Identification of cyst nematodes of agronomic and regulatory concern with PCR-RFLP of ITS1

Allen L. Szalanski, Dezhi D. Sui, T. S. Harris, Thomas O. Powers

Research output: Contribution to journalArticle

87 Citations (Scopus)

Abstract

The first internally transcribed spacer region (ITS1) from cyst nematode species (Heteroderidae) was compared by nucleotide sequencing and PCR-RFLP. European, Asian, and North American isolates of five heteroderid species were examined to assess intraspecific variation. PCR-RFLP patterns of amplified ITS1 DNA from pea cyst nematode, Heterodera goettingiana, from Northern Ireland were identical with patterns from Washington State. Sequencing demonstrated that ITS1 heterogeneity existed within individuals and between isolates, but did not result in different restriction patterns. Three Indian and two U.S. isolates of the corn cyst nematode, Heterodera zeae, were compared. Sequencing detected variation among ITS1 clones from the same individual, between individuals, and between isolates. PCR-RFLP detected several restriction site differences between Indian and U.S. isolates. The basis for the restriction site differences between isolates from India and the U.S. appeared to be the result of additional, variant ITS1 regions amplified from the U.S. isolates, which were not found in the three India isolates. PCR-RFLP from individuals of the U.S. isolates created a composite pattern derived from several ITS1 types. A second primer set was specifically designed to permit discrimination between soybean (H. glycines) and sugar beet (H. schachtii) cyst nematodes. Fok I digestion of amplified product from soybean cyst nematode isolates displayed a uniform pattern, readily discernible from the pattern of sugar beet and clover cyst nematode (H. trifolii).

Original languageEnglish (US)
Pages (from-to)255-267
Number of pages13
JournalJournal of Nematology
Volume29
Issue number3
StatePublished - Sep 1 1997

Fingerprint

cyst nematodes
restriction fragment length polymorphism
sugar beet
Heterodera zeae
Heterodera goettingiana
Heterodera trifolii
Heteroderidae
India
Heterodera glycines
Northern Ireland
peas
digestion
nucleotides
soybeans
clones
corn
DNA

Keywords

  • Clover cyst nematode
  • Corn cyst nematode
  • Diagnosis
  • Heterodera glycines
  • Heterodera goettingiana
  • Heterodera schachtii
  • Heterodera trifolii
  • Heterodera zeae
  • Molecular nematology
  • Nematode PCR-RFLP
  • Pea cyst nematode
  • Regulatory nematology
  • rDNA variation

ASJC Scopus subject areas

  • Agronomy and Crop Science

Cite this

Identification of cyst nematodes of agronomic and regulatory concern with PCR-RFLP of ITS1. / Szalanski, Allen L.; Sui, Dezhi D.; Harris, T. S.; Powers, Thomas O.

In: Journal of Nematology, Vol. 29, No. 3, 01.09.1997, p. 255-267.

Research output: Contribution to journalArticle

Szalanski, Allen L. ; Sui, Dezhi D. ; Harris, T. S. ; Powers, Thomas O. / Identification of cyst nematodes of agronomic and regulatory concern with PCR-RFLP of ITS1. In: Journal of Nematology. 1997 ; Vol. 29, No. 3. pp. 255-267.
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AB - The first internally transcribed spacer region (ITS1) from cyst nematode species (Heteroderidae) was compared by nucleotide sequencing and PCR-RFLP. European, Asian, and North American isolates of five heteroderid species were examined to assess intraspecific variation. PCR-RFLP patterns of amplified ITS1 DNA from pea cyst nematode, Heterodera goettingiana, from Northern Ireland were identical with patterns from Washington State. Sequencing demonstrated that ITS1 heterogeneity existed within individuals and between isolates, but did not result in different restriction patterns. Three Indian and two U.S. isolates of the corn cyst nematode, Heterodera zeae, were compared. Sequencing detected variation among ITS1 clones from the same individual, between individuals, and between isolates. PCR-RFLP detected several restriction site differences between Indian and U.S. isolates. The basis for the restriction site differences between isolates from India and the U.S. appeared to be the result of additional, variant ITS1 regions amplified from the U.S. isolates, which were not found in the three India isolates. PCR-RFLP from individuals of the U.S. isolates created a composite pattern derived from several ITS1 types. A second primer set was specifically designed to permit discrimination between soybean (H. glycines) and sugar beet (H. schachtii) cyst nematodes. Fok I digestion of amplified product from soybean cyst nematode isolates displayed a uniform pattern, readily discernible from the pattern of sugar beet and clover cyst nematode (H. trifolii).

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