期刊
MOLECULAR PLANT-MICROBE INTERACTIONS
卷 14, 期 4, 页码 536-544出版社
AMER PHYTOPATHOLOGICAL SOC
DOI: 10.1094/MPMI.2001.14.4.536
关键词
expressed sequence tags; functional genomics; host-parasite interactions; nematode feeding sites; parasitism genes
Secretions from the esophageal gland cells of plant-parasitic nematodes play critical roles in the nematode-parasitic Cycle. A novel method to isolate cDNA encoding putative nematode secretary proteins was developed that utilizes:mRNA for reverse transcription-polymerase chain reaction derived from microaspiration of the esophageal gland cell contents of parasitic stages of the soybean cyst nematode Heterodera glycines, The resulting H, glycines gland cell cDNA was cloned into the pRK18 vector, and plasmid DNA was transformed into a mutated yeast host for specific selection of cDNA inserts that encode proteins with functional signal peptides, Of the 223 cDNA clones recovered from selection in yeast, 97% of the clones encoded-a predicted signal peptide. Fourteen unique cDNA clones hybridized to genomic DNA of H, glycines on Southern blots and, among them, nine cDNA clones encoded putative extracellular proteins, as predicted by PSORT Il computer analysis. Four cDNA clones hybridized to transcripts within the dorsal esophageal gland cell of parasitic stages of H. glycines, and in situ hybridization within H. glycines was not detected for eight cDNA clones. The protocol provides a direct means to isolate potential plant-parasitic nematode esophageal gland secretory protein genes.
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