الفهرس 
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Abstract
The present investigation was conducted to study the following:
First, the phylogenetic relation-ships between ten Egyptian Hordeum
vulgare L varieties depending on agro-morphological criteria, molecular
genetic markers (RAPD-PCR, ISSR-PCR and microsatellite SSR-PCR)
and protein banding patterns electrophoresis, also determination of the
viable marker which has potential power in discrimination and
identification of cultivars and landraces, also identification of landraces
in the present study.
Second, study the effect of salt stress by NaCl treatments on analysis of
variance of agro-morphological traits to identify which varieties are more
tolerant to salinity. Also, study the effect of salinity on soluble protein
contents and proline concentrations in leaves of the studied varieties.
Third, study the effect of salinity in expression of betaine aldehyde
dehydrogenase gene. Also this gene was isolated and it was alignmented
in GenBank to identify it.
The seeds of seven cultivars of Hordeum vulgaries L. were used,
Giza 123, Giza 126, Giza 127, Giza 128, Giza 129, Giza 130 and Giza
2000 were obtained from Agricultural Research Center, Giza, Egypt and
three landraces were obtained from Egyptian National GenBank,
Agricultural Research Center, Giza, Egypt, under code number (11557,
11580 and 113737), these landraces are collected from Sinai, El-Aresh
(El-Kheroba, El-Sheikh Zuwaid) and Ras Sedr (Wadi Sedr).
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161
The obtained results could be summarized as follows:
I-Agro-morphological Criteria studies:
1- Ten studied Hordeum vulgare L variedies were discriminated and
identified to determine the phylogenetic relationships between them
depending on analysis of variance of studied Agro-morphological
traits (branch number/plant, shoot length (cm), root length (cm),
node number/plant, spike length (cm), spike weight (gm), no. of
grains/spike, grain weight /spike (gm) and grain weight/plant (gm))
The result of agro-morphological phenogram by UPGMA indicated
that the agro-morphological phenogram is completely similar to the
pedigree of studied varieties.
2- from the results obtained from studing the effect of different
concentrations of NaCl (control, 9000, 12000 and 15000 ppm) on
studied agro-morphological traits of ten studied barley varieties, we
concluded that Giza 2000 cultivar and Wadi Sedr landrace are
considered more tolerance varieties between studied varieties while
Giza 129 cultivar considered a sensitive cultivar.
II- Molecular genetic marker studies
Classification technique based on the use of molecular markers like
RAPD, ISSR and SSR provide a much more accurate and powerful
means of analyzing genetic relationship
The result of RAPD, ISSR and SSR fingerprinting patterns revealed
different levels of polymorphism among the ten varieties. This
corresponds to a level of polymorphism of 41.77%, 62.02% and
61.22% respectively this reflect that ISSR markers are more
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162
discriminating than SSR and RAPD to evaluate the genetic diversity
and relationship among studied varieties.
The phenogram obtained from the collective data of three markers
classified the three landraces in separate cluster with Giza 2000 this
indicated that there are a similarity distance between studied landraces
and Giza 2000.
III- gene expression studies under salinity and BADH gene isolation
1- We studied the protein electrophoresis banding patterns on ten
studied varieties to determine the phylogenetic relationships
between them by phenogram of UPGMA, from result, we found
that the studied landraces grouped into a separate cluster and had a
high similarity between them, then the three studied landraces had
high similarity with Giza 2000, also Giza 128, Giza 129 and Giza
130 have the same similarity degree.
2- from studying the effect of salinity on protein electrophoresis
banding pattern of ten studied varieties at three sampling time
points (sampling after 24 hours, 1st week and 2nd week salt
treatment). from results we found that a maximum polymorphism
was observed at third time point (sampling after 2nd week salt
treatments), but showed the lowest variation. While sampling after
24 hours and 1st week salt treatment showed a maximum variation.
Also the bands intensity shown maximum intensity at first time
point (sampling after 24 hours), while showed minimum intensity
at third time point (sampling after 2nd week salt treatments). Also,
the proteins of Mw 28 KDa and Mw 49KDa disappeared in
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162
sensitive varieties (Giza 129) and appeared in tolerant varieties
after 24 hour and 1st week from salt treatments respectively.
3- Proline is a type of proteins play an important role in a biotic stress
as draught and salinity so we determined the concentration of
proline formed in studied varieties under different concentration of
NaCl, from results we found that Giza 2000 and Wadi Sedr formed
a high concentration of proline under salt stress this explain why
this two varieties are tolerant to salt stress while Giza 129
(sensitive cultivars) form low concentration from proline.
4- In the present investigation, BADH-1 gene expression was
determined by Real Time PCR, then this gene was isolated from
genomic DNA and cDNA from the most tolerant barley (Giza 2000
and Wadi Sedr) and sensitive barley Giza 129 cultivar, then
sequencing of this gene was performed and it was aligned in
GenBank. This nucleotide sequences were translated into amino
acid sequences then these amino acid sequences were aligned in
GenBank, also the 3D of BADH-1 enzyme were determined.
These isolated genes were submitted in GenBank under accession
number KX433169, KX342849, KX342850, KX342851,
KX433170 and KX433171.
from Agro-morphological, molecular and biochemical studies, the
three studied landraces could be discriminated and identified, they are
more similar to Giza 2000 cultivars. While from studing the effect of
salinity on three landraces, we found that Wadi Sedr landrace is near
to Giza 2000 cultivar.
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162
from this study, we recommended cultivating the more tolerant
varieties (Giza 2000 and Wadi Sedr) on salin soil within the salinity
range we used in this studies