Factors affecting cross protection of fusarium wilt of pigeon pea by soilborne nonpathogenic fungi

Preinoculation of pigeon pea (Cajanus cajan (L.) Millsp.) seedlings with soilborne fungi nonpathogenic to pigeon pea, namely,Fusarium oxysporum f.sp.niveum; F. oxysporum f.sp.ciceris; F. solani f.sp.pisi; andCephalosporium sacchari, before challenge inoculation with the pathogenFusarium udum, or simultaneous inoculation of the seedlings with nonpathogen and pathogen was effective in controlling wilt of pigeon pea to a great extent. Inoculation with the nonpathogens before the challenge inoculation was more effective than simultaneous inoculation and gave up to 81.6% protection. The higher the spore concentration of the nonpathogenic fungi, the better was the degree of protection. The shorter the period was (in the range of 2–7 days) between nonpathogen inoculation and challenge inoculation, the greater was the degree of protection. Injury to the roots resulted in a decrease of the effect. Maximum disease reduction was obtained when the seedlings were kept at 20-30°C before inoculation; at 5°C and also at 40°C, protection was much less.     

A high-throughput glasshouse bioassay to detect crown rot resistance in wheat germplasm

A high-throughput and reliable seedling bioassay to screen wheat germplasm for crown rot resistance was developed. Single wheat seedlings were grown in square seedling punnets in a glasshouse and inoculated with a monoconidial Fusarium pseudograminearum isolate 10 days after emergence. The punnets were laid horizontally on their side and a 10- µ L inoculum droplet placed on the stem base. Seedlings were incubated at near-saturated relative humidity, and crown rot severity was assessed 35 days after inoculation. Studies on the duration of incubation period, inoculum concentration and temperature were carried out to optimize these parameters. Seedling growth at 25/15(±5)°C in a glasshouse and 48-h incubation at near-saturated RH in darkness gave the best results. When crown rot resistance rankings of 16 Australian cultivars from the bioassay were compared with their field performance, Spearman's rank correlation coefficient was highly significant. This indicated that the seedling bioassay mimicked field resistance to crown rot in adult plants. A bootstrap resampling analysis showed little or no improvement in the coefficient of variation with an increasing number of replications, indicating a high level of precision and reproducibility. By detecting small but consistent differences in crown rot severity, the bioassay proved effective in large-scale screening for partial resistance: already over 1400 wheat genotypes have been screened. The high degree of precision makes this an invaluable tool in the understanding of pathogen aggressiveness, host specialization and parasitic fitness.     

Genetic diversity of Australian Fusarium graminearum and F. pseudograminearum

Genotypic diversity in Fusarium pseudograminearum and F. graminearum from Australia and the relationship between diversity and pathogen aggressiveness for head blight and/or crown rot of wheat were examined. Amplified fragment length polymorphism (AFLP) analysis revealed a high level of genotypic diversity within each species. Sixty-three of the 149 AFLP loci were significantly different between the two species and 70 of 72 F. pseudograminearum and 56 of 59 F. graminearum isolates had distinct haplotypes. When head blight and crown rot severity data from a recently published work on isolates representing the entire range of aggressiveness were used, only the genotypic diversity of F. pseudograminearum was significantly associated with its aggressiveness for the two diseases. Cluster analyses clearly demonstrated the polyphyletic structures that exist in both pathogen populations. The spatial diversity within F. graminearum was high within a single field, while frequent gene flow ( N _m ∼ 14) and a low fixation index ( G _st = 0·03) were recorded among F. pseudograminearum isolates from the adjacent states of New South Wales and Queensland. The differences in population structure between the heterothallic F. pseudograminearum (teleomorph G. coronicola ) and the homothallic F. graminearum (teleomorph G. zeae ) were not as pronounced as expected given their contrasting mating systems. Neither species was panmictic or strictly clonal. This points to sexual recombination in F. pseudograminearum , suggesting that ascospores of G. coronicola may also play a role in its biology and epidemiology.     

Impact of environmental factors on infestation of tea leaves byHelopeltis theivora, and associated changes in flavonoid flavor components and enzyme activities

Attack of tea plant (Camellia sinensis (L.) O. Kuntze) leaves by the tea mosquito bugHelopeltis theivora (Hemiptera: Miridae) was positively correlated to temperature and rainfall, and partially to humidity, as determined in 12 varieties during the period 2000–2002. The insect attack was severe during the months of May–September, which had high temperature and rainfall, and led to severe loss of biomass due to curling and drying up of the leaves. The biochemical response of these 12 varieties of tea to attack by the insect was determined with special reference to oxidative enzymes and flavonoid flavor components. Insect attack led to an increase in the activities of the oxidative enzymes peroxidase, ascorbate peroxidase and polyphenol oxidase. Activities of phenyl alanine ammonia lyase generally decreased as a result of insect attack, which was significant in the United Planters Association of South India (UPASI) varieties. A significant decrease in polyphenols was also obtained in UPASI varieties. HPLC analysis of catechins revealed a decrease in some of the catechins in the infected leaves. Analysis of theaflavins from infusion of healthy andHelopeltis-infested tea leaves revealed no changes. http://www.phytoparasitica.org posting Dec. 19, 2004.     

Eicosapentaenoic acid enrichment from sardine oil by argentation chromatography

Eicosapentaenoic acid (EPA) derived from chemically hydrolyzed sardine oil was concentrated by urea fractionation using methanol at different temperatures (2, 4, and 6 degrees C) and urea/fatty acid ratios (2:1, 3:1, and 4:1 w/w) and purified by argentation neutral alumina column chromatography. The individual fatty acids were determined as fatty acid methyl esters (FAME) by gas-liquid chromatography and gas chromatography-mass spectroscopy as FAME and N-acyl pyrrolidides. In the mass fragmentation pattern of FAME, the base peak was assigned to be the 1-methoxyethenol moiety (m/z = 74) obtained by McLafferty rearrangement. Formation of the cyclic tropylium ion (m/z = 91) in fatty acids with four or more double bonds was apparent in FAME-PUFAs. The base peak of N-acyl pyrrolidides was the McLafferty rearrangement ion, 1-(pyrrolidin-1-yl)ethenol (m/z = 113). The highest concentration of EPA (47.78%) was obtained at the crystallization temperature of 4 degrees C with a urea/fatty acid ratio of 4:1 (w/w) with 93.74% yield. After complexation of saturated and less unsaturated fatty acids by urea complexation, argentation chromatography resulted in an EPA of high purity (99.6%) with an overall recovery of 54.09% using 50% diethyl ether/n-hexane as eluting solvent. The peroxide (POV) and thiobarbituric acid (TBS) values were found to be highest (4.0 mequiv of O2/kg and 5.2 mg of malondialdehyde/kg, respectively) during urea fractionation at the higher crystallization temperature (6 degrees C) and higher urea/fatty acid ratio (4:1). Keywords: Sardine oil; eicosapentaenoic acid (EPA); fatty acid methyl esters (FAME); urea fractionation; argentation column chromatography.