Asparagus racemosus bZIP transcription factor-regulated squalene epoxidase (ArSQE) promotes germination and abiotic stress tolerance in transgenic tobacco.

A. racemosus is a rich source of pharmacologically active steroidal saponins. Most of the studies are related to its chemistry and pharmacology, but the pathway involved in the biosynthesis of steroidal saponin is not much emphasized. Squalene epoxidase acts as a rate-limiting enzyme in this biosynthesis. In this study, we have selected root specific squalene epoxidase ArSQE from A. racemosus for its characterization. ArSQE was able to complement ergosterol auxotrophy in erg1 yeast mutants. Mutants were sensitive to the antifungal drug terbinafine, whereas ArSQE complementation made them tolerant to the same drug. ArSQE plays a significant role in early germination in transgenic tobacco. The transgenic tobacco seedlings overexpressing ArSQE were tolerant to terbinafine and abiotic stress. Expression analysis of transcripts in ArSQE transgenic lines suggests that it mostly affects ABA, GA, stress, and sterol related functions in transgenic tobacco. Further, root specific MeJA responsive A. racemosus bZIP transcription factors (TFs), ArTGA1 and ArTGA2, were identified that bind to MeJA responsive cis-element present in the promoter region of ArSQE. Characterization of ArSQE of A. racemosus provides new information about its regulation through MeJA responsive bZIP TF along with its role in the development and abiotic stress response in transgenic tobacco.

Effect of a fungus, Hypoxylon spp., on endophytes in the roots of Asparagus.

The fungal isolate Hypoxylon spp. (Sj18) was isolated from the root of pecan. It might have effects on the plant's stress tolerance and endophytic community. Inoculation experiments were carried out on the roots of Asparagus with normal and inactivated Sj18, and the diversity and community structure of endophytes in the root of inoculated Asparagus were studied. It was found that Sj18 fungi affected the endophytic community of Asparagus roots. From being a low-abundance genus, the salt-tolerant bacterium Halomonas became the dominant genus. In order to verify that Sj18 can improve salt tolerance, Arabidopsis thaliana was inoculated with Sj18 in a salt tolerance test. The result showed that A. thaliana grew better in a high salt environment after inoculation with Sj18. Sj18 changed the microbe diversity, community composition and structure of endophytes in the roots of Asparagus, which increased the bacterial diversity. A total of 16 phyla and 184 genera of bacteria were detected. However, the diversity of fungi decreased.

Synergetic effects of 5-aminolevulinic acid and Ascophyllum nodosum seaweed extracts on Asparagus phenolics and stress related genes under saline irrigation.

Salinity is one of the major agricultural problems that may threat food security and limit the agricultural lands expansion worldwide. Exploring novel tools controlling saline conditions and increase valuable secondary metabolites in the horticultural crops might have outstanding results that serve humanity in the current century. The current study explores the effects of weekly seaweed extracts (7 mL L-1) and/or 5-aminolevulinic acid (3, 5 and 10 ppm) sprays on Asparagus aethiopicus plants subjected to saline stress conditions (2000 and 4000 ppm) for 6 weeks in two consecutive seasons of 2016 and 2017. Under saline conditions, there were stimulatory synergetic effects of seaweed extracts (SWE) and 5-aminolevulinic acid (ALA) on branch length and number of treated plants. Similar increases were also found in fresh and the dry weight of treated plants compared to control. These morphological improvements associated with increased accumulation of specific phenols (robinin, rutin, apigein, chlorogenic acid and caffeic acid) as revealed by High-Performance Liquid Chromatography with Diode-Array Detection (HPLC-DAD). There were increases in the antioxidant activities of leaf extracts, chlorophyll content and sugars and proline accumulation. The transpiration and photosynthetic rates as well as the stomatal conductance were enhanced. The morphological and physiological improvements associated with increased expression of several genes responsible for water management (ANN1, ANN2 and PIP1), secondary metabolite production (P5CS1 and CHS) and antioxidants accumulation (APX1 and GPX3) in plants. Our findings indicate that SWE + ALA had stimulatory synergetic effects on the growth and secondary metabolites of A. aethiopicus subjected to saline conditions. Several mechanisms are involved in such effects including gas exchange control, sugar buildup, increasing non-enzymatic and enzymatic antioxidants control of reactive oxygen species accumulation as well as transcriptional and metabolic regulation of environmental stress.

Sucrose delays senescence and preserves functional compounds in Asparagus officinalis L.

The high metabolic rate of harvested asparagus spears (Asparagus officinalis L.) causes rapid deterioration. To extend shelf life, we investigated the effect of sucrose treatment on asparagus during storage. Asparagus spears were treated with 3%, 5%, and 10% sucrose and stored at 2 °C for 20 h. Cellular respiration decreased, but other processes were unaltered by exogenous sucrose. The overall appearance of asparagus treated with 3% sucrose and stored at 2 °C for 18 days was rated as good and excellent, unlike that of untreated spears. Asparagus treated with sucrose maintained firmness for 15 days, while untreated spears lost firmness and showed increased water-soluble pectin content during storage. Carbohydrate levels were also higher in sucrose-treated than in control samples. Transcript levels of cell-wall-related genes, including xyloglucan endotransglycosylase (XET)1, XET2, and peroxidase (prx)1, prx2, and prx3 were upregulated by sucrose. Cyanidin 3-O-rutinoside and rutin levels immediately increased upon addition of sucrose and remained high relative to the control during storage. Thus, sucrose modulates asparagus cell wall components and maintains the functionality of important compounds during storage, thus effectively prolonging shelf life.

Evaluation of mercury stress in plants from the Almadén mining district by analysis of phytochelatins and their Hg complexes.

To evaluate plant response to Hg stress, glutathione, phytochelatins, and their Hg complexes were analyzed using HPLC with amperometric detection in samples of Asparagus acutifolius grown in the Almadén mining district (Ciudad Real, Spain), one of the most Hg-contaminated sites in the world. Soils of the Almadén mining district, and specifically from the Almadenejos zone, are highly contaminated, with some zones having values above 4,000 µg Hg g(-1) soil. Although soils have an extremely high concentration of mercury, generally less than 2% is available for plants, as is shown by various soil extractions simulating bioavailability. In plants, Hg concentration increases depending on the content of Hg in soils. In addition, Hg levels in roots are higher than in aerial parts, which is a strategy of plants for protecting their more sensitive aerial parts from the deleterious effects of metal stress. The total content of phytochelatins (PCs) and their complexes are directly related with the amount of mercury in soils. These findings highlight the important role of thiol compounds and their metal complexes in capturing and fixing Hg from soils, giving plants the capacity to deal with the heavy metal toxicity of polluted soils.

Micropropagation of Asparagus by in vitro shoot culture.

Asparagus officinalis is most extensively studied species within the genus Asparagus, which is well known as garden asparagus. This species is dioecious with unisexual flowers, which means that generative propagation gives roughly equal number of male and female plants. Male plants are high yielders and preferred commercially over female plants. Tissue culture techniques could efficiently promote vegetative propagation of male plants and pave the way for efficient plant breeding.This chapter describes an efficient micropropagation protocol for developing rapid growing in vitro Asparagus shoot cultures. The source of explants, inoculation, and shoot proliferation, followed by shoot propagation, rooting, and acclimatization is described. The optimal medium for Asparagus micropropagation described in this chapter is composed of MS macro- and microelements and a combination of auxins and cytokinins. Plant growth regulators NAA, kinetin, and BA were used in various concentrations. Three different media representing the whole micropropagation protocol of Asparagus are described; medium for shoot initiation, medium for shoot multiplication, and medium for root formation. By in vitro propagation of Asparagus, root initiation is difficult, but can be promoted by adding growth retardant ancymidol which also greatly promotes shoot development and suppresses callus formation.

The effect of plant growth-promoting rhizobacteria on asparagus seedlings and germinating seeds subjected to water stress under greenhouse conditions.

Plant growth-promoting rhizobacteria (PGPR) can have positive effects on vigour and productivity, especially under stress conditions. In asparagus (Asparagus officinalis L.) field culture, seeds are planted in high-density nurseries, and 1-year-old crowns are transplanted to production fields. Performance can be negatively affected by water stress, transplant shock, and disease pressure on wounded roots. PGPR inoculation has the potential to alleviate some of the stresses incurred in the production system. In this study, the effects of PGPR (Pseudomonas spp.) treatment were determined on 3-week-old greenhouse-grown seedlings and germinating seeds of 2 asparagus cultivars. The pots were irrigated to a predetermined level that resulted in optimum growth or the plants were subjected to drought or flooding stress for 8 weeks. The cultivars responded differently to PGPR: single inoculations of seedlings enhanced growth of 'Guelph Millennium' under optimum conditions and 'Jersey Giant' seedlings under drought stress. Seed inoculations with PGPR resulted in a positive response only for 'Guelph Millennium', for which both single or multiple inoculations enhanced plant growth under drought stress.

Host-plant finding by the asparagus fly, Plioreocepta poeciloptera (Diptera: Tephritidae), a monophagous, monovoltine tephritid.

The role of various olfactory and visual stimuli was studied in host-plant finding by the asparagus fly Plioreocepta poeciloptera (Schrank), a monophagous monovoltine tephritid causing serious damage to asparagus spears. Volatiles released by asparagus plants were extracted by diethyl ether after cryotrapping concentration, and identified by gas chromatography-mass spectrometry. Twelve of the 13 compounds identified were tested using electroantennography to measure the response of the fly. Behavioural response was analysed using two different flight tunnels according to circadian rhythm, age and sex of adults, presence of the plant and of different coloured lures, presence of a male congener, or exposure to four pure asparagus odour compounds that elicited responses in electroantennography, i.e. hexanal, (E)-2-hexenal, (Z)-2-hexen-1-ol and decanal. Data showed that males locate the host plant more quickly than females. Females are attracted mainly by the blend of plant odour and male pheromone. Both sexes respond to a complex of stimuli only during the afternoon. These findings will be helpful in developing new and effective approaches to control this pest insect.

Heritability and expected selection response for yield traits in blanched asparagus.

Despite the continuous breeding that has been conducted with asparagus (Asparagus officinalis L.) since the beginning of the last century, there is little information on parameters for predicting direct and indirect selection response. Yield traits for blanched asparagus production were studied along a two-year period in a half-sib family population planted in Zavalla, Argentina. Half-sib family mean heritability values were low for total yield and marketable spear number (0.31 and 0.35), intermediate for marketable yield and total spear number (0.55 and 0.64), and relatively high for spear diameter and spear weight (0.75 and 0.74). An average increase in marketable yield of 15.9% is expected after each cycle of selection of the top 5% of the families. Total yield failed to express significant genetic correlations with any of the yield components; meanwhile marketable yield showed highly significant relations with market spear number (0.96) and spear weight (0.89). Indirect selection response over yield components (CRx) failed to be advantageous over direct selection (Rx), since the ratio CRx/Rx was always equal or below unity.

Plant regeneration after long-term callus culture in clones of Asparagus officinalis L.

Callus growth and plant regeneration from long-term callus cultures were studied in two elite clones of Asparagus officinalis cv. Argenteuil, to establish a suitable protocol for a prospective in vitro selection program. Callus initiation and growth was evaluated on MS medium with 3% sucrose, 0.9% agar, 1 mg x l(-1) kinetin, and three levels of 2,4-D. The highest callus relative growth was obtained on medium with 1.5 mg x l(-1) 2,4-D and 1 mg x l(-1) kinetin. Shoot primordia (SP) induction from > 18-months-old calluses was evaluated on several media; the highest percentage of SP induction (89%) and average number of SP per callus (8.6) were obtained with clone "265" on MS medium with 5 mg x l(-1) 2iP, 1 mg x l(-1) IAA, 3% sucrose and 0.9% agar. The highest percentage of root induction (100%) was achieved with clone '265' on MS medium with 0.1 mg x l(-1) kinetin, 0.1 mg x l(-1) NAA, 1.32 mg x l(-1) ancymidol, 7% glucose and 0.8% agar. Important medium x genotype interactions were detected, pointing to the need of adjusting this and other in vitro protocols for specific asparagus genotypes.

Sex ratio of some long-lived dioecious plants in a sand dune area.

In dioecious plants the fraction of males among flowering plants in the field (the secondary sex ratio) is the result of the fraction of males in the seeds (the primary sex ratio) and the subsequent survival and age at first reproduction of the two genders. It has been assumed that survival and age at first reproduction are the main determinants of biased secondary sex ratio but, especially for long-lived perennials, few data are available. We address this issue for natural populations of four long-lived perennials in a dune area. In Asparagus officinale and Bryonia dioica, the secondary sex ratio was unbiased. In Salix repens the secondary sex ratio was female-biased (0.337). Hippophae rhamnoides populations were male-biased; the average sex ratio of flowering plants was 0.658, while the fraction of males varied between 0.39 near the sea to 0.84 at the inland side of the dunes. The primary sex ratio was estimated by germinating seeds and growing plants under favourable conditions with minimal mortality. In S. repens the primary sex ratio in seeds was variable among mother plants and was, on average, female-biased (0.289). This is close to the secondary sex ratio, suggesting that the female bias already originates in the seed stage. In Hippophae rhamnoides the primary sex ratio was slightly male-biased (0.564). We argue that in this species, apart from the primary sex ratio, higher mortality and a later age at first reproduction for females contribute to the strong male bias among flowering plants in the field.

Shrub spatial aggregation and consequences for reproductive success.

To link spatial patterns and ecological processes, we analysed the distribution of two shrub species (one large and dominant, the other smaller) and estimated the reproductive consequences of their distribution for the smaller species. We tested the significance of the spatial distribution pattern of the two shrubs by second-order bivariate point pattern analysis (Ripley's K function). Performance of Asparagus albus, the smaller shrub, was measured as (1) survival of transplanted seedlings in two contrasting habitats: patches of the dominant shrub (Ziziphus lotus), and open interspaces; and (2) reproductive output of plants naturally occurring in both habitats. The two species were significantly aggregated. Transplanted Asparagus albus seedlings had higher survival rates in patches than in the open. Plants produced more flowers, fruits, and showed a higher mass of seeds when living in aggregates than when isolated. The mechanisms responsible for this facilitative effect seem to be related to soil enrichment in patches. These results suggest that the spatial aggregation of species can be indicative of a positive interaction among them, directly affecting fitness of at least one of the species. Facilitation, by inducing variations in the reproductive performance may play a major role in the demography and dynamics of plant populations.

Estimates of heritability in a blanched asparagus population.

To estimate the heritability values of characters frequently used as selective criteria, 32 half-sib families obtained from selected plants of three populations of the asparagus variety Argenteüil were evaluated in a randomized complete block design. The following characters were measured: days to emergence of the first spear, number and diameter of spears, number of stalks, plant height and average weight. The values of realized heritability were estimated and were compared with those obtained by the parent-offspring regression method. Phenotypic correlation coefficients between the different variables were significant. The values of realized heritability for most of the variables were moderate to high (between 0.18 and 0.68), except for days to emergence; lower values were obtained by the regression method. As there was a high degree of heritability, additive genetic factors contributed significantly to the genetic variance, which would allow the selection of phenotypically superior plants for asparagus improvement projects.

Echo planar magnetic resonance imaging of anisotropic diffusion in asparagus stems.

MR diffusion-weighted imaging (DWI) uses the signal loss associated with the random thermal motion of water molecules in the presence of magnetic field gradients to derive a number of parameters that reflect the translational mobility of the water molecules in tissues. In highly organized but asymmetric structures, this mobility may be affected by the obstacles present and this in a direction-dependent way. Important examples of this are white brain matter and the stem of certain plants, both containing fibrous components where diffusion of water molecules across fibers is much more restricted than along the fibers. Diffusion that exhibits such directional dependence is said to be anisotropic, and diffusion tensor magnetic resonance imaging allows localized characterization of this behavior. Interpretation of the information obtained in terms of the underlying tissue structure is often hampered by the complexity of factors that can produce the observed behavior. A phantom that exhibits well-defined anisotropic diffusion and yields sufficient signal can help the experimental testing of the relevant methods and models. In this paper, we have used a phantom consisting of asparagus stems as a test object for assessing the value of the acquisition and postprocessing techniques commonly used in the clinic for this kind of investigation. Because of its strongly fibrous and cylindrically symmetric morphology, exhibiting a well-defined sub-classification of cells on the basis of size and shape, asparagus allows a relatively simple interpretation of the results obtained in the diffusion experiments. Our experiments show that the known structural information about the main cell types encountered correlates well with the behavior patterns of the diffusion parameters.