A relationship between photosynthesis and translocation in plants stressed by ionizing radiation

Since previous investigations have shown that low levels of ionizing radiation can induce a reduction in the rates of apparent photosynthesis and in the magnitude of photoassimilated l4C exported out of a leaf, the present studies were designed and conducted to determine the relationship, if any, between the radiation effects on these two physiological processes. The experiments were particularly designed to determine if the radiation-induced reduction in export is the result of the reduction in photosynthesis and hence availability of materials for translocation or the result of a reduction in the amount of energy available for the vein loading process. This study has shown that the radiation-induced reduction in l4C export out of a leaf is likely related to a loss of energy available for the vein loading process rather than a reduction in the supply of materials available for export due to reduced C02 uptake. The process of photophosphorylation was shown to be reduced by exposure to radiation to an extent similar to the reduction in the export of l4C which was also observed. Both of these processes returned to their pre-irradiation rates 120 minutes following radiatruon exposure. The rate of photosynthetic C02 uptake was also reduced by radiation exposur~ howeve~ this process did not return to the control level nor was the extent of reduction as large as observed for photophosphorylation and photoassimilate export. The observed relationship between the reductions of export and photoph~sphorylation pointed to the utilization of photosynthetically produced ATP in the vein loading process. The radiation-induced reduction in the export of l4C was observed at the highest light intensity used in this study which would also imply the involvement of the photophosphorylation process as an energy seurce for vein loading. The lack of radiation-induced reduction in export at low light intensities was interpreted as being due to the utilization of respiratory derived ATP, a process known to be insensitive to radiation at the levels used in this study, as the energy source for the vein loading process. Studies using plants not stressed by radiation showed that there was an increase in export of 14C with higher light intensities. In summary, the data has been interpreted as showing that at high light intensities the ATP, produced by photophosphorylation, is available for use in the vein loading process. The site of ATP utilization could not be determined from the data obtained in this study but possible sites have been indicated from the work done by other physiologists and are discussed in the thesis.

Trading nitrogen for carbon: Nitrogen and carbon translocation in a plant/fungal (Metarhizium spp.) symbiosis

While nitrogen is critical for all plants, they are unable to utilize organically bound nitrogen in soils. Therefore, the majority of plants obtain useable nitrogen through nitrogen fixing bacteria and the microbial decomposition of organic matter. In the majority of cases, symbiotic microorganisms directly furnish plant roots with inorganic forms of nitrogen. More than 80% of all land plants form intimate symbiotic relationships with root colonizing fungi. These common plant/fungal interactions have been defined largely through nutrient exchange, where the plant receives limiting soil nutrients, such as nitrogen, in exchange for plant derived carbon. Fungal endophytes are common plant colonizers. A number of these fungal species have a dual life cycle, meaning that they are not solely plant colonizers, but also saprophytes, insect pathogens, or plant pathogens. By using 15N labeled, Metarhizium infected, wax moth larvae (Galleria mellonella) in soil microcosms, I demonstrated that the common endophytic, insect pathogenic fungi Metarhizium spp. are able to infect living soil borne insects, and subsequently colonize plant roots and furnish ts plant host with useable, insect-derived nitrogen. In addition, I showed that another ecologically important, endophytic, insect pathogenic fungi, Beauveria bassiana, is able to transfer insect-derived nitrogen to its plant host. I demonstrated that these relationships between various plant species and endophytic, insect pathogenic fungi help to improve overall plant health. By using 13C-labeled CO2, added to airtight plant growth chambers, coupled with nuclear magnetic resosnance spectroscopy, I was able to track the movement of carbon from the atmosphere, into the plant, and finally into the root colonized fungal biomass. This indicates that Metarhizium exists in a symbiotic partnership with plants, where insect nitrogen is exchanged for plant carbon. Overall these studies provide the first evidence of nutrient exchange between an insect pathogenic fungus and plants, a relationship that has potentially useful implications on plant primary production, soil health, and overall ecosystem stability.

Seasonal effects of foliar application of phosphonate on phosphonate translocation, in vitro pollen viability and pollen germination in `Hass' avocado (Persea americana Mill.)

Phosphonate fungicides are used widely in the control of diseases caused by Phytophthora cinnamomi Rands. For the most part phosphonate is seen as a safe to use on crops with phytotoxicity rare. However, recent research has shown that phosphonate has detrimental effects on the floral biology of some indigenous Australian plants. Since phosphonate fungicides are regularly used for the control of Phytophthora root rot in avocados, research was carried out to study the translocation of phosphonate fungicide in 'Hass' trees and any effects on their floral biology. Field-grown trees were sprayed with 0, 0.06 or 0.12 M mono-dipotassium phosphonate (pH 7.2) at summer flush maturity, floral bud break or anthesis. Following treatment, phosphonic acid concentrations were determined in leaves, roots, inflorescence rachi and flowers and in vitro pollen germination and pollen tube growth studied. Phosphonic acid concentration in the roots and floral parts was related to their sink strength at the respective times of application with concentration in roots highest (36.9.mg g±1) after treatment at summer flush maturity and in flowers (234.7 mg g±1) after treatment during early anthesis. Phosphonate at >0.03 M was found to be significantly phytotoxic to in vitro pollen germination and pollen tube growth. However, this rate gave a concentration far in excess of that measured in plant tissues following standard commercial applications of mono-dipotassium phosphonate fungicide. There was a small effect on pollen germination and pollen tube growth when 0.06 and 0.12 M mono-dipotassium phosphonate was applied during early anthesis. However, under favourable pollination and fruit set conditions it is not expected to have commercial impact on tree yield. However, there may be detrimental commercial implications from phosphonate sprays at early anthesis if unfavourable climatic conditions for pollination and fruit set subsequently occur. A commercial implication from this study is that phosphonic acid root concentrations can be elevated and maintained with strategic foliar applications of phosphonate fungicide timed to coincide with peaks in root sink strength. These occur at the end of the spring and summer flushes when shoot growth is relatively quiescent. Additional foliar applications may be advantageous in under high disease-pressure situations but where possible should be timed to minimize overlap with other significant growth events in the tree such as rapid inflorescence, and fruit development and major vegetative flushing.

Uptake and translocation of inorganic and methylated arsenic species by plants

The uptake and translocation into shoots of arsenate, methylarsonate (MA), and dimethylarsinate (DMA) by 46 different plant species were studied. The plants (n = 3 per As species) were exposed for 24 h to 1 mg of As per litre under identical conditions. Total arsenic was measured in the roots and the shoots by acid digestion and inductively coupled plasma mass spectrometry from which, besides total As values, root absorption factors and shoot-to-root transfer factors were calculated. As uptake into the root for the different plant species ranged from 1.2 to 95 (mu g of As per g of dry weight) for As-V, from 0.9 to 44 for MA(V) and from 0.8 to 13 for DMA(V), whereas in shoots the As concentration ranged from 0.10 to 17 for As-V, 0.1 to 13 for MA(V), and 0.2 to 17 for DMA(V). The mean root absorption factor for As-V (1.2 to 95%) was five times higher than for DMA(V) (0.8 to 13%) and 2.5 times higher than for MA(V) (0.9 to 44%). Although the uptake of arsenic in the form of As-V was significantly higher than that of MA(V) and DMA(V), the translocation of the methylated species was more efficient in most plant species studied. Thus, an exposure of plants to DMA(V) or MA(V) can result in higher arsenic concentrations in the shoots than when exposed to As-V. Shoot-to-root transfer factors (TFs) for all plants varied with plant and arsenic species. While As-V had a median TF of 0.09, the TF of DMA(V) was nearly a factor of 10 higher (0.81). The median TF for MA(V) was in between (0.30). Although the TF for MA(V) correlates well with the TF for DMA(V), the plants can be separated into two groups according to their TF of DMA(V) in relation to their TF of As-V. One group can immobilise DMA(V) in the roots, while the other group translocates DMA(V) very efficiently into the shoot. The reason for this is as yet unknown.

Uptake, translocation and transformation of arsenate and arsenite in sunflower (Helianthus annuus):formation of arsenic-phytochelatin complexes during exposure to high arsenic concentrations

The aim of the study was to determine the time-dependent formation of arsenic-phytochelatin (As-PC) complexes in the roots, stems and leaves of an arsenic-nontolerant plant (Helianthus annuus) during exposure to 66 mol l(-1) arsenite (As(III)) or arsenate (As(V)). We used our previously developed method of simultaneous element-specific (inductively coupled plasma mass spectrometry, ICP-MS) and molecular-specific (electrospray-ionization mass spectrometry, ES-MS) detection systems interfaced with a suitable chromatographic column and eluent conditions, which enabled us to identify and quantify As-PC complexes directly. Roots of As-exposed H. annuus contained up to 14 different arsenic species, including the complex of arsenite with two (gamma-Glu-Cys)(2)-Gly molecules [As((III))-(PC(2))(2)], the newly identified monomethylarsonic phytochelatin-2 or (gamma-Glu-Cys)(2)-Gly CH(3)As (MA((III))-PC(2)) and at least eight not yet identified species. The complex of arsenite with (gamma-Glu-Cys)(3)-Gly (As((III))-PC(3)) and the complex of arsenite with glutathione (GSH) and (gamma-Glu-Cys)(2)-Gly (GS-As((III))-PC(2)) were present in all samples (roots, stems and leaves) taken from plants exposed to As. The GS-As((III))-PC(2) complex was the dominant complex after 1 h of exposure. As((III))-PC(3) became the predominant As-PC complex after 3 h, binding up to 40% of the As present in the exposed plants. No As-PC complexes were found in sap (mainly xylem sap from the root system), in contrast to roots, stems and leaves, which is unequivocal evidence that As-PC complexes are not involved in the translocation of As from root to leaves of H. annuus.

Distribution and Translocation of Selenium from Soil to Grain and Its Speciation in Paddy Rice (Oryza sativa L.)

Selenium, an essential micronutrient for humans, is insufficient in dietary intake for millions of people worldwide. Rice as the most popular staple food in the world is one of the dominant selenium (Se) sources for people. The distribution and translocation of Se from soil to grain were investigated in a Se-rich environment in this study. The Se levels in soils ranged widely from 0.5 to 47.7 mg kg(-1). Selenium concentration in rice bran was 1.94 times higher than that in corresponding polished rice. The total Se concentrations in the rice fractions were in the following order: straw > bran > whole grain > polished rice > husk. Significant linear relationships between different rice fractions were observed with each other, and Se in the soil has a linear relationship with different rice fractions as well. Se concentration in rice can easily be predicted by soil Se concentrations or any rice fractions and vice versa according to their linear relationships. In all rice samples for Se speciation, SeMet was the major Se species, followed by MeSeCys and SeCys. The average percentage for SeMet (82.9%) and MeSeCys (6.2%) was similar in the range of total Se from 2.2 to 8.4 mg kg(-1) tested. The percentage of SeCys decreased from 6.3 to 2.8%, although its concentration elevated with the increase in total Se in rice. This could be due to the fact that SeCys is the precursor for the formation of other organic Se compounds. The information obtained may have considerable significance for assessing translocation and accumulation of Se in plant.

An arsenic-contaminated field trial to assess the uptake and translocation of arsenic by genotypes of rice

Compared to other cereals, rice has particular strong As accumulation. Therefore, it is very important to understand As uptake and translocation among different genotypes. A field study in Chenzhou city, Hunan province of China, was employed to evaluate the effect of arsenic-contaminated soil on uptake and distribution in 34 genotypes of rice (including unpolished rice, husk, shoot, and root). The soil As concentrations ranged from 52.49 to 83.86 mg kg-1, with mean As concentration 64.44 mg kg-1. The mean As concentrations in rice plant tissues were different among the 34 rice genotypes. The highest As concentrations were accumulated in rice root (196.27-385.98 mg kg-1 dry weight), while the lowest was in unpolished rice (0.31-0.52 mg kg-1 dry weight). The distribution of As in rice tissue and paddy soil are as follows root » soil > shoot > husk > unpolished rice. The ranges of concentrations of inorganic As in all of unpolished rice were from 0.26 to 0.52 mg kg-1 dry weight. In particular, the percentage of inorganic As in the total As was more than 67 %, indicating that the inorganic As was the predominant species in unpolished rice. The daily dietary intakes of inorganic As in unpolished rice ranged from 0.10 to 0.21 mg for an adult, and from 0.075 to 0.15 mg for a child. Comparison with tolerable daily intakes established by FAO/WHO, inorganic As in most of unpolished rice samples exceeded the recommended intake values. The 34 genotypes of rice were classified into four clusters using a criteria value of rescaled distance between 5 and 10. Among the 34 genotypes, the genotypes II you 416 (II416) with the lowest enrichment of As and the lowest daily dietary intakes of inorganic As could be selected as the main cultivar in As-contaminated field.

Investigation of organic xenobiotic transfers, partitioning and processing in air-soil-plant systems using a microcosm apparatus. Part I:Microcosm development

A microcosm system was developed to investigate transfers of organic xenobiotics in air-soil-plant systems. This was validated using ¹⁴C labelled 1,2-dichlorobenzene (DCB) as a model compound. Trapping efficiency was 106 ± 3% for volatile compounds and 93.0 ± 2.2% for carbon dioxide in a blank microcosm arrangement. Recovery of 1,2-dichlorobenzene spiked to grassed and unplanted soils was > 90% after 1 week. The predominant DCB loss process was volatilisation with no evidence for mineralisation over 1 week and 20-30% of the added spike remained in soil. Although there was no evidence for root uptake and translocation of added label, foliar uptake of soil volatilised compound was detected. The microcosm showed good potential for study of ¹⁴C labelled and unlabelled organic xenobiotic transfers in air-soil-plant systems with single plants and also intact planted cores.

Changes in the magnitude and pattern of translocation of photoassimilated ¹CO in soybean plants following an acute exposure to gamma radiation /

Young soybean plants (Glycine ~. L. cultivar Harosoy '63), grown under controlled conditions, were exposed to gamma radiation on a single occasion. One hour following exposure to 3,750 rads, the mature trifoliate leaf of the soybean plant was isolated in a closed system and permitted to photoassimilate approximately 1-5 pCi of 14C02 for 15 minutes. After an additional 45 minute-period, the plant was sacrificed and the magnitude of translocation and distribution pattern of 14C determined. In the non-irradiated plants 18~ of the total 14C recovered was outside the fed leaf blades and of this translocated 14c, 28~ was above the node of the fed leaf, 38~ in the stem below the node, 28~ in the roots and 7~ in the petiole. As well, in the irradiated plants, a smaller per cent (6~) of the total 14 C recovered was exported out of the source leaf blades. Of this translocated 14c , a smaller per cent (20~) was found in the apical region above the node of the source leaf and a higher per cent (45~) was recovered from the stem below the node and in the petiole (11~). The per cent of exported 14 C recovered from the root was unaffected by the radiation. Replacement of the shoot apex with 20 ppm IAA immediately following irradiation, only J partially increased the magnitude of translocation but did completely restore the pattern of distribution to that observed in the non-irradiated plants. From supplementary studies showing a radiationinduced reduction of photosynthetic rates in the source leaf and a reduction of the cumulative stem and leaf lengths in the apical sink region, the observed effects of radiation on the translocation process have been correlated to damage incurred by the source and sink regions. These data suggest that the reduction in the magnitude of translocation is the result of damage to both the source and sink regions rather than the phloem conducting tissue itself, whereas the change in the pattern of translocation is probably the result of a reduced rate of 14C-assimilate movement caused by a radiation-induced decrease of sink metabolism, especially the decrease in the metabolism of the apical sink.

Antidiabetic activity of Vaccinium vitis-idaea, a medicinal plant from the traditional pharmacopeia of the James Bay Cree

L’incidence du diabète chez les premières nations du Canada est plus de trois fois celle du reste du pays, dû, en partie, aux traitements culturellement inappropriés. Notre projet vise à traiter le diabète chez ces populations à partir de leur pharmacopée de médicine traditionnelle afin d’améliorer l’acceptation des traitements. En utilisant une approche ethnobotanique, notre équipe a identifié 17 plantes médicinales utilisées pour traiter des symptômes du diabète par les Cris d'Eeyou Istchee (Baie James, Québec). Parmi eux, l'extrait éthanolique de baies de Vaccinium vitis-idaea a montré un effet stimulateur sur le transport du glucose dans les cellules musculaires squelettiques et les adipocytes en culture. Le but de cette thèse était d’élucider les mécanismes par lesquels cet extrait exerce ses effets anti-hyperglycémiants, d’identifier ses principes actifs et de confirmer in vivo, son efficacité. Les résultats démontrent que V.vitis a augmenté le transport du glucose dans les cellules musculaires en cultures, C2C12 et L6 et a stimulé la translocation des transporteurs GLUT4 dans les cellules L6. L'extrait a également inhibé la respiration dans les mitochondries isolées du foie du rat. Cet effet est semblable à celui de la metformine et en lien avec la production du stress métabolique et l'activation de l'AMPK. De plus, la voie de signalisation de l’insuline ne semble pas être impliquée dans le mécanisme d’action de V. vitis. Le fractionnement guidé par la stimulation du transport du glucose a mené à l'isolation des principes actifs; la quercétine, la quercétine-3-O-galactoside, et la quercétine-3-O-glucoside. Comparable à l'extrait brut, ses composés ont stimulé la voie AMPK. Cependant, la quércetine était la seule à inhiber la respiration mitochondriale. Pour valider l'effet de V.vitis in vivo, l'extrait (1% dans l'eau de boisson) a été administré aux souris KKAy pendant 10 jours. La glycémie et le poids corporel ont été significativement réduits par V.vitis. Ces effets ont été associés à une diminution de la prise alimentaire, ce qui suggère que V.vitis diminue l'appétit. L'étude pair-fed a confirmé que les effets de V.vitis sont, majoritairement, dû à la réduction de l’appétit. De plus, V.vitis a augmenté la teneur en GLUT4 dans le muscle squelettique, a stimulé la iv phosphorylation de l'ACC et a augmenté les niveaux de PPAR-α dans le foie des souris KKAy. Ces effets se voient être additifs à l’effet anorexigène de V. vitis. Au cours du fractionnement bioguidé de l’extrait, l’ester méthylique de l'acide caféique (CAME), un produit formé lors de la procédure du fractionnement, a démontré un effet stimulateur puissant sur le transport du glucose dans les celules C2C12 et donc un potentiel anti-diabétique. Pour identifier d'autres acides caféique active (AC) et pour élucider leurs relations structure-activité et structure-toxicité, vingt dérivés AC ont été testés. Outre CAME, quatre composés ont stimulé le transport du glucose et ont activé l'AMPK suite au stress métabolique résultant d'un découplage de la phosphorylation oxydative mitochondriale. L’activité nécessite une fonction d’AC intacte dépourvu de groupements fortement ionisés et ceci était bien corrélée avec la lipophilicite et la toxicité. Les résultats de cette thèse soutiennent le potentiel thérapeutique de V. vitis, ses composés actifs ainsi que de la famille de l’AC et pour la prévention et le traitement du diabète.