Kajian variasi genetik dan hubungan ke atas mimosa sp. dan lima genera tertentu famili zingiberaceae melalui kaedah biologi molekul
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Date
2002
Authors
Sul' Ain, Mohd Dasuki (Haji)
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Abstract
Two molecular biology methods i.e SDS-PAGE and RAPD-PCR were used to
evaluate variation and determine relationship among Mimosa sp. and five selected
genus of ginger in Penang. The three species of Mimosa sp. investigated were M
pudica, M pigra and M invisa. Through SDS-PAGE, the overall bands that were
different among the three species indicating a high variability. Protein variation from
the flowers were better as compared to that from the leaves. The existence of certain
molecular weight and unique bands can be used as a diagnostic tool for species
determination. Three species of J!imosa sp. showed a high variability among each
others. Random amplified polymorphic DNA (RAPD) analysis was used to evaluate
the inter-generic relationships among Mimosa sp. Among 30 primers tested, only
eleven arbitrary 1 O-rner primer gave the best result. Six from operon kit-C and five
from operon kit-Z. All the primers detected polymorphism irr all the three species of
Mimosa. A total of 92 bands were amplified with 82.61% of RAPDs were
polymorphic. The total polymorphism at inter-generic level of more than 80% among
three Mimosa sp. The interspecies polymorphism among two species of Mimosa sp.,
varied from 56% to 73%. The genetic relationships among the species were analyzed
by cluster analysis of Nei's genetic distance data according to the unweighted pair
group method with arithmetic averages (UPGMA). The dendrogram was constructed
by cluster analysis of distance matrix data. The genetic distance showed that M
pudica was closely related toM pigra (3.095) compared to 1\1. invisa (3.591). From
the dendrogram, relationships among the Mimosa sp. were determined. M pudica was
in the same cluster with M pigra while M invisa in the different cluter.
SDS-PAGE was used to detect variation and relationships among five selected genus
of Zingiberaceae, i.e Alpinia, Curcuma, Costus, Etlingera and Zingiber. Through
SDS-PAGE, a total of 194 bands were produced by 21 species of ginger that selected.
SDS-P AGE result showed variation of protein bands. The molecular weight was
important and as introduction or identification of variety, species or genus. The
existance of unique bands and certain molecular weights were used to differentiate
and detect variation among five genus of ginger. The sharing of bands and molecular
weight showed, a close relationship among species or genus. Random amplified
polymorphic DNA (RAPD) analysis was used to study the inter and intra-generic
relationships from five genus of Zingiberaceae. Twenty one species were analyzed
with 40 random decamer primers using the Polymerase Chain Reaction (PCR). Ten
arbitrary 1 O-rner gave the best result. Nine primers detected polymorphism in all 21
species, while only one produced monomorphic amplification profiles. A total of 92
bands were amplified. At the inter-generic level, about 95% of RAPDs were
polymorphic and others were monomorphic. The polymorphism at the intra-generic
level, varied from 28% to 85%. Genetic distance among species was calculated with
'RAPDistance' method by Armstrong. The phylogenetic relationships among the
species were analyzed using PAUP 3 .1.1 (Phylogenetic Analysis Using Parsimony),
for a maximum parsimony and the "Neighbour-Joining Pairwise" distance method,
calculated based on the Tamura-Nei algorithm. A phylogram was constructed and
was compared with the morphological schemes of relationships in the Zingiberaceae.
The implicati<:>ns of these results for taxonomic purposes were briefly discussed.
Description
Keywords
Variasi genetik , Lima genera , Famili zingiberaceae