Acid Biosynthesis In Plants Research Articles

Glyphosate residues in soil and phosphate fertilizer affect foliar endophytic microbial community composition and phytohormone levels in potato

Glyphosate, the active ingredient of glyphosate-based herbicide (GBH) controls the growth of weeds by inhibiting shikimate pathway, thereby interrupting amino acid biosynthesis in plants. However, several microbes have shikimate pathway and the action of glyphosate on these non-target organisms are ignored. Along with other agrochemicals such as phosphate fertilizers, the action of GBH is further complicated, often varying their mode of action depending on soil type or plant species. To address the impact of GBH and phosphate fertilizer, we simulated agricultural application of GBH and phosphate fertilizer in a field study, investigating the composition of endophytic microbial communities and correlation of phytohormone concentrations with the microbial diversity of potato (Solanum tuberosum). In leaves, glyphosate residues in soil from GBH treatment alone and in combination with phosphate significantly shifted bacterial community whereas phosphate alone and in combination with glyphosate significantly altered the composition of fungal community. There were no significant changes in microbial communities in roots and tubers. Plants treated with GBH showed higher ratios of potentially glyphosate-resistant bacteria, with Xanthomonadaceae and Moraxallaceae being more abundant. Additionally, phytohormone concentrations showed various correlations with bacterial and fungal diversity in different treatments. The study highlights the impact of GBH residues in soil, particularly in combination with phosphate fertilizers on the composition of plant-associated microbial communities. Together with changes in phytohormone concentrations, plant health may be affected. Moreover, future studies could provide insights to whether these agrochemicals influence plant microbiome, leading to changes in phytohormones or vice-versa.

Expression of malic enzyme reveals subcellular carbon partitioning for storage reserve production in soybeans.

Central metabolism produces amino and fatty acids for protein and lipids that establish seed value. Biosynthesis of storage reserves occurs in multiple organelles that exchange central intermediates including two essential metabolites, malate, and pyruvate that are linked by malic enzyme. Malic enzyme can be active in multiple subcellular compartments, partitioning carbon and reducing equivalents for anabolic and catabolic requirements. Prior studies based on isotopic labeling and steady-state metabolic flux analyses indicated malic enzyme provides carbon for fatty acid biosynthesis in plants, though genetic evidence confirming this role is lacking. We hypothesized that increasing malic enzyme flux would alter carbon partitioning and result in increased lipid levels in soybeans. Homozygous transgenic soybean plants expressing Arabidopsis malic enzyme alleles, targeting the translational products to plastid or outside the plastid during seed development, were verified by transcript and enzyme activity analyses, organelle proteomics, and transient expression assays. Protein, oil, central metabolites, cofactors, and acyl-acyl carrier protein (ACPs) levels were quantified overdevelopment. Amino and fatty acid levels were altered resulting in an increase in lipids by 0.5-2% of seed biomass (i.e. 2-9% change in oil). Subcellular targeting of a single gene product in central metabolism impacts carbon and reducing equivalent partitioning for seed storage reserves in soybeans.

Genetic and biochemical strategies for regulation of L-ascorbic acid biosynthesis in plants through the L-galactose pathway.

Vitamin C (L-ascorbic acid, AsA) is an essential compound with pleiotropic functions in many organisms. Since its isolation in the last century, AsA has attracted the attention of the scientific community, allowing the discovery of the L-galactose pathway, which is the main pathway for AsA biosynthesis in plants. Thus, the aim of this review is to analyze the genetic and biochemical strategies employed by plant cells for regulating AsA biosynthesis through the L-galactose pathway. In this pathway, participates eight enzymes encoded by the genes PMI, PMM, GMP, GME, GGP, GPP, GDH, and GLDH. All these genes and their encoded enzymes have been well characterized, demonstrating their participation in AsA biosynthesis. Also, have described some genetic and biochemical strategies that allow its regulation. The genetic strategy includes regulation at transcriptional and post-transcriptional levels. In the first one, it was demonstrated that the expression levels of the genes correlate directly with AsA content in the tissues/organs of the plants. Also, it was proved that these genes are light-induced because they have light-responsive promoter motifs (e.g., ATC, I-box, GT1 motif, etc.). In addition, were identified some transcription factors that function as activators (e.g., SlICE1, AtERF98, SlHZ24, etc.) or inactivators (e.g., SlL1L4, ABI4, SlNYYA10) regulate the transcription of these genes. In the second one, it was proved that some genes have alternative splicing events and could be a mechanism to control AsA biosynthesis. Also, it was demonstrated that a conserved cis-acting upstream open reading frame (5'-uORF) located in the 5'-untranslated region of the GGP gene induces its post-transcriptional repression. Among the biochemical strategies discovered is the control of the enzyme levels (usually by decreasing their quantities), control of the enzyme catalytic activity (by increasing or decreasing its activity), feedback inhibition of some enzymes (GME and GGP), subcellular compartmentation of AsA, the metabolon assembly of the enzymes, and control of AsA biosynthesis by electron flow. Together, the construction of this basic knowledge has been establishing the foundations for generating genetically improved varieties of fruits and vegetables enriched with AsA, commonly used in animal and human feed.

Functional analysis of the dehydratase domains of the PUFA synthase from Emiliania huxleyi in Escherichia coli and Arabidopsis thaliana

BackgroundPolyunsaturated fatty acid (PUFA) synthase is a multi-domain mega-enzyme that effectively synthesizes a series of PUFAs in marine microorganisms. The dehydratase (DH) domain of a PUFA synthase plays a crucial role in double bond positioning in fatty acids. Sequencing results of the coccolithophore Emiliania huxleyi (E. huxleyi, Eh) indicated that this species contains a PUFA synthase with multiple DH domains. Therefore, the current study, sought to define the functions of these DH domains (EhDHs), by cloning and overexpressing the genes encoding FabA-like EhDHs in Escherichia coli (E. coli) and Arabidopsis thaliana (A. thaliana).ResultsA complementation test showed that the two FabA-like DH domains could restore DH function in a temperature-sensitive (Ts) mutant. Meanwhile, overexpression of FabA-like EhDH1 and EhDH2 domains increased the production of unsaturated fatty acids (UFAs) in recombinant E. coli by 43.5–32.9%, respectively. Site-directed mutagenesis analysis confirmed the authenticity of active-site residues in these domains. Moreover, the expression of tandem EhDH1-DH2 in A. thaliana altered the fatty acids content, seed weight, and germination rate.ConclusionsThe two FabA-like DH domains in the E. huxleyi PUFA synthase function as 3-hydroxyacyl-acyl carrier protein dehydratase in E. coli. The expression of these domains in E. coli and A. thaliana can alter the fatty acid profile in E. coli and increase the seed lipid content and germination rate in A. thaliana. Hence, introduction of DH domains controlling the dehydration process of fatty acid biosynthesis in plants might offer a new strategy to increase oil production in oilseed plants.

The influence of exogenous gibberellic acid (GA3) and 24-epibrassinolide (24-EpiBL) on seed germination and the expression of genes involved in GA and BR synthesis/signalling in pepper (Capsicum annuum L.)

Gibberellins (GAs) and brassinosteroids (BRs) are the plant hormones involved in various physiological processes including seed germination. In this study, the effects of exogenous gibberellic acid (GA3) and 24-epibrassinolide (24-EpiBL) treatments on the expression of key genes involved in GA and BR syntheis/signalling during seed germination were investigated in pepper (Capsicum annuum L).
 The expressions of BES1 and BRI1 involved in BR synthesis/signalling pathway as well as GA3OX1 and GA20OX1 associated with gibberellic acid biosynthesis in plants were determined. Exogenous GA3 treatments increased BES1 expression and the highest increase was determined with 10⁻⁸ M BR + 100 µM GA3 (P<0.05). On the contrary, the expression of BRI1 gene was significantly decreased by 10-8 M BR + 100 µM GA3 (P<0.05). The expression of GA3OX1 gene was induced with BR and GA3 treatments (P<0.05). GA20OX1 gene expression was generally higher compared to the expression of GA3OX1 and significantly increased by the GA3 treatments. Our findings are expected to bring an insight to the influence of BRs during seed germination together with the expression of associated genes.

Genetic Variation for Seed Oil Biosynthesis in Soybean

Soybean is an important oilseed crop which supplies both industrial processes and animal feed as well as having a number of food uses. The high yields of oil from soybean seed, along with large planting acreage, significant production infrastructure, and its capacity to fix nitrogen, make it the second most valuable crop in the USA and fourth in the world. In this review, we discuss the current understanding of the genetics of fatty acid biosynthesis in plants, based on soybean and other species. We describe recent biotechnological approaches to improve or alter oil content and oil profile in soybean. We provide a list of the presently understood genetic variants, both natural and induced by mutagenesis, in the known soybean fatty acid biosynthesis genes, including four mutations not previously described. We discuss the implications of the increasing amount of genomic data available for composition improvement, using the known fatty acid biosynthesis genes as a case study, and describe potentially new oil gene mutants based on genome sequence data.

Knockdown of the plastid-encoded acetyl-CoA carboxylase gene uncovers functions in metabolism and development.

De novo fatty acid biosynthesis in plants relies on a prokaryotic-type acetyl-CoA carboxylase (ACCase) that resides in the plastid compartment. The enzyme is composed of four subunits, one of which is encoded in the plastid genome, whereas the other three subunits are encoded by nuclear genes. The plastid gene (accD) encodes the β-carboxyltransferase subunit of ACCase and is essential for cell viability. To facilitate the functional analysis of accD, we pursued a transplastomic knockdown strategy in tobacco (Nicotiana tabacum). By introducing point mutations into the translational start codon of accD, we obtained stable transplastomic lines with altered ACCase activity. Replacement of the standard initiator codon AUG with UUG strongly reduced AccD expression, whereas replacement with GUG had no detectable effects. AccD knockdown mutants displayed reduced ACCase activity, which resulted in changes in the levels of many but not all species of cellular lipids. Limiting fatty acid availability caused a wide range of macroscopic, microscopic, and biochemical phenotypes, including impaired chloroplast division, reduced seed set, and altered storage metabolism. Finally, while the mutants displayed reduced growth under photoautotrophic conditions, they showed exaggerated growth under heterotrophic conditions, thus uncovering an unexpected antagonistic role of AccD activity in autotrophic and heterotrophic growth.

Functionality and DNA-damage properties of blood cells in lactating cows exposed to glyphosate contaminated feed at different feed energy levels

ABSTRACTGlyphosate (GL) inhibits the aromatic amino acid biosynthesis in plants and is worldwide the most used non-selective herbicide. Less is known about in vivo effects of GL contaminated feedstuffs on the health of dairy cows. The aim of this study was to examine the effects of GL residues in feed at different concentrate feed proportions (CFP) on haematology, immunological and redox parameters and on DNA-damage of blood cells in lactating dairy cows. During a 16-trial, 61 German Holstein cows (207 ± 49 d in milk; mean ± SD) were fed the same ration in week 0. Afterwards, they were assigned to either a group receiving a GL contaminated or a group receiving an uncontaminated total mixed ration (CON). Each group was subdivided into a “low concentrate” group (LC) and a “high concentrate” group (HC) with an energy content of 6.63 MJ NEL and 7.18 MJ NEL/kg dry matter, respectively. The diets were offered for ad libitum consumption. Blood samples were taken at weeks 0, 4, 8, 12 and 16. All blood samples were analysed for white and red blood cell counts. T-cell subpopulations, oxidative burst capability of leukocytes, apoptosis rate, phagocytic activity, activities of superoxide dismutase and glutathione peroxidase, the total non-enzymatic antioxidative capacity, viability and stimulation capacity of peripheral blood mononuclear cells and micronucleus- and comet assay on bovine leukocytes were measured only in week 16. The average individual GL intake of groups CON, GLLC and GLHC was 1.2 µg, 112.6 µg and 132.8 µg per kg body weight and day, respectively. GL contamination did not affect any of the tested parameters whereas CFP and time-influenced leukocytes, granulocytes, red blood cells, haemoglobin, haematocrit, mean corpuscular volume, mean corpuscular haemoglobin and CD4+ T-cells in an interactive manner characterised by a time-dependent increase in HC groups. It can be concluded that GL and GL in combination with different CFP showed no influence on any of the tested endpoints, whereas CFP and time influenced most parameters in an interactive manner.

The impact and toxicity of glyphosate and glyphosate-based herbicides on health and immunity

Glyphosate, or N-phosphomethyl(glycine), is an organophosphorus compound and a competitive inhibitor of the shikimate pathway that allows aromatic amino acid biosynthesis in plants and microorganisms. Its utilization in broad-spectrum herbicides, such as RoundUp®, has continued to increase since 1974; glyphosate, as well as its primary metabolite aminomethylphosphonic acid, is measured in soils, water, plants, animals and food. In humans, glyphosate is detected in blood and urine, especially in exposed workers, and is excreted within a few days. It has long been regarded as harmless in animals, but growing literature has reported health risks associated with glyphosate and glyphosate-based herbicides. In 2017, the International Agency for Research on Cancer (IARC) classified glyphosate as “probably carcinogenic” in humans. However, other national agencies did not tighten their glyphosate restrictions and even prolonged authorizations of its use. There are also discrepancies between countries’ authorized levels, demonstrating an absence of a clear consensus on glyphosate to date. This review details the effects of glyphosate and glyphosate-based herbicides on fish and mammal health, focusing on the immune system. Increasing evidence shows that glyphosate and glyphosate-based herbicides exhibit cytotoxic and genotoxic effects, increase oxidative stress, disrupt the estrogen pathway, impair some cerebral functions, and allegedly correlate with some cancers. Glyphosate effects on the immune system appear to alter the complement cascade, phagocytic function, and lymphocyte responses, and increase the production of pro-inflammatory cytokines in fish. In mammals, including humans, glyphosate mainly has cytotoxic and genotoxic effects, causes inflammation, and affects lymphocyte functions and the interactions between microorganisms and the immune system. Importantly, even as many outcomes are still being debated, evidence points to a need for more studies to better decipher the risks from glyphosate and better regulation of its global utilization.

Enhancing oil production in Arabidopsis through expression of a ketoacyl-ACP synthase domain of the PUFA synthase from Thraustochytrium

BackgroundPlant seed oil is an important bioresource for human food and animal feed, as well as industrial bioproducts. Therefore, increasing oil content in seeds has been one of the primary targets in the breeding programs of oilseed crops. Thraustochytrium is a marine protist that can produce a high level of very long-chain polyunsaturated fatty acids (VLCPUFAs) using a PUFA synthase, a polyketide synthase-like fatty acid synthase with multiple catalytic domains. Our previous study showed that a KS domain from the synthase could complement an Escherichia coli mutant defective in β-ketoacyl-ACP synthase I (FabB) and increase the total fatty acid production. In this study, this KS domain from the PUFA synthase was further functionally analyzed in Arabidopsis thaliana for the capacity of oil production.ResultsThe plastidial expression of the KS domain could complement the defective phenotypes of a KASI knockout mutant generated by CRISPR/Cas9. Seed-specific expression of the domain in wild-type Arabidopsis significantly increased seed weight and seed oil, and altered the unsaturation level of fatty acids in seeds, as well as promoted seed germination and early seedling growth.ConclusionsThe condensation process of fatty acid biosynthesis in plants is a limiting step, and overexpression of the KS domain from a PUFA synthase of microbial origin offers a new strategy to increase oil production in oilseed plants.

Chlorogenic acid-mediated chemical defence of plants against insect herbivores.

Chlorogenic acid is one of the most abundant beneficial polyphenols in plants and is well known as a nutritional antioxidant in plant-based foods. Apart from its dietary antioxidant activity, it has been proved to be an efficient defence molecule against a broad range of insect herbivores. In the last two decades, several reports have shown the effectiveness of chlorogenic acid in insect growth deterrence. The pathway for chlorogenic acid biosynthesis in plants was previously elucidated, and metabolic engineering of the principal pathway showed high chlorogenic acid production in tomato plants. Herbivore-mediated induction of chlorogenic acid biosynthesis was also demonstrated both at metabolite and transcript level, although herbivore-mediated molecular regulation of chlorogenic acid biosynthesis is not yet fully elucidated. In this communication, we present our views on the efficacy of chlorogenic acid as an anti-herbivore defence molecule in plants and also discuss its future outlook.

Cyclopropane fatty acid biosynthesis in plants: phylogenetic and biochemical analysis of Litchi Kennedy pathway and acyl editing cycle genes.

This report describes the most extensive known gene discovery study from an oilseed that produces cyclopropane fatty acids, a novel industrial feedstock. Nature contains hundreds of examples of plant species that accumulate unusual fatty acids in seed triacylglycerols (TAG). Although lipid metabolic genes have been cloned from several exotic plant species, the underlying mechanisms that control the production of novel TAG species are still poorly understood. One such class of unusual fatty acids contain in-chain cyclopropane or cyclopropene functionalities that confer chemical and physical properties useful in the synthesis of lubricants, cosmetics, dyes, coatings, and other types of valuable industrial feedstocks. These cyclopropyl fatty acids, or CPFAs, are only produced by a small number of plants, primarily in the order Malvidae. Litchi chinensis is one member of this group; its seed oil contains at least 40mol% CPFAs. Several genes, representing early, middle, and late steps in the Litchi fatty acid and TAG biosynthetic pathways have been cloned and characterized here. The tissue-specific and developmental transcript expression profiles and biochemical characteristics observed indicate which enzymes might play a larger role in Litchi seed TAG biosynthesis and accumulation. These data, therefore, provide insights into which genes likely represent the best targets for either silencing or overexpression, in future metabolic engineering strategies aimed at altering CPFA content.

Overexpression of a cytosolic NADP+-isocitrate dehydrogenase causes alterations in the vascular development of hybrid poplars.

Cytosolic NADP+-isocitrate dehydrogenase (ICDH) is one of the major enzymes involved in the production of 2-oxoglutarate for amino acid biosynthesis in plants. In most plants studied, ICDH is encoded by either one gene or a small gene family, and the protein sequence has been highly conserved during evolution, suggesting it plays different and essential roles in metabolism and differentiation. To elucidate the role of ICDH in hybrid poplar (Populus tremula x P. alba), transgenic plants overexpressing the Pinus pinaster gene were generated. Overexpression of ICDH resulted in hybrid poplar (Populus tremula × P. alba) trees with higher expression levels of the endogenous ICDH gene and higher enzyme content than control untransformed plants. Transgenic poplars also showed an increased expression of glutamine synthetase (GS1.3), glutamate decarboxylase (GAD) and other genes associated with vascular differentiation. Furthermore, these plants exhibited increased growth in height, longer internodes and enhanced vascular development in young leaves and the apical region of stem. Modifications in amino acid and organic acid content were observed in young leaves of the transgenic lines, suggesting an increased biosynthesis of amino acids for building new structures and also for transport to other sink organs, as expanding leaves or young stems. Taken together, these results support an important role of ICDH in plant growth and vascular development.

Structural Biology of Aromatic Amino Acid Biosynthesis in Plants: The Prephenate Branch Point

Plants are chemists that have spent the last 425 million years evolving unique secondary metabolites to communicate with their surroundings, defend themselves against pathogens, insects, and herbivores, and grow and develop. Many secondary metabolites are biosynthesized from the aromatic amino acids tryptophan, tyrosine, and phenylalanine, and the shikimate and aromatic amino acid biosynthetic pathways link primary and specialized metabolism. Prephenate aminotransferase (PAT) is a key enzyme in aromatic amino acid biosynthesis because its product, arogenate, is the immediate precursor for both tyrosine and phenylalanine. Interestingly, the enzyme has a dual role in metabolism and can also function as a classical aspartate aminotransferase. AtPAT is pyridoxal 5′‐phosphate‐dependent (PLP) enzyme belonging to the AAT‐like aspartate aminotransferase domain and follows a ping‐pong bi‐bi reaction mechanism through Schiff‐base formation to a catalytic K306 in the active site. Each subunit of the AtPAT dimer consists of a large domain and a small domain that belong to the α‐β class of protein fold. Despite the structural similarities to aspartate aminotransferases, the molecular basis of prephenate binding specificity remains elusive. To gain an understanding of the enzyme's chemistry and substrate selectivity, a 2.5 Å x‐ray structure of PAT from the model plant Arabidopsis thaliana (thale cress) was solved in complex with the PLP. To our knowledge, this is the first structure of a bona fide prephenate aminotransferase. Amino acid sequence comparisons of all known functional PATs shows that T84 and K169 are highly conserved; however, some enzymes annotated as PAT display sequence variations at these positions. Biochemical analysis of T84V, K169V, and T84V/K169V PAT reveals that the double mutant gained tyrosine aminotransferase activity and was able to convert 4‐hydroxyphenylpyruvate to tyrosine. T84 is on the opposite side of the active site pocket, while the amine backbone of K169 hydrogen bonds to PLP. We hypothesize that Thr84 and Lys169 are important for substrate recognition in the active site and will use these mutants in future crystallographic experiments. Overall, our structural studies will assist in understanding the role of this bi‐functional enzyme in the context of aromatic amino acid biosynthesis in plants.Support or Funding InformationThis material is based upon work supported by the National Science Foundation Graduate Research Fellowship under Grant No. DGE‐1143954. Any opinion, findings, and conclusions or recommendations expressed in this material are those of the authors and do not necessarily reflect the views of the National Science Foundation. Results shown in this report are derived from work performed at Argonne National Laboratory, Structural Biology Center at the Advanced Photon Source. Argonne is operated by UChicago Argonne, LLC, for the U.S. Department of Energy, Office of Biological and Environmental Research under contract DE‐AC02‐06CH11357.

Sunflower (Helianthus annuus) fatty acid synthase complex: β-hydroxyacyl-[acyl carrier protein] dehydratase genes.

Two sunflower hydroxyacyl-[acyl carrier protein] dehydratases evolved into two different isoenzymes showing distinctive expression levels and kinetics' efficiencies. β-Hydroxyacyl-[acyl carrier protein (ACP)]-dehydratase (HAD) is a component of the type II fatty acid synthase complex involved in 'de novo' fatty acid biosynthesis in plants. This complex, formed by four intraplastidial proteins, is responsible for the sequential condensation of two-carbon units, leading to 16- and 18-C acyl-ACP. HAD dehydrates 3-hydroxyacyl-ACP generating trans-2-enoyl-ACP. With the aim of a further understanding of fatty acid biosynthesis in sunflower (Helianthus annuus) seeds, two β-hydroxyacyl-[ACP] dehydratase genes have been cloned from developing seeds, HaHAD1 (GenBank HM044767) and HaHAD2 (GenBank GU595454). Genomic DNA gel blot analyses suggest that both are single copy genes. Differences in their expression patterns across plant tissues were detected. Higher levels of HaHAD2 in the initial stages of seed development inferred its key role in seed storage fatty acid synthesis. That HaHAD1 expression levels remained constant across most tissues suggest a housekeeping function. Heterologous expression of these genes in E. coli confirmed both proteins were functional and able to interact with the bacterial complex 'in vivo'. The large increase of saturated fatty acids in cells expressing HaHAD1 and HaHAD2 supports the idea that these HAD genes are closely related to the E. coli FabZ gene. The proposed three-dimensional models of HaHAD1 and HaHAD2 revealed differences at the entrance to the catalytic tunnel attributable to Phe166/Val1159, respectively. HaHAD1F166V was generated to study the function of this residue. The 'in vitro' enzymatic characterization of the three HAD proteins demonstrated all were active, with the mutant having intermediate K m and V max values to the wild-type proteins.

Species-specific differences in synthesis of flavonoids and phenolic acids under increasing periods of enhanced blue light

Species-specific differences in synthesis of flavonoids and phenolic acids were studied under lengthening periods of enhanced blue light in a greenhouse experiment in Northern Finland in the autumn of 2012. The aim was to compare red- and blue- weighted light spectra in relation to biosynthesis of the compounds. The species studied were red leaf lettuce and basil. There were five treatments for these inter-lighting LED manipulations using traditional high pressure sodium lamps as background light sources. Two treatments were exposed for the entire experimental period with (1) red- and (2) blue- weighted light for 48 days. The other three treatments were initiated with red- weighted light, but after each subsequent 12 day sub-period, one red-weighted treatment was switched to blue, resulting in the following treatments: 48, 36, 24, 12 and 0 days under enhanced blue light. Flavonoid and phenolic acid biosynthesis in plants were found to be species dependent. The most abundant compound in red leaf lettuce was cichoric acid (a dicaffeoyltartaric acid) while rosmaric acid (an ester of caffeic acid and 3,4-dihydroxyphenyllactic acid) dominated in basil. Other compounds detected also varied between the species. Red leaf lettuce was much more responsive to supplemental blue light. Based on these results, it is suggested that both blue and red light may be needed to regulate the accumulation of phenolics in basil. Some of the compounds detected accumulated continuously as a function of the spent time under supplemental blue light in red leaf lettuce, but not in basil.

Target-Site Point Mutations Conferring Resistance to ACCase-Inhibiting Herbicides in Smooth Barley (Hordeum glaucum) and Hare Barley (Hordeum leporinum)

Acetyl coenzyme A carboxylase (ACCase)-inhibiting herbicides affect fatty acid biosynthesis in plants and are widely used to control smooth and hare barley in dicot crops in Australia. Recently, growers have experienced difficulty in controlling smooth and hare barley with herbicides from this mode of action. Dose–response experiments conducted on five suspected resistant populations confirmed varying levels of resistance to quizalofop and haloxyfop. The level of resistance in these populations was greater than 27-fold to quizalofop and greater than 15-fold to haloxyfop. The quizalofop dose required to reduce shoot biomass by 50% (GR50) for the resistant populations varied from 52.6 to 111.9 g ha−1, and for haloxyfop from 26.5 to 71.3 g ha−1. Sequencing the CT domain of the ACCase gene from resistant plants of different populations confirmed the presence of previously known mutations Ile1781Leu and Gly2096Ala. Amino acid substitution at the 2096 position conferred a greater level of resistance to haloxyfop than the substitution at the 1781 position. This study documents the first known case of field-evolved target-site resistance to ACCase-inhibiting herbicides in Australian populations of smooth barley.

Transcriptional regulation of fatty acid production in higher plants: Molecular bases and biotechnological outcomes

Acyl lipids are essential constituents found in every plant cell, where they fulfill diverse biological functions. Requirements for acyl chains depend on the cell type considered and vary greatly, implying a tight regulation of de novo fatty acid production in the plastids so that supply fits demand. Data generated by extensive transcriptome analyses carried out in various plant species and the characterization of the WRINKLED1 transcription factor in the model plant Arabidopsis thaliana have first established the importance of transcriptional regulation for modulating the rate of fatty acid production. The fatty acid biosynthetic pathway is indeed subjected to a system of global transcriptional control that coordinates the expression of most genes encoding enzymes of the pathway. Although much remains to be elucidated, the framework of the regulatory system controlling fatty acid biosynthesis in plants is coming into focus.Practical applications: Fatty acids constitute the most abundant form of reduced carbon chains available from nature and plant oils represent the main renewable resource of these fatty acids. Plant oils therefore represent a highly valuable agricultural commodity, the demand for which is increasing rapidly. They are primarily used for food and feed, but they are increasingly being utilized as renewable sources of industrial feedstock and fuel. Knowledge regarding the regulation of fatty production is extensively exploited in the frame of genetic engineering for oilseed crop improvement.Fatty acids constitute the most abundant form of reduced carbon chains available from nature and plant oils represent the main renewable resource of these fatty acids. Plant oils therefore represent a highly valuable agricultural commodity, the demand for which is increasing rapidly. They are primarily used for food and feed, but they are increasingly being utilized as renewable sources of industrial feedstock and fuel. Knowledge regarding the regulation of fatty production is extensively exploited in the frame of genetic engineering for oilseed crop improvement. Data generated by extensive transcriptome analyses carried out in various plant species and the characterization of the WRINKLED1 transcription factor in the model plant Arabidopsis thaliana have established the importance of transcriptional regulation for modulating the rate of fatty acid production. The fatty acid biosynthetic pathway is indeed subjected to a system of global transcriptional control that coordinates the expression of most genes encoding enzymes of the pathway. Recent advance suggest that WRINKLED1 activity may be modulated at the post‐translational level through homo‐dimerization, phosphorylation, and/or ubiquitination.

A Soybean Acyl Carrier Protein, GmACP, Is Important for Root Nodule Symbiosis

Legumes (members of family Fabaceae) establish a symbiotic relationship with nitrogen-fixing soil bacteria (rhizobia) to overcome nitrogen source limitation. Single root hair epidermal cells serve as the entry point for bacteria to infect the host root, leading to development of a new organ, the nodule, which the bacteria colonize. In the present study, the putative role of a soybean acyl carrier protein (ACP), GmACP (Glyma18g47950), was examined in nodulation. ACP represent an essential cofactor protein in fatty acid biosynthesis. Phylogenetic analysis of plant ACP protein sequences showed that GmACP was classified in a legume-specific clade. Quantitative reverse-transcription polymerase chain reaction analysis demonstrated that GmACP was expressed in all soybean tissues but showed higher transcript accumulation in nodule tissue. RNA interference-mediated gene silencing of GmACP resulted in a significant reduction in nodule numbers on soybean transgenic roots. Fluorescent protein-labeled GmACP was localized to plastids in planta, the site of de novo fatty acid biosynthesis in plants. Analysis of the fatty acid content of root tissue silenced for GmACP expression, as determined by gas chromatography-mass spectrometry, showed an approximately 22% reduction, specifically in palmitic and stearic acid. Taken together, our data provide evidence that GmACP plays an important role in nodulation.

Isolation, characterization and expression analysis of the genes—GhAOS, GhAOC and GhOPR3: Encoding the key enzymes involved in jasmonic acid biosynthesis in Gladiolus hybridus

Jasmonic acid (JA) plays a regulatory role in plant development such as tuber and corm formation and their responses to environmental stresses. The action of JA in regulating plant growth and stress responses often requires the elevation of endogenous levels by de novo synthesis. Allene oxide synthase (AOS), allene oxide cyclase (AOC) and 12-oxo-phytodienoic acid reductase 3 (OPR3) are the key enzymes involved in jasmonic acid biosynthesis in plants. Here, we isolated the AOS, AOC and OPR3 genes (GhAOS, GhAOC and GhOPR3) in full length from in vitro developing corms of Gladiolus hybridus. The subcellular localization analysis indicated that both GhAOS and GhAOC were chloroplast localization proteins whereas GhOPR3 was localized within the peroxisome. Real-time quantitative PCR showed that GhAOS, GhAOC and GhOPR3 genes were expressed constitutively in all organs with different levels having a relatively higher level in corms and cormels. Additionally, we observed that MJ treatments contributed to increasing the amount of three genes, mRNA level and endogenous MJ content thus promoting corm formation and enlargement. These results revealed that GhAOS, GhAOC and GhOPR3 were genes of significant importance and their expression contributed to JA biosynthesis thus playing an important role in corm formation and enlargement.