and that IAA14 interacted with ARF7 and ARF19 in yeasts. These results strongly suggest that mIAA14-GR directly inactivates ARF7/ARF19 functions, thereby blocking LR formation. Post-embryonic expression of mIAA14-GR under the SCARECROW promoter, which is expressed in the specific cell lineage during LR primordium formation, caused disorganized LR development. This indicates that normal auxin signaling in LR primordia, which involves the unknown ARFs and Aux/IAAs, is necessary for the establishment of LR primordium organization. Thus, our data show that tissue-specific expression of a stabilized Aux/IAA protein allows analysis of tissue-specific auxin responses in LR development by inactivating ARF functions.     

Multiple AUX/IAA�CARF modules regulate lateral root formation: the role of Arabidopsis SHY2/IAA3-mediated auxin signalling

In Arabidopsis thaliana, lateral root (LR) formation is regulated by multiple auxin/indole-3-acetic acid (Aux/IAA)–AUXIN RESPONSE FACTOR (ARF) modules: (i) the IAA28–ARFs module regulates LR founder cell specification; (ii) the SOLITARY-ROOT (SLR)/IAA14–ARF7–ARF19 module regulates nuclear migration and asymmetric cell divisions of the LR founder cells for LR initiation; and (iii) the BODENLOS/IAA12–MONOPTEROS/ARF5 module also regulates LR initiation and organogenesis. The number of Aux/IAA–ARF modules involved in LR formation remains unknown. In this study, we isolated the shy2-101 mutant, a gain-of-function allele of short hypocotyl2/suppressor of hy2 (shy2)/iaa3 in the Columbia accession. We demonstrated that the shy2-101 mutation not only strongly inhibits LR primordium development and emergence but also significantly increases the number of LR initiation sites with the activation of LATERAL ORGAN BOUNDARIES-DOMAIN16/ASYMMETRIC LEAVES2-LIKE18, a target gene of the SLR/IAA14–ARF7–ARF19 module. Genetic analysis revealed that enhanced LR initiation in shy2-101 depended on the SLR/IAA14–ARF7–ARF19 module. We also showed that the shy2 roots contain higher levels of endogenous IAA. These observations indicate that the SHY2/IAA3–ARF-signalling module regulates not only LR primordium development and emergence after SLR/IAA14–ARF7–ARF19 module-dependent LR initiation but also inhibits LR initiation by affecting auxin homeostasis, suggesting that multiple Aux/IAA–ARF modules cooperatively regulate the developmental steps during LR formation.     

Gibberellin Acts through Jasmonate to Control the Expression of MYB21, MYB24, and MYB57 to Promote Stamen Filament Growth in Arabidopsis

Precise coordination between stamen and pistil development is essential to make a fertile flower. Mutations impairing stamen filament elongation, pollen maturation, or anther dehiscence will cause male sterility. Deficiency in plant hormone gibberellin (GA) causes male sterility due to accumulation of DELLA proteins, and GA triggers DELLA degradation to promote stamen development. Deficiency in plant hormone jasmonate (JA) also causes male sterility. However, little is known about the relationship between GA and JA in controlling stamen development. Here, we show that MYB21, MYB24, and MYB57 are GA-dependent stamen-enriched genes. Loss-of-function of two DELLAs RGA and RGL2 restores the expression of these three MYB genes together with restoration of stamen filament growth in GA-deficient plants. Genetic analysis showed that the myb21-t1 myb24-t1 myb57-t1 triple mutant confers a short stamen phenotype leading to male sterility. Further genetic and molecular studies demonstrate that GA suppresses DELLAs to mobilize the expression of the key JA biosynthesis gene DAD1, and this is consistent with the observation that the JA content in the young flower buds of the GA-deficient quadruple mutant ga1-3 gai-t6 rga-t2 rgl1-1 is much lower than that in the WT. We conclude that GA promotes JA biosynthesis to control the expression of MYB21, MYB24, and MYB57. Therefore, we have established a hierarchical relationship between GA and JA in that modulation of JA pathway by GA is one of the prerequisites for GA to regulate the normal stamen development in Arabidopsis.     

Genome-wide analysis of Aux/IAA genes in Vitis vinifera: cloning and expression profiling of a grape Aux/IAA gene in response to phytohormone and abiotic stresses

Auxin is one of the most important phytohormones involved in plant growth and development. Here, we identified a total of 26 Aux/IAA genes displaying high sequence identity within the conserved domains I, II, III, and IV by screening the grapevine genome proteome 12× database. The Vitis vinifera Aux/IAA proteins can be classified into two groups (A and B) on the basis of their phylogenetic relationships. A search for cis-regulatory elements in the promoter sequences of VvAux/IAA genes revealed that the majority of these proteins may be regulated by light, phytohormones, and abiotic stresses. We also report the isolation and expression analysis of the cDNA of VvAux/IAA4, the most highly expressed VvAux/IAA gene from V. vinifera cv. Sultanine, according to ESTs in the NCBI database. The VvAux/IAA4 gene contains a full-length open reading frame of 1,080 bp, and its predicted amino acid sequence is highly similar to those of Aux/IAA proteins from other plants, including the presence of an AuxIAA/ARF dimerization motif in the C-terminal region. The expression of VvAux/IAA4 was found to be elevated during berry development, and slowly decrease from the initiation of ripening to the overripening stage. VvAux/IAA4 was highly expressed in young leaves and roots, and expressed at low levels in pollen and tendrils. Finally, the expression of VvAux/IAA4 was rapidly induced in response to NAA treatment, but was decreased by salt, drought, and SA treatments. Our results provide evidence of crosstalk between phytohormone and abiotic stresses, and support a role for auxin in stress responses.     

Contrasting modes of diversification in the Aux/IAA and ARF gene families

The complete genomic sequence for Arabidopsis provides the opportunity to combine phylogenetic and genomic approaches to study the evolution of gene families in plants. The Aux/IAA and ARF gene families, consisting of 29 and 23 loci in Arabidopsis, respectively, encode proteins that interact to mediate auxin responses and regulate various aspects of plant morphological development. We developed scenarios for the genomic proliferation of the Aux/IAA and ARF families by combining phylogenetic analysis with information on the relationship between each locus and the previously identified duplicated genomic segments in Arabidopsis. This analysis shows that both gene families date back at least to the origin of land plants and that the major Aux/IAA and ARF lineages originated before the monocot-eudicot divergence. We found that the extant Aux/IAA loci arose primarily through segmental duplication events, in sharp contrast to the ARF family and to the general pattern of gene family proliferation in Arabidopsis. Possible explanations for the unusual mode of Aux/IAA duplication include evolutionary constraints imposed by complex interactions among proteins and pathways, or the presence of long-distance cis-regulatory sequences. The antiquity of the two gene families and the unusual mode of Aux/IAA diversification have a number of potential implications for understanding both the functional and evolutionary roles of these genes.     

Light Promotes Protein Stability of Auxin Response Factor 7

Light is an environmental signaling, whereas Aux/IAA proteins and Auxin Response Factors (ARFs) are regulators of auxin signalling. Aux/IAA proteins are unstable, and their degradation dependents on 26S ubiquitin-proteasome and is promoted by Auxin. Auxin binds directly to a SCF-type ubiquitin-protein ligase, TIR1, facilitates the interaction between Aux/IAA proteins and TIR1, and then the degradation of Aux/IAA proteins. A few studies have reported that some ARFs are also unstable proteins, and their degradation is also mediated by 26S proteasome. In this study, by using of antibodies recognizing endogenous ARF7 proteins, we found that protein stability of ARF7 was affected by light. By expressing MYC tagged ARF activators in protoplasts, we found that degradation of ARF7 was inhibited by 26 proteasome inhibitors. In addition, at least ARF5 and ARF19 were also unstable proteins, and degradation of ARF5 via 26S proteasome was further confirmed by using stable transformed plants overexpressing ARF5 with a GUS tag.     

Light controls stamen elongation via cryptochromes,phytochromes and COP1 through HY5 and HYH

In Arabidopsis, stamen elongation, which ensures male fertility, is controlled by the auxin response factor ARF8, which regulates the expression of the auxin repressor IAA19. Here, we uncover a role for light in controlling stamen elongation. By an extensive genetic and molecular analysis we show that the repressor of light signaling COP1, through its targets HY5 and HYH, controls stamen elongation, and that HY5 – oppositely to ARF8 – directly represses the expression of IAA19 in stamens. In addition, we show that in closed flower buds, when light is shielded by sepals and petals, the blue light receptors CRY1/CRY2 repress stamen elongation. Coherently, at flower disclosure and in subsequent stages, stamen elongation is repressed by the red and far‐red light receptors PHYA/PHYB. In conclusion, different light qualities – sequentially perceived by specific photoreceptors – and the downstream COP1–HY5/HYH module finely tune auxin‐induced stamen elongation and thus male fertility.     

Aboveground Whitefly Infestation Modulates Transcriptional Levels of Anthocyanin Biosynthesis and Jasmonic Acid Signaling-Related Genes and Augments the Cope with Drought Stress of Maize

Up to now, the potential underlying molecular mechanisms by which maize (Zea mays L.) plants elicit defense responses by infestation with a phloem feeding insect whitefly [Bemisia tabaci (Genn.)] have been barely elucidated against (a)biotic stresses. To fill this gap of current knowledge maize plants were infested with whitefly and these plants were subsequently assessed the levels of water loss. To understand the mode of action, plant hormone contents and the stress-related mRNA expression were evaluated. Whitefly-infested maize plants did not display any significant phenotypic differences in above-ground tissues (infested site) compared with controls. By contrast, root (systemic tissue) biomass was increased by 2-fold by whitefly infestation. The levels of endogenous indole-3-acetic acid (IAA), jasmonic acid (JA), and hydrogen peroxide (H₂O₂) were significantly higher in whitefly-infested plants. The biosynthetic or signaling-related genes for JA and anthocyanins were highly up-regulated. Additionally, we found that healthier plants were obtained in whitefly-infested plants under drought conditions. The weight of whitefly-infested plants was approximately 20% higher than that of control plants at 14 d of drought treatment. The drought tolerance-related genes, ZmbZIP72, ZmSNAC1, and ZmABA1, were highly expressed in the whitefly-infected plants. Collectively, our results suggest that IAA/JA-derived maize physiological changes and correlation of H₂O₂ production and water loss are modulated by above-ground whitefly infestation in maize plants.     

烟草TTG2蛋白质对NPRI介导的抗病防卫反应与ARF8影响的生长发育进行交叉调控的机制

NPR1蛋白是水杨酸信号和系统获得性抗性的转录调节因子,它的功能受蛋白质降解酶体CUL3．E3的控制。植物的发育主要受生长素信号通路的控制,生长素反应因（ARF）参与生长素信号转导转录调控。植物转录因于NPR1和ARF8分别在蛋白质降解酶体CUL3-E3与CUL1-E3控制下,调控抗病防卫与生长发育。烟草TTG2促进ARF8从细胞质向细胞核转运及其转录调控作用,因此促进生长发育;相反,TTG2把NPR1扣留在细胞质,阻止它对防卫反应基因的转录调控作用,从而抑制抗病性。TTG2与NPR1或ARF8并不直接互作,说明存在协助因子。     

Two Alternatively Spliced Isoforms of the Arabidopsis SR45 Protein Have Distinct Roles during Normal Plant Development

The serine-arginine-rich (SR) proteins constitute a conserved family of pre-mRNA splicing factors. In Arabidopsis (Arabidopsis thaliana), they are encoded by 19 genes, most of which are themselves alternatively spliced. In the case of SR45, the use of alternative 3′ splice sites 21 nucleotides apart generates two alternatively spliced isoforms. Isoform 1 (SR45.1) has an insertion relative to isoform 2 (SR45.2) that replaces a single arginine with eight amino acids (TSPQRKTG). The biological implications of SR45 alternative splicing have been unclear. A previously described loss-of-function mutant affecting both isoforms, sr45-1, shows several developmental defects, including defects in petal development and root growth. We found that the SR45 promoter is highly active in regions with actively growing and dividing cells. We also tested the ability of each SR45 isoform to complement the sr45-1 mutant by overexpression of isoform-specific green fluorescent protein (GFP) fusion proteins. As expected, transgenic plants overexpressing either isoform displayed both nuclear speckles and GFP fluorescence throughout the nucleoplasm. We found that SR45.1-GFP complements the flower petal phenotype, but not the root growth phenotype. Conversely, SR45.2-GFP complements root growth but not floral morphology. Mutation of a predicted phosphorylation site within the alternatively spliced segment, SR45.1-S219A-GFP, does not affect complementation. However, a double mutation affecting both serine-219 and the adjacent threonine-218 (SR45.1-T218A + S219A-GFP) behaves like isoform 2, complementing the root but not the floral phenotype. In conclusion, our study provides evidence that the two alternatively spliced isoforms of SR45 have distinct biological functions.     

Foliar spray of Auxin/IAA modulates photosynthesis,elemental composition,ROS localization and antioxidant machinery to promote growth of Brassica juncea

Auxins (Aux) are primary growth regulators that regulate almost every aspect of growth and development in plants. It plays a vital role in various plant processes besides controlling the key aspects of cell division, cell expansion, and cell differentiation. Considering the significance of Aux, and its potential applications, a study was conducted to observe the impact of indole acetic acid (IAA), a most active and abundant form of Aux on Brassica juncea plants growing under natural environmental conditions. Different concentrations (0, 10⁻¹⁰, 10⁻⁸, 10⁻⁶ M) of IAA were applied once in a day at 25-day stage of growth for 5 days, consecutively. Various parameters (growth, photosynthetic, biochemical, oxidative biomarkers and nutrient composition) were assessed at different days after sowing (DAS). Scanning electron microscopy (SEM) of leaf stomata, reactive oxygen species (ROS) localization in leaf and roots, and confocal microscopy were also conducted. The results revealed that all the IAA concentrations were effective in growth promotion and ROS reduction, however, the 10⁻⁸ M of IAA exhibited the maximum improvement in all the above mentioned parameters as compared to the control.