BIOCHEMICAL AND MOLECULAR ANALYSIS OF CMS, MAINTANER AND RESTORER LINES IN PEARLMILLET (Pennisetum glaucum (L.) R.Br.) 1725
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Date
2013-02
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JAU,JUNAGADH
Abstract
Pearlmillet (Pennisatum. glauccum [L.] R.Br.), commonly known
as pearl, cat tail, spiked millet is a member of the poaeeae family and
has a relatively small diploid genome (2n=2x=14) with a DNA content
of lC=2.36pg (Martel et al, 1997). It is the staple food and fodder crop
of millions of people living on the most marginal agricultural lands of
sub-Saharan Africa and the Indian subcontinent.
The present investigation on "Biochemical and molecular
analysis of CMS, maintaner and restorer lines in pearlmillet
[Pennisetum glaucum (L.) R.Br.)" was planned with three main
objectives, (1) To study molecular markers (ISSR and SSR) in different
pearl millet CMS, maintaner and restorer lines. (2) To study the
protein profiling of different pearl millet CMS, maintainer and restorer
lines. (3) To study isoenzymatic patterns in different pearl millet CMS
maintainer and restorer lines.
Ten ISSR primers produced 31 allelcs with 95.5% polymorphism
with an average of 2.8 alleles per primer. 100% polymorphism was
m'>'T<RAcr
observed with the ISSR primers viz., ISSR-1, ISSR-2, ISSR-3, ISSR-5,
ISSR-6, ISSR-7, ISSR-8 and ISSR-9. The PIC varied from 0.387 (ISSR
3) to 0.929 (ISSR-5) with an average of 0.715 per primer. Genetic
Similarity ranged from 36% to 93% with an average similarity of
55.5%. The phylogenetic tree constructed by UPGMA method
generated two main clusters, cluster 1 consisted of four genotypes and
11 consisted of fourteen genotypes.
Twelve SSR primers out of fifteen generated total 14 alleles out
of which 11 allclcs were polymorphic with an average of 0.91 alleles
per primers and average polymorphism of 75%. Highest (100%)
polymorphism was observed with the SSR primers viz., lCMP-3027,
lCMP-3039, lCMP-3063, lCMP-3081, ICMP-88, lCMP-3093, ICMP-
3050, PGIRD-43 and PGIRD-50. The PIC ranged from 0.000 to 0.452
with an average of 0.052 and genetic similarity varied from 28% to
100%. Eighteen genotypes were grouped into two main clusters and
the groupings of genotypes were sub clustered.
The pooled study of molecular marker through ISSR and SSR
revealed a dendrogram consisted of two clusters, I and 11 with an
average similarity of 54%. The similarity coefficient ranged from 34%
to 89%. The highest (89%) similarity was between 95444-B and
J-2290 while the lowest (34%) similarity was between 95222-A and
J-2454. Cluster I consisted of sixteen genotypes while, cluster II
contains two genotypes, 9522-A and J-2372.
The present study indicated that the ISSR seems to be more
effective than SSR as they gave higher percentage of polymorphic loci
and greater range of genetic diversity among 18 pearlmillet genotypes
Protein analysis was done at seedlings stage of 9 DAG The
protein profile generated 9 bands with highest Rm value of 0 515
I80e„.ymes pallcrns for u» charaaerization „r
P«.rlm,lle, genMypes, Thy is„enpy„,e analysis was carrlad oul at 9
msn:mc^
DAG. The maximum numbers of four bands were observed on Native-
PAGE for esterase while, three bands, two bands and one band were
visualized by peroxidase, o-amylase and catalase, respectively.
Thus, DNA fingerprinting and genotype identification through
molecular markers resulted in developing highly diversified
dendrogram techniques are more precise than biochemical markers.
The data revealed that molecular techniques are more precise and
more accurate and can be used for genetic diversity analysis and
developing highly diversified dendrogram of eighteen pearlmillet
genotypes. The information gathered here would be helpful for
genomic mapping studies and for the development of pearlmillet
genotypes with wider and diverse genetic background to enhance crop
productivity.
Ill
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BIOTECHNOLOGY