Seishi Akino

Comparison of Chinese and Japanese A1 isolates of Phytophthora infestans

The mating type, glucose-6-phosphate isomerase (Gpi) and peptidase (Pep) genotypes, RG57 fingerprint, and mitochondrial DNA (mtDNA) haplotype of Chinese isolates of Phytophthora infestans collected in Hebei and Gansu in 1996 were compared with those of Japanese isolates collected during 1997–2000. The Chinese isolates were divided into four genotypes, one of which was identical to the dominant Japanese genotype, A1-A (mating type A1; Gpi 100/100; Pep 100/100; RG57 100010001100110100011001110: 1–25, 14a, and 24a; and mtDNA haplotype IIa). Comparison of the genotypes with reported data revealed that some completely and partially identical genotypes occur in Russia and parts of Europe. The other two A1 genotypes and one A2 genotype were also detected in Gansu (Gpi 100/100, Pep 100/100, and mtDNA haplotype Ia), which were regarded as unique to this region.

Phytophthora infestans : a review of past and current studies on potato late blight

Phytophthora infestans is the causal agent of potato late blight. Genotypes of Japanese populations of P. infestans have been classified as US-1, JP-1, JP-2, JP-3, and JP-4 based on analyses of DNA polymorphisms. These populations may have been introduced to Japan by the migration of P. infestans from other countries and by domestic changes produced through sexual and asexual propagation. Resistance to late blight has been an ongoing desire of potato farmers in Japan and elsewhere. Recurrent backcrossing of Solanum demissum to varieties of S. tuberosum has been used to transfer late blight resistance. Many varieties carry the R1 gene, whereas others carry R2, R3, and/or R4. However, R genes provided only transient resistance to late blight. New races rapidly overcome R-gene-mediated resistance. The R genes of potato generally encode receptors that recognize secretory effector (Avr) proteins produced by P. infestans. These effector proteins induce robust resistance in potato varieties containing R genes, while they suppress resistance in potato varieties lacking R genes. Conserved molecules from Phytophthora species such as fatty acids, glucans, and elicitins also act as elicitors in Solanaceae species. These P. infestans-derived elicitors induce defensive reactions, including the accumulation of phytoalexins and hypersensitive cell death. A future challenge will be to combine our accumulated knowledge with that from other scientific fields to develop a disease management approach for late blight.

Common Spear Rot of Oil Palm in Indonesia

Common spear rot (CSR), which is also known as crown disease, was first reported in Indonesia in the 1920s. It has caused considerable losses in young oil palm plantings, and yet the pathogenic agent has remained elusive. Symptomatic spear leaves were collected from oil palm plantations and farm plots in South Sumatra, North Sumatra, and Bangka-Belitung, Indonesia. Of the 14 different fungi isolated, Fusarium incarnatum, F. solani, an undescribed Fusarium sp., and Ceratocystis paradoxa were isolated most frequently from diseased leaf tissue. F. incarnatum and the undescribed Fusarium sp. were also frequently isolated from healthy leaf tissue, along with Pestalotiopsis microspora and Curvularia affinis. Ceratocystis paradoxa was never isolated from healthy leaf tissue. Kochs postulate experiments showed that C. paradoxa was able to infect wounded oil palm leaves causing a symptom of extensive rotting similar to that found in the field. Although isolated less frequently and less virulent than C. paradoxa, F. sacchari was also capable of causing lesions on succulent wounded, inoculated leaves. For both C. paradoxa and F. sacchari, the disease severity index was greater when the oil palm leaves appeared to have more succulent growth. Likewise, other Fusarium species and other nonfusarial fungi that were usually not pathogenic were weakly virulent on palms with more succulent growth. These findings confirm that C. paradoxa is one pathogen that is associated with CSR of oil palm in Indonesia.

Identification and pathogenicity of Pythium species causing damping-off of kidney bean

Five Pythium species (Pythium irregulare, P. mamillatum, P. myriotylum, P. spinosum and P. ultimum var. ultimum) were isolated from the hypocotyls and roots of kidney bean plants with damping-off from a commercial field and from experimental plots that have undergone either continuous cropping with kidney bean or rotational cropping with arable crops. In inoculation tests, all five Pythium species were pathogenic to kidney bean. This is the first report of damping-off of kidney bean caused by Pythium species; we named this disease damping-off of kidney bean.

New multilocus genotypes of Phytophthora infestans in Japan

Twelve isolates of Japanese Phytophthora infestans, which differed from the major genotypes US-1, JP-1, JP-2, and JP-3, were analyzed for RG57 fingerprints, mtDNA haplotypes, two allozyme genotypes, and mating types. Genotypes JP-1.1, JP-2.1, JP-2.2, JP-3.1, and JP-4 were newly defined. JP-1.1 and JP-2.1 were isolated discontinuously from potato fields in several years, and JP-1.1 was found in Hokkaido and Kagoshima. These results show that some minor genotypes can overwinter and disperse from their original site.

Soft rot of root chicory in Hokkaido and its causal bacteria

In August 2010, bacterial soft rot was found on root chicory (Cichorium intybus var. sativum) in Hokkaido, Japan. Severely infected plants in fields were discolored, had wilted foliage, and black necrosis of petioles near the crown. Wilted leaves subsequently collapsed and died, forming a dry, brown or black rosette. The root and crown became partially or wholly soft-rotted. Slimy masses on infected areas of roots, turned dark brown or black. Gram-negative, rod-shaped, peritrichously flagellated, facultatively anaerobic bacteria were exclusively isolated from rotted roots, and typical symptoms were reproduced after inoculation with the strains. The bacteria were identified as Dickeya dianthicola, Pectobacterium carotovorum subsp. carotovorum, and Pectobacterium carotovorum subsp. odoriferum based on further bacteriological characterization and the sequence analysis of the malate dehydrogenase gene and 16S rRNA gene. These bacteria should be included with the previously reported Dickeya (=Erwinia) chrysanthemi in Saitama Prefecture, Japan, as causal pathogens of bacterial wilt of chicory.

Use of microsatellite marker Pi26 for rapid discrimination of five Japanese genotypes of Phytophthora infestans

Microsatellite markers were tested to rapidly discriminate five Japanese genotypes (US-1, JP-1, JP-2, JP-3, and JP-4) of Phytophthora infestans. Collected from 1958 to 2007, 111 isolates of Japanese P. infestans were examined using a fluorescent-labeled primer and capillary electrophoresis. Microsatellite marker Pi26 generated specific products for each genotype without any differences in terms of isolation area or year for a particular genotype. The Pi26 marker is a powerful tool for obtaining information on the structure of Japanese populations of P. infestans.

N605ab, a specific molecular marker for Phytophthora infestans, distinguishes three genotypes in Japan

Random amplified polymorphic DNA (RAPD) analysis using the OPG-06 primer generated specific patterns for Japanese genotypes US-1, JP-1, and a new A1 (JP-2, JP-3, and JP-4) of Phytophthora infestans. N605, a specific RAPD fragment, was cloned and sequenced. PCR primers BD1/BD2 were constructed based on the N605 sequence and were used to clarify the genotypes. PCR products using the BD1/BD2 primers (N605ab marker) easily distinguished the new A1 from US-1 and JP-1. This technique provides a simple and effective method for rapid genotype discrimination that can be used in ecological experiments and forecasts for the occurrence of late blight.

Gene mpl1, activated during mating in Phytophthora infestans : similar to genes encoding pectate lyases

Clones of genes activated in mating cultures of A1 and A2 mating-type strains of Phytophthora infestans were isolated using the cDNA-representational difference analysis subtraction method. Clone cET58 was selected based on its accumulation in mating cultures and then was used as a probe to isolate cDNA clone cET58L2 from a cDNA library that was constructed from mycelia grown under mating conditions. Sequence analysis revealed that cET58L2 was 1043 bp long and contained a complete open reading frame of 789 bp. The amino acid sequence of the putative protein was similar to a pectate lyase, PLD, of Fusarium solani f. sp. pisi. The central region of the predicted protein was highly similar to the sequence of other pectate lyases. The gene from which the cDNA clones were derived was designated mpl1. A probe corresponding to the protein-coding region of mpl1 was prepared (probe p58L) for Northern and Southern analyses. The maximum rate of oogonia increase and mpl1 transcript accumulation reached a maximum after 5 days in mating culture. More than 13 genes with sequences similar to that of mpl1 were found in the genome, revealing mpl1 to be a multicopy gene. The mpl1 may be a pectate lyase gene that is activated in P. infestans during mating.

Enhanced virulence of Fusarium species associated with spear rot of oil palm following recovery from osmotic stress

ABSTRACT Fusarium spp., which are common inhabitants of oil palm leaves, are weak pathogens of common spear rot (CSR). We investigated the influence of osmotic stress on the growth, virulence, and activity of cell wall-degrading enzymes of CSR fungi, using potato dextrose agar (PDA) supplemented with KCl or sucrose (hyperosmotic medium). Hyperosmotic stress significantly inhibited mycelial growth, but growth rapidly recovered when mycelia were transferred to control medium. When inoculated into oil palm spear leaflets, Fusarium sp., and F. incarnatum precultured on 1.0 and 1.5 M KCl-hyperosmotic medium induced lesions that were two to four times larger than those in non-stressed cultures, suggesting enhanced virulence of the weak pathogens. Lesion size was not greatly affected in hyperosmotic cultures of moderately virulent F. sacchari. No activity of pectin lyase was detected in liquid cultures of the Fusarium isolates. All isolates except F. incarnatum BT48 secreted polygalacturonase (PG), which was active in both liquid cultures and inoculated leaves. Significantly increased PG activity (5–32-fold) was observed on leaves inoculated with hyperosmotic cultures of Fusarium sp. and F. sacchari. These findings suggest that Fusarium sp., F. incarnatum, and F. sacchari exhibit an adaptive physiological plasticity to hyperosmotic stress that results in enhanced virulence.