Citation
Rajinder Singh
(2005)
Molecular Markers and Their Applications in the Construction of Genetic Linkage Maps and Analysis of Monogenic and Quantitative Traits in Oil Palm.
UNSPECIFIED thesis, Universiti Putra Malaysia.
Abstract
Molecular breeding is well suited to a perennial crop, like oil palm, in which the
economic products are not produced until several years after planting. The use of
DNA markers for selection in such crops could greatly reduce the number of
breeding cycles needed. As such, the primary aim of this project was to map the oil
palm genome by using restriction fragment length polymorphism (RFLP), amplified
fragment length polymorphism (AFLP) and microsatellite (SSR) markers, in order
to provide markers for selection in the breeding programme. A progeny derived
from the selfing of a tenera guineensis palm (palm Tl28), which was segregating for
the economically important monogenic characters of shell thickness and fruit color,
was used for linkage map construction. A total of 523 segregating markers (117
RFLP, 22 SSR and 384 AFLP) were mapped to 17 linkage groups. In the current
linkage map for the T128 palm, we have successfully mapped both the shell (Sh)
gene and the fruit color (Vir) gene. Two RFLP markers mapped close to the Vir gene
at a distance of 3 and 4 centiMorgans (eM), respectively. These two RFLP markers were tested by using seven other independent crosses segregating for this trait. The
markers could predict the trait with about 95% certainty. The Sh gene was located
about 8 cM from the nearest marker (EAGG/MCTT-250, an AFLP marker). The
AFLP marker could distinguish the pisifera palms (absence of shell) from the dura
(thick shell) and tenera palms (thin shell) in the mapping family evaluated. These
markers are useful tools for application in a molecular breeding programme for oil
palm. Analysis was also extended to include mapping of quantitative traits (QTLs)
associated with oil quality. Oil quality is determined by the fatty acid composition
(FAC) in oil palm. By using the genome wide threshold levels calculated
independently for each of the traits, significant QTLs were identified for myristic
acid (CI4:0), palmitic acid (CI6:0), palmitoleic acid (CI6:1) and stearic acid
(CI8:0). In the attempt at mapping additional QTLs associated with oil quality, an
interspecific cross between a Colombian oleifera (UP1026) and the Tl28 palm was
also examined. A map consisting of 412 markers (302 AFLP, 83 RFLP and 27 SSR)
in 18 linkage groups was generated at a LOD score of 5.0. At a genome wide
significance threshold of P<O.OI and P<0.05, significant QTLs were detected for
iodine value (IV), C14:0, CI6:0, C16:1 and CI8:0. There were common markers
revealing significant linkages for the same trait in both the mapping families
analyzed. Such tagging of markers to economically important traits will aid in
expediting the production of elite planting materials with greater precision. To
facilitate oil palm genome analysis leading to physical mapping and the
identification of molecular markers associated with QTL and map based cloning, we
attempted to develop the tools and techniques needed to construct a bacterial
artificial chromosome (BAC) library for oil palm. A suitable method to purify and
prepare the single copy vector (pBeloBACll) for BAC transformation was established. The proper partial digestion conditions of oil palm megabase DNA for
BAC library construction were also determined. Several oil palm BAC clones were
successfully identified. Hybridization of these BAC clones with oil palm DNA as
probe confirmed the presence of oil palm DNA in those clones. In map based
cloning efforts, cosmid libraries (with cloning capacity of up to 50 kb) are useful
tools to complement BAC libraries. For this reason we constructed a cosmid library
for oil palm. The library contains 65,000 clones with insert size ranging from 30 kb
to 42 kb. Hybridization of randomly selected cosmid clones with oil palm DNA also
confirmed the presence of oil palm DNA in these clones.
Download File
Additional Metadata
Actions (login required)
![View Item View Item](/style/images/action_view.png) |
View Item |