There are approximately 60 species in the genus Amaranthus, of which seven are used as grains, leafy vegetables, or ornamentals. The majority of the remaining species are considered important weeds. A new fungal species, Phomopsis amaranthicola, isolated from stem and leaf lesions on an Amaranthus sp. plant, was found to be pathogenic to 22 species of Amaranthus tested. The efficacy of this fungus was tested in field trials using one or two postemergent applications of the fungus consisting of two concentrations of conidia or mycelial suspensions. Species tested for susceptibility in the field included Amaranthus hybridus, A. lividus, A. viridus, A. spinosus, and a triazine-resistant A. hybridus. The cumulative disease incidence data for each treatment within each species were plotted versus time using regression for lifetime data. Plant mortality was recorded 2, 4, and 6 weeks after inoculation. There were significant differences between the treatment effects in the control plots versus the plots treated with P. amaranthicola. The highest level of control was obtained in the first trial when the fungus was applied at 6 × 107 conidia per ml. Final mortality of all species, except A. hybridus, reached 100% in inoculated plots 25 days earlier than in noninoculated control plots. Conidial suspensions were more effective in controlling the species in the second trial than were mycelial suspensions. Spread of the pathogen to noninoculated control plots was faster in the second year than in other years. High levels of plant mortality were achieved in plots of A. spinosus, A. lividus, and A. viridis. A. hybridus and the triazine-resistant A. hybridus were not effectively controlled in the second year due to the advanced stage of plant growth (8 to 10 true leaves) at the time of pathogen application. Results confirmed that P. amaranthicola is an effective biocontrol agent of some of the Amaranthus spp. tested.

Growth and sporulation of Dactylaria higginsii were quantified on complex agar media containing biological materials (group 1) and chemically defined agar media (group 2), as well as on grains, and the inoculum produced on these various substrates was tested for virulence on Cyperus rotundus. The fungus grew well between 25 and 30°C on potato dextrose agar (PDA), with 27°C being the optimum temperature. Generally, conidial production was highly variable and lower on complex agar media than on chemically defined media. Addition of purple nutsedge leaves to PDA did not increase colony growth or conidial production when compared with una-mended PDA. Conidial production was lowest on brown rice compared with white rice or white rice with nutsedge leaves. Peak production on grain media occurred from day 12 in test 1 (2.4 × 106 spores/g of grain) and on day 16 in test 2 (2.5 × 106 spores/g of grain). Germination rate of conidia produced on white rice was 50% compared with the near 100% germination of conidia produced on PDA or on white rice amended with potato dextrose broth (PDB). Conidia produced on white rice or PDA, when tested fresh or after two washings, were less virulent on C. rotundus than conidia from white rice amended with PDB. After four washings, conidia from all three media produced the same level of disease severity. White rice supplemented with PDB and PDA in trays were suitable for mass production of conidia of D. higginsii.

Eight polymers capable of forming aqueous gels were compared for their capacity to retain hydration over time, to promote spore germination, and to prolong the viability of germinated spores (= germlings) ofAlternaria cassiae, a bioherbicide agent for sicklepod. When compared at a standard 0.1% w/w (gel/water) concentration, the eight gels retained hydration for 6 d with no significant differences among them in the rate of dehydration. The best concentration of each gel that yielded 95 to 100% spore germination within 6 h after hydration was then chosen, and the gels were compared at these concentrations to determine the duration of effectiveness of the gels. The effectiveness was rated on the basis of the proportions of alive germlings versus germinated spores and alive germlings versus total spores, determined with the aid of a fluorescent vital stain. Based on these two parameters, the most effective gel was Kelzan® xanthan gum. However, all gels supported > 50% alive germlings over a period of 1 wk, suggesting that the addition of any of these polymers to the inoculum suspension should enable the fungal propagules to remain moist for a prolonged period, benefit from the high ambient moisture to improve germination, and promote disease development. Accordingly, seven of these gels were tested for their ability to enhance pathogenicity of a mycelial inoculum ofA. eichhorniae, a bioherbicide agent for waterhyacinth. Gellan gum and Kelgin®-HV were most effective in promoting disease, followed by Evergreen® 500 polyacrylamide, and Kelgin®-LV, Metamucil®, Kelzan® xanthan gum, and N-Gel™ were no better than the control inoculum without any gel. Thus, the gels may have differential effects on different fungi and inoculum types. Nonetheless, the results confirm the utility and feasibility of hydrophilic gels as formulating materials for bioherbicides.

We are studying the possibility of altering the virulence and host range of a phytopathogen by transferring and expressing certain genes from the soil-dwelling saprophyte, Streptomyces hygroscopicus, in a plant pathogen model, Xanthomonas campestris pv. campestris (XCC). The genes, referred to herein as the “bialaphos genes,” encode the production of bialaphos, a potent glutamine-synthetase-inhibiting herbicide. This cluster of genes was originally isolated from several biosynthetically blocked mutants of S. hygroscopicus and constructed into a plasmid vector, pBG9. We have transferred a fragment of the gene cluster into pLAFR3, a plasmid that functions in both Escherichia coli and XCC and contains a tetracycline resistance marker. The resulting plasmid, named pIL-1, was used to transform E. coli and was incorporated into XCC by conjugation. The transfer of the fragment was confirmed by Southern analysis. The genes were maintained in XCC for about 47 generations in the absence of selection for tetracycline, and no changes in cultural phenotypes were seen in the transformed XCC (XCC/pIL-1). The XCC/pIL-1 cells were pathogenic to their natural hosts cabbage and broccoli, but induced an altered hypersensitive response in the nonhosts bean, pepper, sunflower, and tobacco. The pathogenic host-reaction, induced by the parent XCC, XCC/pLAFR3, and XCC/pIL-1, was a typical black rot disease in inoculated leaves of the two hosts. The nonhost reaction on the nonhost leaves was necrotic hypersensitivity, induced by XCC and XCC/pLAFR3, or the inhibition of hypersensitivity accompanied by only chlorosis at sites inoculated with XCC/pIL-1. We hypothesize that the altered hypersensitivity phenotype may be due to the transformed XCC becoming more compatible with the nonhosts, a step toward acquiring nonhost-virulence, or due to disruption of the normal expression of the hypersensitivity and pathogenicity genes in the transformed XCC. More work is needed to confirm that the introduced genes are being expressed in XCC. With further understanding, this approach may provide a useful model to study host range, virulence, and strain improvement of plant pathogens for biological control of weeds.