Bioherbicides can be defined as plant pathogens, phytotoxins derived from pathogens or other microorganisms, augmentatively applied to control weeds. Although many pathogens with bioherbicidal potential have been discovered, most lack sufficient aggressiveness to overcome weed defenses to achieve adequate control. Plants use various physical and biochemical mechanisms to defend against pathogen infectivity, including callose deposition, hydroxyproline-rich glycoprotein accumulation, pathogenesis-related proteins (PR-proteins), phytoalexin production, lignin and phenolic formation, and free radical generation. Some herbicides, plant growth regulators, specific enzyme inhibitors, and other chemicals can alter these defenses. Various pathogens also produce chemical suppressors of plant defenses. Secondary plant metabolism is a major biochemical pathway related to several defense processes. Increased activity of a key enzyme of this pathway, phenylalanine ammonia-lyase (PAL), is often a response to pathogen attack, as demonstrated in two weeds and their associated bioherbicidal pathogens: Alternaria cassiae on sicklepod and A. crassa on jimsonweed. Weakening of physical and biochemical defenses, and lowering of resistance to pathogen attack, may result from reduced production of phenolics, lignin, and phytoalexins caused by herbicides and other chemicals that affect cuticular component biosynthesis and/or key aspects of secondary plant metabolism. Potent PAL inhibitors [aminooxyacetic acid, α-aminooxy-β-phenylpropionic acid, and (l-amino-2-phenylethyl)phosphonic acid] have some regulatory action on secondary plant metabolism and pathogenicity. Various herbicides and other chemicals dramatically affect extractable PAL activity levels and/or substantially alter PAL product production. Some non-pathogenic organisms can alter herbicide efficacy, and some herbicides influence disease development in plants. Research has shown some synergistic interactions of microbes and chemicals with relevance to weed control. Further research on pathogen interactions with agrochemicals (or other chemicals/regulators) could result in increased efficacy of pathogen-herbicide combinations, reduction of herbicide and pathogen levels required for weed control, and expanded pathogen host range.

Colletotrichum gloeosporioides f. sp. malvae (C.g.m.) was evaluated under greenhouse conditions for round-leaved mallow control in combination with the following herbicides: bromoxynil plus MCPA (1:1), diclofop, imazethapyr, metribuzin, sethoxydim, and 2,4-DB. Treatments consisted of C.g.m. and herbicide applied alone, tank mixed, and C.g.m. applied 48 h before, and 1, 24, and 72 h after herbicide. Split applications with bromoxynil plus MCPA, imazethapyr, metribuzin, (C.g.m. before or after) and sethoxydim (C.g.m. before) enhanced round-leaved mallow control over C.g.m. applied alone. However, diclofop or 2,4-DB tank mixed or applied in a split application with C.g.m. did not improve round-leaved mallow control over C.g.m. applied alone. Bromoxynil plus MCPA, metribuzin, 2,4-DB, and imazethapyr may be tank mixed with C.g.m. without significantly reducing round-leaved mallow control, unlike sethoxydim or diclofop which inhibited C.g.m. development. Dew period did not influence the effectiveness of bromoxynil plus MCPA applied 30 h before C.g.m., but round-leaved mallow control increased with increasing dew period for metribuzin applied 30 h before C.g.m.

Sodium alginate was used to prepare pelletized formulations for each of five fungi. Aqueous mixtures of 1.0% (w/v) sodium alginate and homogenized mycelia of Alternaria cassiae Jurair & Khan, Alternaria macrospora Zimm., Fusarium lateritium Nees ex Fr., Colletotrichum malvarum (A. Braun & Casp.) Southworth, or a Phyllosticta sp. were pelletized by dropwise additions of each mycelial-alginate mixture into 0.25 M CaCl2. Abundant conidia were produced on the pellets 24 to 48 h after the pellets were spread into trays and exposed 10 min/12 h to 275-W sunlamps. These conidia germinated readily (90 to 100%) and readily infected the respective host plants. Each liter of mycelium plus growth medium from submerged liquid cultures produced 4 L of the mycelial-alginate mixture. Each liter of the mycelial-alginate mixture produced approximately 18 g of air-dried formulation. When 10% (w/v) clay was incorporated into the pellets, each liter of the mycelial-alginate mixture produced approximately 118 g of air-dried formulation. The pelletized fungi sporulated readily following storage at 4 or 25 C for 6 to 8 months. This method of pelletization is potentially useful for the formulation of inoculum of fungi used as mycoherbicides, for the mass production of pycnidium-forming fungi, and for the production of inoculum for host-plant resistance studies.

Spraying with tank mix combinations of the fungus Colletotrichum coccodes and the plant growth regulator thidiazuron increased mortality, decreased biomass and height of velvetleaf, and increased soybean yield in field experiments. The velvetleaf isolate of C. coccodes was compatible with thidiazuron, and tank mix combinations consistently improved control over either component alone. In the moist 1986 season the fungus and the growth regulator interacted synergistically to increase velvetleaf mortality. In the drier 1987 season no significant interaction was detected between the two components.

Teliospores of Sphacelotheca holci Jack. [= S. cruenta (Kuhn.) Potter] infected johnsongrass [Sorghum halepense (L.) Pers. # SORHA) systemically after inoculation of cut stems in greenhouse studies. A preliminary host range test in the greenhouse revealed that no infection occurred on four varieties of sorghum (S. vulgare L.) and three cultivars of sudangrass (S. Sudanese L.). Infected johnsongrass plants grown in the greenhouse exhibited reduced vegetative growth and altered morphological characteristics compared to healthy johnsongrass plants. The aboveground biomass and lateral rhizome expansion of smutted johnsongrass field transplants were significantly lower than that of uninfected plants. Significant correlations between fraction of johnsongrass stems infected and plant weight, and basal area of plants were observed in field studies. Corn (Zea mays L.) and alfalfa (Medicago sativa L.) grown in competition with johnsongrass, and johnsongrass infected with S. holci reduced the aboveground biomass and rhizome length of johnsongrass plants significantly, compared to that of uninfected johnsongrass plants grown alone in greenhouse studies. Uninfected johnsongrass plants also exhibited higher plant weight than infected plants when both were grown in competition with alfalfa, regardless of the alfalfa planting density in the greenhouse or when measured after 3 months in field trials.

A method is described for the large-scale production of a granular formulation of Alternaria macrospora Zimm. The granular preparation consisted of spores, mycelia, and vermiculite. This formulation of A. macrospora, applied preemergence or postemergence, controlled spurred anoda (Anoda cristata Schlecht.) plants in greenhouse and field tests. The pathogen did not significantly affect emergence or dry weight production of cotton (Gossypium hirsutum L. ‘Stoneville 213’) seedlings. The fungicides PCNB (pentachloronitrobenzene) and ETMT [5-ethoxy-3-(trichloromethyl)-1,2,4-thiadiazole] did not significantly affect spurred anoda control with A. macrospora. The method for production and formulation of A. macrospora was modified to produce spores and granular formulations of other Fungi Imperfecti: Drechslera sp. and Fusarium spp. (Moniliales), Colletotrichum spp. (Melanconiales), and Phyllosticta sp. (Sphaeropsidales).

Field plot experiments were conducted to examine the interactions between tall fescue grass (Festuca arundinacea Schreb.), musk thistle (Carduus nutans L. = thoermeri Weinmann # CRUNU), and two thistle weevils Trichosirocalus horridus (Panzer) and Rhinocyllus conicus Froelich. Restriction of musk thistle growth was greatest when the weevils were allowed to feed on the musk thistles competing with tall fescue. Significant reductions were found in total musk thistle seeds per plant, root weight, flower buds per plant, stem dry weight, seeds per head, root crown diameter, stem height, rosette diameter, and head diameter. Seed weight and viability were not reduced. Of the three stress factors (tall fescue and the two Carduus thistle weevils), tall fescue had the greatest impact. When musk thistle seeds were planted in 1-yr-old tall fescue, germination was low. Seeds that germinated did not grow more than four Leaves and none of the thistles developed to the reproductive stage. A 1-year-old tall fescue stand effectively prevented musk thistle reproduction. Thus, tall fescue helped suppress musk thistle growth more quickly than the use of Carduus thistle weevils alone without competitive vegetation. Dry weight of tall fescue grass was lower in musk thistle-infested plots than in the thistle-free plots.

A method is described to produce inoculum of Alternaria cassiae Jurair and Khan for biocontrol studies. Approximately 8 g of a conidial preparation containing 1 × 108 conidia/g were produced per liter of growth medium. In greenhouse studies the effectiveness of A. cassiae as a biocontrol agent for sicklepod (Cassia obtusifolia L.) was affected by dew-period temperature, dew-period duration, inoculum concentration, and the stage of growth of sicklepod seedlings at the time of inoculation. Optimum environmental conditions included at least 8 h of free moisture at 20 to 30C. Spray mixtures containing approximately 5 × 104 or more conidia/ml gave maximum control when sicklepod seedlings were sprayed to runoff. Sicklepod seedlings in the cotyledon to first-leaf stage were most susceptible to the pathogen.

The fungus Colletotrichum coccodes (Wallr.) Hughes was isolated from diseased plants of eastern black nightshade (Solanum ptycanthum Dun. ♯ SOLPT) growing in the field. In greenhouse experiments, the isolate was highly pathogenic to eastern black nightshade and less so to seven other weedy species within the Solanaceae. No disease symptoms were observed on seedlings of tomato (Lycopersicon esculentum Mill.), potato (Solanum tuberosum L.), or 10 other species inoculated with the isolate. Dew periods of 16 h or longer were required for maximum disease development. This requirement could limit the practical use of this pathogen as a my coherbicide.

Greenhouse experiments were conducted to evaluate the effect of the fungus Dactylaria higginsii on the interference of Cyperus rotundus (purple nutsedge) with Lycopersicon esculentum (tomato ‘Agroset'). Cyperus rotundus plants established from tubers were planted at initial densities equivalent to 40, 80, 160, and 320 tubers m−2 with L. esculentum in 35-cm-diam pots. Cyperus rotundus plants were inoculated by spraying with a spore suspension of D. higginsii containing 0.5% Metamucil as a carrier. The treatments were Metamucil only, 104 conidia ml−1 + Metamucil, or 106 conidia ml−1 + Metamucil. In the absence of D. higginsii, C. rotundus at densities of 80, 160, and 320 tubers m−2 reduced L. esculentum yield by 14, 68, and 70%, respectively. In contrast, yield of L. esculentum treated with D. higginsii at 106 conidia ml−1 was equal to that in the weed-free control. The rate of disease increase (r G) was higher in treatments with 106 conidia ml−1 (r G = 0.126 to 0.136) compared to 104 conidia ml−1 (r G = 0.046 to 0.050). At 106 conidia ml−1D. higginsii reduced the interference from C. rotundus, controlled C. rotundus better, and increased L. esculentum yield compared to the weedy checks.

In four tests conducted in 2 years, Alternaria cassiae Jurair and Khan effectively controlled sicklepod (Cassia obtusifolia L. ♯ CASOB) as indicated by stand counts of the weed and by soybean [Glycine max (L.) Merr.] yields. The number of sicklepod seedlings was reduced more than 90% within 14 days after foliar applications of A. cassiae conidia were banded (46 cm) at rates of 4.2 and 2.1 kg/ha (9.4 and 4.7 kg/ha broadcast, respectively). One banded application of 0.04 or 0.4 kg/ha (0.09 or 0.9 kg/ha broadcast, respectively) did not satisfactorily control sicklepod. These results indicate that sicklepod control may be enhanced by multiple applications of A. cassiae.

Puccinia obtegens (Link) Tul., an autoecious rust pathogen, is a potential biological control agent of Canada thistle [Cirsium arvense (L.) Scop.]. Ten ecotypes of Canada thistle were inoculated with uredospores of P. obtegens and sporulation was observed on all ecotypes. Infection types varied among and within ecotypes, indicating that host-resistance is one factor limiting rust infection. No correlation was found between Canada thistle susceptibility to the rust and host plant ecotype classification, stomatal density, amount of leaf pubescence, or spore germinability on leaf surfaces.

The forage selection and grazing behavior of sheep in leafy spurge (Euphorbia esula L. ♯3 EPHES)-infested pastures was examined in a 3-month grazing study conducted during the summer, 1981. No definite preference for or avoidance of leafy spurge was detected. An initial 1- to 3-week avoidance was observed, followed by a steady increase in leafy spurge consumption. A maximum intake of 40 to 50% of the diet per animal was observed. Weight gain by ewes in pastures infested with leafy spurge was not significantly different from weight gain by ewes in pastures free of leafy spurge. Results from these studies indicate that leafy spurge may be classified as a forage for sheep. Sheep will consume a significant amount of vegetation while suffering neither harmful internal effects nor a significant loss of body weight. Sheep can be considered an effective biological control agent for leafy spurge.

In greenhouse studies, Fusarium lateritium Nees ex Fr. was effective in suppressing the growth of velvetleaf (Abutilon theophrasti Medic. # ABUTH) and prickly sida (Sida spinosa L. # SIDSP). Sprays containing conidia of the fungus and fungus-infested sodium alginate-kaolin clay granules controlled both weeds in corn (Zea mays L.), cotton (Gossypium hirsutum L.), and soybeans [Glycine max (L.) Merr.] without causing a reduction in dry weight of the crops. In field tests, postemergence foliar applications of the fungus controlled velvetleaf and prickly sida 40 and 80%, respectively, in 1982, but controlled only 27 and 34% of velvetleaf and prickly sida, respectively, in 1983. Preemergence applications of the fungus in the granule formulation controlled velvetleaf and prickly sida 46 and 59%, respectively, in 1982. In 1983, this treatment controlled the weeds 35 and 38%, respectively.

Plant growth, stem numbers per plant, receptacle diameters, number of Rhinocyllus conicus Froelich (Coleoptera:Curculionidae) larval cells per seed head, and mortality of the weevil within cells were monitored in infestations of the thistles Carduus thoermeri Weinmann and C. macrocephalus Deft. Both species of thistle provided similar seed head area for weevil development, and no differential effects on the biology of R. conicus were detected.

An isolate of Fusarium lateritium (Nees) emend. Snyder and Hansen was associated with naturally occurring disease of spurred anoda [Anoda cristata (L.) Schlecht.] and prickly sida (Sida spinosa L.) plants. In greenhouse inoculation studies, the Fusarium isolate was pathogenic to healthy spurred anoda and prickly sida plants. The disease symptoms were characterized by leaf, stem, and root lesions. The stem lesions enlarged with time and formed cankers that often girdled the stems and killed inoculated plants. In host-range studies, velvetleaf (Abutilon theophrasti Medic.), Venice mallow (Hibiscus trionum L.), okra [Abelmoschus esculentus (L.) Moench], and hollyhock [Althaea rosea (L.) Cav.] were also susceptible to the pathogen. This is the first report of these six species as hosts for this pathogen. Corn (Zea mays L.), cotton (Gossypium hirsutum L. and G. barbadense L.), soybean [Glycine max (L.) Merr.], and 18 other representative crop and weed species in eight families were resistant to the pathogen. F. lateritium may be a useful biological herbicide for susceptible malvaceous weeds.

Field research was conducted in Arkansas to determine the effect of selected rice pesticides on the microbial herbicide Colletotrichum gloeosporioides (Penz.) Sacc. f. sp. aeschynomene (henceforth designated C.g.a.) and to integrate C.g.a. into control programs for northern jointvetch [Aeschynomene virginica (L.) B.S.P. # AESVI] in rice (Oryza sativa L.). Water suspensions of dry-formulated spores of C.g.a. at 1.9 × 1011 spores/ha were applied midseason in sequence with normal use rates of other pesticides. Carbofuran (2,3-dihydro-2,2-dimethyl-7-benzofuranyl methyl-carbamate) at 0.56 kg ai/ha, propanil (3’4’-dichloropropio-anilide) at 2.2 kg ai/ha, acifluorfen {5-[2-chloro-4-(trifluoromethyl)phenoxy]-2-nitrobenzoic acid} at 0.14 kg ai/ha, or fentin hydroxide (triphenyltin hydroxide) at 0.56 kg ai/ha did not reduce disease development of C.g.a. on northern jointvetch. Sequential applications of benomyl [methyl 1-(butylcarbamoyl)-2-benzimidazolecarbamate] at 0.56 kg ai/ha reduced activity of the fungus on northern jointvetch when fungicide applications began 1 week after C.g.a. application but had no effect on mycoherbicide activity when applications began 2 weeks after C.g.a.

The combined influence of plant competition and defoliation on development of Canada thistle was investigated in a 2-yr field study. Plant competition was induced by seeding tall fescue and crown vetch. Artificial defoliation was used to simulate various levels of leaf removal by insects. Both defoliation and induced competition reduced biomass of Canada thistle but their impact varied with environmental conditions. Defoliation had a greater detrimental influence than induced competition on thistle biomass in a dry year when growth of the plant competitors was suppressed. In a wet year, induced competition was more important in suppressing Canada thistle than defoliation, and moderate levels of defoliation (25%), applied once when the thistles were 12 to 15 cm in diam, stimulated root weight within the top 20 cm of soil. Reduction in thistle biomass increased with level of defoliation and was greatest when defoliation was applied repeatedly at 14-d intervals in the presence of induced competition. Crown vetch showed very little growth in one season and tall fescue was the primary source of competition for the thistles. The results confirm the hypothesis that combined stresses can substantially reduce biomass development of Canada thistle plants.

Puccinia canaliculata (Schw.) Lagerh. from Salisbury, MD, identified in greenhouse tests as specific to yellow nutsedge, was field evaluated for biological control of yellow nutsedge over a 2-yr period. Treatments included all combinations of P. canaliculata and 0.3 or 0.6 kg ai/ha benzaton. Disease development was reduced 50% by treatment with bentazon, compared with P. canaliculata alone. P. canaliculata reduced tuber recovery by 44%. Phytotoxicity was significantly higher from the rust treatments than controls. Treatment with P. canaliculata did not affect the number of shoots/m2.

During 1992, the Research Committee of the Weed Science Society of America (WSSA) prepared a list of research needs for the discipline of weed science. The identified needs fit well with the recommendations of the Future Directions for Weed Science Symposium held in Washington D.C. on April 15, 1993, thus are provided as a supplement to the Symposium publication. The Research Committee concluded weed science research should focus on increasing knowledge related to the economic and biological impact of weeds across a wide variety of environments (crop production, forestry, right-of-way and aquatic). An increased understanding of the biology, ecology and genetics of weeds is needed to optimize their management. Research on control of weeds with biological agents and natural products should be conducted with emphasis on optimizing performance in the field environment. The development of weed populations resistant to chemical and biological control agents should be studied so the occurrence of such populations can be minimized. Because herbicides will continue to be the primary basis of weed management for the foreseeable future, research is needed to optimize herbicide application technology and to minimize their impact on the environment, particularly related to surface and groundwater contamination. Finally, new and reliable methods to detect herbicide residues in water, soil and vegetation are needed.