Constitutive expression of a phenylalanine ammonia-lyase gene from Stylosanthes humilis in transgenic tobacco leads to enhanced disease resistance but impaired plant growth

Transgenic tobacco plants expressing a phenylalanine ammonia-lyase cDNA (ShPAL), isolated from Stylosanthes humilis, under the control of the 35S promoter of the cauliflower mosaic virus were produced to test the effect of high level PAL expression on disease resistance. The transgenic plants showed up to eightfold PAL activity and were slowed in growth and flowering relative to non-transgenic controls which have segregated out the transgene. The expression of the ShPAL transgene and elevated PAL levels were correlated and stably inherited. In T-1 and T-2 tobacco plants with increased PAL activity, lesion expansion was significantly reduced by up to 55% on stems inoculated with the Oomycete pathogen Phytophthora parasitica pv. nicotianae, Lesion area was significantly reduced by up to 50% on leaves inoculated with the fungal pathogen Cercospora nicotianae. This study provides further evidence that PAL has a role in plant defence. (C) 2002 Elsevier Science Ltd. All rights reserved.

Feeding ecology of a carnivorous bladderwort (Utricularia uliginosa, Lentibulariaceae)

The terrestrial carnivorous bladderwort, Utricularia uliginosa Vahl. (Lentibulariaceae) was studied to determine the species assemblage present in traps of these plants in situ across four sites over 15 months. The immediate soil environment was also sampled to determine the fauna present, and to compare the fauna present in traps with the fauna in the environment. The soil fauna consisted of 10 taxon types, which occupied either pelagic, epibenthic or interstitial microhabitats. All were found in traps of U. uliginosa, with the main prey being interstitial taxa followed by epibenthic and occasionally pelagic taxa. Numbers of individuals of the two most abundant soil taxa (nematodes, Elaphoidella) varied independently across the four sites over the 15 months of the study, as did numbers of Elaphoidella in the traps of U. uliginosa. Numbers of nematodes in the traps of U. uliginosa showed significant differences among sites, but not differences among times. Comparison of the trap fauna with the soil fauna revealed differences in relative abundance between soil samples and trap samples for two of the three taxa examined. There was an under-representation of nematodes in the traps relative to numbers in surrounding soil. There was an over-representation of the copepod Elaphoidella in the traps of U. uliginosa relative to numbers in soil at some of the times of sampling. Acarina were equally abundant in soil and trap samples. The patterns observed for Elaphoidella and nematodes may be due to selectivity in trapping by U. uliginosa, and/or differences in digestibility of the prey. Elaphoidella individuals were found to be attracted to U. uliginosa in a behavioural experiment. This may contribute to the over-representation of Elaphoidella in the traps of U. uliginosa in the field.

Molecular rates parallel diversification contrasts between carnivorous plant sister lineages

In the carnivorous plant family Lentibulariaceae, the bladderwort lineage (Utricularia and Genlisea) is substantially more species-rich and morphologically divergent than its sister lineage, the butterworts (Pinguicula). Bladderworts have a relaxed body plan that has permitted the evolution of terrestrial, epiphytic, and aquatic forms that capture prey in intricately designed suction bladders or corkscrew-shaped lobster-pot traps. In contrast, the flypaper-trapping butterworts maintain vegetative structures typical of angiosperms. We found that bladderwort genomes evolve significantly faster across seven loci (the trnL intron, the second trnL exon, the trnL-F intergenic spacer, the rps16 intron, rbcL, coxI, and 5.8S rDNA) representing all three genomic compartments. Generation time differences did not show a significant association. We relate these findings to the contested speciation rate hypothesis, which postulates a relationship between increased nucleotide substitution and increased cladogenesis. (C) 2002 The Willi Hennig Society.

Callus induction and plant regeneration from broccoli (Brassica oleracea var. italica) for transformation

We evaluated the efficiency of callus induction and plantlet regeneration from hypocotyl explants of broccoli (Brassica oleracea var. italica). The cultivars were ‘Marathon’, ‘Greenbelt’, and ‘Shogun’. Transformation success was not affected by the presence of tobacco feeder-cell layers on the culture media. The frequency of shoot regeneration was greater from 10-d-old hypocotyls than from 14-d-old hypocotyls. Both ‘Marathon’ and ‘Greenbelt’ had higher potentials for tissue regeneration than did ‘Shogun’. We found that for transformation selection, the optimum concentration was either 50 mg/L kanamycin or 100 mg/L genetkin.

Simulation of insect movement with respect to plant architecture and morphogenesis

Developments in computer and three dimensional (3D) digitiser technologies have made it possible to keep track of the broad range of data required to simulate an insect moving around or over the highly heterogeneous habitat of a plant's surface. Properties of plant parts vary within a complex canopy architecture, and insect damage can induce further changes that affect an animal's movements, development and likelihood of survival. Models of plant architectural development based on Lindenmayer systems (L-systems) serve as dynamic platforms for simulation of insect movement, providing ail explicit model of the developing 3D structure of a plant as well as allowing physiological processes associated with plant growth and responses to damage to be described and Simulated. Simple examples of the use of the L-system formalism to model insect movement, operating Lit different spatial scales-from insects foraging on an individual plant to insects flying around plants in a field-are presented. Such models can be used to explore questions about the consequences of changes in environmental architecture and configuration on host finding, exploitation and its population consequences. In effect this model is a 'virtual ecosystem' laboratory to address local as well as landscape-level questions pertinent to plant-insect interactions, taking plant architecture into account. (C) 2002 Elsevier Science B.V. All rights reserved.

Partial automation of database processing of simulation outputs from L-systems models of plant morphogenesis

Models of plant architecture allow us to explore how genotype environment interactions effect the development of plant phenotypes. Such models generate masses of data organised in complex hierarchies. This paper presents a generic system for creating and automatically populating a relational database from data generated by the widely used L-system approach to modelling plant morphogenesis. Techniques from compiler technology are applied to generate attributes (new fields) in the database, to simplify query development for the recursively-structured branching relationship. Use of biological terminology in an interactive query builder contributes towards making the system biologist-friendly. (C) 2002 Elsevier Science Ireland Ltd. All rights reserved.

Structure and function of plant toxins (with emphasis on cystine knot toxins)

Plant toxins are substances produced and secreted by plants to defend themselves against predators. In a broad sense, this includes all substances that have a toxic effect on targeted organisms, whether they are microbes, other plants, insects, or higher animals. Plant toxins have a diverse range of structures, from small organic molecules through to proteins. This review gives an overview of the various classes of plant toxins but focuses on an interesting class of protein-based plant toxins containing a cystine knot motif. This structural motif confers exceptional stability on proteins containing it and is associated with a wide range of biological activities. The biological activities and structural stability offer many potential applications in the pharmaceutical and agricultural fields. One particularly exciting prospect is in the use of protein-based plant toxins as molecular scaffolds for displaying pharmaceutically important bioactivities. Future applications of plant toxins are likely to involve genetic engineering techniques and molecular pharming approaches.

Zeolite/rock phosphate - a novel slow release phosphorus fertiliser for potted plant production

A glasshouse study was undertaken to determine if the zeolite mineral clinoptilolite from an Australian deposit in combination with rock phosphate (RP) could significantly enhance the uptake of P by sunflowers. The zeolite/RP combination was intended to act as an exchange-fertiliser, with Ca2+ exchanging onto the zeolite in response to plant uptake of nutrient cations (NH4+ or K) enhancing the dissolution of the RP. A reactive RP (Sechura) and a relatively non-reactive RP (Duchess) were examined. Zeolite was used in Ca2+-, K+- and NH4+-saturated forms at ratios of 3.5:1 and 7:1 with RP; Ca2+-zeolite was considered the control, with exchange-induced dissolution possible from K+-and NH4+-zeolite, The zeolite/RP mixture was applied as a vertical band adjacent to the sunflower seedling. In addition, N was supplied as urea in an effort to determine if RP dissolution resulted from H+ release by nitrification. Phosphorus supply from the zeolite/RP system was compared with an available P source (KH2PO4). The experiment clearly demonstrated greatly enhanced plant uptake of P from RP when applied in combination with NH4-zeolite, though the P uptake was lower than that from the soluble P source. The zeolite/RP interaction was much more effective with the reactive R-P than the non-reactive material, Within the NH4+-zeolite/RP band, root proliferation was greatly increased, as would be expected in an exchange-fertiliser system. The K+-zeolite system did not produce a significantly greater yield than the Ca2+-zeolite control, probably because adequate K+ supply from the basal application reduced uptake within the zeolite/RP band, thus reducing the extent of exchange-induced dissolution. Nevertheless, increased root proliferation within the band was observed, implying that exchange-induced dissolution may also be possible from this system. The zeolite/RP system offers the considerable advantage of P release in response to plant demand and is unique in this regard. (C) 2002 Elsevier Science B.V. All rights reserved.

Yield response of rice (Oryza sativa L.) genotypes to different types of drought under rainfed lowlands - Part 3. Plant factors contributing to drought resistance

A series of experiments were conducted in drought-prone northeast Thailand to examine the magnitude of yield responses of diverse genotypes to drought stress environments and to identify traits that may confer drought resistance to rainfed lowland rice. One hundred and twenty eight genotypes were grown under non-stress and four different types of drought stress conditions. Under severe drought conditions, the maintenance of PWP of genotypes played a significant role in determining final grain yield. Because of their smaller plant size (lower total dry matter at anthesis) genotypes that extracted less soil water during the early stages of the drought period, tended to maintain higher PWP and had a higher fertile panicle percentage, filled grain percentage and final grain yield than other genotypes. PWP was correlated with delay in flowering (r = -0.387) indicating that the latter could be used as a measure of water potential under stress. Genotypes with well-developed root systems extracted water too rapidly and experienced severe water stress at flowering. RPR which showed smaller coefficient of variation was more useful than root mass density in identifying genotypes with large root system. Under less severe and prolonged drought conditions, genotypes that could achieve higher plant dry matter at anthesis were desirable. They had less delay in flowering, higher grain yield and higher drought response index, indicating the importance of ability to grow during the prolonged stress period. Other shoot characters (osmotic potential, leaf temperature, leaf rolling, leaf death) had little effect on grain yield under different drought conditions. This was associated with a lack of genetic variation and difficulty in estimating trait values precisely. Under mild stress conditions (yield loss less than 50%), there was no significant relationship between the measured drought characters and grain yield. Under these mild drought conditions, yield is determined more by yield potential and phenotype than by drought resistant mechanisms per se. (C) 2002 Elsevier Science B.V. All rights reserved.

Impact numbers: measures of risk factor impact on the whole population from case-control and cohort studies

Objective: To describe new measures of risk from case-control and cohort studies, which are simple to understand and relate to numbers of the population at risk. Design: Theoretical development of new measures of risk. Setting: Review of literature and previously described measures. Main results: The new measures are: (1) the population impact number (PIN), the number of those in the whole population among whom one case is attributable to the exposure or risk factor (this is equivalent to the reciprocal of the population attributable risk),- (2) the case impact number (CIN) the number of people with the disease or outcome for whom one case will be attributable to the exposure or risk factor (this is equivalent to the reciprocal of the population attributable fraction); (3) the exposure impact number (EIN) the number of people with the exposure among whom one excess case is attributable to the exposure (this is equivalent to the reciprocal of the attributable risk); (4) the exposed cases impact number (ECIN) the number of exposed cases among whom one case is attributable to the exposure (this is equivalent to the reciprocal of the aetiological fraction). The impact number reflects the number of people in each population (the whole population, the cases, all those exposed, and the exposed cases) among whom one case is attributable to the particular risk factor. Conclusions: These new measures should help communicate the impact on a population, of estimates of risk derived from cohort or case-control studies.

Impact numbers in health policy decisions

Objective: To outline the major methodological issues appropriate to the use of the population impact number (PIN) and the disease impact number (DIN) in health policy decision making. Design: Review of literature and calculation of PIN and DIN statistics in different settings. Setting: Previously proposed extensions to the number needed to treat (NNT): the DIN and the PIN, which give a population perspective to this measure. Main results: The PIN and DIN allow us to compare the population impact of different interventions either within the same disease or in different diseases or conditions. The primary studies used for relative risk estimates should have outcomes, time periods and comparison groups that are congruent and relevant to the local setting. These need to be combined with local data on disease rates and population size. Depending on the particular problem, the target may be disease incidence or prevalence and the effects of interest may be either the incremental impact or the total impact of each intervention. For practical application, it will be important to use sensitivity analyses to determine plausible intervals for the impact numbers. Conclusions: Attention to various methodological issues will permit the DIN and PIN to be used to assist health policy makers assign a population perspective to measures of risk.

Using crop simulation to generate genotype by environment interaction effects for sorghum in water-limited environments

Multi-environment trials (METs) used to evaluate breeding lines vary in the number of years that they sample. We used a cropping systems model to simulate the target population of environments (TPE) for 6 locations over 108 years for 54 'near-isolines' of sorghum in north-eastern Australia. For a single reference genotype, each of 547 trials was clustered into 1 of 3 'drought environment types' (DETs) based on a seasonal water stress index. Within sequential METs of 2 years duration, the frequencies of these drought patterns often differed substantially from those derived for the entire TPE. This was reflected in variation in the mean yield of the reference genotype. For the TPE and for 2-year METs, restricted maximum likelihood methods were used to estimate components of genotypic and genotype by environment variance. These also varied substantially, although not in direct correlation with frequency of occurrence of different DETs over a 2-year period. Combined analysis over different numbers of seasons demonstrated the expected improvement in the correlation between MET estimates of genotype performance and the overall genotype averages as the number of seasons in the MET was increased.

Predicting plant leaf area production: shoot assimilate accumulation and partitioning, and leaf area ratio, are stable for a wide range of sorghum population densities

Predicting plant leaf area production is required for modelling carbon balance and tiller dynamics in plant canopies. Plant leaf area production can be studied using a framework based on radiation intercepted, radiation use efficiency (RUE) and leaf area ratio (LAR) (ratio of leaf area to net above-ground biomass). The objective of this study was to test this framework for predicting leaf area production of sorghum during vegetative development by examining the stability of the contributing components over a large range of plant density. Four densities, varying from 2 to 16 plants m(-2), were implemented in a field experiment. Plants were either allowed to tiller or were maintained as uniculm by systematic tiller removal. In all cases, intercepted radiation was recorded daily and leaf area and shoot dry matter partitioning were quantified weekly at individual culm level. Up to anthesis, a unique relationship applied between fraction of intercepted radiation and leaf area index, and between shoot dry weight accumulation and amount of intercepted radiation, regardless of plant density. Partitioning of shoot assimilate between leaf, stem and head was also common across treatments up to anthesis, at both plant and culm levels. The relationship with thermal time (TT) from emergence of specific leaf area (SLA) and LAR of tillering plants did not change with plant density. In contrast, SLA of uniculm plants was appreciably lower under low-density conditions at any given TT from emergence. This was interpreted as a consequence of assimilate surplus arising from the inability of the plant to compensate by increasing the leaf area a culm could produce. It is argued that the stability of the extinction coefficient, RUE and plant LAR of tillering plants observed in these conditions provides a reliable way to predict leaf area production regardless of plant density. Crown Copyright (C) 2002 Published by Elsevier Science B.V. All rights reserved.

A rapid PCR protocol for marker assisted detection of heterozygotes in segregating generations involving 1BL/1RS translocation and normal wheat lines

A rapid and reliable polymerase chain reaction (PCR)-based protocol was developed for detecting zygosity of the 1BL/1RS translocation in hexaploid wheat. The protocol involved a multiplex PCR with 2 pairs of oligonucleotide primers, rye-specific Ris-1 primers, and consensus 5S intergenic spacer (IGS) primers, and digestion of the PCR products with the restriction enzyme, MseI. A small piece of alkali-treated intact leaf tissue is used as a template for the PCR, thereby eliminating the necessity for DNA extraction. The test is simple, highly sensitive, and rapid compared with the other detection systems of 1BS1RS heterozygotes in hexaploid wheat. PCR results were confirmed with AFLP analyses. Diagnostic tests for 1BL/1RS translocation based on Sec-1-specific ELISA, screening for chromosome arm 1RS controlled rust resistance locus Yr9, and the PCR test differed in their ability to detect heterozygotes. The PCR test and rust test detected more heterozygotes than the ELISA test. The PCR test is being used to facilitate S1 family recurrent selection in the Germplasm Enhancement Program of the Australian Northern Wheat Improvement Program. A combination of the PCR zygosity test with other markers currently being implemented in the breeding program makes this test economical for 1BL/1RS characterisation of S1 families.