Evidence for resistance in wheat cultivars grown in sand culture to the take-all pathogen, Gaeumannomyces graminis var. tritici

A laboratory method to inoculate seedlings uniformly with Gaeumannomyces graminis var. tritici is described. Resistance is defined via the rate of hyphal entry into the vascular tissue of host seedlings, and is measured by direct observation and by early stelar lesion development in seminal roots. The two scores for resistance are compared and evaluated, for infection with an isolate of low virulence. Evidence was obtained for resistance in the roots of wheat seedlings to G. graminis.     

Studies on the spread of Gaeumannomyces graminis var. tritici in wheat.

The effect of cultivations on the spread of Gaeumannomyces graminis var. tritici from a line source of inoculum, consisting of naturally infected stubble and roots, was recorded in the field over 2 yr. With the aid of cultivations, spread in the first wheat crop occurred frequently to a distance of 0·9 m and occasionally to a distance of 2·5 m. However, the following wheat crop was uniformly infected, probably as a result of a rapid build-up of background inoculum in the first wheat, so that any spread by cultivations was masked.     

Disease in wheat genotypes naturally infected with Gaeumannomyces graminis var. tritici

Wheat genotypes consisting of seven homozygous lines and 40 segregate families were studied at two sites naturally infested with the take-all pathogen, Gaeumannomyces graminis var. tritici. The numbers of seminal and coronal roots infected with G. graminis and other root pathogens were recorded. The genotypes differed in infection with G. graminis, with little evidence of genotype × environment interactions. The incidence of G. graminis and Rhizoctonia solani was negatively associated, but the association did not greatly influence differences between wheats in infection with G. graminis. The distribution of R. solani was negatively associated with the severity of take-all at only one site. Of symptoms of infection with G. graminis, wheat genotypes differed most in incidence of deadheads, but differences were not consistent over environments, and were associated with earliness of maturity. Wheats differed more in expression of disease than in infection with G. graminis. The course of disease was deduced from associations between the incidence of pathogens and components of plant growth and yield. G. graminis was the dominant pathogen at both sites, and caused a yield loss of 0–15% at one site, and an average 62% loss at the other. More components of yield were affected where disease was most severe.     

小麦全蚀病菌致病性遗传规律研究*

利用弱致病性白化突变菌株与强致病菌株进行有性重组,研究了小麦全蚀病菌(禾顶囊壳小麦变种,Gaeumannomyces gramimis var. tritici)致病性的遗传规律。结果表明,以接种株地上部干重为致病性指标时,该菌对小麦的致病性为数量遗传性状,估计控制致病性的基因数为5 ~ 7个。弱致病菌株与强致病菌株杂交F1代致病性由弱至强呈连续分布,子代致病性的平均水平接近种亲代致病性的平均值。致病性的遗传力为54.36% ~ 71.00%,平均为62.32%。致病性表型易受环境因素的影响。菌落颜色与致病性无明显相关。     

The use of ammonium fertilisers to reduce the severity of take-all (Gaeumannomyces graminis var. tritici) on wheat in Western Australia

In field experiments in Western Australia ammonium fertilisers significantly reduced the severity of root damage to wheat ( Triticum aestivum ) caused by Gaeumannomyces graminis var. tritici. A series of response lines depicting the effect of sources and rates of nitrogen on take-all are presented. In general, the higher the rate of ammonium nitrogen used, the greater the reduction in the severity of take-all. Ammonium sulphate and ammonium sulphate/mono-ammonium phosphate mixture were more effective in reducing disease than di-ammonium phosphate, urea or ammonium nitrate. Sodium nitrate did not reduce take-all.     

Colonization of barley roots by endophytic fungi and their reduction of take-all caused by Gaeumannomyces graminis var. tritici

Fungal root endophytes obtained from natural vegetation were tested for antifungal activity in dual culture tests against the root pathogen Gaeumannomyces graminis var. tritici. Fifteen isolates, including Acremonium blochii, Acremonium furcatum, Aspergillus fumigatus, Cylindrocarpon sp., Cylindrocarpon destructans, Dactylaria sp., Fusarium equiseti, Phoma herbarum, Phoma leveillei, and a sterile mycelium, selected based on the dual culture test, were inoculated on barley roots in growth tubes under axenic conditions, both in the absence and presence of G. graminis var. tritici. All isolates colonized the rhizosphere and very often the root cortex without causing disease symptoms and without affecting plant growth. Eight isolates significantly reduced the symptoms caused by G. graminis var. tritici, and 6 of them reduced its presence in the roots.     

Purification and Characterization of a Secreted Laccase of Gaeumannomyces graminis var. tritici

We purified a secreted fungal laccase from filtrates of Gaeumannomyces graminis var. tritici cultures induced with copper and xylidine. The active protein had an apparent molecular mass of 190 kDa and yielded subunits with molecular masses of 60 kDa when denatured and deglycosylated. This laccase had a pI of 5.6 and an optimal pH of 4.5 with 2,6-dimethoxyphenol as its substrate. Like other, previously purified laccases, this one contained several copper atoms in each subunit, as determined by inductively coupled plasma spectroscopy. The active enzyme catalyzed the oxidation of 2,6-dimethoxyphenol (K_m = 2.6 × 10⁻⁵ ± 7 × 10⁻⁶ M), catechol (K_m = 2.5 × 10⁻⁴ ± 1 × 10⁻⁵ M), pyrogallol (K_m = 3.1 × 10⁻⁴ ± 4 × 10⁻⁵ M), and guaiacol (K_m = 5.1 × 10⁻⁴ ± 2 × 10⁻⁵ M). In addition, the laccase catalyzed the polymerization of 1,8-dihydroxynaphthalene, a natural fungal melanin precursor, into a high-molecular-weight melanin and catalyzed the oxidation, or decolorization, of the dye poly B-411, a lignin-like polymer. These findings indicate that this laccase may be involved in melanin polymerization in this phytopathogen’s hyphae and/or in lignin depolymerization in its infected plant host.     

Occurrence of Phialophora radicicola var. graminicola and Gaeumannomyces graminis var. tritici on roots of wheat in field crops

Assessments of Phialophora radicicola var. graminicola (PRG) and Gaeumannomyces graminis var. tritici (GGT) were made by culturing and by direct microscopic examination of pieces of seminal roots from 16 winter wheat crops grown in different cropping sequences and with different phosphate manuring. PRG occurred on all wheat crops, but was abundant only on wheat after grass, where it seemed to delay the onset of damaging take-all by 1 yr. Delayed occurrence of take-all by phosphate fertiliser was not related to differences in populations of PRG. Wheat grown in ‘take-all decline’ soils had only small amounts of PRG, indicating that the development and the decline of take-all epidemics may be influenced by different biological control mechanisms; breaking sequences of wheat crops by 1 yr grass leys might harness the advantages of both mechanisms.     

Purification and characterization of a secreted laccase of Gaeumannomyces graminis var. tritici.

We purified a secreted fungal laccase from filtrates of Gaeumannomyces graminis var. tritici cultures induced with copper and xylidine. The active protein had an apparent molecular mass of 190 kDa and yielded subunits with molecular masses of 60 kDa when denatured and deglycosylated. This laccase had a pI of 5.6 and an optimal pH of 4.5 with 2,6-dimethoxyphenol as its substrate. Like other, previously purified laccases, this one contained several copper atoms in each subunit, as determined by inductively coupled plasma spectroscopy. The active enzyme catalyzed the oxidation of 2, 6-dimethoxyphenol (Km = 2.6 x 10(-5) +/- 7 x 10(-6) M), catechol (Km = 2.5 x 10(-4) +/- 1 x 10(-5) M), pyrogallol (Km = 3.1 x 10(-4) +/- 4 x 10(-5) M), and guaiacol (Km = 5.1 x 10(-4) +/- 2 x 10(-5) M). In addition, the laccase catalyzed the polymerization of 1, 8-dihydroxynaphthalene, a natural fungal melanin precursor, into a high-molecular-weight melanin and catalyzed the oxidation, or decolorization, of the dye poly B-411, a lignin-like polymer. These findings indicate that this laccase may be involved in melanin polymerization in this phytopathogen's hyphae and/or in lignin depolymerization in its infected plant host.     

Changes in population structure of the soilborne fungus Gaeumannomyces graminis var. tritici during continuous wheat cropping

A method was developed to assess the genetic structure of Gaeumannomyces graminis var. tritici (Ggt) populations and test the hypothesis of an association between disease level in the field with changes in pathogen populations. A long-term wheat monoculture experiment, established since 1994, generated different take-all epidemics with varying the number of wheat crop successions in the 1999-2000 cropping season. Genetic polymorphism in Ggt populations was investigated over natural, local epidemics. Four populations of 30 isolates were isolated from necrotic wheat roots in a first, third, fourth, and sixth wheat crop in the same year. Each Ggt isolate was characterized with RAPD (Random Amplification Polymorphism DNA) markers and AFLP (Amplified Fragment Length Polymorphism) fingerprinting. Seventeen multilocus genotypes based on the combination of RAPD and AFLP markers were identified among all these populations. The 120 isolates were divided into two main groups, G1 and G2, according to bootstrap values higher than 86%, except for an unique isolate from the third wheat crop. Within each group, populations ranged between 93 and 100% similarity. Both groups included isolates collected from the first, third, fourth or sixth wheat crop. However, G1 group profiles dominated amongst isolates sampled in the first and the sixth wheat crops, whereas G2 group profiles largely dominated amongst isolates collected from the third and fourth wheat crops. Aggressiveness of group G2 (38%) was significantly greater than that of G1 (29.5%). These results suggest that changes in Ggt population structure occur during continuous wheat cropping. The distinction of two Ggt groups provides a simple basis for further spatio-temporal analysis of Ggt population during polyetic take-all decline.     

Introduced Pseudomonas fluorescens VUPf5 as an important biocontrol agent for controlling Gaeumannomyces graminis var. tritici the causal agent of take-all disease in wheat

About 900 bacterial strains obtained from plants rhizosphere from different areas of Iran and biocontrol ability were widely survey in vitro and greenhouse conditions against Gaeumannomyces graminis var. tritici the causal agent of take-all disease. Finally the best strain (VUPf5 strain) in the additional tests biocontrol ability in vitro and greenhouse conditions was selected for next studies. VUPf5 suppressed of take-all disease 85%. Based on biochemical and morphological tests, this isolate is belonging to the species Pseudomonas fluorescens. This isolate significantly produced the secondary metabolites such as siderophores, hydrogen cyanide, protease, phenazine and volatile metabolites.     

Effects of take-all (Gaeumannomyces graminis var. tritici) on crop N uptake and residual mineral N in soil at harvest of winter wheat

Background and aims

Take-all, caused by the fungus Gaeumannomyces graminis var. tritici, is the most damaging root disease of wheat. A severe attack often leads to premature ripening and death of the plant resulting in a reduction in grain yield and effects on grain quality (Gutteridge et al. in Pest Manag Sci 59:215–224, 2003). Premature death of the plant could also lead to inefficient use of applied nitrogen (Macdonald et al. in J Agric Sci 129(2):125–154, 1997). The aim of this study was to determine crop N uptake and the amount of residual mineral N in the soil after harvest where different severities of take-all had occurred.

Methods

Plant and soil samples were taken at anthesis and final harvest from areas showing good and poor growth (later confirmed to be caused by take-all disease) in three winter wheat crops grown on the same soil type on Rothamsted Farm in SE England in 1995, 2007 and 2008 (harvest sampling only). All crops received fertiliser N in spring at recomended rates (190–200?kg?N ha^?1). On each ocassion crops were assessed for severity of take-all infection (TAR) and crop N uptakes and soil nitrate plus ammonium (SMN) was determined. Grain yields were also measured.

Results

Grain yields (at 85% dry matter) of crops with moderate infection (good crops) ranged from 4.3 to 13.0?t ha^?1, compared with only 0.9–4.5?t ha^?1 for those with severe infection (poor crops). There were significant (P?<?0.05) negative relationships between crop N uptake and TAR at anthesis and final harvest. At harvest, good crops contained 129–245?kg?N ha^?1 in grain, straw and stubble, of which 85–200?kg?N ha^?1 was in the grain. In contrast, poor crops contained only 46–121?kg?N ha^?1, of which only 22–87?kg?N ha^?1 was in the grain. Positive relationships between SMN and TAR were found at anthesis and final harvest. The SMN in the 0–50?cm layer following harvest of poor crops was significantly (P?<?0.05) greater than that under good crops, and most (73–93%) was present as nitrate.

Conclusions

Localised patches of severe take-all infection decreased the efficiency with which hexaploid wheat plants recovered soil and fertiliser derived N, and increased the subsequent risk of nitrate leaching. The risk of gaseous N losses to the atmosphere from these areas may also have been enhanced.     

Effects of sowing date and volunteers on the infectivity of soil infested with Gaeumannomyces graminis var. tritici and on take-all disease in successive crops of winter wheat

In a field experiment on winter wheat, take‐all on plants and the infectivity of the soil were studied in crop sequences with different combinations of sowing dates. Take‐all was negligible in the first wheat crop, but thereafter the mean disease intensity (measured using a take‐all rating, TAR, with a maximum of 300) was 108, 190, 118 and 251 in the second to fifth successive crops. In each growing season, the disease differed amongst sequences and built up more rapidly and was more intense on plants sown in mid‐September than on plants sown in mid‐October. In late‐sown plots, where volunteers had been present during the mid‐September to mid‐October period, take‐all reached an intensity intermediate between that in early‐sown plots and that in late‐sown plots that had been kept free of volunteers. Volunteers, therefore, partially offset the expected beneficial effect of decreased disease with later sowing. Differences in take‐all amongst sequences were most pronounced in the second wheat crop and early sowing of the previous wheat increased intensity of disease. In the following (third) crop, differences in disease intensity amongst sequences were smaller. Soil infectivity (measured by seedling bioassay after harvest) built up progressively from a low level after the first crop to peak after the third crop. In this build‐up phase, soil infectivity estimates were always numerically greater after harvest of early‐sown treatments than after later‐sown treatments, although never significant at P= 0.05. The greatest difference (P= 0.06) was recorded in October before sowing of the third crop, where the comparison was between soil after two previous early sowings and soil after two previous later sowings and control of volunteers. In the same autumn, presence of green cover (i.e. volunteers) was associated with a smaller loss of soil infectivity between harvest and later sowing than occurred in an absence of green cover. In 2^nd–4^th crops, where comparisons were available and mean TARs indicated moderate levels of take‐all, sowing later had no yield benefit, despite more take‐all and greater soil infectivity associated with early sowing. Important considerations for the management of crops at risk of take‐all are 1) choosing appropriate sowing dates to minimize take‐all or to encourage take‐all decline and 2) controlling volunteers and weed hosts where crops are sown late to minimise take‐all.     

Incidence of Gaeumannomyces graminis var. tritici and K deficiency increase rhizospheric denitrification

Wheat inoculated with the root pathogen Gaeumannomyces graminis var. tritici (Ggt) was grown in quartz silt at two levels of potassium nutrition. While in plants well supplied with K the incidence of Ggt did not affect plant growth, it reduced shoot and root weight of K deficient plants. Denitrification, measured by the acetylene inhibition technique and expressed as N₂O/mg root weight, was increased either by low K nutrition or by Ggt infection. Highest denitrification in the rhizosphere of plants was found with a combination of both, K deficiency and Ggt attack.     

小麦全蚀病菌原生质体制备的条件及再生菌株的致病性

分别利用崩溃酶、溶壁酶、纤维素酶和蜗牛酶酶解小麦全蚀病菌,进行原生质体的制备试验。结果显示,4种酶均能消化该菌细胞壁,获得一定数量的原生质体;产生原生质体效率最高的是溶壁酶,该酶在浓度为16 mg/mL时产生的原生质体数量最多,最佳的酶解时间为2~3 h,最适作用温度为28℃。制备的原生质体可以再生并与原始出发菌株具有相同的致病能力。