DNA fingerprinting of self-Compatible and self-Incompatible genotypes of sunflower (Helianthus Annuuns L.)
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Date
2005
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Publisher
CCSHAU
Abstract
Sunflower (Helianthus annuus L.) ranks second among oilseed crops in the
world after soybean. Sunflower is protrandrous. Seed set in sunflower is a
complex phenomenon and one of the mean to overcome this problem is by
identifying self fertile and open pollinating lines. The present study was
undertaken to identify molecular marker closely linked to self compatible and
self incompatible gene(s) in sunflower using RAPD analysis.
A total of 26 genotypes (13 self compatible + 13 self incompatible) of
sunflowers were used. Protocols were optimized for DNA extraction and PCR
amplification of self compatible and self incompatible genotypes of sunflower
using RAPD markers.
DNA was isolated using CTAB method with some modifications and among
different genotypes, CMS 338 (C)A gave highest quantity (1034.5 μg/ml) of DNA
whereas genotypes Acc-1445-6 gave lowest amount (317.5 μg/ml) DNA. Quality of
DNA was tested by agorase gel electrophoresis and UV spectrophotometer. A single
discrete band of high molecular weight showed that DNA was pure and free from
contaminants. Optimum PCR amplification was observed on all DNA samples when
reaction mixture contained genomic DNA (100 ng), MgCl2 (1.5 mM), Taq DNA
polymerase (3 units), dNTPs (200 M), 10X Taq DNA polymerase buffer (1 μl),
primer (10 μM) and annealing temperature of 400C.
A total of fifty four random primers were screened and thirty eight primers
produced polymorphism. These thirty eight primers were screened with self
compatible and self incompatible bulks. Primer OPA-9 and OPE-2 produced a unique
DNA band in self incompatible and self compatible bulks, respectively. Marker OPA-9
produced unique band in all the self incompatible genotypes except genotypes IHT-298
and HB-342. This marker is 7.6 cM away from the genes for self incompatible.
Similarly, marker OPE-2 produced unique band in all the self compatible genotypes
except IB-43, Acc-1351-3, HB-105. This marker is about 11.53 cM away from the
genes for self compatibility. Marker OPA-9 produced a unique band in twelve self
compatible genotypes. Marker OPE-2 produced unique band in eleven self
incompatible genotypes.
These markers may be proved useful in marker assisted selection for self
compatible and self incompatible traits, respectively.