Elevated fruit nitrogen impairs oil biosynthesis in olive (Olea europaea L.).

Oil in fruits and seeds is an important source of calories and essential fatty acids for humans. This specifically holds true for olive oil, which is appreciated for its superior nutritional value. Most olive orchards are cultivated to produce oil, which are the outcome of fruit yield and oil content. Little information is available on the effect of nitrogen (N) on olive fruit oil content. The response of olive trees to different rates of N was therefore studied in soilless culture (3 years) and commercial field (6 years) experiments. In both experiments, fruit N level and oil biosynthesis were negatively associated. Fruit N increased in response to N fertilization level and was inversely related to fruit load. The negative correlation between fruit N and oil content was more pronounced under high fruit load, indicating sink limitation for carbon. These results agree with those reported for oilseed crops for which a trade-off between oil and protein was proposed as the governing mechanism for the negative response to elevated N levels. Our results suggest that the protein/oil trade-off paradigm cannot explain the noticeable decrease in oil biosynthesis in olives, indicating that additional mechanisms are involved in N-induced inhibition of oil production. This inhibition was not related to the soluble carbohydrate levels in the fruit, which were comparable regardless of N level. These results emphasize the importance of balanced N nutrition in oil-olive cultivation to optimize production with oil content.

Steroidal alkaloids defence metabolism and plant growth are modulated by the joint action of gibberellin and jasmonate signalling.

Steroidal glycoalkaloids (SGAs) are protective metabolites constitutively produced by Solanaceae species. Genes and enzymes generating the vast structural diversity of SGAs have been largely identified. Yet, mechanisms of hormone pathways coordinating defence (jasmonate; JA) and growth (gibberellin; GA) controlling SGAs metabolism remain unclear. We used tomato to decipher the hormonal regulation of SGAs metabolism during growth vs defence tradeoff. This was performed by genetic and biochemical characterisation of different JA and GA pathways components, coupled with in vitro experiments to elucidate the crosstalk between these hormone pathways mediating SGAs metabolism. We discovered that reduced active JA results in decreased SGA production, while low levels of GA or its receptor led to elevated SGA accumulation. We showed that MYC1 and MYC2 transcription factors mediate the JA/GA crosstalk by transcriptional activation of SGA biosynthesis and GA catabolism genes. Furthermore, MYC1 and MYC2 transcriptionally regulate the GA signalling suppressor DELLA that by itself interferes in JA-mediated SGA control by modulating MYC activity through protein-protein interaction. Chemical and fungal pathogen treatments reinforced the concept of JA/GA crosstalk during SGA metabolism. These findings revealed the mechanism of JA/GA interplay in SGA biosynthesis to balance the cost of chemical defence with growth.

Stimuli-Free Transcuticular Delivery of Zn Microelement Using Biopolymeric Nanovehicles: Experimental, Theoretical, and In Planta Studies.

This paper reports one-step synthesis of polysaccharide-based nanovehicles, capable of transporting ionic zinc via plant cuticle without auxiliary stimulation. Delivery of highly hydrophilic nutritive microelements via the hydrophobic cuticle of plant foliage is one of the major challenges in modern agriculture. In traditional nutrition via roots, up to 80% of microelements permeate to soil and get wasted; therefore, foliar treatment is an environmentally and economically preferable alternative. Carboxymethyl cellulose (CMC) was modified to amphiphilic N-octylamide-derivative (CMC-8), which spontaneously self-assemble to nanovehicles. It was found that hydrophobic substituents endow a biopolymer with unexpected affinity toward a hydrophilic payload. CMC-8 nanovehicles effectively encapsulated ionic zinc (ZnSO4) and delivered it upon foliar application to pepper (Capsicum annuum) and tomato (Solanum lycopersicum) plants. Zinc uptake and translocation in plants were monitored by SEM-EDS and fluorescence microscopic methods. In planta monitoring of the carrier was done by labeling nanovehicles with fluorescent carbon dots. Three-dimensional (3-D) structural modeling and conformational dynamics explained the CMC-8 self-assembly mechanism and zinc coordination phenomenon upon introduction of hydrophobic substituents.

The metabolic reserves, carbohydrate balance and nutritional status of jojoba (Simmondsia chinensis), in relation to its annual cycle and fruit load.

Jojoba (Simmondsia chinensis (Link) Schneider) holds high industrial value and an extended cultivation trend. Despite its increased importance, there is a lack of fundamental information about its metabolic reserves and development. Our objective was to characterise metabolite allocation and fluctuations in the carbohydrate and nutrient balance of jojoba plants, as affected by fruit load and the plant's annual cycle. Metabolite profiles were performed for each organ. Soluble carbohydrates (SC) and starch concentrations were surveyed in underground and aboveground organs of high-yield and fruit-removed plants. Simultaneously, nitrogen, potassium and phosphorus were determined in the leaves to evaluate the plant's nutritional status. We found that sucrose and pinitol were the most abundant sugars in all jojoba organs. Each sugar had a 'preferred' organ: glucose was accumulated mainly in the leaves, sucrose and pinitol in woody branches, and fructose in the trunk wood. We found that fruit load significantly influenced the carbohydrate levels in green branches, trunk wood and thin roots. The phenological stage strongly affected the SC-starch balance. Among the examined minerals, only the leaf potassium level was significantly influenced by fruit load. We conclude that jojoba's nutrient and carbohydrate balance is affected by fruit load and the phenological stage, and describe the organ-specific metabolic reserves.

It takes two: Reciprocal scion-rootstock relationships enable salt tolerance in 'Hass' avocado.

Commercial avocado orchards typically consist of composite trees. Avocado is salt-sensitive, suffering from substantial growth and production depreciation when exposed to high sodium and chloride levels. Salt ions penetrate the roots and are subsequently transferred to the foliage. Hence, understanding distinct physiological responses of grafted avocado plant organs to salinity is of great interest. We compared the ion, metabolite and lipid profiles of leaves, roots and trunk drillings of mature 'Hass' scion grafted onto two different rootstocks during gradual exposure to salinity. We found that one rootstock, VC840, did not restrict the transport of irrigation solution components to the scion, leading to salt accumulation in the trunk and leaves. The other rootstock, VC152, functioned selectively, moderating the movement of toxic ions to the scion organs by accumulating them in the roots. The leaves of the scion grafted on the selective rootstock acquired the standard level of essential minerals without being exposed to excessive salt concentrations. However, this came with an energetic cost as the leaves transferred carbohydrates and storage lipids downward to the rootstock organs, which became a strong sink. We conclude that mutual scion-rootstock relationships enable marked tolerance to salt stress through selective ion transport and metabolic modifications.

Potassium and storage root development: focusing on photosynthesis, metabolites and soluble carbohydrates in cassava.

The linkage between K and the development of storage roots in root crops is partially understood, hence this experiment determined some of the mechanisms involved in cassava. The effects of 10, 40, 70, 100, 150 and 200 mg K l-1 fertigation on photosynthetic attributes, soluble carbohydrates, starch, metabolites, growth and yield were studied in a greenhouse. Storage root yield, number of storage roots, stomatal conductance and net photosynthesis reached maximum at 150 mg K l-1 . However, soluble carbohydrates and starch in the leaves significantly declined with an increasing concentration of K solution, similarly to the trend of glycerol in the leaves. Conversely, malic acid, citric acid and propionic acid gradually increased reaching maximum at 150, 150 and 70 mg K l-1 respectively. Combined, these results suggest that sugars were transported from the leaves to a stronger sink - the bulking storage roots. This and the increase of intermediate metabolites of tricarboxylic acid cycle provided the energy required for the bulking process and the development of the storage roots. Although the measured parameters indirectly link K to storage root development, they nonetheless form a basis for studies on direct interactions.

Excessive nitrogen impairs hydraulics, limits photosynthesis, and alters the metabolic composition of almond trees.

Horticulture nitrogen (N) runoffs are major environmental and health concerns, but current farming practices cannot detect ineffective N applications. Hence, we set to recognize high N conditions and characterize their effects on the physiology of almond trees grown in drainage lysimeters. Water and nutrients mass balances exhibited that N benefitted almond trees in a limited range (below 60 mg N L-1 in irrigation), while higher N conditions (over a 100 mg N L-1) reduced evapotranspiration (ET) by 50% and inherently constrained N uptake. Respectively, whole-tree hydraulic conductance reduced by 37%, and photosynthesis by 17%, which implied that high N concentrations could damage trees. Through gas-chromatography, we realized that high N conditions also affected components of the citric acid cycle (TCA) and carbohydrates availability. Such changes in the metabolic composition of roots and leaves probably interfered with N assimilation and respiration. It also determined the proportions between N and starch in almond leaves, which formed a new index (N:ST) that starts at 0.4 in N deficiency and reaches 0.6-0.8 in optimal N conditions. Importantly, this index continues to increase in higher N conditions (as starch reduces) and essentially indicates to excessive N applications when it exceeds 1.1.

Auxin Response Dynamics During Wild-Type and entire Flower Development in Tomato.

The plant hormone auxin is a major regulator of plant development and response to environmental cues. Auxin plays a particularly central role in flower development, but the knowledge of its role of flower development in crop plants with fleshy fruits, such as tomato, is still scarce. Mutations in the Aux/IAA gene ENTIRE/Indole Acetic Acid 9 (E/IAA9) lead to the precocious development of young gynoecia into parthenocarpic fruits. Here, we compared the distribution of the auxin response sensor DR5::VENUS and the auxin efflux transporter PIN1 between the wild type and entire during successive stages of flower and fruit development. Up-regulation of the DR5::VENUS signal in the shoot apical meristem (SAM) was observed upon the transition to flowering, implicating a possible role for auxin in the transition from a vegetative SAM into an inflorescence meristem. DR5::VENUS was expressed in all initiating floral organs. Additionally, DR5::VENUS was highly expressed during gametogenesis, in both male and female organs, and in the developing seeds during embryogenesis. DR5::VENUS is expressed in functional cell layers such as the anther stomium and tapetum, suggesting that auxin plays a role in flower organ development and function. The entire mutation affected DR5::VENUS expression patterns during inflorescence formation and flower organ development, which correlated with phenotypic alterations. We also show dynamic distribution and localization of the auxin transporter PIN1 during flower and fruit organ development. These results emphasize the dynamic auxin response in inflorescence and flower development and suggest multiple roles of auxin in these processes.

Phosphorous Nutritional Level, Carbohydrate Reserves and Flower Quality in Olives.

The olive tree is generally characterized by relatively low final fruit set consequential to a significant rate of undeveloped pistils, pistil abortion, and flower and fruitlet abscission. These processes are acknowledged to be governed by competition for resources between the developing vegetative and reproductive organs. To study the role of phosphorus (P) nutritional level on reproductive development, trees were grown under four levels of P for three years in large containers. Phosphorus nutritional level was positively related to rate of reproductive bud break, inflorescence weight, rate of hermaphrodite flowers, pistil weight, fruitlet persistence, fruit set and the consequential total number of fruits. The positive impact of P nutrition on the productivity parameters was not related to carbohydrate reserves or to carbohydrate transport to the developing inflorescence. Phosphorous deficient trees showed significant impairment of assimilation rate, and yet, carbohydrates were accumulated in inflorescences at levels comparable to or higher than trees receiving high P. In contrast to female reproductive organs, pollen viability was consistently higher in P deficient trees, possibly due to the enhanced carbohydrate availability. Overall, the positive effect of P on female reproductive development was found to be independent of the total carbohydrate availability. Hence, P is speculated to have a direct influence on reproductive processes.

AUXIN RESPONSE FACTOR 2 Intersects Hormonal Signals in the Regulation of Tomato Fruit Ripening.

The involvement of ethylene in fruit ripening is well documented, though knowledge regarding the crosstalk between ethylene and other hormones in ripening is lacking. We discovered that AUXIN RESPONSE FACTOR 2A (ARF2A), a recognized auxin signaling component, functions in the control of ripening. ARF2A expression is ripening regulated and reduced in the rin, nor and nr ripening mutants. It is also responsive to exogenous application of ethylene, auxin and abscisic acid (ABA). Over-expressing ARF2A in tomato resulted in blotchy ripening in which certain fruit regions turn red and possess accelerated ripening. ARF2A over-expressing fruit displayed early ethylene emission and ethylene signaling inhibition delayed their ripening phenotype, suggesting ethylene dependency. Both green and red fruit regions showed the induction of ethylene signaling components and master regulators of ripening. Comprehensive hormone profiling revealed that altered ARF2A expression in fruit significantly modified abscisates, cytokinins and salicylic acid while gibberellic acid and auxin metabolites were unaffected. Silencing of ARF2A further validated these observations as reducing ARF2A expression let to retarded fruit ripening, parthenocarpy and a disturbed hormonal profile. Finally, we show that ARF2A both homodimerizes and interacts with the ABA STRESS RIPENING (ASR1) protein, suggesting that ASR1 might be linking ABA and ethylene-dependent ripening. These results revealed that ARF2A interconnects signals of ethylene and additional hormones to co-ordinate the capacity of fruit tissue to initiate the complex ripening process.

Quinclorac resistance: a concerted hormonal and enzymatic effort in Echinochloa phyllopogon.

BACKGROUND: Quinclorac (3,7-dichloro-quinoline-carboxylic acid) is a selective herbicide widely used to control annual grasses and certain broadleaf weeds. Echinochloa phyllopogon (Stapf) Koss. is the most noxious grass weed in California rice fields and has evolved resistance to multiple herbicides with different modes of action. A quinclorac-resistant (R) E. phyllopogon biotype found in a Sacramento Valley rice field where quinclorac has never been applied was investigated. RESULTS: Resistant to susceptible (S) GR(50) (herbicide rate for 50% growth reduction) ratios ranged from 6 to 17. The cytochrome P450 inhibitor malathion (200 mg L(-1)) caused R plants to become as quinclorac susceptible as S plants. Quinclorac rapidly (6 HAT) stimulated ethylene formation in S plants, but only marginally in R plants. Malathion pretreatment did not reduce ethylene formation by quinclorac-treated S and R plants. Activity of ß-cyanoalanine synthase (ß-CAS) in tissue extracts was 2-3-fold greater in R than in S plants, and incubation of shoot extracts with 1 mM malathion reduced ß-CAS activity by 40% in both biotypes. CONCLUSION: Resistance to quinclorac in R E. phyllopogon involved at least two mechanisms: (a) insensitivity along the response pathway whereby quinclorac induces ethylene production; (b) enhanced ß-CAS activity, which should enable greater HCN detoxification following quinclorac stimulation of ethylene biosynthesis. This unveils new resistance mechanisms for this multiple-resistant biotype widely spread throughout California rice fields.

Differential oxidative metabolism and 5-ketoclomazone accumulation are involved in Echinochloa phyllopogon resistance to clomazone.

Echinochloa phyllopogon (late watergrass) is a major weed of California rice (Oryza sativa) that has evolved cytochrome P450-mediated metabolic resistance to different herbicides with multiple modes of action. E. phyllopogon populations from Sacramento Valley rice fields have also recently shown resistance to the herbicide clomazone. Clomazone is a proherbicide that must be metabolized to 5-ketoclomazone, which is the active compound that inhibits deoxyxylulose 5-phosphate synthase, a key enzyme of the nonmevalonate isoprenoid pathway. This study evaluated the differential clomazone metabolism within strains of the same species to investigate whether enhanced oxidative metabolism also confers clomazone resistance in E. phyllopogon. Using reverse-phase liquid chromatography-tandem mass spectrometry techniques in the multireaction monitoring mode, we elucidated that oxidative biotransformations are involved as a mechanism of clomazone resistance in this species. E. phyllopogon plants hydroxylated mostly the isoxazolidinone ring of clomazone, and clomazone hydroxylation activity was greater in resistant than in susceptible plants. The major clomazone metabolites resulted from monohydroxylation and dihydroxylation of the isoxazolidinone ring. Resistant plants accumulated 6- to 12-fold more of the monohydroxylated metabolite than susceptible plants, while susceptible plants accumulated 2.5-fold more of the phytotoxic metabolite of clomazone, 5-ketoclomazone. Our results demonstrate that oxidative metabolism endows multiple-herbicide-resistant E. phyllopogon with cross-resistance to clomazone through enhanced herbicide degradation and lower accumulation of the toxic metabolite in resistant versus susceptible plants.

Mechanism of resistance to penoxsulam in late watergrass [ Echinochloa phyllopogon (Stapf) Koss.].

Late watergrass [ Echinochloa phyllopogon (Stapf.) Koss.] is a major weed of California rice that has evolved P450-mediated metabolic resistance to multiple herbicides. Resistant (R) populations are also poorly controlled by the recently introduced herbicide penoxsulam. Ratios (R/S) of the R to susceptible (S) GR(50) (herbicide rate for 50% growth reduction) ranged from 5 to 9. Although specific acetolactate synthase (ALS) activity was 1.7 higher in R than in S plants, the enzyme in R plants was about 6 times more susceptible to the herbicide. R plants exhibited faster (2.8 times) oxidative [(14)C]-penoxsulam metabolism than S plants 24 h after treatment. Addition of malathion (P450 inhibitor) enhanced herbicide phytotoxicity and reduced penoxsulam metabolism in R plants. Tank mixtures with thiobencarb (can induce P450) antagonized penoxsulam toxicity in R plants, suggesting penoxsulam may be broken down by a thiobencarb-inducible enzyme. These results suggest E. phyllopogon resistance to penoxsulam is mostly due to enhanced herbicide metabolism, possibly via P450 monooxidation.

Responses to clomazone and 5-ketoclomazone by Echinochloa phyllopogon resistant to multiple herbicides in Californian rice fields.

BACKGROUND: Late watergrass [Echinochloa phyllopogon (Stapf.) Koss.] is a major weed of Californian rice that has evolved P450-mediated metabolic resistance to multiple herbicides. Resistant (R) populations are also poorly controlled by the recently introduced herbicide clomazone. The authors assessed whether this cross-resistance was also P450 mediated, and whether R plants also had reduced sensitivity to photooxidation. Understanding mechanism(s) of resistance facilitates the design of herbicide management strategies to delay resistance evolution. RESULTS: Ratios (R/S) of R to susceptible (S) GR(50) were near 2.0. [(14)C]Clomazone uptake was similar in R and S plants. Clomazone and its metabolite 5-ketoclomazone reduced chlorophyll and carotenoids in S more than in R plants. The P450 inhibitors disulfoton and 1-aminobenzo-triazole (ABT) safened clomazone in R and S plants. Disulfoton safened 5-ketoclomazone only in S plants, while ABT synergized 5-ketoclomazone mostly against S plants. Paraquat was more toxic in S than in R plants. CONCLUSION: Cross-resistance to clomazone explains failures to control R plants in rice fields, and safening by P450 inhibitors suggests that oxidative activation of clomazone is needed for toxicity to E. phyllopogon. Clomazone resistance requires mitigation of 5-ketoclomazone toxicity, but P450 detoxification may not significantly confer resistance, as P450 inhibitors poorly synergized 5-ketoclopmazone in R plants. Responses to paraquat suggest research on mechanisms to mitigate photooxidation in R and S plants is needed.

Glyphosate-induced anther indehiscence in cotton is partially temperature dependent and involves cytoskeleton and secondary wall modifications and auxin accumulation.

Yield reduction caused by late application of glyphosate to glyphosate-resistant cotton (Gossypium hirsutum; GRC) expressing CP4 5-enol-pyruvylshikmate-3-P synthase under the cauliflower mosaic virus-35S promoter has been attributed to male sterility. This study was aimed to elucidate the factors and mechanisms involved in this phenomenon. Western and tissue-print blots demonstrated a reduced expression of the transgene in anthers of GRC compared to ovules of the same plants. Glyphosate application to GRC grown at a high temperature regime after the initiation of flower buds caused a complete loss of pollen viability and inhibition of anther dehiscence, while at a moderate temperature regime only 50% of the pollen grains were disrupted and anther dehiscence was normal. Glyphosate-damaged anthers exhibited a change in the deposition of the secondary cell wall thickenings (SWT) in the endothecium cells, from the normal longitudinal orientation to a transverse orientation, and hindered septum disintegration. These changes occurred only at the high temperature regime. The reorientation of SWT in GRC was accompanied by a similar change in microtubule orientation. A similar reorientation of microtubules was also observed in Arabidopsis (Arabidopsis thaliana) seedlings expressing green fluorescent protein tubulin (tubulin alpha 6) following glyphosate treatment. Glyphosate treatment induced the accumulation of high levels of indole-3-acetic acid in GRC anthers. Cotton plants treated with 2,4-dichlorophenoxyacetic acid had male sterile flowers, with SWT abnormalities in the endothecium layer similar to those observed in glyphosate-treated plants. Our data demonstrate that glyphosate inhibits anther dehiscence by inducing changes in the microtubule and cell wall organization in the endothecium cells, which are mediated by auxin.