Expression Of Allene Oxide Synthase Research Articles

Trigalactosyldiacylglycerol 3 protein orthologs are required for basal disease resistance in Nicotiana benthamiana.

Phosphatidic acid plays an important role in Nicotiana benthamiana immune responses against phytopathogenic bacteria. We analyzed the contributions of endoplasmic reticulum-derived chloroplast phospholipids, including phosphatidic acid, to the resistance of N. benthamiana against Ralstonia solanacearum. Here, we focused on trigalactosyldiacylglycerol 3 (TGD3) protein as a candidate required for phosphatidic acid signaling. On the basis of Arabidopsis thaliana TGD3 sequences, we identified two putative TGD3 orthologs in the N. benthamiana genome, NbTGD3-1 and NbTGD3-2. To address the role of TGD3s in plant defense responses, we created double NbTGD3-silenced plants using virus-induced gene silencing. The NbTGD3-silenced plants showed a moderately reduced growth phenotype. Bacterial growth and the appearance of bacterial wilt disease were accelerated in NbTGD3-silenced plants, compared with control plants, challenged with R. solanacearum. The NbTGD3-silenced plants showed reduced both expression of allene oxide synthase that encoded jasmonic acid biosynthetic enzyme and NbPR-4, a marker gene for jasmonic acid signaling, after inoculation with R. solanacearum. Thus, NbTGD3-mediated endoplasmic reticulum-chloroplast lipid transport might be required for jasmonic acid signaling-mediated basal disease resistance in N. benthamiana.

Overexpression of Arabidopsis OPR3 in Hexaploid Wheat (Triticum aestivum L.) Alters Plant Development and Freezing Tolerance.

Jasmonates are plant hormones that are involved in the regulation of different aspects of plant life, wherein their functions and molecular mechanisms of action in wheat are still poorly studied. With the aim of gaining more insights into the role of jasmonic acid (JA) in wheat growth, development, and responses to environmental stresses, we have generated transgenic bread wheat plants overexpressing Arabidopsis 12-OXOPHYTODIENOATE REDUCTASE 3 (AtOPR3), one of the key genes of the JA biosynthesis pathway. Analysis of transgenic plants showed that AtOPR3 overexpression affects wheat development, including germination, growth, flowering time, senescence, and alters tolerance to environmental stresses. Transgenic wheat plants with high AtOPR3 expression levels have increased basal levels of JA, and up-regulated expression of ALLENE OXIDE SYNTHASE, a jasmonate biosynthesis pathway gene that is known to be regulated by a positive feedback loop that maintains and boosts JA levels. Transgenic wheat plants with high AtOPR3 expression levels are characterized by delayed germination, slower growth, late flowering and senescence, and improved tolerance to short-term freezing. The work demonstrates that genetic modification of the jasmonate pathway is a suitable tool for the modulation of developmental traits and stress responses in wheat.

Jasmonic acid is a downstream component in the modulation of somatic embryogenesis by Arabidopsis Class 2 phytoglobin.

Previous studies have shown that the beneficial effect of suppression of the Arabidopsis phytoglobin 2 gene, PGB2, on somatic embryogenesis occurs through the accumulation of nitric oxide (NO) within the embryogenic cells originating from the cultured explant. NO activates the expression of Allene oxide synthase (AOS) and Lipoxygenase 2 (LOX2), genes encoding two key enzymes of the jasmonic acid (JA) biosynthetic pathway, elevating JA content within the embryogenic tissue. The number of embryos in the single aos1-1 mutant and pgb2-aos1-1 double mutant declined, and was not rescued by increasing levels of NO stimulating embryogenesis in wild-type tissue. NO also influenced JA responses by up-regulating PLANT DEFENSIN 1 (PDF1) and JASMONATE-ZIM-PROTEIN (JAZ1), as well as down-regulating MYC2. The NO and JA modulation of MYC2 and JAZ1 controlled embryogenesis. Ectopic expression of JAZ1 or suppression of MYC2 promoted the formation of somatic embryos, while repression of JAZ1 and up-regulation of MYC2 reduced the embryogenic performance. Sustained expression of JAZ1 induced the transcription of several indole acetic acid (IAA) biosynthetic genes, resulting in higher IAA levels in the embryogenic cells. Collectively these data fit a model integrating JA in the PGB2 regulation of Arabidopsis embryogenesis. Suppression of PGB2 increases JA through NO. Elevated levels of JA repress MYC2 and induce JAZ1, favoring the accumulation of IAA in the explants and the subsequent production of somatic embryos.

Two closely related members of Arabidopsis 13‐lipoxygenases (13‐LOXs), LOX3 and LOX4, reveal distinct functions in response to plant‐parasitic nematode infection

The responses of two closely related members of Arabidopsis 13-lipoxygenases (13-LOXs), LOX3 and LOX4, to infection by the sedentary nematodes root-knot nematode (Meloidogyne javanica) and cyst nematode (Heterodera schachtii) were analysed in transgenic Arabidopsis seedlings. The tissue localization of LOX3 and LOX4 gene expression using β-glucuronidase (GUS) reporter gene constructs showed local induction of LOX3 expression when second-stage juveniles reached the vascular bundle and during the early stages of plant-nematode interaction through gall and syncytia formation. Thin sections of nematode-infested knots indicated LOX3 expression in mature giant cells, and high expression in neighbouring cells and those surrounding the female body. LOX4 promoter was also activated by nematode infection, although the GUS signal weakened as infection and disease progressed. Homozygous insertion mutants lacking LOX3 were less susceptible than wild-type plants to root-knot nematode infection, as reflected by a decrease in female counts. Conversely, deficiency in LOX4 function led to a marked increase in females and egg mass number and in the female to male ratio of M. javanica and H. schachtii, respectively. The susceptibility of lox4 mutants was accompanied by increased expression of allene oxide synthase, allene oxide cyclase and ethylene-responsive transcription factor 4, and the accumulation of jasmonic acid, measured in the roots of lox4 mutants. This response was not found in lox3 mutants. Taken together, our results reveal that LOX4 and LOX3 interfere differentially with distinct metabolic and signalling pathways, and that LOX4 plays a major role in controlling plant defence against nematode infection.

Differential expression of allene oxide synthase (AOS), and jasmonate relationship with ethylene biosynthesis in seed and mesocarp of developing peach fruit

To better understand the physiological role of jasmonates (JAs) during fruit development and ripening, and relationships with ethylene, the time course of transcript accumulation of the key enzyme in JA biosynthesis, allene oxide synthase (AOS), was analyzed during S1–S4 fruit developmental stages, until ripening, in the mesocarp and seed of peach ( Prunus persica var. laevis Gray), cv. ‘Stark Red Gold’. In the mesocarp, transcript amounts of PpAOS1 were the highest in S1–S2, then decreased to almost undetectable levels at the transition S3/S4, to rise slightly at harvest. In parallel, JA concentration was measured in the mesocarp during development; maximum jasmonic acid accumulation was observed in S1, the lowest levels in S2 and S3 and a new rise in S4. In the seed, an AOS1 transcript was detected whose accumulation differed in the embryo as compared to cotyledons during seed growth. The reciprocal JA/ethylene relationships were investigated in fruit at different physiological stages by exogenously supplying methyl jasmonate (MJ). The effects of treatments with 0.88 mM MJ on transcript levels of PpAOS1 and ethylene biosynthetic genes PpACS1, PpACO1 and PpACO2 were evaluated in the mesocarp and seed of detached nectarines at an early (S1) and late (S4) developmental stage. Results show that JA biosynthesis appears to be developmentally regulated in seed and mesocarp of nectarine fruit and that the response to MJ depends upon the fruit physiological stage.

Expression of allene oxide synthase and allene oxide cyclase in the interactions between pea and fungal pathogens

Allene oxide synthase (AOS) and allene oxide cyclase (AOC) are enzymes in the octadecanoid pathway that lead to jasmonate (jasmonic acid and methyljasmonic acid) biosynthesis. Pea AOS and AOC cDNAs were isolated by the reverse transcriptase-mediated polymerase chain reaction method with degenerated primers. Treatment of pea epicotyls with suppressor alone or suppressor plus elicitor induced an increase in AOS mRNA 5 h after treatment. AOS was also induced by a virulent pathogen, Mycosphaerella pinodes, 12 h after inoculation but not by a nonpathogen, Ascochyta rabiei, or the fungal elicitor. On the other hand, AOC mRNA accumulation was not significantly affected by treatment with elicitor or suppressor or by inoculations with M. pinodes or A. rabiei. Previously, we observed that gene expression of oxophytodienoic acid reductase (OPR), an enzyme in the octadecanoid pathway, was induced in the pea by suppressor treatment and inoculation of a compatible pathogen. Thus, it is likely that, in addition to OPR, activation of AOS gene expression contributes to the establishment of compatibility.

Cytochrome P450-dependent metabolism of oxylipins in tomato. Cloning and expression of allene oxide synthase and fatty acid hydroperoxide lyase.

Allene oxide synthase (AOS) and fatty acid hydroperoxide lyase (HPL) are plant-specific cytochrome P450s that commit fatty acid hydroperoxides to different branches of oxylipin metabolism. Here we report the cloning and characterization of AOS (LeAOS) and HPL (LeHPL) cDNAs from tomato (Lycopersicon esculentum). Functional expression of the cDNAs in Escherichia coli showed that LeAOS and LeHPL encode enzymes that metabolize 13- but not 9-hydroperoxide derivatives of C(18) fatty acids. LeAOS was active against both 13S-hydroperoxy-9(Z),11(E),15(Z)-octadecatrienoic acid (13-HPOT) and 13S-hydroperoxy-9(Z),11(E)-octadecadienoic acid, whereas LeHPL showed a strong preference for 13-HPOT. These results suggest a role for LeAOS and LeHPL in the metabolism of 13-HPOT to jasmonic acid and hexenal/traumatin, respectively. LeAOS expression was detected in all organs of the plant. In contrast, LeHPL expression was predominant in leaves and flowers. Damage inflicted to leaves by chewing insect larvae led to an increase in the local and systemic expression of both genes, with LeAOS showing the strongest induction. Wound-induced expression of LeAOS also occurred in the def-1 mutant that is deficient in octadecanoid-based signaling of defensive proteinase inhibitor genes. These results demonstrate that tomato uses genetically distinct signaling pathways for the regulation of different classes of wound responsive genes.