Isolation and characterization of novel cyclotides from Viola hederaceae - Solution structure and anti-HIV activity of vhl-1, a leaf-specific expressed cyclotide

Based on a newly established sequencing strategy featured by its efficiency, simplicity, and easy manipulation, the sequences of four novel cyclotides (macrocyclic knotted proteins) isolated from an Australian plant Viola hederaceae were determined. The three-dimensional solution structure of V. hederaceae leaf cyclotide-1 ( vhl-1), a leaf-specific expressed 31-residue cyclotide, has been determined using two-dimensional H-1 NMR spectroscopy. vhl-1 adopts a compact and well defined structure including a distorted triple-stranded β- sheet, a short 310 helical segment and several turns. It is stabilized by three disulfide bonds, which, together with backbone segments, form a cyclic cystine knot motif. The three-disulfide bonds are almost completely buried into the protein core, and the six cysteines contribute only 3.8% to the molecular surface. A pH titration experiment revealed that the folding of vhl-1 shows little pH dependence and allowed the pK(a) of 3.0 for Glu(3) and ∼ 5.0 for Glu(14) to be determined. Met(7) was found to be oxidized in the native form, consistent with the fact that its side chain protrudes into the solvent, occupying 7.5% of the molecular surface. vhl-1 shows anti-HIV activity with an EC50 value of 0.87 μ m.

Effect of seasonal variation and storage temperature on leaf chlorophyll fluorescence and vase life of cut roses

Low temperature injury (LTI) of roses (Rosa hybrida L.) is difficult to assess by visual observation. Relative chlorophyll fluorescence (CF; F-v/F-m) is a non-invasive technique that provides an index of stress effects on photosystem 11 (PS 11) activity. This instrumental technique allows determination of the photosynthetic efficiency of plant tissues containing chloroplasts, such as rose leaves. In the present study, pre- and Post-Storage measurements of F-v/F-m were carried out to assess LTI in 'First Red' and 'Akito' roses harvested year round. Relationships between the pre-harvest environment conditions of temperature, relative humidity and photon flux density (PFD), F-v/F-m, and, vase life duration after storage are reported. After harvest, roses were stored at 1, 5 and 10 degrees C for 10 days. Non-stored roses were the control treatment. F-v/F-m ratios were reduced following storage, suggesting LTI of roses. However, reductions in F-v/F-m were not closely correlated with reduced vase life duration and were seasonally dependent. Only during winter experiments was F-v/F-m of roses stored at 1 degrees C significantly (P <= 0.001) lower compared to F-v/F-m of non-stored control roses and roses stored at 5 and 10 degrees C. Thus, the fall of F-v/F-m was due to an interaction of growing season and storage at 1 degrees C. Vase lives of roses grown during winter were significantly (P <= 0.001) shorter compared to roses grown during summer. Length of vase life was intermediate for roses grown during autumn and spring. Because of the lack of correlation between F-v/F-m and post-storage vase life it is concluded that the CF parameter F-v/F-m is nota practical index for assessing LTI in cold-stored roses. Higher PFD and temperature in summer were positively and significantly correlated with maintenance of post-storage FvIF ratios and longer vase life. It is suggested that shorter vase lives and lower post-storage F-v/F-m values after storage at 1 degrees C are consequences of reduced photosynthesis and smaller carbohydrate pools in winter-harvested roses. (c) 2004 Elsevier B.V All rights reserved.

Effect of nitrogen supply on leaf appearance, leaf growth, leaf nitrogen economy and photosynthetic capacity in maize (Zea mays L.)

Leaf area growth and nitrogen concentration per unit leaf area, N-a (g m(-2) N) are two options plants can use to adapt to nitrogen limitation. Previous work indicated that potato (Solanum tuberosum L.) adapts the size of leaves to maintain Na and photosynthetic capacity per unit leaf area. This paper reports on the effect of N limitation on leaf area production and photosynthetic capacity in maize, a C4 cereal. Maize was grown in two experiments in pots in glasshouses with three (0.84-6.0 g N pot(-1)) and five rates (0.5-6.0 g pot(-1)) of N. Leaf tip and ligule appearance were monitored and final individual leaf area was determined. Changes with leaf age in leaf area, leaf N content and light-saturated photosynthetic capacity, P a,, were measured on two leaves per plant in each experiment. The final area of the largest leaf and total plant leaf area differed by 16 and 29% from the lowest to highest N supply, but leaf appearance rate and the duration of leaf expansion were unaffected. The N concentration of expanding leaves (N-a or %N in dry matter) differed by at least a factor 2 from the lowest to highest N supply. A hyperbolic function described the relation between P-max and N-a. The results confirm the 'maize strategy': leaf N content, photosynthetic capacity, and ultimately radiation use efficiency is more sensitive to nitrogen limitation than are leaf area expansion and light interception. The generality of the findings is discussed and it is suggested that at canopy level species showing the 'potato strategy' can be recognized from little effect of nitrogen supply on radiation use efficiency, while the reverse is true for species showing the 'maize strategy' for adaptation to N limitation. (c) 2004 Elsevier B.V. All rights reserved.

Ecology of Leptocoris Hahn (Hemiptera: Rhopalidae) Soapberry Bugs in Australia

Soapberry bugs are worldwide seed predators of plants in the family Sapindaceae. Australian sapinds are diverse and widespread, consisting of about 200 native trees and shrubs. This flora also includes two introduced environmental weeds, plus cultivated lychee (Litchi chinensis Sonn.), longan (Dimocarpus longan Lour.) and rambutan (Nephelium lappaceum L.). Accordingly, Australian soapberry bugs may be significant in ecology, conservation and agriculture. Here we provide the first account of their ecology. We find five species of Leptocoris Hahn in Australia, and list sapinds that do and do not serve as reproductive hosts. From museum and field records we map the continental distributions of the insects and primary hosts. Frequency of occupation varies among host species, and the number of hosts varies among the insects. In addition, differences in body size and beak length are related to host use. For example, the long-beaked Leptocoris tagalicus Burmeister is highly polyphagous in eastern rainforests, where it occurs on at least 10 native and non-native hosts. It aggregates on hosts with immature fruit and commences feeding before fruits dehisce. Most of its continental range, however, matches that of a single dryland tree, Atalaya hemiglauca F. Muell., which has comparatively unprotected seeds. The taxon includes a smaller and shorter-beaked form that is closely associated with Atalaya, and appears to be taxonomically distinct. The other widespread soapberry bug is the endemic Leptocoris mitellatus Bergroth. It too is short-beaked, and colonises hosts phenologically later than L. tagalicus, as seeds become more accessible in open capsules. Continentally its distribution is more southerly and corresponds mainly to that of Alectryon oleifolius Desf. Among all host species, the non-native environmental weeds Cardiospermum L. and Koelreuteria Laxm. are most consistently attacked, principally by L. tagalicus. These recent host shifts have biocontrol implications. In contrast, the sapinds planted as fruit crops appear to be less frequently used at present and mainly by the longer-beaked species.

Confirmation of the existence of alloparasitoids in nature - host relationships of an Australian Coccophagus species that parasitizes mealy bugs

Heteronomous hyperparasitoids are parasitic wasps with sex-related host relationships that are unique to a group of genera in the chalcidoid family Aphelinidae. Females are primary parasitoids of various sedentary bugs (mainly, scale insects, mealy bugs, and whiteflies). Males, in contrast, are hyperparasitic, and they frequently develop at the expense of female conspecifics. Alloparasitoids constitute a special category of heteronomous hyperparasitoids, for their males never develop through female conspecifics. The existence of alloparasitic host relationships and the utility of the category 'alloparasitoid' have both been questioned. Here, we present results that confirm the existence of the alloparasitic way of life among heteronomous aphelinids. We investigated an undescribed species of Coccophagus (Hymenoptera: Aphelinidae), an Australian parasitoid that attacks the introduced lantana mealy bug, Phenacoccus parvus Morrison (Homoptera: Pseudococcidae), in Queensland. A year-long field survey regularly returned large numbers of female Coccophagus spec. near gurneyi individuals from P. parvus (total n = 4212), but only few males (n = 11). Males emerged from samples only when the encyrtid parasitoid Anagyrus diversicornis (Howard) (Hymenoptera: Encyrtidae) was present in samples in relatively high numbers. Laboratory oviposition tests confirmed that A. diversicornis is a male host and showed that males do not develop at the expense of conspecific females. Other studies show that males are attracted in numbers to virgin females held in cages above mealy bug-infested Lantana montevidensis (Spreng.) Briq. (Verbenaceae) in the field, demonstrating that they are common in the population as a whole. This confirms that the males need hosts other than conspecific females and that their usual hosts are present outside of the lantana/P. parvus system. The implications of these results for developing a realistic classification of heteronomous host relationships are discussed.

Genotypic consistency in the expression of leaf water potential in rice (Oryza sativa L.)

Early work has shown variation in the grain yield of rice cultivars grown under water stress conditions to be associated with the plant water status, mainly with the maintenance of high leaf water potential (LWP) at flowering and grain filling stage. Considerable variation for LWP among rice varieties has been recorded. The present work was designed to investigate genotypic consistency in water potential within the plant and under canopy manipulation to vary plant water requirement. In a glasshouse experiment, with six rice genotypes, a consistent water potential gradient from stem base to leaf tip has been observed. Leaf tip water potential has been found as the minimum LWP that can be recorded at any time of stress. Genotypes with similar canopy size could maintain different levels of LWP under stress conditions. In a field experiment, with four selected lines, four canopy sizes and two canopy mixture treatments were introduced prior to the imposition of control, mild and severe water stress conditions. It was found that the line differences in LWP and relative water content (RWC) were expressed under both mild and severe stress conditions, regardless of canopy size, tiller number and whether they were mixed with another line with different capacity to maintain LWP. Although there were some differences among canopy size treatments for radiation interception in three water conditions, canopy manipulation (plant size) within a line did not affect the expression of LWP and hence genotypic variation in LWP was maintained. Under both glasshouse and field conditions, lines that maintained high LWP had larger xylem diameter and stem areas than those that had low LWP. The results indicated that the size of the vascular bundles could influence the maintenance of plant water relations under water deficit. (c) 2005 Elsevier B.V. All rights reserved.

Is the distribution of Fiji leaf gall in Australian sugarcane explained by variation in the vector Perkinsiella saccharicida?

Fiji leaf gall (FLG) is an important virally induced disease in Australian sugarcane. It is confined to southern canegrowing areas, despite its vector, the delphacid planthopper Perkinsiella saccharicida, occurring in all canegrowing areas of Queensland and New South Wales. This disparity between distributions could be a result of successful containment of the disease through quarantine and/or geographical barriers, or because northern Queensland populations of Perkinsiella may be poorer vectors of the disease. These hypotheses were first tested by investigating variation in the ITS2 region of the rDNA fragment among eastern Australian and overseas populations of Perkinsiella. The ITS2 sequences of the Western Australian P. thompsoni and the Fijian P. vitiensis were distinguishable from those of P. saccharicida and there was no significant variation among the 26P. saccharicida populations. Reciprocal crosses of a northern Queensland and a southern Queensland population of P. saccharicida were fertile, so they may well be conspecific. Single vector transmission experiments showed that a population of P. saccharicida from northern Queensland had a higher vector competency than either of two southern Queensland populations. The frequency of virus acquisition in the vector populations was demonstrated to be important in the vector competency of the planthopper. The proportion of infected vectors that transmitted the virus to plants was not significantly different among the populations tested. This study shows that the absence of FLG from northern Queensland is not due to a lack of vector competency of the northern population of P. saccharicida.

Nutrient deficiencies in greater yam and their effects on leaf nutrient concentrations

This paper describes the first systematic study of nutritional deficiencies of greater yam (Dioscorea alata). Yam plants (cv. 'Mahoa'a') were propagated from tuber discs and grown in nutrient solution, with nutrients supplied following a modified programmed nutrient-addition method. After an establishment period of four weeks, deficiencies of nitrogen (N), phosphorus (P), potassium (K), calcium (Ca), magnesium (Mg), sulfur (S), iron (Fe), boron (B), manganese (Mn), copper (Cu), zinc (Zn), and molybdenum (Mo) were induced by omitting the relevant nutrient from the solution. Foliar symptoms were recorded photographically. Notably, deficiencies of the mobile macronutrients failed to induce senescence of oldest leaves, while vine growth and younger leaves were affected. Leaf blades of the main stem were sampled in sequence and analyzed chemically, providing the distribution of each nutrient from youngest to oldest leaves in both adequately supplied and deficient plants. The nutrient-concentration profiles, together with the visible symptoms, indicated that little remobilization of mobile macronutrients had occurred. For both macro- and micronutrients, young leaves gave the best separation of nutrient concentrations between well-nourished and deficient plants.

Cutting management of multipurpose tree legumes: effects on green herbage production, leaf retention and water-use-efficiency during the dry season in Timor, Indonesia

Edible herbage production and water-use-efficiency of three tree legumes (Leucaena leucocephala cv. Tarramba, L. pallida x L. leucocephala (KX2) and Gliricidia sepium), cut at different times of the year (February, April, June and uncut) were compared in a semi-arid area of Timor Island, Indonesia. Cutting in the early and mid dry-season (April and June) resulted in higher total leaf production (P< 0.05) and water-use-efficiency (P< 0.05), than cutting late in the wet-season (February) or being left uncut. For the leucaena treatments removing leaf in the early to mid dry-season reduced transpiration, saving soil water for subsequent regrowth as evidenced by the higher relative water contents of leaves from these treatments. This cutting strategy can be applied to local farming conditions to increase the supply of feed for livestock during the dry season.

Towards developing reliable models of leaf area on grapevines (Vitis vinifera L.)

A field study in three vineyards in southern Queensland (Australia) was carried out to develop predictive models for individual leaf area and shoot leaf area of two cultivars (Cabernet Sauvignon and Shiraz) of grapevines (Vitis Vinifera L.). Digital image analysis was used to measure leaf vein length and leaf area. Stepwise regressions of untransformed and transformed models consisting of up to six predictor variables for leaf area and three predictor variables for shoot leaf area were carried out to obtain the most efficient models. High correlation coefficients were found for log10 transformed individual leaf and shoot leaf area models. The significance of predictor variables in the models varied across vineyards and cultivars, demonstrating the discontinuous and heterogeneous nature of vineyards. The application of this work in a grapevine modeling environment and in a dynamic vineyard management context are discussed. Sample sizes for quantification of individual leaf areas and areas of leaves on shoots are proposed based on target margins of errors of sampled data.