Synergistic Derepression of Gibberellin Signaling by Removing RGA and GAI Function in Arabidopsis thaliana

Genetics ◽  
2001 ◽  
Vol 159 (2) ◽  
pp. 777-785
Author(s):  
Alyssa Dill ◽  
Tai-ping Sun
Keyword(s):  
Arabidopsis Thaliana ◽  
Feedback Inhibition ◽  
Stem Elongation ◽  
Dominant Role ◽  
Signal Transduction Pathway ◽  
Loss Of Function ◽  
Wild Type ◽  
Ga Signaling ◽  
Mutant Phenotypes ◽  
Redundant Functions

Abstract RGA and GAI are negative regulators of the gibberellin (GA) signal transduction pathway in Arabidopsis thaliana. These genes may have partially redundant functions because they are highly homologous, and plants containing single null mutations at these loci are phenotypically similar to wild type. Previously, rga loss-of-function mutations were shown to partially suppress defects of the GA-deficient ga1-3 mutant. Phenotypes rescued include abaxial trichome initiation, rosette radius, flowering time, stem elongation, and apical dominance. Here we present work showing that the rga-24 and gai-t6 null mutations have a synergistic effect on plant growth. Although gai-t6 alone has little effect, when combined with rga-24, they completely rescued the above defects of ga1-3 to wild-type or GA-overdose phenotype. However, seed germination and flower development defects were not restored. Additionally, rga-24 and rga-24/gai-t6 but not gai-t6 alone caused increased feedback inhibition of expression of a GA biosynthetic gene in both the ga1-3 and wild-type backgrounds. These results demonstrate that RGA and GAI have partially redundant functions in maintaining the repressive state of the GA-signaling pathway, but RGA plays a more dominant role than GAI. Removing both RGA and GAI function allows for complete derepression of many aspects of GA signaling.

scholarly journals Goα Regulates Volatile Anesthetic Action in Caenorhabditis elegans

Genetics ◽  
2001 ◽  
Vol 158 (2) ◽  
pp. 643-655 ◽  
Author(s):  
Bruno van Swinderen ◽  
Laura B Metz ◽  
Laynie D Shebester ◽  
Jane E Mendel ◽  
Paul W Sternberg ◽  
...  
Keyword(s):  
Caenorhabditis Elegans ◽  
Novel Mutation ◽  
Volatile Anesthetic ◽  
Loss Of Function ◽  
Wild Type ◽  
Α Subunit ◽  
C Elegans ◽  
Missense Mutant ◽  
Anesthetic Action ◽  
Mutant Phenotypes

Abstract To identify genes controlling volatile anesthetic (VA) action, we have screened through existing Caenorhabditis elegans mutants and found that strains with a reduction in Go signaling are VA resistant. Loss-of-function mutants of the gene goa-1, which codes for the α-subunit of Go, have EC50s for the VA isoflurane of 1.7- to 2.4-fold that of wild type. Strains overexpressing egl-10, which codes for an RGS protein negatively regulating goa-1, are also isoflurane resistant. However, sensitivity to halothane, a structurally distinct VA, is differentially affected by Go pathway mutants. The RGS overexpressing strains, a goa-1 missense mutant found to carry a novel mutation near the GTP-binding domain, and eat-16(rf) mutants, which suppress goa-1(gf) mutations, are all halothane resistant; goa-1(null) mutants have wild-type sensitivities. Double mutant strains carrying mutations in both goa-1 and unc-64, which codes for a neuronal syntaxin previously found to regulate VA sensitivity, show that the syntaxin mutant phenotypes depend in part on goa-1 expression. Pharmacological assays using the cholinesterase inhibitor aldicarb suggest that VAs and GOA-1 similarly downregulate cholinergic neurotransmitter release in C. elegans. Thus, the mechanism of action of VAs in C. elegans is regulated by Goα, and presynaptic Goα-effectors are candidate VA molecular targets.

scholarly journals Induced Systemic Resistance in Arabidopsis thaliana Against Pseudomonas syringae pv. tomato by 2,4-Diacetylphloroglucinol-Producing Pseudomonas fluorescens

2012 ◽  
Vol 102 (4) ◽  
pp. 403-412 ◽  
Author(s):  
David M. Weller ◽  
Dmitri V. Mavrodi ◽  
Johan A. van Pelt ◽  
Corné M. J. Pieterse ◽  
Leendert C. van Loon ◽  
...  
Keyword(s):  
Arabidopsis Thaliana ◽  
Pseudomonas Fluorescens ◽  
Induced Resistance ◽  
Pseudomonas Syringae ◽  
Induced Systemic Resistance ◽  
Signal Transduction Pathway ◽  
Systemic Resistance ◽  
Wild Type ◽  
Challenge Inoculation ◽  
Dependent Signal

Pseudomonas fluorescens strains that produce the polyketide antibiotic 2,4-diacetylphloroglucinol (2,4-DAPG) are among the most effective rhizobacteria that suppress root and crown rots, wilts, and damping-off diseases of a variety of crops, and they play a key role in the natural suppressiveness of some soils to certain soilborne pathogens. Root colonization by 2,4-DAPG-producing P. fluorescens strains Pf-5 (genotype A), Q2-87 (genotype B), Q8r1-96 (genotype D), and HT5-1 (genotype N) produced induced systemic resistance (ISR) in Arabidopsis thaliana accession Col-0 against bacterial speck caused by P. syringae pv. tomato. The ISR-eliciting activity of the four bacterial genotypes was similar, and all genotypes were equivalent in activity to the well-characterized strain P. fluorescens WCS417r. The 2,4-DAPG biosynthetic locus consists of the genes phlHGF and phlACBDE. phlD or phlBC mutants of Q2-87 (2,4-DAPG minus) were significantly reduced in ISR activity, and genetic complementation of the mutants restored ISR activity back to wild-type levels. A phlF regulatory mutant (overproducer of 2,4-DAPG) had ISR activity equivalent to the wild-type Q2-87. Introduction of DAPG into soil at concentrations of 10 to 250 μM 4 days before challenge inoculation induced resistance equivalent to or better than the bacteria. Strain Q2-87 induced resistance on transgenic NahG plants but not on npr1-1, jar1, and etr1 Arabidopsis mutants. These results indicate that the antibiotic 2,4-DAPG is a major determinant of ISR in 2,4-DAPG-producing P. fluorescens, that the genotype of the strain does not affect its ISR activity, and that the activity induced by these bacteria operates through the ethylene- and jasmonic acid-dependent signal transduction pathway.

scholarly journals Usp14 is required for spermatogenesis and ubiquitin stress responses in Drosophila melanogaster

2019 ◽  
Author(s):  
Levente Kovács ◽  
Ágota Nagy ◽  
Margit Pál ◽  
Peter Deák
Keyword(s):  
Male Sterility ◽  
Stress Responses ◽  
Expression Patterns ◽  
Loss Of Function ◽  
Wild Type ◽  
Cellular Processes ◽  
Protein Conjugates ◽  
Covalently Linked ◽  
Mutant Phenotypes

ABSTRACTDeubiquitinating (DUB) enzymes free covalently linked ubiquitins from ubiquitin-ubiquitin and ubiquitin-protein conjugates, and thereby maintain the equilibrium between free and conjugated ubiquitins and regulate ubiquitin-mediated cellular processes. The present genetic analyses of mutant phenotypes demonstrate that loss of Usp14 function results in male sterility, with defects in spermatid individualization and reduced testicular free monoubiquitin levels. These phenotypes were rescued by germline specific overexpression of wild type Usp14. Synergistic genetic interactions with Ubi-p63E and cycloheximide sensitivity suggest that ubiquitin shortage is a primary cause of male sterility. In addition, Usp14 is predominantly expressed in testes in Drosophila, and differential expression patterns may be causative of testis-specific loss of function Usp14 phenotypes. Collectively, these results suggest a major role of Usp14 in maintaining normal steady state free monoubiquitin levels during the later stages of Drosophila spermatogenesis.

Phytochrome contributes to blue-light-mediated stem elongation and flower initiation in mature Arabidopsis thaliana plants

2021 ◽  
Author(s):  
Yun Kong ◽  
Youbin Zheng
Keyword(s):  
Arabidopsis Thaliana ◽  
Blue Light ◽  
Stem Elongation ◽  
Growth Trait ◽  
Stem Length ◽  
Photon Flux ◽  
Flower Initiation ◽  
Shade Avoidance ◽  
Wild Type ◽  
Avoidance Responses

To examine whether phytochromes contribute to blue-light-mediated stem elongation, plant phenotypic responses were investigated in wild type Arabidopsis thaliana (Col-0), and its quintuple phytochrome (phyA phyB phyC phyD phyE) mutant plants under the following light treatments: (1) R, a pure red light from 660-nm LED; (2) B, a pure blue light from 455-nm LED; (3) BR, a impure blue light from LED combination of 94% B and 6% R; and (4) BRF, another impure blue light from LED combination of BR and 6 µmol m−2 s−1 of FR (735 nm). A photosynthetic photon flux density of ≈100 μmol m−2 s−1 was provided for all the light treatments. The calculated phytochrome photoequilibrium was 0.89, 0.50, 0.69, and 0.60 for R, B, BR, and BRF, respectively, indicating a higher phytochrome activity under R and BR than B and BRF. After 18 days of light treatment, B or BRF increased main stem length in wild-type plants compared with R, but BR had an inhibition effect similar to R. Also, B and BRF relative to R or BR induced earlier flowering and reduced leaf size in wild type plants, showing typical shade-avoidance responses. In phytochrome-deficient mutant plants, the above shade-avoidance responses were inhibited under B or BRF. However, hypocotyl length, a growth trait characterizing the de-etiolation stage, was reduced under B, BR and BRF vs. R regardless of phytochrome absence. These findings suggest that for mature Arabidopsis plants, phytochrome plays a role in blue-light-mediated stem elongation and the associated shade-avoidance responses.

Aquaporin AtTIP5;1 as an essential target of gibberellins promotes hypocotyl cell elongation in Arabidopsis thaliana under excess boron stress

2018 ◽  
Vol 45 (3) ◽  
pp. 305 ◽  
Author(s):  
Yongqi Pang ◽  
Jintong Li ◽  
Bishu Qi ◽  
Mi Tian ◽  
Lirong Sun ◽  
...  
Keyword(s):  
Arabidopsis Thaliana ◽  
Cell Elongation ◽  
Stem Elongation ◽  
Hypocotyl Elongation ◽  
Wild Type ◽  
Downstream Target ◽  
Della Proteins ◽  
High Concentrations ◽  
Tonoplast Aquaporin ◽  
Ga Biosynthesis

Aquaporins play essential roles in growth and development including stem elongation in plants. Tonoplast aquaporin AtTIP5;1 has been proposed to positively regulate hypocotyl elongation under high concentrations of boron (high-B) in Arabidopsis thaliana (L.) Heynh. However, the mechanism underlying this process remains unanswered. Here, we show that paclobatrazol, an inhibitor of GA biosynthesis, significantly suppressed the hypocotyl cell elongation of wild-type (WT) seedlings, and more strongly suppressed that of AtTIP5;1 overexpressors under high-B stress. Two AtTIP5;1 null mutants displayed arrested elongation of cells in the upper part of hypocotyls compared with the WT in the presence of high-B or GA3. Moreover, paclobatrazol treatment completely inhibited the increases in AtTIP5;1 transcripts induced by high-B, whereas GA3 application upregulated AtTIP5;1 expression in the WT. In addition, treatment with high-B remarkably elevated the expression levels of GA3ox1, GA20ox1 and GA20ox2 – key biosynthesis genes of GAs – in WT seedlings. The GA3 and GA4 content also increased in WT seedlings grown in MS medium containing high-B. Additionally, application of high-B failed to enhance AtTIP5;1 expression in the double mutant rga-24–gai-t6 of DELLA genes. Together, these results suggest that AtTIP5;1 is an essential downstream target of GAs. High-B induces the accumulation of GAs, which activates AtTIP5;1 through modulation of the DELLA proteins Repressor of ga1–3 and GA-insensitive, further promoting hypocotyl elongation in A. thaliana.

scholarly journals Arabidopsis Phototropins Participate in the Regulation of Dark-Induced Leaf Senescence

2021 ◽  
Vol 22 (4) ◽  
pp. 1836
Author(s):  
Aleksandra Eckstein ◽  
Joanna Grzyb ◽  
Paweł Hermanowicz ◽  
Piotr Zgłobicki ◽  
Justyna Łabuz ◽  
...  
Keyword(s):  
Arabidopsis Thaliana ◽  
Dominant Role ◽  
Photosynthetic Pigment ◽  
Photochemical Activity ◽  
Wild Type ◽  
Delayed Senescence ◽  
Photosynthetic Genes ◽  
Cellular Components ◽  
Senescence Associated Genes

Senescence is the final stage of plant development, affecting individual organs or the whole organism, and it can be induced by several environmental factors, including shading or darkness. Although inevitable, senescence is a complex and tightly regulated process, ensuring optimal remobilization of nutrients and cellular components from senescing organs. Photoreceptors such as phytochromes and cryptochromes are known to participate in the process of senescence, but the involvement of phototropins has not been studied to date. We investigated the role of these blue light photoreceptors in the senescence of individually darkened Arabidopsis thaliana leaves. We compared several physiological and molecular senescence markers in darkened leaves of wild-type plants and phototropin mutants (phot1, phot2, and phot1phot2). In general, all the symptoms of senescence (lower photochemical activity of photosystem II, photosynthetic pigment degradation, down-regulation of photosynthetic genes, and up-regulation of senescence-associated genes) were less pronounced in phot1phot2, as compared to the wild type, and some also in one of the single mutants, indicating delayed senescence. This points to different mechanisms of phototropin operation in the regulation of senescence-associated processes, either with both photoreceptors acting redundantly, or only one of them, phot1, playing a dominant role.

scholarly journals Golgi-localized TMN1/EMP12 functions in the deposition of rhamnogalacturonan II and I for cell growth in Arabidopsis

2021 ◽  
Author(s):  
Akihiko Hiroguchi ◽  
Shingo Sakamoto ◽  
Nobutaka Mitsuda ◽  
Kyoko Miwa
Keyword(s):  
Cell Growth ◽  
Cell Walls ◽  
Stem Elongation ◽  
Primary Root ◽  
Loss Of Function ◽  
Wild Type ◽  
Pectic Polysaccharides ◽  
Rhamnogalacturonan I ◽  
Rhamnogalacturonan Ii ◽  
Dependent Growth

Abstract Appropriate pectin deposition in cell walls is important for cell growth in plants. Rhamnogalacturonan II (RG-II) is a portion of pectic polysaccharides; its borate crosslinking is essential for maintenance of pectic networks. However, the overall process of RG-II synthesis is not fully understood. To identify a novel factor for RG-II deposition or dimerization in cell walls, we screened Arabidopsis mutants with altered boron (B)-dependent growth. The mutants exhibited alleviated disorders of primary root (PR) and stem elongation, and fertility under low B conditions, but reduced PR lengths under sufficient B conditions. Altered PR elongation was associated with cell elongation changes caused by loss of function in TMN1 (Transmembrane Nine 1) /EMP12, which encodes a Golgi-localized membrane protein of unknown function that is conserved among eukaryotes. Mutant leaf and root dry weights were lower than those of wild-type plants, regardless of B conditions. In cell walls, TMN1 mutations reduced contents of B, RG-II specific 2-keto-3-deoxy monosaccharides, and rhamnose largely derived from rhamnogalacturonan I (RG-I), suggesting reduced RG-II and RG-I. Together, our findings demonstrate that TMN1 is required for the deposition of RG-II and RG-I for cell growth and we also find that pectin levels modulate plant growth under low B conditions.

scholarly journals PUCHI Regulates Giant Cell Morphology During Root-Knot Nematode Infection in Arabidopsis thaliana

2021 ◽  
Vol 12 ◽  
Author(s):  
Reira Suzuki ◽  
Mizuki Yamada ◽  
Takumi Higaki ◽  
Mitsuhiro Aida ◽  
Minoru Kubo ◽  
...  
Keyword(s):  
Arabidopsis Thaliana ◽  
Cell Wall ◽  
Genetic Regulation ◽  
Three Dimensional ◽  
Acid Synthesis ◽  
Giant Cells ◽  
Loss Of Function ◽  
Wild Type ◽  
Root Knot Nematode ◽  
In The Wild

Parasitic root-knot nematodes transform the host’s vascular cells into permanent feeding giant cells (GCs) to withdraw nutrients from the host plants. GCs are multinucleated metabolically active cells with distinctive cell wall structures; however, the genetic regulation of GC formation is largely unknown. In this study, the functions of the Arabidopsis thaliana transcription factor PUCHI during GC development were investigated. PUCHI expression was shown to be induced in early developing galls, suggesting the importance of the PUCHI gene in gall formation. Despite the puchi mutant not differing significantly from the wild type in nematode invasion and reproduction rates, puchi GC cell walls appeared to be thicker and lobate when compared to the wild type, while the cell membrane sometimes formed invaginations. In three-dimensional (3D) reconstructions of puchi GCs, they appeared to be more irregularly shaped than those in the wild type, with noticeable cell-surface protrusions and folds. Interestingly, the loss-of-function mutant of 3-KETOACYL-COA SYNTHASE 1 showed GC morphology and cell wall defects similar to those of the puchi mutant, suggesting that PUCHI may regulate GC development via very long chain fatty acid synthesis.

scholarly journals The conserved PFT1 tandem repeat is crucial for proper flowering in Arabidopsis thaliana

2014 ◽  
Author(s):  
Pauline Rival ◽  
Maximilian O Press ◽  
Jacob Bale ◽  
Tanya Grancharova ◽  
Soledad F Undurraga ◽  
...  
Keyword(s):  
Arabidopsis Thaliana ◽  
Transgenic Plants ◽  
Tandem Repeat ◽  
Allelic Diversity ◽  
Selective Constraint ◽  
Dependent Manner ◽  
Functional Requirements ◽  
Loss Of Function ◽  
Wild Type ◽  
Genomic Regions

It is widely appreciated that short tandem repeat (STR) variation underlies substantial phenotypic variation in organisms. Some propose that the high mutation rates of STRs in functional genomic regions facilitate evolutionary adaptation. Despite their high mutation rate, some STRs show little to no variation in populations. One such STR occurs in the Arabidopsis thaliana gene PFT1 (MED25), where it encodes an interrupted polyglutamine tract. Though the PFT1 STR is large (~270 bp), and thus expected to be extremely variable, it shows only minuscule variation across A. thaliana strains. We hypothesized that the PFT1 STR is under selective constraint, due to previously undescribed roles in PFT1 function. We investigated this hypothesis using plants expressing transgenic PFT1 constructs with either an endogenous STR or with synthetic STRs of varying length. Transgenic plants carrying the endogenous PFT1 STR generally performed best across adult PFT1-dependent traits. In stark contrast, transgenic plants carrying a PFT1 transgene lacking the STR entirely phenocopied a pft1 loss-of-function mutant for flowering time phenotypes, and were generally hypomorphic for other traits, establishing the functional importance of this domain. Transgenic plants carrying various synthetic constructs occupied the phenotypic space between wild-type and pft1-loss-of-function mutants. By varying PFT1 STR length, we discovered that PFT1 can act as either an activator or repressor of flowering in a photoperiod-dependent manner. We conclude that the PFT1 STR is constrained to its approximate wild-type length by its various functional requirements. Our study implies that there is strong selection on STRs not only to generate allelic diversity, but also to maintain certain lengths pursuant to optimal molecular function.

scholarly journals DAO1 catalyzes temporal and tissue-specific oxidative inactivation of auxin in Arabidopsis thaliana

2016 ◽  
Vol 113 (39) ◽  
pp. 11010-11015 ◽  
Author(s):  
Jun Zhang ◽  
Jinshan Ella Lin ◽  
Chinchu Harris ◽  
Fernanda Campos Mastrotti Pereira ◽  
Fan Wu ◽  
...  
Keyword(s):  
Acetic Acid ◽  
Plant Growth ◽  
Expression Patterns ◽  
Loss Of Function ◽  
Wild Type ◽  
Iaa Oxidase ◽  
Oxidative Inactivation ◽  
Mutant Phenotypes

Tight homeostatic regulation of the phytohormone auxin [indole-3-acetic acid (IAA)] is essential to plant growth. Auxin biosynthetic pathways and the processes that inactivate auxin by conjugation to amino acids and sugars have been thoroughly characterized. However, the enzyme that catalyzes oxidation of IAA to its primary catabolite 2-oxindole-3-acetic acid (oxIAA) remains uncharacterized. Here, we show that DIOXYGENASE FOR AUXIN OXIDATION 1 (DAO1) catalyzes formation of oxIAA in vitro and in vivo and that this mechanism regulates auxin homeostasis and plant growth. Null dao1-1 mutants contain 95% less oxIAA compared with wild type, and complementation of dao1 restores wild-type oxIAA levels, indicating that DAO1 is the primary IAA oxidase in seedlings. Furthermore, dao1 loss of function plants have altered morphology, including larger cotyledons, increased lateral root density, delayed sepal opening, elongated pistils, and reduced fertility in the primary inflorescence stem. These phenotypes are tightly correlated with DAO1 spatiotemporal expression patterns as shown by DAO1pro:β-glucuronidase (GUS) activity and DAO1pro:YFP-DAO1 signals, and transformation with DAO1pro:YFP-DAO1 complemented the mutant phenotypes. The dominant dao1-2D mutant has increased oxIAA levels and decreased stature with shorter leaves and inflorescence stems, thus supporting DAO1 IAA oxidase function in vivo. A second isoform, DAO2, is very weakly expressed in seedling root apices. Together, these data confirm that IAA oxidation by DAO1 is the principal auxin catabolic process in Arabidopsis and that localized IAA oxidation plays a role in plant morphogenesis.

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