Taketo Uehara

PCR-based cloning of two Ξ-1,4-endoglucanases from the root-lesion nematode Pratylenchus penetrans

Polymerase chain reaction (PCR) with degenerate primers was performed on cDNA from Pratylenchus penetrans to amplify a conserved region in β-1,4-endoglucanase genes. Two distinct genomic DNA fragments that encoded partial β-1,4-endoglucanase sequences were isolated. Two corresponding full-length cDNAs, designated Pp-eng-1 and Pp-eng-2, were obtained by 3′ and 5′ rapid amplification of cDNA ends. Pp-eng-1 and Pp-eng-2 consisted of 1521 nucleotides encoding 455 amino acids and 1179 nucleotides encoding 328 amino acids, respectively. The two cloned genes encoded family 5 β-1,4-endoglucanases. The predicted protein PP-ENG-1 contained a cellulose-binding domain (CBD), and the predicted protein PP-ENG-2 lacked a CBD.

Morphological and molecular characterisation of Heterodera koreana (Vovlas, Lamberti & Choo, 1992) Mundo-Ocampo, Troccoli, Subbotin, Del Cid, Baldwin & Inserra, 2008 (Nematoda: Heteroderidae) from bamboo in Japan

The Korean cyst nematode, Heterodera koreana , was recorded for the first time from Japan and characterised morphologically, morphometrically and molecularly. In total, 41 populations were detected from soil samples collected from the rhizosphere of four bamboo species in Japan: 31 populations from moso bamboo ( Phyllostachys edulis ), seven from madake ( P. bambusoides ), two from henon bamboo ( P. nigra var. henonis ) and one from fish pole bamboo ( P. aurea ). The morphology and morphometrics of the Japanese population were in agreement with those of the original description of H. koreana from South Korea and other subsequent descriptions from China and Iran, with the exception of some minor differences. The results of the phylogenetic analyses of the D2-D3 expansion segments of 28S rRNA gene and ITS rRNA gene sequences confirmed the species identification and phylogenetic relationship of H. koreana with other Heterodera species. The COI mtDNA gene sequences were obtained for the first time for H. koreana . Three COI haplotypes found in Japanese H. koreana populations showed a characteristic geographical distribution in Japan.

Characterisation of populations of Heterodera trifolii Goffart, 1932 (Nematoda: Heteroderidae) in Japan and their phylogenetic relationships with closely related species

The Japanese populations of the clover cyst nematode, Heterodera trifolii , were characterised morphologically, morphometrically and molecularly. The morphology and morphometrics of six Japanese populations of H. trifolii were in congruence with those of previous descriptions of this species from Germany, Italy, Japan, New Zealand, Russia, The Netherlands, UK and USA. The results of the phylogenetic analyses of the D2-D3 expansion segments of 28S rRNA gene and the COI mtDNA gene sequences discriminated H. trifolii from most of the other closely related species of the Schachtii group. The intraspecific sequence differences in the Schachtii group were much higher in COI than in D2-D3. The H. trifolii populations displayed very low levels of intraspecific variations (up to 0.3%) in the COI mtDNA gene sequences. The phylogenetic relationships of H. trifolii with other Heterodera species, as inferred from the analyses of the ITS rRNA gene, D2-D3 of 28S rRNA gene and COI mtDNA gene sequences, were determined.