Isozyme, protein, and RAPD markers within a half-sib family of buffelgrass segregating for apospory

David L. Gustine, Robert T. Sherwood, Yannis Gounaris, David Robert Huff

Research output: Contribution to journalArticle

15 Citations (Scopus)

Abstract

Isolation of genes controlling apomixis would be useful to plant breeders for fixing hybrid vigor. A single gene codes for aposporous apomixis in buffelgrass [Pennisetum ciliare (L.) Link]. This study was undertaken to assess the feasibility of using isozyme, protein, and random amplified polymorphic DNA (RAPD) markers to detect apospory-linked sequences within a segregating half-sib population. Sexual plant B-2s, five sexual and three aposporous progeny of sexual B-2s, and cultivar Higgins were studied. Floret and leaf proteins were separated by starch gel electrophoresis, and enzymes in the gel were stained to detect isozyme polymorphisms. Of 22 isozyme systems tested, 12 showed polymorphisms but none cozegregated with apomixis. Two-dimensional polyacrylamide gel electrophoresis was used to separate steady state proteins of pistils at meiotic and post-meiotic stages. This technique revealed ≃12% polymorphism within 308 spots, but none of the spots cosegregated with reproductive mode. Genomic DNA was screened for RAPD markers with 111 10-mer random primers and polymerase chain reaction. Of 569 markers identified, 87% were polymorphic. One marker cosegregated with sexual lines, but none cosegregated with aposporous lines. Analysis of molecular variance examination of the B-2s parent and the eight half-sib progeny (Higgins excluded) showed that on the basis of 404 RAPD markers, the apasporous and sexual groups were not significantly different. No RAPD markers were tightly linked with apospory. Additional screening of new primers will allow identification of markers for the gene in full-sib families.

Original languageEnglish (US)
Pages (from-to)723-727
Number of pages5
JournalCrop Science
Volume36
Issue number3
DOIs
StatePublished - Jan 1 1996

Fingerprint

random amplified polymorphic DNA technique
apomixis
isozymes
genetic markers
genetic polymorphism
proteins
Cenchrus ciliaris
leaf protein
plant breeders
starch gels
two-dimensional gel electrophoresis
pistil
florets
heterosis
gel electrophoresis
polyacrylamide gel electrophoresis
genes
polymerase chain reaction
gels
screening

All Science Journal Classification (ASJC) codes

  • Agronomy and Crop Science

Cite this

Gustine, David L. ; Sherwood, Robert T. ; Gounaris, Yannis ; Huff, David Robert. / Isozyme, protein, and RAPD markers within a half-sib family of buffelgrass segregating for apospory. In: Crop Science. 1996 ; Vol. 36, No. 3. pp. 723-727.
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abstract = "Isolation of genes controlling apomixis would be useful to plant breeders for fixing hybrid vigor. A single gene codes for aposporous apomixis in buffelgrass [Pennisetum ciliare (L.) Link]. This study was undertaken to assess the feasibility of using isozyme, protein, and random amplified polymorphic DNA (RAPD) markers to detect apospory-linked sequences within a segregating half-sib population. Sexual plant B-2s, five sexual and three aposporous progeny of sexual B-2s, and cultivar Higgins were studied. Floret and leaf proteins were separated by starch gel electrophoresis, and enzymes in the gel were stained to detect isozyme polymorphisms. Of 22 isozyme systems tested, 12 showed polymorphisms but none cozegregated with apomixis. Two-dimensional polyacrylamide gel electrophoresis was used to separate steady state proteins of pistils at meiotic and post-meiotic stages. This technique revealed ≃12{\%} polymorphism within 308 spots, but none of the spots cosegregated with reproductive mode. Genomic DNA was screened for RAPD markers with 111 10-mer random primers and polymerase chain reaction. Of 569 markers identified, 87{\%} were polymorphic. One marker cosegregated with sexual lines, but none cosegregated with aposporous lines. Analysis of molecular variance examination of the B-2s parent and the eight half-sib progeny (Higgins excluded) showed that on the basis of 404 RAPD markers, the apasporous and sexual groups were not significantly different. No RAPD markers were tightly linked with apospory. Additional screening of new primers will allow identification of markers for the gene in full-sib families.",
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Isozyme, protein, and RAPD markers within a half-sib family of buffelgrass segregating for apospory. / Gustine, David L.; Sherwood, Robert T.; Gounaris, Yannis; Huff, David Robert.

In: Crop Science, Vol. 36, No. 3, 01.01.1996, p. 723-727.

Research output: Contribution to journalArticle

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T1 - Isozyme, protein, and RAPD markers within a half-sib family of buffelgrass segregating for apospory

AU - Gustine, David L.

AU - Sherwood, Robert T.

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N2 - Isolation of genes controlling apomixis would be useful to plant breeders for fixing hybrid vigor. A single gene codes for aposporous apomixis in buffelgrass [Pennisetum ciliare (L.) Link]. This study was undertaken to assess the feasibility of using isozyme, protein, and random amplified polymorphic DNA (RAPD) markers to detect apospory-linked sequences within a segregating half-sib population. Sexual plant B-2s, five sexual and three aposporous progeny of sexual B-2s, and cultivar Higgins were studied. Floret and leaf proteins were separated by starch gel electrophoresis, and enzymes in the gel were stained to detect isozyme polymorphisms. Of 22 isozyme systems tested, 12 showed polymorphisms but none cozegregated with apomixis. Two-dimensional polyacrylamide gel electrophoresis was used to separate steady state proteins of pistils at meiotic and post-meiotic stages. This technique revealed ≃12% polymorphism within 308 spots, but none of the spots cosegregated with reproductive mode. Genomic DNA was screened for RAPD markers with 111 10-mer random primers and polymerase chain reaction. Of 569 markers identified, 87% were polymorphic. One marker cosegregated with sexual lines, but none cosegregated with aposporous lines. Analysis of molecular variance examination of the B-2s parent and the eight half-sib progeny (Higgins excluded) showed that on the basis of 404 RAPD markers, the apasporous and sexual groups were not significantly different. No RAPD markers were tightly linked with apospory. Additional screening of new primers will allow identification of markers for the gene in full-sib families.

AB - Isolation of genes controlling apomixis would be useful to plant breeders for fixing hybrid vigor. A single gene codes for aposporous apomixis in buffelgrass [Pennisetum ciliare (L.) Link]. This study was undertaken to assess the feasibility of using isozyme, protein, and random amplified polymorphic DNA (RAPD) markers to detect apospory-linked sequences within a segregating half-sib population. Sexual plant B-2s, five sexual and three aposporous progeny of sexual B-2s, and cultivar Higgins were studied. Floret and leaf proteins were separated by starch gel electrophoresis, and enzymes in the gel were stained to detect isozyme polymorphisms. Of 22 isozyme systems tested, 12 showed polymorphisms but none cozegregated with apomixis. Two-dimensional polyacrylamide gel electrophoresis was used to separate steady state proteins of pistils at meiotic and post-meiotic stages. This technique revealed ≃12% polymorphism within 308 spots, but none of the spots cosegregated with reproductive mode. Genomic DNA was screened for RAPD markers with 111 10-mer random primers and polymerase chain reaction. Of 569 markers identified, 87% were polymorphic. One marker cosegregated with sexual lines, but none cosegregated with aposporous lines. Analysis of molecular variance examination of the B-2s parent and the eight half-sib progeny (Higgins excluded) showed that on the basis of 404 RAPD markers, the apasporous and sexual groups were not significantly different. No RAPD markers were tightly linked with apospory. Additional screening of new primers will allow identification of markers for the gene in full-sib families.

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