Characterization of two class II chitinase genes from peanut and expression studies in transgenic tobacco plants

Two different genes encoding class II chitinases from peanut (Arachis hypogaea L. cv. NC4), A.h.Chi2;1 and A.h.Chi2;2, have been cloned. In peanut cell suspension cultures, mRNA levels of A.h.Chi2;2 increased after ethylene or salicylate treatment and in the presence of conidia from Botrytis cinerea. The second gene, A.h.Chi2;1, was only expressed after treatment with the fungal spores. Transgenic tobacco plants containing the complete peanut A.h.Chi2;1 gene exhibited essentially the same expression pattern in leaves as observed in peanut cell cultures. Expression characteristics of transgenic tobacco carrying a promoter-GUS fusion of A.h.Chi2;1 are described.     

Genotypic variation in zinc efficiency and resistance to crown rot disease (Fusarium graminearum Schw. Group 1) in wheat

A crown rot disease in wheat caused by the fungusFusarium graminearum Schw. Group 1 is a widespread problem in chronically Zn-deficient Australian soils. A link between crown rot and Zn deficiency was established by Sparrow and Graham (1988). This paper reports a test of a further hypothesis, that wheat genotypes more efficient at extracting zinc from low-zinc soils are more resistant to infection by this pathogen. Three wheat cultivars (Excalibur, Songlen and Durati) of differential Zn efficiency were tested at three zinc levels (0.05, 0.5 and 2.0 mg Zn kg⁻¹ of soil) and three levels ofF. graminearum S. Group 1 inoculum (0.1 g and 0.3 g kg⁻¹ live chaff-inoculum and control having 0.1 g kg⁻¹ dead chaff inoculum). Six weeks after sowing dry matter production of shoots and roots was decreased byFusarium inoculation at 0.05 mg and 0.5 mg kg⁻¹ applied Zn.Fusarium inoculum at 0.1 g was as effective as 0.3 g kg⁻¹ for infection and decreasing dry matter. The infection at the basal part of culm decreased significantly by increasing the rate of Zn application. Excalibur, a Zn-efficient cultivar (tolerant to Zn deficiency) produced significantly more shoot and root dry matter, and showed less disease infection compared with Zn-inefficient cultivars (Durati and Songlen) at low (0.05 mg Zn kg⁻¹ soil) and medium (0.5 mg Zn kg⁻¹ soil) Zn fertilization rates. Higher rate of Zn fertilization (2.0 mg Zn kg⁻¹ soil) reduced the disease level in Durati to the level of Excalibur but the disease level of Songlen was still high, indicating its high Zn requirement and or sensitivity to crown rot. The data on Zn uptake show that Excalibur, being Zn-efficient, was able to scavenge enough Zn from Zn-deficient soil, we suggest that besides sustaining growth Excalibur was able to build and maintain resistance to the pathogen; inefficient cultivars needed extra Zn fertilization to achieve performance comparable to that of Excalibur. The present study indicates that growing Zn-efficient cultivars of wheat along with judicious use of Zn fertilizer in Zn-deficient areas where crown rot is a problem may sustain wheat production by reducing the severity of the disease as well as by increasing the plant vigour through improved Zn nutrition. ei]Section editor: R Rodriques-Kalana     

Suppression of fungal β-glucan-induced plant defence in soybean (Glycine max L.) by cyclic 1,3-1,6-β-glucans from the symbiont Bradyrhizobium japonicum

The production of antimicrobial phytoalexins is one of the best-known inducible defence responses following microbial infection of plants or treatment with elicitors. In the legume soybean (Glycine max L.), 1,3-1,6--glucans derived from the fungal pathogen Phytophthora sojae have been identified as potent elicitors of the synthesis of the phytoalexin, glyceollin. Recently it has been reported that during symbiotic interaction between soybean and the nitrogen-fixing bacterium Bradyrhizobium japonicum USDA 110 the bacteria synthesize cyclic 1,3-1,6--glucans. Here we demonstrate that both the fungal and the bacterial -glucans are ligands of -glucan-binding sites which are putative receptors for the elicitor signal compounds in soybean roots. Whereas the fungal -glucans stimulate phytoalexin synthesis at low concentrations, the bacterial cyclic 1,3-1,6--glucans appear to be inactive even at relatively high concentrations. Competition studies indicate that increasing concentrations of the bacterial 1,3-1,6--glucans progressively inhibit stimulation of phytoalexin synthesis in a bioassay induced by the fungal 1,3-1,6--glucans. Another type of cyclic -glucan, a 1,2--glucan from Rhizobium meliloti, that does not nodulate on soybean, seems to be inactive as elicitor and as ligand of the -glucan-binding sites. These results may indicate a novel mechanism for a successful plant-symbiont interaction by suppressing the plant's defence response.Abbreviations HG-APEA 1-[2-(4-aminophenyl)ethyl]amino-l-[hexaglucosyl]deoxyglucitol - HG-AzPEA l-[2-(4-azidophenyl)-ethyl]amino-l-[hexaglucosyl]deoxyglucitol - IC₅₀ concentration for half-maximal displacement We thank Ines Arlt for excellent technical assistance. This work was supported by the Deutsche Forschungsgemeinschaft (SFB 369), the Bundesministerium für Bildung, Wissenschaft, Forschung und Technologie, Fonds der Chemischen Industrie (J.E.), and USDA CSRS NRI Competitive Research grant 93373059233 (A.A.B.).     

Identification and characterization of bacterial endophytes of rice

We isolated seven different bacteria from rice seedlings grown from surface sterilized seeds. Three were associated with the rice seed husk and the other four were growing endophytically within the seed. Microscopic studies revealed that the endophytes were concentrated in the root stele region. Some of the bacteria exhibited strong anti-fungal activity against Rhizoctonia solani, Pythium myriotylum, Guamannomyces graminis and Heterobasidium annosum.     

Paracoccidioides brasiliensis antigen batches from the same isolate show immunological and biochemical differences

We investigated the occurrence of antigenic and biochemical variability among Paracoccidioides brasiliensis antigen batches prepared according to the same protocol. Initially (experiment #1), we analyzed two antigen lots of two human isolates (Bt1 & Bt2), cultured in two media (PYG: bactopeptone, yeast extract, glucose; MMM: McVeigh & Morton medium) in SDS-PAGE and in two immunological tests (imunodiffusion-ID and footpad swelling test-FPT). Afterwards (experiment #2), we compared the antigenic profile of three antigen batches from three human isolates (Bt1, Bt2 & Bt3) by two-dimensional immunoelectrophoresis (2 D-IEP) against a reference system for P. brasiliensis antigens. In experiment #1, there were important intra- and inter-strain antigenic differences between batches of the fungal isolates cultured on both media. The block titration of the antigen batches for the immunological tests revealed correlation between protein concentration and biological activity in ID and no correlation in FPT. In experiment #2, the reference system for P. brasiliensis showed 26 antigen peaks. There were important differences between batches prepared from the same isolate and between batches from different isolates. Our data suggested the occurrence of instability in the synthesis of antigenic components by a same P. brasiliensis isolate, under controlled incubation conditions.     

Tahiti's native flora endangered by the invasion of Miconia calvescens DC. (Melastomataceae)

Abstract. The native flora of tropical oceanic islands is known to be particularly susceptible both to displacement and extinction, following the invasion of alien organisms. Miconia calvescens DC. (Melastomataceae), first introduced to Tahiti (French Polynesia, South Pacific Ocean) in 1937 as an ornamental plant, now covers over two-thirds of the Island. As it forms dense monotypic stands which have progressively overwhelmed the native forests, this plant pest is a direct threat to the rich Tahitian indigenous flora. Between 40 and 50 species of the 107 species endemic to Tahiti are thought to be on the verge of extinction. M. calvescens was finally declared a'noxious species in French Polynesia'in 1990. Without efficient control efforts and effective endangered plant conservation and protection legislation, M. calvescens could cause Tahiti and all the high islands of French Polynesia to become ecological deserts.     

Fungal succession in the early decomposition process of pine cones on the floor ofPinus densiflora forests

The fungal succession on pine cones on the floor ofPinus densiflora forest was investigated in the early decomposition process (within ca. 30% decrease in dry weight). The fungal flora was examined by both washing and surface-sterilization methods on artificially placed cones and naturally fallen cones. The decomposition rates of artificially placed cones were 0.081–0.082 yr^–1. On withered cones still attached to the tree,Pestalotiopsis spp. were dominant. These fungi also occurred with higher frequencies after cones had lain on the floor and on cones in the L and FH horizons.Xylaria sp. andPhomopsis sp., which seem to colonize the interior of the tissue, occurred with higher frequencies on the cones on the tree, but their occurrence frequencies decreased after cones had lain on the forest floor. Conversely,Mortierella spp. andTrichoderma spp. newly occurred or their occurrence frequencies increased on lying cones. Of these,Trichoderma koningii increased rapidly and showed high occurrence frequencies.Thysanophora penicillioides, which prefers coniferous substrates, showed higher occurrence frequencies in the early stages of lying on the forest floor. On cones lying on the floor, the fungal flora did not significantly change during the investigation period.     

Drought response of a native and introduced Hawaiian grass

The alien grass, Pennisetum setaceum, dominates many of the lowland arid regions that once supported native Heteropogon contortus grassland on the island of Hawaii. Response to drought in a glasshouse was compared between these C₄ grasses to test if success as an invader is related to drought tolerance or plasticity for traits that confer drought tolerance. Pennisetum produced 51% more total biomass, allocated 49% more biomass to leaves, and had higher net photosynthetic rates (P _n) on a leaf area basis than Heteropogon. Plants of both species under drought produced less total biomass and increased their allocation to roots compared to well-watered plants, but there was no difference between the two species in the magnitude of these responses. The decline in P _n with decreasing leaf water potential (₁) was greater for Pennisetum compared to Heteropogon. Plasticity in the response of P _n to ₁, osmotic potentials, and the water potentials at turgor loss in response to drought were not different between the two species. Stomata were more responsive to w in Heteropogon than in Pennisetum and for well-watered plants compared to droughted plants. Plasticity for the stomatal response to w, however, was not different between the species. There was no evidence that the alien, Pennisetum, had greater plasticity for traits related to drought tolerance compared to the native, Heteropogon. Higher P _n and greater biomass allocation to leaves resulted in greater growth for Pennisetum compared to Heteropogon and may explain the success of Pennisetum as an invader of lowland arid zones on Hawaii.     

Preliminary caged‐field trial,using the fungal pathogen Zoophthora radicans brefeld (zygomycetes: entomophthorales) against the diamondback moth Plutella xylostella L. (lepidoptera: Yponomeutidae) in the UK

Zoophthora radicans Brefeld was tested for control of Plutella xylostella L. in a caged‐field trial. No infection was seen in control cages, but between 36% and 68% of larvae in live samples, and 38% and 55% of larvae at the final harvest, were infected in inoculated cages, suggesting the biological control potential of this fungus.     

The impact of hybrids between genetically modified crop plants and their related species: biological models and theoretical perspectives

Forces affecting the rate of spread and increase of hybrids between genetically modified crop plants and their related species remain qualitatively similar, irrespective of whether genetic modification was achieved using traditional methods, those of biotechnology or as a result of the natural evolutionary process. However, the precise magnitude of the forces and, consequently, the likely environmental impact of such hybrids, may depend strongly on the nature of the gene or genes introduced into the native species. While many classes of transgenes are similar to those manipulated by conventional breeding techniques or evolution, biotechnology offers the potential to introduce genes into crops which are novel both from the point of view of function and origin. The qualitative similarity between transgenes and the products of conventional or evolutionary modification suggests that a historical view of the environmental impact of hybrids between traditionally produced crops or exotic species and their relatives would be of use in estimating the probable fate of hybrids containing transgenes in the environment. However, with certain classes of transgenes for which there are no existing analogues, there will need to be greater care in assessing the possible risks associated with release into the environment.