Glomeromycotan mycorrhizal fungi from tropical Australia III. Measuring diversity in natural and disturbed habitats

Aims and Background

The aim was to investigate the diversity and distribution of Glomeromycotan fungi forming arbuscular mycorrhizal associations (AMF) in undisturbed and disturbed habitats in the vicinity of Kakadu National Park in tropical Australia. This is a tropical region with a 7–9 month dry season and a monsoonal wet season. Complimentary methods of fungus detection were used to investigate the diversity and relative dominance of AMF at a regional scale.

Methods

Soils were sampled from 32 sites, representing eucalypt savanna woodlands, wetlands, sandstone escarpment, rainforest, and disturbed mine waste rock dumps (overburden or spoil). Populations of AMF were identified and quantified using spores from soil. Morphology patterns of fungi colonising bait plant roots were examined and isolates were obtained by four complimentary pot-culturing methods.

Results

Different methods of detecting fungi produced different answers about which AMF were most important in the tested soils. In particular, spore surveys apparently underestimated the importance of Glomus species and overestimated the activity of Acaulospora species with numerous small spores, while calculated spore biovolumes overestimated the importance of Scutellospora and Gigaspora species with large spores, relative to inoculum levels of these fungus categories measured in bioassays. Spore surveys revealed 15 species of fungi and 8 additional fungi were recovered from the same soil samples using pot-culture isolation methods. Pot-cultures were especially important for detecting Glomus species that had high inoculum levels, but rarely produced spores in soils. Spores of AMF increased in abundance as vegetation developed in mine habitats reaching a peak that was higher than in undisturbed plant communities. Spore numbers (but not biovolumes) were well correlated with bioassay measurements of inoculum levels.

Conclusions

Most AMF species were widespread, but several were restricted to disturbed habitats or wetland soils. Undisturbed sites had a substantially higher diversity of AMF than partially vegetated mine waste rock dumps. It is recommended that AMF population surveys should not be based entirely on spore occurrence data, to avoid overlooking important fungi that sporulate infrequently. These fungi could be detected by bioassays or pot culture isolation from soil. Major variations in the detectability of AMF correspond to different life history strategies and can mask variations in their abundance.     

Tissue Culture Studies of Tomato (Lycopersicon esculentum)

Tomato is a major vegetable crop that has achieved tremendous popularity over the last century. It is grown in almost every country of the world. Development of protocols for in vitro selection can provide new advances for the production of stress tolerant cultivars. Techniques have been optimised for the production of haploids and somatic hybrids. Attempts have also been made to transfer the higher regenerative ability of wild varieties to cultivated tomatoes. Although, some information is available on the morphogenesis of tomato, the techniques have not been developed to a level at which they can be utilised in large-scale multiplication of commercially important cultivars. The morphogenesis response seems to be highly dependent PGRs used in the media, which is again cultivar and genotypic specific. Somatic embryogenesis in tomato is still at its infancy, and efficient procedures for large-scale production via somatic embryogenesis are yet to be developed. Genetic stability of the tissue culture raised tomato plants also needs to be addressed. The use of a combination of molecular and conventional breeding techniques could be the option for the development of cultivars resistant to biotic and abiotic stresses. This paper reviews the advances made in various aspects of tissue culture in tomato. It also discusses the issues that still need to be addressed to utilise the full potential of plant tissue culture techniques in genetic improvement and mass propagation of tomato.     

Ex Vitro Rooting of Micropropagated Shoots of Stackhousia Tryonii

Micropropagated shoots of Stackhousia tryonii were exposed (individually or in combination) to indole-3-acetic acid (IAA), indole-3-butyric acid (IBA), and 1-naphthalene acetic acid (NAA) at concentrations 1, 2 or 4 g dm^–3 with the view to induce rooting under ex vitro conditions. The treated microshoots were grown in a mist room for four weeks and assessed for survival, rooting percentage, number of roots and root length. The results showed that IBA at 2 g dm^–3 was most effective in inducing roots. Mixing of two or more auxins markedly reduced rooting percentage indicating antagonistic effects. The results demonstrated the potential of combining ex vitro rooting and hardening in one step, with view to reducing costs of multiplying plants via micropropagation.     

Bacterial Quorum Sensing Molecule N-3-Oxo-Dodecanoyl-L-Homoserine Lactone Causes Direct Cytotoxicity and Reduced Cell Motility in Human Pancreatic Carcinoma Cells

In spite of chemotherapeutic and surgical advances, pancreatic cancer continues to have a dismal prognosis. Metastasis due to tumor cell migration remains the most critical challenge in treating pancreatic cancer, and conventional chemotherapy is rarely curative. In the quest for more novel molecules to fight this disease, we tested the hypothesis that the Pseudomonas aeruginosa quorum sensing signal molecule N-3-oxo-dodecanoyl-L-homoserine lactone (O-DDHSL) would be cytotoxic to and reduce mobility of pancreatic carcinoma cells (Panc-1 and Aspc-1). Results showed a decrease in cell viability from apoptosis, diminished colony formation, and inhibition of migration of the evaluated pancreatic carcinoma cell lines. Also, cell viability decreased in the presence of O-DDHSL when cells were grown in matrigel basement membrane matrix. While messenger RNA for IQGAP-1 decreased in Panc-1 and HPDE cells upon exposure to O-DDHSL, no change was observed in Aspc-1 cells. Cofilin mRNA expression was found to be increased in both HPDE and Panc-1 cells with marginal decrease in Aspc-1 cells. RhoC, a Rho-family GTPase involved in cell motility, increased in the presence of O-DDHSL, suggesting a possible compensatory response to alteration in other migration associated genes. Our results indicate that O-DDHSL could be an effective biomolecule in eukaryotic systems with multimodal function for essential molecular targeting in pancreatic cancer.     

In vitro propagation of Stackhousia tryonii Bailey (Stackhousiaceae): a rare and serpentine-endemic species of central Queensland, Australia

Stackhousia tryonii Bailey, a rare species whichhyperaccumulates nickel and with a potential to be exploited inphytoremediation/phytomining, is difficult to propagate via seeds. This studyinvestigated the development of a micropropagation protocol for the productionof large stocks of S. tryonii. Disinfested shoot tips andnodal buds were precultured on Gamborg's (B₅) basal medium toobtain aseptic shoots for the optimisation of the protocol. 6-Benzyl aminopurine(BAP) at 1.0 mg l^–1 produced the highest number ofshoots per explant in B₅ medium. Comparison betweenB₅ and MS media showed similar responses, but with marked influenceof BAP concentration on shoot numbers. Transfer of shoots from MS(multiplication) medium to MS medium supplemented with indole-3-acetic acid(IAA) and indole-3-butyric acid (IBA), individually or in combination, indicatedthat a combination of IAA and IBA (0.75 mg l^–1each) is required to produce roots on young shoots (75%) compared to IBA(15–45%) or IAA (0–10%) alone. This study demonstrated that by usingthis protocol, a high multiplication rate (up to 18 shoots per explant) could be produced within 4 weeks, andthey can be readily hardened (98% survival) in a glasshouse by transplantingthem into a potting mixture of sand and perlite (4:1).     

Ascorbic acid prevents high glucose-induced apoptosis in human brain pericytes

High glucose concentrations due to diabetes increase apoptosis of vascular pericytes, impairing vascular regulation and weakening vessels, especially in brain and retina. We sought to determine whether vitamin C, or ascorbic acid, could prevent such high glucose-induced increases in pericyte apoptosis. Culture of human microvascular brain pericytes at 25 mM compared to 5 mM glucose increased apoptosis measured as the appearance of cleaved caspase 3. Loading the cells with ascorbate during culture decreased apoptosis, both at 5 and 25 mM glucose. High glucose-induced apoptosis was due largely to activation of the receptor for advanced glycation end products (RAGE), since it was prevented by specific RAGE inhibition. Culture of pericytes for 24 h with RAGE agonists also increased apoptosis, which was completely prevented by inclusion of 100 μM ascorbate. Ascorbate also prevented RAGE agonist-induced apoptosis measured as annexin V binding in human retinal pericytes, a cell type with relevance to diabetic retinopathy. RAGE agonists decreased intracellular ascorbate and GSH in brain pericytes. Despite this evidence of increased oxidative stress, ascorbate prevention of RAGE-induced apoptosis was not mimicked by several antioxidants. These results show that ascorbate prevents pericyte apoptosis due RAGE activation. Although RAGE activation decreases intracellular ascorbate and GSH, the prevention of apoptosis by ascorbate may involve effects beyond its function as an antioxidant.     

Model-based optimal PEEP in mechanically ventilated ARDS patients in the Intensive Care Unit

Background

Swallowing accelerometry has been suggested as a potential non-invasive tool for bedside dysphagia screening. Various vibratory signal features and complementary measurement modalities have been put forth in the literature for the potential discrimination between safe and unsafe swallowing. To date, automatic classification of swallowing accelerometry has exclusively involved a single-axis of vibration although a second axis is known to contain additional information about the nature of the swallow. Furthermore, the only published attempt at automatic classification in adult patients has been based on a small sample of swallowing vibrations.

Methods

In this paper, a large corpus of dual-axis accelerometric signals were collected from 30 older adults (aged 65.47 ± 13.4 years, 15 male) referred to videofluoroscopic examination on the suspicion of dysphagia. We invoked a reputation-based classifier combination to automatically categorize the dual-axis accelerometric signals into safe and unsafe swallows, as labeled via videofluoroscopic review. From these participants, a total of 224 swallowing samples were obtained, 164 of which were labeled as unsafe swallows (swallows where the bolus entered the airway) and 60 as safe swallows. Three separate support vector machine (SVM) classifiers and eight different features were selected for classification.

Results

With selected time, frequency and information theoretic features, the reputation-based algorithm distinguished between safe and unsafe swallowing with promising accuracy (80.48 ± 5.0%), high sensitivity (97.1 ± 2%) and modest specificity (64 ± 8.8%). Interpretation of the most discriminatory features revealed that in general, unsafe swallows had lower mean vibration amplitude and faster autocorrelation decay, suggestive of decreased hyoid excursion and compromised coordination, respectively. Further, owing to its performance-based weighting of component classifiers, the static reputation-based algorithm outperformed the democratic majority voting algorithm on this clinical data set.

Conclusion

Given its computational efficiency and high sensitivity, reputation-based classification of dual-axis accelerometry ought to be considered in future developments of a point-of-care swallow assessment where clinical informatics are desired.     

Tolerance of Australian tropical and subtropical Acacias to acid soil

Two glasshouse experiments were conducted to refine screening procedures and to rank 36 native Acacia species for acid-soil tolerance. The first experiment showed that nitrogen fixation was more sensitive to soil acidity than plant growth per se. This suggests that, in screening experiments where acid soil tolerant rhizobia are not available, inorganic nitrogen should be supplied in determining the species' intrinsic ability to tolerate soil acidity and to avoid confounding sensitivity to acid soil with ineffectiveness of Rhizobium/Bradyrhizobium. The second experiment enabled us to rank 36 species into 4 classes of differing tolerance to acid soil. This experiment also indicated that the ranking order of species differed with the soil type, suggesting the need to use more than one soil type in screening experiments in order to obtain a better and more reliable ranking of species for tolerance to acid soil.     

Comparative Performance of Micropropagated and Seed-Grown Tomato Plants

Morphological, physiological, fruit yield and quality related traits were compared between the seed-grown and tissue-cultured plants of tomato (Lycopersicon esculentum Mill.) cv. Red Coat in a greenhouse. No significant differences were observed for any of the traits studied except for the number of leaves and branches, which were higher in the seed-grown plants than in tissue-cultured plants at the later stages of growth. No phenotypic abnormality of the tissue-cultured plants was observed suggesting that genetic fidelity of tissue cultured plants can be maintained if appropriate plant growth regulators are used with fewer member of subcultures in the multiplication medium.     

Correlation between yield and biochemical parameters in the mulberry silkworm,Bombyx mori L

A detailed study was carried out on six biochemical parameters and four yield attributes using multiple regression analysis to investigate their relationship in the mulberry silkworm,Bombyx mori. The study generated new information on the importance of digestive amylase activity for the survival of the silkworm and revealed the inability of other enzymes to affect this relationship. Data also substantiate the observations made earlier on the genetic variability of amylase in the mulberry silkworm. Analyses extend the positive role of alkaline phosphatase and invertase in the expression of the other yield traits studied and indicate the definite possibility of using biochemical markers for silkworm breeding.