Detoxification of Benzoxazolinone Allelochemicals from Wheat by Gaeumannomyces graminis var. tritici, G. graminis var. graminis, G. graminis var. avenae, and Fusarium culmorum

The ability of phytopathogenic fungi to overcome the chemical defense barriers of their host plants is of great importance for fungal pathogenicity. We studied the role of cyclic hydroxamic acids and their related benzoxazolinones in plant interactions with pathogenic fungi. We identified species-dependent differences in the abilities of Gaeumannomyces graminis var. tritici, Gaeumannomyces graminis var. graminis, Gaeumannomyces graminis var. avenae, and Fusarium culmorum to detoxify these allelochemicals of gramineous plants. The G. graminis var. graminis isolate degraded benzoxazolin-2(3H)-one (BOA) and 6-methoxy-benzoxazolin-2(3H)-one (MBOA) more efficiently than did G. graminis var. tritici and G. graminis var. avenae. F. culmorum degraded BOA but not MBOA. N-(2-Hydroxyphenyl)-malonamic acid and N-(2-hydroxy-4-methoxyphenyl)-malonamic acid were the primary G. graminis var. graminis and G. graminis var. tritici metabolites of BOA and MBOA, respectively, as well as of the related cyclic hydroxamic acids. 2-Amino-3H-phenoxazin-3-one was identified as an additional G. graminis var. tritici metabolite of BOA. No metabolite accumulation was detected for G. graminis var. avenae and F. culmorum by high-pressure liquid chromatography. The mycelial growth of the pathogenic fungi was inhibited more by BOA and MBOA than by their related fungal metabolites. The tolerance of Gaeumannomyces spp. for benzoxazolinone compounds is correlated with their detoxification ability. The ability of Gaeumannomyces isolates to cause root rot symptoms in wheat (cultivars Rektor and Astron) parallels their potential to degrade wheat allelochemicals to nontoxic compounds.     

Occurrence of Ophiobolus graminis var. Avenae on wheat crops in the field

Ophiobolus graminis var. Avenae has been found on oat crops in ten counties in Scotland. It has also been shown to attack wheat crops in the same districts.     

Carbohydrate Uptake and Metabolism of Ophiobolus graminis

The carbohydrate nutrition of Ophiobolus graminis, the cause of the take-all disease of wheat, was investigated in growth and respiration experiments. In a synthetic medium, d-mannitol was the only carbohydrate of thirteen studied which the fungus could not use for growth. However, the fungus was found to take up mannitol by an active mechanism, which was stopped by 2,4-dinitrophenol. Di- and trisaccharides were hydrolyzed extracellularly, and the monosaccharides were assimilated at different rates.     

The effects of formalin, nabam, irrigation and nitrogen on Heterodera avenae Woll., Ophiobolus graminis Sacc. and the growth of spring wheat

In 1964, nabam (sodium ethylene bisdithiocarbamate), water and three amounts of nitrogen fertilizer were applied to spring wheat on soil treated and untreated with formalin. The experiment lasted for 3 years during which there were eight different formalin, no–formalin sequences. The nabam and irrigation treatments were discontinued when it was found they did not affect the principal pathogens present, the cereal cyst nematode Heterodera avenae and the take–all fungus Ophiobolus graminis. Formalin increased grain and straw yields in the year in which it was applied but led to increased H. avenae populations which adversely affected the succeeding crop. Formalin controlled O. graminis in the year it was applied except on land treated for the first time in 1966. H. avenae seemed to be the main check to growth until about June, and O. graminis later. At the end of the experiment, total grain yields and nematode soil populations were greatest in the plots treated with formalin each year and least in those never treated with formalin. Yield loss from either O. graminis or H. avenae alone could not be assessed because formalin usually controlled both during the season in which it was applied and both were present in untreated plots. However, in 1965, some comparisons of the effects of each pathogen were possible when one occurred in the presence of differing amounts of the other. A doubling of total grain yield over 3 years was accompanied by an eightfold increase in H. avenae in sequences of continuous formalin or formalin 1964 and 1965, whereas yield increases caused by extra nitrogen were not matched by such a big increase in H. avenae. This suggests that formalin might be affecting H. avenae through factors other than increased plant size and vigour, which in themselves would tend to encourage larger nematode populations. In the absence of formalin, H. avenae soil populations either fell or failed to increase.     

Control of Gaeumannomyces graminis Var. tritici in Wheat by Isolates of the G. graminis var. graminis/Phialophora sp. (Lobed Hyphopodia) Complex under Field Conditions

In an experiment carried out under field conditions, wheat inoculated with Gaeumanmyces graminis var. tritics had less take-all and yielded significantly more grain when simultaneously inoculated with G. graminis var. graminis and a lobed hyphopodiate phialophora sp. then when unprotected with these fungi.     

Host and environmental effects on post-penetration development of Puccinia graminis avenae and P. coronata avenae

After inoculation of Avena sterilis and the oat cultivars Algerian and Garry with Puccinia graminis avenae the time required for the eruption of pustules of the rust was markedly less at 30–35 than at 20–25 ^oC. In addition, the area of pustules and number of uredospores produced were significantly greater at 30–35 than at 20 ^oC. Peaks of uredospore production occurred between 12 and 22 days after inoculation. In comparable experiments, the time required for pustules of P. coronata avenae to erupt, and the size of pustules, were relatively insensitive to change of temperature, although weight of uredospores produced was greater at 20 than at 30 ^oC. Peaks of uredospore production occurred between 14 and 18 days after inoculation. Both rusts showed straight-line relationships between pustule area and number of uredospores produced. The percentage of infection foci that developed into pustules was similar with both rusts and on all the oat cultivars examined. Both rusts produced susceptible reaction types on all the hosts tested. Pustules of P. graminis avenae were smaller and fewer and generation time longer on cv. Garry than on cv. Algerian or Avena sterilis and the numbers of pustules per unit of inoculum of both rusts were greatest on Algerian, least on Garry. It is suggested that these quantitative differences in phases of the infection process contribute towards the ‘slow-rusting’ reaction of cv. Garry.     

Effect of Ophiobolus graminis infection on the growth of wheat and barley

Glasshouse experiments are reported, in which the development of young wheat and barley plants was examined following inoculation with Ophiobolus graminis (Sacc.) Sacc. The dry weight, leaf area, tiller number and water content of the shoot were reduced by infection. Reductions were equally severe in wheat and barley. The seminal root system of both was severely attacked and its growth retarded. Inoculated plants, however, translocated a greater proportion of their total assimilates to the root system and produced more adventitious roots than healthy plants. As a result there was an increase in both the number and proportion of healthy roots on these plants following the initial infection of their root systems. This effect was more pronounced in barley than in wheat. It is suggested that this may in part account for the reported relative tolerance of barley to take-all attack under field conditions.     

Host and environmental effects on the penetration of oats by Puccinia graminis avenae and Puccinia coronata avenae

The frequency of penetration from appressoria of Puccinia graminis avenae and P. coronata avenae varied among Avena species and between oat cultivars, although both rusts produced susceptible infection type pustules on the cultivars tested. Penetration on cv. Garry was significantly less than that on the Avena species (A. barbata, A.fatua and A. sterilis) studied and penetration of these Avena species was significantly less than on the cvs Algerian and Fulmark. When the rusts were allowed to develop into pustules on seedlings which had been inoculated with fixed amounts of inoculum, there was a direct relationship between number of pustules produced and penetration frequency. The effects of temperature, light and dew period on penetration from appressoria of ‘single race’ and ‘mixed race’ inocula was also studied on these cultivars and species. Penetration by P. graminis avenae was greatest at 30–35 °C and at light intensities of 5625 lux and above, whereas that by P. coronata avenae was greatest at 20 °C and was unaffected by artificial light intensities up to n 250 lux. Maximal penetration by P. graminis avenae and P. coronata avenae was observed after inoculated plants had been exposed to dew periods of 16 and 12 h respectively. Some penetration was observed after a dew period of 8 h. The time taken for each rust to attain maximum penetration varied from 36 to 52 h after inoculation, depending on the environment, and was usually less for P. coronata avenae than for P. graminis avenae.     

The effects of soil sterilants on the cereal cyst-nematode (Heterodera avenae Woll.), take-all (Ophiobolus graminis Sacc.) and yields of spring wheat and barley

In a 3-year field experiment on sandy loam at Woburn, methyl bromide, chloropicrin, D-D mixture, dazomet, formalin and mercury salts were applied in the first year only, or in the first and second years, before drilling spring wheat. In the third year, their residual effects on spring barley were measured. All sterilants except mercury decreased cereal cyst-nematode ( Heterodera avenae ) numbers and take-all ( Ophiobolus graminis ) incidence, and increased crop yields in the year they were applied. Dazomet gave the best control of H. avenae in the first year and controlled O. graminis best in both years of application. In the third year, O. graminis increased in all plots previously treated (except after two successive D-D treatments), but H. avenae increased only after formalin. Two formalin applications more than trebled the H. avenae egg count by the end of the third year and depressed the yield of barley. Two successive applications of chloropicrin gave the best nematode control.
Other fungus diseases, Fusarium foot rot, Pythium root rot, eye-spot ( Cercosporella herpotrichoides Fron.) and sharp eye spot ( Rhizoctonia solani Kühn), were uncommon and sterilants had no detectable effects on them. The largest aggregate straw yield (3-year total) was obtained from two applications of dazomet, but the best yields after treatment in the first year only were given by D-D and methyl bromide.     

A scanning electron microscope study of interactions between micro-organisms andGaeumannomyces graminis (Syn.Ophiobolus graminis) on wheat roots

Roots of wheat grown in unsterilized sand inoculated withGaeumannomyces graminis (Sacc.) von Arx and Olivier were examined by scanning electron microscopy. Healthy roots had a mucilaginous covering and were sparsely colonized by bacteria, but asG. graminis colonized the roots the mucilage disappeared and the numbers of bacteria on the surface increased. Lysis of the hyphae occurred, apparently caused by bacteria that colonized the hyphae. Inoculation of wheat in axenic culture with a strain ofPseudomonas fluorescens that was antagonistic toG. graminis in agar gave some protection against the pathogen; lysis of hyphae was observed where protection occurred.     

Effects of nitrogen and glucose on saprophytic survival of Ophiobolus graminis in buried straw

Calcium nitrate prolonged the saprophytic survival of Ophiobolus graminis (Sacc.) Sacc. in artificially colonized straws buried in soil in the laboratory, whether supplied to the soil (at 12·5 or 100 mg nitrogen/100 g soil) or to the straws before colonization (at 0·5 or 1·0 g nitrogen/100 g straw). Glucose (at 2·5 g/100 g soil, and at 10 g/100 g straw) shortened survival. When straws colonized in the presence of 0·5 g nitrogen/100 g straw were buried in soil supplied with 14·1 mg nitrogen/100 g soil, the level of soluble soil nitrogen reached equilibrium at 2–4mg/100g soil; this allowed rapid straw decomposition and, although the added soil nitrogen prolonged survival in straws that remained undecomposed, it also accelerated substrate exhaustion. Addition of 100 mg nitrogen/100 g soil was supra-optimal for survival: although some nitrogen was necessary for maximum survival, the equilibrium concentration of soluble nitrogen (24–56 mg/100 g soil) was high enough in this case to have an inhibitory effect in addition.     

Infection process of Gaeumannomyces graminis var. graminis on the roots and culms of rice

Crown sheath rot, caused by the ascomycete Gaeumannomyces graminis var. graminis that infects the root and the base of the culm of rice, causes early grains maturation, tiller death and reduced yield. As a paucity of information exists in the literature on the rice‐G. graminis var. graminis interaction at the microscopic level, this study aimed to gain novel insights into the infection process of this pathogen in the root and culm of rice using both light and scanning electron microscopy. In the roots, the fungus initially colonized the epidermal, exodermal and sclerenchyma cells. At 15 days after inoculation (dai), fungal hyphae colonized the cortex and clusters of perithecia were observed in the roots. At 20 dai, the fungus reached the central cylinder, and an intense fungal colonization at the base of the culm was observed that resulted in the formation of a mycelial mat on both adaxial and abaxial surfaces of the leaf sheaths. At 25 dai, fungal growth was noticed in the parenchyma cells, vascular bundles and airspaces. Perithecia emerged through the base of prophyllum and from the first leaf sheath at 30 dai. The results of this study provide new insights into the infection process of G. graminis var. graminis in rice and may help to find better control measures in reducing crown sheath rot development.