Variation in oil palm (Elaeis guineensis Jacq.) tissue culture-derived regenerants revealed by AFLPs with methylation-sensitive enzymes |
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Authors: | M Matthes R Singh S-C Cheah A Karp |
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Institution: | (1) ACR-Long Ashton Research Station, Department of Agricultural Sciences, University of Bristol, Long Ashton, Bristol, BS 41 9AF, UK e-mail: angela.karp@bbrc.ac.uk Fax: +44 1275 394007, GB;(2) Palm Oil Research Institute of Malaysia, 6, Persiaran Institusi, Bandar Baru Bangi, 43000 Kajang, Selangor, Malaysia, MY |
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Abstract: | Tissue culture-derived plants of oil palm (Elaeis guineensis Jacq.) can develop abnormal flowers in which stamen primordia are converted into carpel-like tissues (mantled fruit). This
abnormality can be heritable; individual palms may show variation in mantling and reversion to the normal phenotype over time
has been observed. Four sets of ortets (mother plant used as tissue source) and ramets (regenerated plants) were compared
using standard amplified fragment length polymorphism (AFLP) analysis and AFLPs using methylation-sensitive enzymes. No polymorphisms
were found when standard AFLPs were produced with ten different primer combinations. In contrast, when methylation-sensitive
AFLPs were used, polymorphisms were detectable. Polymorphisms appeared as new bands in the ramets, suggesting that a reduction
in methylation had occurred during tissue culture. The highest number of polymorphic bands (0.3%) was obtained when HpaII was used as the restriction endonuclease, indicating that the loss of methylation had occurred most frequently at the internal
C within the HpaII recognition sequence 5’-CCGG-3’. Conversion of nine of the polymorphic bands into probes for Southern analysis confirmed
that these were not due to partial digestion of the AFLP templates and showed that the majority were single-copy sequences.
The exceptions were fragments showing homology to 25S ribosomal RNA genes and the chalcone synthase gene family. Examination
of the Southern blots suggested that most of the single-copy sequences were partially de-methylated, and one example was found
in which de-methylation affected only one allele. No polymorphism was consistently different between normal and abnormal clones
in all the sets. This suggests that, whilst this method is an effective way of detecting variation in tissue culture-derived
plants, different approaches will be required to identify the causal basis of the mantled fruit abnormality.
Received: 25 May 2000 / Accepted: 28 August 2000 |
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Keywords: | Oil palm Tissue culture Regeneration Somaclonal variation Methylation AFLP |
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