Dataset Information

Molecular Characterization of Arginine Kinases in the Soybean Cyst Nematode (Heterodera glycines).

ABSTRACT: Arginine kinase (AK) is a phosphagen kinase that plays a key role in energy mobilization in invertebrates. Alignment of expressed sequence tags (ESTs) for soybean cyst nematode (SCN) (Heterodera glycines) produced two separate contiguous sequences (contigs) and three singletons encoding peptides with high similarity to AKs. One contig, Hg-AK1, had 244 ESTs in the alignment whereas the other, Hg-AK2, had only three; nonetheless, the consensus sequence for Hg-AK1 was missing much of the 5' end. Polymerase chain reaction (PCR) was used to prepare clones that were then sequenced to obtain full-length sequences for both Hg-AK1 and Hg-AK2. Hg-AK1 has an open reading frame of 1080 nucleotides (nt) encoding a protein of 360 amino acids (aa) with a predicted molecular weight of 40 kDa. The open reading frame for Hg-AK2 is 1221 nt, 407 aa, and 46 kDa with a 71% aa identity with Hg-AK1. Semi-quantitative reverse transcriptase polymerase chain reaction (RT-PCR) indicated that Hg-AK1 and Hg-AK2 are expressed constitutively throughout the SCN life cycle. Phylogenetic analysis of peptide sequences for near full-length nematode contigs and other AKs in the Swisspro database indicates that the nematode AKs evolved from a single gene after divergence of insects and nematodes.

SUBMITTER: Matthews BF

PROVIDER: S-EPMC2620652 | biostudies-other | 2003 Sep

REPOSITORIES: biostudies-other

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Publications

Molecular Characterization of Arginine Kinases in the Soybean Cyst Nematode (Heterodera glycines).

Matthews B F BF Macdonald M H MH Thai V K VK Tucker M L ML

Journal of nematology 20030901 3

Arginine kinase (AK) is a phosphagen kinase that plays a key role in energy mobilization in invertebrates. Alignment of expressed sequence tags (ESTs) for soybean cyst nematode (SCN) (Heterodera glycines) produced two separate contiguous sequences (contigs) and three singletons encoding peptides with high similarity to AKs. One contig, Hg-AK1, had 244 ESTs in the alignment whereas the other, Hg-AK2, had only three; nonetheless, the consensus sequence for Hg-AK1 was missing much of the 5' end. Po ...[more]

PMID: 19262758

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Project description:The soybean cyst nematode (SCN), Heterodera glycines, causes economically significant damage to soybeans (Glycine max) in many parts of the world. The cysts of this nematode can remain quiescent in soils for many years as a reservoir of infection for future crops. To investigate bacterial communities associated with SCN cysts, cysts were obtained from eight SCN-infested farms in southern Ontario, Canada, and analyzed by culture-dependent and -independent means. Confocal laser scanning microscopy observations of cyst contents revealed a microbial flora located on the cyst exterior, within a polymer plug region and within the cyst. Microscopic counts using 5-(4,6-dichlorotriazine-2-yl)aminofluorescein staining and in situ hybridization (EUB 338) indicated that the cysts contained (2.6 +/- 0.5) x 10(5) bacteria (mean +/- standard deviation) with various cellular morphologies. Filamentous fungi were also observed. Live-dead staining indicated that the majority of cyst bacteria were viable. The probe Nile red also bound to the interior polymer, indicating that it is lipid rich in nature. Bacterial community profiles determined by denaturing gradient gel electrophoresis analysis were simple in composition. Bands shared by all eight samples included the actinobacterium genera Actinomadura and STREPTOMYCES: A collection of 290 bacteria were obtained by plating macerated surface-sterilized cysts onto nutrient broth yeast extract agar or on actinomycete medium. These were clustered into groups of siblings by repetitive extragenic palindromic PCR fingerprinting, and representative isolates were tentatively identified on the basis of 16S rRNA gene sequence. Thirty phylotypes were detected, with the collection dominated by Lysobacter and Variovorax spp. This study has revealed the cysts of this important plant pathogen to be rich in a variety of bacteria, some of which could presumably play a role in the ecology of SCN or have potential as biocontrol agents.

Dataset Information

Molecular Characterization of Arginine Kinases in the Soybean Cyst Nematode (Heterodera glycines).

Publications

Molecular Characterization of Arginine Kinases in the Soybean Cyst Nematode (Heterodera glycines).

Similar Datasets

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