Distinct cis-elements in the Asparagus officinalis asparagine synthetase promoter respond to carbohydrate and senescence signals

The Asparagus officinalis L. asparagine (Asn) synthetase (AS) promoter was analysed for elements responding to carbohydrate and senescence signals. Transgenic Arabidopsis thaliana L. plants containing deletion constructs of the –1958 bp AS promoter linked to the β-glucuronidase (GUS) reporter gene (AS::GUS) were analysed by measuring GUS specific activity. Inclusion of sucrose (Suc), glucose (Glc) or fructose (Fru) in plant media repressed levels of GUS activity in –1958AS::GUS plants, regardless of the light environment, with increases in GUS found 1 d after incubation on Suc-lacking media. Hexokinase is likely to be involved in the signal pathway, as Suc, Glc, Fru, 2-deoxy-d-glucose and mannose were more effective repressors than 3-O-methylglucose, and the hexokinase inhibitor mannoheptulose reduced repression. Plants containing AS::GUS constructs with deletions that reduced the promoter to less than –405 bp did not show low sugar induction. AS::GUS activity was significantly higher in excised leaves induced to senesce by dark storage for 24 h, compared to fresh leaves, for lines containing at least –640 bp of the AS promoter but not those with –523 bp or smaller promoter fragments. Fusion of the –640 to –523 bp region to a –381AS::GUS construct generated a promoter that retained senescence induction but lacked low sugar induction. Alignment of this region to the 33-bp senescence-related sequence of the Arabidopsis and Brassica napus L. SAG12 promoters identified the sequence TTGCACG as being conserved in all the promoters, and which may be an important senescence-responsive element.

Caractérisation moléculaire de la biodiversité des Fusarium associés à la fusariose de l'asperge (Asparagus officinalis L.) au Québec

Mémoire numérisé par la Direction des bibliothèques de l'Université de Montréal.

Caractérisation moléculaire de la biodiversité des Fusarium associés à la fusariose de l'asperge (Asparagus officinalis L.) au Québec

Mémoire numérisé par la Direction des bibliothèques de l'Université de Montréal.

Effects of aqueous extract from Asparagus officinalis L. roots on hypothalamic-pituitary-gonadal axis hormone levels and the number of ovarian follicles in adult rats

Background: Asparagus is a plant with high nutritional, pharmaceutical, and industrial values. Objective: The present study aimed to evaluate the effect of aqueous extract of asparagus roots on the hypothalamic-pituitary-gonadal axis hormones and oogenesis in female rats. Materials and Methods: In this experimental study, 40 adult female Wistar rats were divided into five groups, which consist 8 rats. Groups included control, sham and three experimental groups receiving different doses (100, 200, 400 mg/kg/bw) of aqueous extract of asparagus roots. All dosages were administered orally for 28 days. Blood samples were taken from rats to evaluate serum levels of Gonadotropin releasing hormone (GnRH), follicular stimulating hormone (FSH), Luteinal hormone (LH), estrogen, and progesterone hormones. The ovaries were removed, weighted, sectioned, and studied by light microscope. Results: Dose-dependent aqueous extract of asparagus roots significantly increased serum levels of GnRH, FSH, LH, estrogen, and progestin hormones compared to control and sham groups. Increase in number of ovarian follicles and corpus luteum in groups treated with asparagus root extract was also observed (p<0.05). Conclusion: Asparagus roots extract stimulates secretion of hypothalamic- pituitary- gonadal axis hormones. This also positively affects oogenesis in female rats.

A review of Australian classical biological control of weeds programs and research activities over the past 12 years

Considerable progress has been made towards the successful classical biological control of many of Australia’s exotic weeds over the past decade. Some 43 new arthropod or pathogen agents were released in 19 projects. Effective biological control was achieved in several projects with the outstanding successes being the control of rubber vine, Cryptostegia grandiflora, and bridal creeper, Asparagus asparagoides. Significant developments also occurred in target prioritization, procedures for target and agent approval, funding, infrastructure and cooperation between agencies. Scientific developments included greater emphasis on climate matching, plant and agent phylogeny, molecular diagnostics, agent prioritization and agent evaluation.

Palinologia e taxonomia dos gêneros Elephantopus L., Orthopappus Gleason e Pseudelephantopus Rohr. (Subtribo Elephantopinae, Vernonieae- Asteraceae) ocorrentes no Brasil

A subtribo Elephantopinae tem sido alvo de poucos estudos taxonômicos e palinológicos, necessitando de uma reavaliação em seus limites. Assim, o presente estudo teve como proposta atualizar e ampliar o conhecimento de 13 espécies de Elephantopinae ocorrentes no Brasil através de um acurado estudo palinológico, da anatomia foliar e taxonômico, promovendo a delimitação das espécies dos gêneros Elephantopus (10 espécies) Orthopappus (uma espécie) e Pseudelephantopus (duas espécies)além de oferecer subsídios para posteriores análises filogenéticas. O material botânico utilizado foi obtido através de exsicatas depositadas nos herbários brasileiros e de material coletado em campo. Os grãos de pólen foram tratados pelo método acetolítico, sendo posteriormente mensurados, descritos, e analisados sob microscopia de luz e eletrônica de varredura. As eletromicrografias foram obtidas de grãos de pólen não acetolisados, as análises taxonômicas baseiam-se na metodologia clássica. A anatomia foliar deu-se através da metodologia usual. Os resultados mostram grãos de pólen médios, 3-porados, de exina equinolofada com malhas poligonais organizadas ou não em Elephantopus e Pseudelephantopus, podendo apresentar interrupção na malha poral em Elephantopus. Em Orthopapus os grãos de pólen são 3-colporados, de sexina subequinolofada. A anatomia foliar possibilitou a separação de E. hirtiflorus e E. riparius através da forma ou contorno da nervura principal (planoconvexo), nas demais espécies o contorno é biconvexo. As espécies apresentaram tricomas capitados bisseriados com 8 células, tricomas filamentosos 3-4 celulares, bem como filamentosos de célula distal globosa ou ovóide em E. biflorus, E. micropappus, E. tomentous e Orthopappus angustifolius. Registrou-se variação na ornamentação da cutícula podendo ser lisa ou estriada entre as espécies, bem como a presença de substâncias pécticas na epiderme e drusas nos parênquimas. Em relação aos microcaracteres florais, foram considerados de importância diagnóstica os lóbulos da corola, que variaram entre glandular, glabro e penicilado, este ultimo apenas em E. hirtiflorus, os ápices das anteras que variaram entre obtuso, retuso em E. hirtiflorus e apiculado, lancelolado em E. mollis e E. racemosus, a base da antera variou entre sagitada, caudada lisa em E. riparius e E. tomentosus e caudada em E. riparius. Os macrocaracteres que apresentaram maior valor taxonômico foram a cipsela e o papus, a organização das coflorescências, o número de flores por capítulo e limbo foliar. Neste trabalho aceitou-se a segregação dos três gêneros, com base nos caracteres analisados

大赖草总DNA转化小麦引起的HMW-GS基因变化及大赖草HMW-GS基因的克隆

应用花粉管通道技术将新疆大赖草总DNA导入小麦，用高重序列分析方法，已为大赖草总DNA转入小麦提供了初步的分子证据。在转 化后代中选育出稳定遗传的大穗变异株系，分析表明，这些转化株中蛋白质含量明显增高(13%-17%)。对基因供体新疆大赖草、受 体春麦761、转化株的高分子量谷蛋白亚基(HMW-GS)进行了SDS-PAGE分析，发现这些转化株中HMW-GS发生了很大变化，并在此基础 上，用来自小麦基因组的四对特异引物，以PCR方法扩增供体、受体以及转化株的1Ax、1Bx、1Dx及1Ay、1By、1Dy型HMW-GS全基因 ，比较他们扩增产物的差异，结果表明，受体与转化株在HMW-GS基因1Ax、1Bx位点上的扩增产物差异不大，在HMW-GS基因位点1Dx 和y型基因上的扩增产物有较大差异，说明了受体在基因位点1Dx、1Ay、1By和1Dy上可能发生了多位点插入，可能由于这些基因位 点上的插入引起了转化株的高分子量谷蛋白亚基(HMW-GS)的变化，这就再一次为大赖草总DNA导入提供了直接的分子证据。虽然大 赖草总DNA导入提高了小麦蛋白质的含量，改变了HMW-GS的组成，部分改变了品质评分，但我们感到这些转化株在品质改良方面仍 有很大余地，如何更好地利用新疆大赖草蛋白质的优良特性及避免总DNA导入给转化株带来的不良性状，一个大赖草HMW-GS基因正 被分离及克隆，并准备将其利用于未来的品质育种当中。Total DNA of Leymus racemosus had been transformed into wheat through pollen tube pathway. Analysis of the repeated gene sequence had provided an elementary proof. Some variant cultivars with big ear were screened from their offsprings, and their protein content increased greatly from 13% (receptor)to 17%(transformed). The result from SDS-PAGE analysis of high-molecular-weight glutenin subunits(HMW- GS) respectively in donor(Xinjiang Leymus racemosus), receptor(spring wheat cultivar 761)and transformed wheats, showed the HMW-GS composition changed in the transformed plants. On the basis of the research, Four special pairs primers from wheat(T.aestivum L.) genome were used to amplify complete coding regions of HMW-GS genes on 1Ax、1Bx 、1Dx and 1Ay、1By、1Dy loci of donor、receptor and the big ear transformed cultivars. By comparing amplified PCR products. Faint differences were found among receptor and transformed cultivar's 1Ax、1Bx PCR amplifed products and apparent differences on those of 1Dx、y-typePCR product. We gueseed that there may be some DNA inserts in 1Dx 、1By、1Dy loci resulted in the changes of the HMW-GS among transformed cultivars. This provides second direct molecular witness to the exogenous DNA introduction. Even though the transformed plants have higher protein content, changed HMW-GS composition, partially improved process quality, there still leave much work to improve quality. In order to make full use of the excellent property of Leymus racemosus protein and avoid the disadvantages introducced by total DNA transformation, a HMW-GS gene of Leymus racemosus was being isolated and cloned.

Classificação de hortaliças.

1999

Hardware Software Interface: a Strategy for the implementation of Urban Agriculture

This paper describes how urban agriculture differs from conventional agriculture not only in the way it engages with the technologies of growing, but also in the choice of crop and the way these are brought to market. The authors propose a new model for understanding these new relationships, which is analogous to a systems view of information technology, namely Hardware-Software- Interface.
The first component of the system is hardware. This is the technological component of the agricultural system. Technology is often thought of as equipment, but its linguistic roots are in ‘technis’ which means ‘know how’. Urban agriculture has to engage new technologies, ones that deal with the scale of operation and its context which is different than rural agriculture. Often the scale is very small, and soils are polluted. There this technology in agriculture could be technical such as aquaponic systems, or could be soil-based agriculture such as allotments, window-boxes, or permaculture. The choice of method does not necessarily determine the crop produced or its efficiency. This is linked to the biotic that is added to the hardware, which is seen as the ‘software’.
The software of the system are the ecological parts of the system. These produce the crop which may or may not be determined by the technology used. For example, a hydroponic system could produce a range of crops, or even fish or edible flowers. Software choice can be driven by ideological preferences such as permaculture, where companion planting is used to reduce disease and pests, or by economic factors such as the local market at a particular time of the year. The monetary value of the ‘software’ is determined by the market. Obviously small, locally produced crops are unlikely to compete against intensive products produced globally, however the value locally might be measured in different ways, and might be sold on a different market. This leads to the final part of the analogy - interface.
The interface is the link between the system and the consumer. In traditional agriculture, there is a tenuous link between the producer of asparagus in Peru and the consumer in Europe. In fact very little of the money spent by the consumer ever reaches the grower. Most of the money is spent on refrigeration, transport and profit for agents and supermarket chains. Local or hyper-local agriculture needs to bypass or circumvent these systems, and be connected more directly to the consumer. This is the interface. In hyper-localised systems effectiveness is often more important than efficiency, and direct links between producer and consumer create new economies.

Is GABA involved in regulating plant growth and development? /

Rapid and large accumulation of GABA (y-aminobutyric acid) in response to a number of plant stresses has been well documented. But the role(s) of GABA in plants is not well defined. In recent years, the possibility of GABA involvement in regulating plant growth and development has been raised. In the present study, this possibility was examined. First, to rapidly and accurately determine GABA levels in plant tissues, a spectrometric method for GABA determination was developed based on a commercially available enzyme Gabase. Seventy mM LaCb almost completely removed water-soluble pigments from plant tissues which greatly interfere with the absorbance reading at 340nm. Inactivation of GAD (glutamate decarboxylase) by immediately adding methanol to a frozen plant tissue powder was suggested to prevent GABA production during extraction. The recovery of GABA with this method was approximately 100%. Second, the relationship between GABA levels and hypocotyl elongation in soybean seedlings was analyzed using different approaches to regulate in vivo GABA levels and the elongation of hypocotyls. The following major observations were made. (1) Mechanical stimulation by stroking elevated GABA levels and concurrently induced a rapid and significant reduction in hypocotyl elongation. (2) External GABA was demonstrated to penetrate into the hypocotyls using '*C-GABA. Application of external GABA elevated in vivo GABA levels, but failed to inhibit hypocotyl elongation. (3) LaCla and blue light irradiation caused an inhibition in the elongation of dark-grown hypocotyls, whereas GABA levels were not significantly affected. (4) Ca^was suggested to be involved in the signal transduction pathway leading from mechanical stimulation to GABA production, as indicated by the ability of La'* to inhibit GABA production in stimulated hypocotyls. (5) Bicuculline, saclofen and baclofen (agonists and antagonists of GABA receptors in animals) had no effect on hypocotyl elongation. It might indicate that GABA-binding components which are structurally similar to animal GABA receptors and functionally capable of regulating plant growth may not exist in plants. Therefore, the conclusion was drawn that GABA alone is not sufficient to inhibit hypocotyl elongation. Third, chloride influx in isolated Asparagus cells was enhanced by lOmM GABA during a 3 hour incubation, but the effect was not specific for GABA. Chloride efflux was not influenced by GABA. Both influx and efflux of chloride were significantly inhibited by NPPB, a chloride channel blocker. These results suggest that GABA does not influence the activity of plant chloride channels.

The impact of the Multicolor Asian Lady Beetle (Harmonia axyridis) on wine quality /

The Impact of the Multicolor Asian Lady Beetle (Harmonia axyridis) on Niagara Wine Quality The possible influence of Harmonia axyridis (the Multicolored Asian Lady Beetle) on the sensory properties of wine was investigated. H. axyridis beetles were added to white and red grape musts at a rate of 0, 1 or 10 per L, and a trained panel evaluated the finished wines using flavor-profiling techniques. Significant modification of both wine aroma and flavor characteristics were observed in the 10 beetlelL treatments, with smaller effects noted at the 1 beetlelL rate. Vinification in the presence of H. axyridis gave higher intensity scores for peanut, bell pepper and asparagus aromas and flavors in the white wines, and peanut, asparagus/bell pepper, and earthy/herbaceous aromas and flavors in the red wines. In addition, sweet, acid and bitter tastes were affected in red wines, and a general trend of decreasing fruit and floral intensities with increasing beetle rate was observed in both white and red wines. 15 ngIL Isopropylmethoxypyrazine was added to control wines and sensory profiles similar to high beetle treatments were obtained, supporting the hypothesis that methoxypyrazines from beetles are implicated in the taint. A trained panel evaluated the treated wines after 10 months of aging using the same sensory methods described above. Sensory profiles were very similar. Fennenting in the presence of Harmonia Axyridis (HA) had little influence on the chemical composition of the ftnished wine. The notable exception IS Isopropylmethoxypyrazine content, which was assessed usmg GC-MS analysis and showed increased concentration with increasing beetle nwnber for both white and red wmes. The influence of potential remedial treatments on the sensory properties of white and red wines tainted by Harmonia axyridis were also investigated. Bentonite, activated charcoal, oak chips, de-odorized oak chips, and UV or light irradiation were applied to tainted wine, and these wines evaluated chemically and sensorially. Both white and red wines treated with oak chips had strong oak characteristics, which masked the Harmonia axyridis-associated aroma and flavour attributes. In red wine, asparagus/bell pepper characteristics were decreased by bentonite and charcoal treatments. Only activated charcoal significantly decreased methoxypyrazine levels and only in white wine.

GABA accumulation and the hypersensitive response in isolated mesophyll cells treated with the G protein activator mastoparan

GABA (y-amino butyric acid) is a non-protein amino acid synthesized through the a-decarboxylation of L-glutamate. This reaction is catalyzed by L-glutamate decarboxylase (EC 4.1.1.15), a cytosolic Ca2+/calmodulin-stimulated enzyme. The purpose of this study is to determine whether or not GABA accumulation is associated with the hypersensitive response of isolated Asparagus sprengeri mesophyll cells. The addition of 25 J.lM mastoparan, a G protein activator, to suspensions of isolated asparagus mesophyll cells significantly increased GABA synthesis and cell death. Cell death was assessed using Evan's blue dye and fluorescein diacetate tests for cell viability. In addition, mastoparan stimulated pH-dependent alkalinization of the external medium, and a rapid and large 02 consumption followed by a loss of photosynthetic activity. The rate of 02 consumption and the net decrease in 02 in the dark was enhanced by light. The inactive mastoparan analogue Mas17 was ineffective in stimulating GABA accumulation, medium alkalinization, 02 uptake and cell death. Accumulation of H202 in response tomastoparan was not detected, however, mastoparan caused the cell-dependent degradation of added H202. The pH dependence of mastoparan-stimulated alkalinization suggests cellular electrolyte leakage, while the consumption of 02 corresponds to the oxidative burst in which 02 at the cell surface is reduced to form various active oxygen species. The results are indicative of the "hypersensitive response" of plants to pathogen attack, namely, the death of cells in the locality of pathogen invasion. The data are compatible with a model in which mastoparan triggers G protein activity, subsequent intracellular signal transduction pathway/s, and the hypersensitive response. It is postulated that the physiological elicitation of the hypersensitive response involves G protein signal transduction. The synthesis of GABA during the hypersensitive response has not been documented previously; however the role/s of GABA synthesis in the hypersensitive response, if any, remain unclear.

The production of 4-adminobutyrate in response to treatments reducing cytosolic pH and the regulation of intracellular pH

GABA (4-aminobutyrate) is synthesized through the decarboxylation of LGlu- (L-Glu-+ H+ ---> GABA + C02), and compared to many free amino acids is present in high concentrations in plant cells. GABA levels rise rapidly and dramatically in response to varied stress conditions including anaerobiosis. Recent papers suggest that GABA production and associated H+ consumption are parts of a metabolic pH-stat mechanism which ameliorates the intracellular pH decline associated with anaerobiosis or other treatments. To test this hypothesis GABA production and efflux have been measured in isolated Asparagus sprengeri cells in response to three treatments which potentially cause intracellular acidification. Acid loads were imposed using 60 min of (i) anaerobiosis, (ii) H+/LGlu- cotransport, and (iii) treatment with permeant weak acids (butyric, acetic and propionic). Both intra- and extracellular GABA concentrations increased more than 100% after anaerobiosis, almost 1000% after H+/L-Glu- cotransport (light or dark) and almost 5000/0 after addition of 5 mM butyric acid at pH 5.0. HPLC analysis of amino acids indicates that as GABA concentrations increased in response to butyric acid addition, glutamate concentrations decreased. Time-course studies demonstrated that added butyric acid stimulates GABA production by 2800/0 within 15 seconds. A fluorescent determination of cytosolic pH indicates that addition of butyric or other weak acids resulted in a rapid reduction in cytosolic pH of 0.6 pH units. The half time for the response to butyric acid addition is 2.1 seconds, indicating that the decline in cytosolic pH is rapid enough to account for the rapid stimulation of GABA production. The acid load in response to butyric acid addition was assayed by measurements of 14C-butyric acid uptake. Calculations indicate that GABA production accounted for 45% of the imposed acid load. The biological significance of GABA efflux is not yet understood. The results support the original hypothesis suggesting a role for GABA production in cellular pH regulation.

Methoxypyrazine removal in grape juice: Development of a removal system using odorant and pheromone binding proteins coupled with bentonite fining

Multicoloured Asian Lady Beetles (MALB) and 7-spot Lady Beetles that infect vineyards can secrete alkyl-methoxypyrazines when they are processed with the grapes, resulting in wines containing a taint. The main methoxypyrazine associated with this taint is 3-isopropyl-2-methoxypyrazine (IPMP). The wines are described as having aroma and flavours of peanut butter, peanut shells, asparagus and earthy which collectively, have become known as “ladybug taint”. To date, there are no known fining agents used commercially added to juice or wine that are effective in removing this taint. The goal of this project was to use previously identified proteins with an ability to bind to methoxypyrazines at low pH, and subsequently develop a binding assay to test the ability of these proteins to bind to and remove methoxypyrazines from grape juice. The piglet odorant binding protein (plOBP) and mouse major urinary protein (mMUP) were identified, cloned and expressed in the Pichia pastoris expression system. Protein expression was induced using methanol and the proteins were subsequently purified from the induction media using anion exchange chromatography. The purified proteins were freeze-dried and rehydrated prior to use in the methoxypyrazine removal assay. The expression and purification system resulted in yields of approximately 78% of purified plOBP and 62% of purified mMUP from expression to rehydration. Purified protein values were 87 mg of purified plOPB per litre of induction media and 19 mg of purified mMUP per litre of induction medium. In order to test the ability of the protein to bind to the MPs, an MP removal assay was developed. In the assay, the purified protein is incubated with either IPMP or 3-isobutyl-2-methoxypyrazine (IBMP) for two hours in either buffer or grape juice. Bentonite is then used to capture the protein-MP complex and the bentonite-protein-MP complex is then removed from solution by filtration. Residual MP is measured in solution following the MP removal assay and compared to that in the starting solution by Gas Chromatography Mass Spectrometry (GC/MS). GC/MS results indicated that the mMUP was capable of removing IBMP and IPMP from 300 ng/L in buffer pH 4.0, buffer pH 3.5 and Riesling Juice pH 3.5 down to the limit of quantification of the instrument, which is 6ng/L and 2ng/L for IBMP and IPMP, respectively. The results for the plOBP showed that although it could remove some IBMP, it was only approximately 50-70 ng/L more than bentonite treatment followed by filtration, resulting in approximately 100 ng/L of the MPs being left in solution. pIOBP was not able to remove IPMP in buffer pH 3.5 using this system above that removed by bentonite alone. As well, the pIOBP was not able to remove any additional MPs from Chardonnay juice pH 3.5 above that already removed by the bentonite and filtration alone. The mouse MUP was shown to be a better candidate protein for removal of MPs from juice using this system.