Strategies to obtain stable transgenic plants from non-embryogenic lines: complementation of the nn(1) mutation of the NORK gene in Medicago sativa MN1008

Strategies to introduce genes into non-embryogenic plants for complementation of a mutation are described and tested on tetraploid alfalfa (Medicago sativa). Genes conditioning embryogenic potential, a mutant phenotype, and a gene to complement the mutation can be combined using several different crossing and selection steps. In the successful strategy used here, the M. sativa genotype MnNC-1008(NN) carrying the recessive non-nodulating mutant allele nn(1) was crossed with the highly embryogenic alfalfa line Regen S and embryogenic hybrid individuals were identified from the F1 progeny. After transformation of these hybrids with the wild-type gene (NORK), an F2 generation segregating for the mutation and transgene were produced. Plants homozygous for the mutant allele and carrying the wild-type NORK transgene could form root nodules after inoculation with Sinorhizobium meliloti demonstrating successful complementation of the nn(1) mutation.

Summer dormancy in Dactylis glomerata L.: the influence of season of sowing and a simulated mid-summer storm on two contrasting cultivars

A series of trials to increase understanding of the summer dormancy trait in Dactylis glomerata was conducted. Autumn-sown reproductive and younger, spring-sown plants of 2 drought-resistant cultivars, contrasting for summer dormancy, were established and then tested in summer 2002 under long drought, drought + midsummer storm, or full irrigation. The autumn-sown reproductive plants of cv. Kasbah were summer dormant under all moisture regimes and exhibited the characteristic traits including growth cessation, rapid herbage senescence, and dehydration of surviving organs (-6.7MPa). Cultivar Kasbah used 8% less soil water over the summer and also began to rehydrate its leaf bases from conserved soil water before the drought broke. The non-dormant cv. Medly grew for 10 days longer under drought and whenever moisture was applied; Medly also responded to the storm with a decline in dehydrin expression in leaf bases, whereas no decline occurred in Kasbah, presumably because it remained dormant and therefore much drier. The irrigated, younger, spring-sown swards of cv. Kasbah had restrained growth and produced only about 25% of the herbage of cv. Medly. Drought reduced activity and growth of young plants of both cultivars, but whereas Medly regrew in response to the storm, cv. Kasbah did not, indicating that dormancy, although only partially expressed after spring sowing, was reinforced by summer drought. A longer drought in 2003 caused a 22% loss of the basal cover in cv. Medly, whereas Kasbah fully maintained its sward and therefore produced a higher post-drought autumn yield. This work confirms summer dormancy as a powerful trait for improving persistence over long, dry summers.

Summer dormancy in perennial temperate grasses

Background and Aims Dormancy has been extensively studied in plants which experience severe winter conditions but much less so in perennial herbaceous plants that must survive summer drought. This paper reviews the current knowledge on summer dormancy in both native and cultivated perennial temperate grasses originating from the Mediterranean Basin, and presents a unified terminology to describe this trait. Scope Under severe drought, it is difficult to separate the responses by which plants avoid and tolerate dehydration from those associated with the expression of summer dormancy. Consequently, this type of endogenous (endo-) dormancy can be tested only in plants that are not subjected to moisture deficit. Summer dormancy can be defined by four criteria, one of which is considered optional: (1) reduction or cessation of leaf production and expansion; (2) senescence of mature foliage; (3) dehydration of surviving organs; and (4, optional) formation of resting organs. The proposed terminology recognizes two levels of summer dormancy: (a) complete dormancy, when cessation of growth is associated with full senescence of foliage and induced dehydration of leaf bases; and (b) incomplete dormancy, when leaf growth is partially inhibited and is associated with moderate levels of foliage senescence. Summer dormancy is expressed under increasing photoperiod and temperature. It is under hormonal control and usually associated with flowering and a reduction in metabolic activity in meristematic tissues. Dehydration tolerance and dormancy are independent phenomena and differ from the adaptations of resurrection plants. Conclusions Summer dormancy has been correlated with superior survival after severe and repeated summer drought in a large range of perennial grasses. In the face of increasing aridity, this trait could be used in the development of cultivars that are able to meet agronomic and environmental goals. It is therefore important to have a better understanding of the genetic and environmental control of summer dormancy.

Branching genes are conserved across species. Genes controlling a novel signal in pea are coregulated by other long-distance signals

Physiological and genetic studies with the ramosus (rms) mutants in garden pea (Pisum sativum) and more axillary shoots (max) mutants in Arabidopsis (Arabidopsis thaliana) have shown that shoot branching is regulated by a network of long-distance signals. Orthologous genes RMS1 and MAX4 control the synthesis of a novel graft-transmissible branching signal that may be a carotenoid derivative and acts as a branching inhibitor. In this study, we demonstrate further conservation of the branching control system by showing that MAX2 and MAX3 are orthologous to RMS4 and RMS5, respectively. This is consistent with the longstanding hypothesis that branching in pea is regulated by a novel long-distance signal produced by RMS1 and RMS5 and that RMS4 is implicated in the response to this signal. We examine RMS5 expression and show that it is more highly expressed relative to RMS1, but under similar transcriptional regulation as RMS1. Further expression studies support the hypothesis that RMS4 functions in shoot and rootstock and participates in the feedback regulation of RMS1 and RMS5 expression. This feedback involves a second novel long-distance signal that is lacking in rms2 mutants. RMS1 and RMS5 are also independently regulated by indole-3-acetic acid. RMS1, rather than RMS5, appears to be a key regulator of the branching inhibitor. This study presents new interactions between RMS genes and provides further evidence toward the ongoing elucidation of a model of axillary bud outgrowth in pea.

Plasma steroids and steroid-binding capacity in male semelparous dasyurid marsupials (Phascogale tapoatafa) that survive beyond the breeding season in captivity

The semelparous dasyurids display a unique life history, in that all males die within a few weeks of the completion of the breeding season. Studies of several semelparous species have revealed that the male die-off is stress-related, and accompanied by increased plasma androgen and cortisol levels and decreased corticosteroid binding capacity, resulting in suppression of immune and inflammatory responses. This study examines the endocrine profile of male brush-tailed phascogales (Phascogale tapoatafa) that survive beyond the breeding season in captivity. Plasma cortisol, corticosteroid binding globulin and albumin levels were monitored in both males and females and steroid partitioning calculated. Captive males surviving beyond the breeding season did not show the elevation in plasma cortisol and decrease in corticosteroid binding capacity reported in wild males. Plasma albumin concentrations also remained constant during the sampling period. These data indicate that captive males do not undergo the same stress response described in wild populations. (c) 2006 Elsevier Inc. All rights reserved.

Summer dormancy in Festuca arundinacea Schreb.; the influence of season of sowing and a simulated mid-summer storm on two contrasting cultivars

Due to the shortage of information on summer dormancy in tall fescue (Festuca arundinacea, syn. Lolium arundinaceum), we tested the response of 2 cultivars of differing dormancy expression and growth stage to a range of summer moisture conditions, including full irrigation, drought, and a simulated mid-summer storm and analysed whether traits associated with summer dormancy conferred better survival under severe field drought. Autumn-sown reproductive and younger, spring-sown plants of 2 cultivars, claimed to exhibit contrasting summer dormancy, were established and then tested in summer 2002 under either long drought, drought+ simulated mid-summer storm, or full irrigation. The autumn-sown reproductive plants of cv. Flecha exhibited traits that can be associated with partial summer dormancy since under summer irrigation they reduced aerial growth significantly and exhibited earlier herbage senescence. Moreover, cv. Flecha used 35% less soil water over the first summer. However, the water status of leaf bases of young vegetative tillers of both cultivars was similar under irrigation and also throughout most of the drought (leaf potential and water content maintained over -4MPa and at approx. 1 g H2O/g DM, respectively). The summer-active cv. Demeter did not stop leaf elongation even in drought and produced twice as much biomass as Flecha under irrigation. Cultivar Demeter responded to the simulated storm with a decline in dehydrin expression in leaf bases, whereas no decline occurred in Flecha, presumably because it remained partially dormant. The younger, spring-sown swards of both cultivars had similar biomass production under summer irrigation but whereas Demeter regrew in response to the simulated storm, cv. Flecha did not, indicating that dormancy, although only partially expressed, was reinforced by summer drought. In all trials, cv. Flecha out-yielded Demeter in autumn regrowth. In particular, the severe drought in 2003 caused a 25% loss of the basal cover in cv. Demeter, whereas Flecha fully maintained its sward allowing it to produce a higher post-drought autumn yield. This work links summer dormancy with higher persistence over long, dry summers.

Precipitation and Fire Effects on Flowering of a Rare Prairie Orchid

A small, isolated population of the threatened western prairie fringed orchid (Platanthera praeclara Sheviak & Bowles) occurs at Pipestone National Monument, Minnesota, in a mesic prairie that is periodically burned to control invasive cool-season grasses. During 1995-2004, monitoring counts of flowering orchids in the monument varied considerably for different years. Similar precipitation amounts in the spring and histories of burning suggest that fire and precipitation in the spring were not the causes of the variation. For the eight non-burn years in the monitoring record, we compared the number of flowering plants and the precipitation amounts during six growth stages of the orchid and found a 2-variab1e model (precipitation during senescence/bud development and precipitation in the dormant period) explained 77% of the annual variation in number of flowering plants. We also conducted a fire experiment in early May 2002, the typical prescribed burn period for the monument, and found that the frequency of flowering, vegetative, and absent plants observed in July did not differ between burned and protected locations of orchids. We used the model and forecasts of precipitation in the spring to develop provisional burn decision scenarios. We discussed management implications of the scenarios.

Roost tree characteristics determine use by the white-striped freetail bat (Tadarida australis, Chiroptera : Molossidae) in suburban subtropical Brisbane, Australia

We examined factors affecting roost tree selection by the white-striped freetail bat Tadarida australis (Chiroptera: Molossidae), a large insectivorous bat in suburban Brisbane, Australia. We compared biophysical characteristics associated with 34 roost trees and 170 control trees of similar diameter, height and tree senescence characters. Roost trees used by the white-striped freetail bat had significantly higher numbers of hollows in the trunk and branches (P < 0.003) and were more likely to contain a large trunk cavity with an internal diameter of > 30 cm (P < 0.001) than control trees. These trees also accommodated more species of hollow-using fauna (P = 0.005). When comparing roost trees with control trees of similar diameters and heights, roost trees were on average at a later stage of tree senescence (P < 0.001). None of the roost trees were found in the large forest reserves fringing the Brisbane metropolitan area despite these areas being used for foraging by the white-striped freetail bat. Although all tree locations in this study were in modified landscapes, roost trees tended to be surrounded by groups of trees and undergrowth. Roost trees provide important habitat requirements for hollow-using fauna in suburban, rural and forested environments.

The complexity of p53 stabilization and activation

A number of proteins are activated by stress stimuli but none so spectacularly or with the degree of complexity as the tumour suppressor p53 (human p53 gene or protein). Once stabilized, p53 is responsible for the transcriptional activation of a series of proteins involved in cell cycle control, apoptosis and senescence. This protein is present at low levels in resting cells but after exposure to DNA-damaging agents and other stress stimuli it is stabilized and activated by a series of post-translational modifications that free it from MDM2 (mouse double minute 2 but used interchangeably to denote human also), a ubiquination ligase that ubiquitinates it prior to proteasome degradation. The stability of p53 is also influenced by a series of other interacting proteins. In this review, we discuss the post-translational modifications to p53 in response to different stresses and the consequences of these changes.

Vase treatments containing gibberellic acid do not increase longevity of cut Grevillea 'Sylvia' inflorescences

The longevity of Grevillea 'Sylvia' inflorescences can be very short and is influenced by exposure to ethylene. Gibberellic acid has the potential to delay senescence in some cut flowers by acting as an anti-ethylene treatment. Gibberellic acid was therefore applied to Grevillea 'Sylvia' inflorescences in vase solutions to determine its effects on longevity. Treatments with gibberellic acid did not prolong the longevity of inflorescences or influence 1-aminocyclopropane-1-carboxylic acid concentrations. Treatments at high gibberellic acid concentrations enhanced flower abscission and we therefore conclude that vase-applied gibberellic acid treatments are not suitable for extending the longevity of cut Grevillea 'Sylvia' inflorescences.

Short root mutant of Lotus japonicus with a dramatically altered symbiotic phenotype

Legume plants carefully control the extent of nodulation in response to rhizobial infection. To examine the mechanism underlying this process we conducted a detailed analysis of the Lotus japonicus hypernodulating mutants, har1-1, 2 and 3 that define a new locus, HYPERNODULATION ABERRANT ROOT FORMATION (Har1), involved in root and symbiotic development. Mutations in the Har1 locus alter root architecture by inhibiting root elongation, diminishing root diameter and stimulating lateral root initiation. At the cellular level these developmental alterations are associated with changes in the position and duration of root cell growth and result in a premature differentiation of har1-1 mutant root. No significant differences between har1-1 mutant and wild-type plants were detected with respect to root growth responses to 1-aminocyclopropane1-carboxylic acid, the immediate precursor of ethylene, and auxin; however, cytokinin in the presence of AVG (aminoetoxyvinylglycine) was found to stimulate root elongation of the har1-1 mutant but not the wild-type. After inoculation with Mesorhizobium loti, the har1 mutant lines display an unusual hypernodulation (HNR) response, characterized by unrestricted nodulation (hypernodulation), and a concomitant drastic inhibition of root and shoot growth. These observations implicate a role for the Har1 locus in both symbiotic and non-symbiotic development of L. japonicus, and suggest that regulatory processes controlling nodule organogenesis and nodule number are integrated in an overall mechanism governing root growth and development.

Long-distance signaling in nodulation directed by a CLAVATA1-like receptor kinase

Proliferation of legume nodule primordia is controlled by shoot-root signaling known as autoregulation of nodulation (AON). Mutants defective in AON show supernodulation and increased numbers of lateral roots. Here, we demonstrate that AON in soybean is controlled by the receptor-like protein kinase GmNARK (Glycine max nodule autoregulation receptor kinase), similar to Arabidopsis CLAVATA1 (CLV1). Whereas CLV1 functions in a protein complex controlling stem cell proliferation by short-distance signaling in shoot apices, GmNARK expression in the leaf has a major role in long-distance communication with nodule and lateral root primordia.