Regulation of Expression of the prb-1b / ACC Deaminase Gene by UV-8 in Transgenic Tomatoes

Transgenic tomato plants with 1-aminocyclopropane-1-carboxylic acid (ACC) deaminase gene from Enterobacter cloacae UW4 under the control of a pathogenesis-related promoter (prb-1b) from tobacco were challenged by abiotic stresses to determine the expression patterns ofthe transgene. No ACC deaminase RNA or protein was detected by RT-PCR and in western blots prepared from leaf proteins of transgenic plants after wounding or treatment with α-amino butyric acid, xylanase, ethephon, salicylic acid, jasmonic acid, ethylene, or ethylene plus jasmonic acid. However, expression of the ACC deaminase transgene was observed in leaves and roots oftransformed tomato lines exposed to UV light. The UV response required a minimum of 48 h of exposure and was specific to UV-8 light.     

Molecular hydrogen can take part in phytohormone signal pathways in wild rice

Molecular hydrogen (H₂) could be a novel signal in phytohormone signaling pathways in response to biotic and abiotic stresses. Here, we employed two wild rice species (Oryza rufipogon Griff. and O. minuta J. Presl) to test this hypothesis using hydrogen-rich water (HW). The expression differences of phytohormone and hydrogenase genes between conventional rice (Oryza sativa L,) and wild rice were determined by real-time quantitative polymerase chain reaction, and the effects of HW on gene expression of wild rice were detected during three growth stages. Expression of hydrogenase genes, synthesis genes, and receptor genes of salicylic acid (SA), jasmonic acid (JA), and ethylene (ET) signalling pathways was higher in six wild rice types than in conventional rice. Hydrogen-rich water up-regulated expression of two hydrogenase genes, SA, JA, and ET receptor genes and synthesis genes in the seedling stage of wild rice. But this positive regulation by HW was less significant in the vegetative and reproductive stages.     

Acylated monogalactosyl diacylglycerol: prevalence in the plant kingdom and identification of an enzyme catalyzing galactolipid head group acylation in Arabidopsis thaliana

The lipid phase of the thylakoid membrane is mainly composed of the galactolipids mono‐ and digalactosyl diacylglycerol (MGDG and DGDG, respectively). It has been known since the late 1960s that MGDG can be acylated with a third fatty acid to the galactose head group (acyl‐MGDG) in plant leaf homogenates. In certain brassicaceous plants like Arabidopsis thaliana, the acyl‐MGDG frequently incorporates oxidized fatty acids in the form of the jasmonic acid precursor 12‐oxo‐phytodienoic acid (OPDA). In the present study we further investigated the distribution of acylated and OPDA‐containing galactolipids in the plant kingdom. While acyl‐MGDG was found to be ubiquitous in green tissue of plants ranging from non‐vascular plants to angiosperms, OPDA‐containing galactolipids were only present in plants from a few genera. A candidate protein responsible for the acyl transfer was identified in Avena sativa (oat) leaf tissue using biochemical fractionation and proteomics. Knockout of the orthologous gene in A. thaliana resulted in an almost total elimination of the ability to form both non‐oxidized and OPDA‐containing acyl‐MGDG. In addition, heterologous expression of the A. thaliana gene in E. coli demonstrated that the protein catalyzed acylation of MGDG. We thus demonstrate that a phylogenetically conserved enzyme is responsible for the accumulation of acyl‐MGDG in A. thaliana. The activity of this enzyme in vivo is strongly enhanced by freezing damage and the hypersensitive response.     

Colonization of the <Emphasis Type="Italic">Arabidopsis</Emphasis> rhizosphere by fluorescent <Emphasis Type="Italic">Pseudomonas</Emphasis> spp. activates a root-specific,ethylene-responsive <Emphasis Type="Italic">PR-5</Emphasis> gene in the vascular bundle

Plants of which the roots are colonized by selected strains of non-pathogenic, fluorescent Pseudomonas spp. develop an enhanced defensive capacity against a broad spectrum of foliar pathogens. In Arabidopsis thaliana, this rhizobacteria-induced systemic resistance (ISR) functions independently of salicylic acid but requires responsiveness to jasmonic acid and ethylene. In contrast to pathogen-induced systemic acquired resistance (SAR), ISR is not associated with systemic changes in the expression of genes encoding pathogenesis-related (PR) proteins. To identify genes that are specifically expressed in response to colonization of the roots by ISR-inducing Pseudomonas fluorescens WCS417r bacteria, we screened a collection of Arabidopsis enhancer trap and gene trap lines containing a transposable element of the Ac/Ds system and the GUS reporter gene. We identified an enhancer trap line (WET121) that specifically showed GUS activity in the root vascular bundle upon colonization of the roots by WCS417r. Fluorescent Pseudomonas spp. strains P. fluorescens WCS374r and P. putida WCS358r triggered a similar expression pattern, whereas ISR-non-inducing Escherichia coli bacteria did not. Exogenous application of the ethylene precursor 1-aminocyclopropane-1-carboxylate (ACC) mimicked the rhizobacteria-induced GUS expression pattern in the root vascular bundle, whereas methyl jasmonic acid and salicylic acid did not, indicating that the Ds element in WET121 is inserted in the vicinity of an ethylene-responsive gene. Analysis of the expression of the genes in the close vicinity of the Ds element revealed AtTLP1 as the gene responsible for the in cis activation of the GUS reporter gene in the root vascular bundle. AtTLP1 encodes a thaumatin-like protein that belongs to the PR-5 family of PR proteins, some of which possess antimicrobial properties. AtTLP1 knockout mutant plants showed normal levels of WCS417r-mediated ISR against the bacterial leaf pathogen Pseudomonas syringae pv. tomato DC3000, suggesting that expression of AtTLP1 in the roots is not required for systemic expression of ISR in the leaves. Together, these results indicate that induction of AtTLP1 is a local response of Arabidopsis roots to colonization by non-pathogenic fluorescent Pseudomonas spp. and is unlikely to play a role in systemic resistance.     

Hyaluronic acid of Streptococcus sp. as a potent elicitor for induction of systemic resistance against plant diseases

The potential of hyaluronic acid (HA) in inducing systemic resistance to cucumber, tomato and pepper was tested in planta. In the study, HA was found to be a potent agent for suppressing disease caused by Cucumber Mosaic Virus (CMV) (in pepper), Pseudomonas syringae pv. tomato (tomato speck disease), Xanthomonas axonopodis pv. vesicatoria (tomato spot disease), Pseudomonas syringae pv. lachrymans (cucumber angular leaf spot), and Colletotrichum orbiculare (cucumber anthracnose). Disease control was obtained with spraying, injection and drenching of plants with HA. HA did not exhibit direct antimicrobial action against the pathogens tested. Studies carried out in transgenic tobacco indicated that defense genes PR 1a and PDF 1.2 were activated upon treatment with HA, demonstrating salicylic acid (SA) and jasmonic acid (JA) pathways getting activated during defense. Further work is warranted to evaluate the use of HA-mediated disease suppression in crop plants.     

Rice <Emphasis Type="Italic">SERK1</Emphasis> gene positively regulates somatic embryogenesis of cultured cell and host defense response against fungal infection

Here we report on the isolation and characterization of a somatic embryogenesis receptor-like kinase (OsSERK1) gene in rice (Oryza sativa). The OsSERK1 gene belongs to a small subfamily of receptor-like kinase genes in rice and shares a highly conserved gene structure and extensive sequence homology with previously reported plant SERK genes. Though it has a basal level of expression in various rice organs/tissues, as high expression level was detected in rice callus during somatic embryogenesis. Suppression of OsSERK1 expression in transgenic calli by RNA interference resulted in a significant reduction of shoot regeneration rate (from 72% to 14% in the japonica rice Zhonghua11). Overexpression of OsSERK1, however, increased the shoot regeneration rate (from 72% to 86%). Interestingly, OsSERK1 is significantly activated by the rice blast fungus, particularly during the incompatible interaction, and is associated with host cell death in Sekigushi lesion mimic mutants. This gene is also inducible by defense signaling molecules such as salicylic acid, jasmonic acid, and abscisic acid. Furthermore, constitutive overexpression of OsSERK1 in two rice cultivars led to an increase in host resistance to the blast fungus. Our data suggest that OsSERK1 may partially mediate defense signal transduction in addition to its basic role in somatic embryogenesis.     

Growth-promoting bacteria and natural regulators mitigate salt toxicity and improve rapeseed plant performance

Salinity is a major environmental stress that limits plant production and portraits a critical challenge to food security in the world. In this research, the impacts of plant growth–promoting bacteria (Pseudomonas RS-198 and Azospirillum brasilense RS-SP7) and foliar application of plant hormones (salicylic acid 1 mM and jasmonic acid 0.5 mM) on alleviating the harmful effects of salt stress in rapeseed plants (Brassica napus cv. okapi) were examined under greenhouse condition. Salt stress diminished rapeseed biomass, leaf area, water content, nitrogen, phosphorus, potassium, calcium, magnesium, and chlorophyll content, while it increased sodium content, endogenous salicylic and jasmonic acids, osmolyte production, H₂O₂ and O₂^•− generations, TBARS content, and antioxidant enzyme activities. Plant growth, nutrient content, leaf expansion, osmolyte production, and antioxidant enzyme activities were increased, but oxidative and osmotic stress indicators were decreased by bacteria inoculation + salicylic acid under salt stress. Antioxidant enzyme activities were amplified by jasmonic acid treatments under salt stress, although rapeseed growth was not generally affected by jasmonic acid. Bacterial + hormonal treatments were superior to individual treatments in reducing detrimental effects of salt stress. The best treatment in rectifying rapeseed growth under salt stress was combination of Pseudomonas and salicylic acid. This combination attenuated destructive salinity properties and subsequently amended rapeseed growth via enhancing endogenous salicylic acid content and some essential nutrients such as potassium, phosphorus, and magnesium.

     

Auxin,biosynthesis of ethylene and sex expression in cucumber (Cucumis sativus)

Ethylene production, level of 1-aminocyclopropane-1-carboxylic acid (ACC) and activity of the ethylene forming enzyme (EFE) were higher in apices of gynoecious cucumber (Cucumis sativus cv. Alma) as compared to monoecious cucumber (C. sativus cv. Elem). Application of indole-3-acetic acid (IAA) enhanced ethylene and ACC production in both cultivars. The stimulatory effect of IAA was more pronounced in gynoecious apices. Induction of ethylene production and accumulation of ACC resulting from treatment with IAA were effectively blocked by aminoethoxyvinylglycine (AVG). Content of endogenous IAA, measured by an enzyme immunoassay, was lower in gynoecious cucumber as compared to monoecious one. Treatment of gynoecious plants with the antiauxins -(p-chlorophenoxy)isobutyric acid (PCIB) and -naphthaleneacetic acid (-NAA) did not inhibit female sex expression.It appears that although exogenous IAA enhances ACC and ethylene production, endogenous IAA might not have a major role in the control of sex expression in cucumber of the Beit-Alfa type.Prof. Rudich passed away in May 1986.     

Inverse correlation between jasmonic acid and salicylic acid during early wound response in rice

This study presents a kinetic analysis of the response to wounding in rice plants. In particular, jasmonic acid, salicylic acid, and lipoxygenase activity were measured in leaves of wounded rice plants during the early tillering phase. The results show that endogenous jasmonic acid transiently increases to a maximum 30 min after wounding (jasmonic acid burst) and lipoxygenase activity increases after the jasmonic acid burst, but not after the second smaller peak of endogenous jasmonic acid 23 h after wounding. In contrast, endogenous salicylic acid decreases during the jasmonic acid burst, such that the kinetic profiles of jasmonic acid and salicylic acid are inversely correlated during the early response to wounding. It is proposed here that the increase in endogenous jasmonic acid and the decrease in endogenous salicylic acid may contribute for establishing the efficient negative cross-talk between jasmonic acid and salicylic acid signaling pathways during the early response to wounding in rice.     

Stimulatory effect of ethephon,ACC, gibberellin A3 and A4+7 on germination of methyl jasmonate inhibited Amaranthus caudatus L. seeds

Methyl jasmonate (JA-Me) inhibited or retarded germination of Amaranthus caudatus seeds in darkness at 24°C, Ethephon, ACC and gibberellins (GA₃ or GA₄₊₇) partially or completely reversed this inhibition depending on the concentration of JA-Me applied. Both ethephon and the gibberellins were more effective than ACC. Both GA₃ and GA₄₊₇ enhanced the stimulatory effect of ethephon or ACC on germination of seeds inhibited by JA-Me.Abbreviations ACC 1-aminocyclopropane-1-carboxylic acid - JA jasmonic acid - JA-Me methyl jasmonate     

Induction of tomato stress protein mRNAs by ethephon, 2,6-dichloroisonicotinic acid and salicylate

To study the possible involvement of plant hormones in the synthesis of stress proteins in tomato upon inoculation with Cladosporium fulvum, we investigated the induction of mRNAs encoding PR proteins and ethylene biosynthesis enzymes by ethephon, 2,6-dichloroisonicotinic acid (INA) and salicylic acid (SA) by northern blot analysis. Ethephon slightly induced some but not all mRNAs encoding intra- and extracellular PR proteins. INA induced all PR protein mRNAs analysed, except for intracellular chitinase and extracellular PR-4. SA induced all PR protein mRNAs analyzed, except for intracellular chitinase and osmotin. None of the inducers affected the expression of ACC synthase mRNA, whereas all three induced ethylene-forming enzyme (EFE) mRNA.Abbreviations ACC 1-aminocyclopropane-1-carboxylic acid - EFE ethylene-forming enzyme - HR hypersensitive response - INA 2,6-dichloroisonicotinic acid - PR pathogenesis-related - SA salicylic acid - SAR systemic acquired resistance     

Parasitism by Cuscuta pentagona sequentially induces JA and SA defence pathways in tomato

While plant responses to herbivores and pathogens are well characterized, responses to attack by other plants remain largely unexplored. We measured phytohormones and C₁₈ fatty acids in tomato attacked by the parasitic plant Cuscuta pentagona, and used transgenic and mutant plants to explore the roles of the defence‐related phytohormones salicylic acid (SA) and jasmonic acid (JA). Parasite attachment to 10‐day‐old tomato plants elicited few biochemical changes, but a second attachment 10 d later elicited a 60‐fold increase in JA, a 30‐fold increase in SA and a hypersensitive‐like response (HLR). Host age also influenced the response: neither Cuscuta seedlings nor established vines elicited a HLR in 10‐day‐old hosts, but both did in 20‐day‐old hosts. Parasites grew larger on hosts deficient in SA (NahG) or insensitive to JA [jasmonic acid‐insensitive1 (jai1) ], suggesting that both phytohormones mediate effective defences. Moreover, amounts of JA peaked 12 h before SA, indicating that defences may be coordinated via sequential induction of these hormones. Parasitism also induced increases in free linolenic and linoleic acids and abscisic acid. These findings provide the first documentation of plant hormonal signalling induced by a parasitic plant and show that tomato responses to C. pentagona display characteristics similar to both herbivore‐ and pathogen‐induced responses.     

Signalling molecules and blast pathogen attack activates rice OsPR1a and OsPR1b genes: A model illustrating components participating during defence/stress response

Recently the rice (Oryza sativa L.) OsPR1a and OsPR1b genes were primarily characterized against jasmonic acid, ethylene and protein phosphatase 2A inhibitors. The dicot PR1 are recognized as reliable marker genes in defence/stress responses, and we also propose OsPR1 as marker genes in rice, a model monocot crop genus. Therefore, to gain further insight into the expression/regulation of OsPR1 genes, we characterized their activation against signalling molecules such as salicylic acid (SA), abscisic acid (ABA) and hydrogen peroxide (H₂O₂), and the blast pathogen Magnaporthe grisea. Here, we report that SA and H₂O₂ strongly induced the mRNA level of both OsPR1 genes, whereas ABA was found to be moderately effective. These inductions were specific in nature and required a de novo synthesized protein factor. A potential interaction amongst the signalling molecules in modulating the expression of OsPR1 genes was observed. Moreover, a specific induction of OsPR1 expression in an incompatible versus compatible host-pathogen interaction was also found. Finally, based on our present and previous results, a model of OsPR1 expression/regulation has been proposed, which reveals their essential role in defence/stress responses in rice and use as potent gene markers.