University of Kufa

College of Agriculture
Department of Horticulture and Landscape

Morphological Study and some Biochemical

Changes for Water Stress Grape
(Vitis vinifera L.) In vitro

A thesis
Submitted to the Council of the College of Agriculture
at the University of Kufa in Partial Fulfillment of the
Requirements for the Degree of Master in
Horticulture Science
(Tissue Culture)

By
Baqer Sejad Mohmod

Supervised by
Asst. Prof. Dr Muslim Abd Ali Abdulhussein
2011

Abstract
This study was conducted at Plant Tissue Culture Laboratory /
Horticulture Department / Agriculture College / Kufa University and
Agro biotechnology Laboratory - Faculty of Agriculture , University
Putra Malaysia(UPM), which included performance of three factorial
experiments with complete random design(CRD).
The first one was for studying the effect of plant part explants
(node, leaf) and medium supplemented with growth regulators (2,4-D,
NAA+BA) in callus induction of two types of grape (Vitis vinifera L.)
cultivars (Amber Queen and Cardinal) where the callus growth, volume
and fresh weight were regarded as an indicators for these effects.
The second and third experiments were designed to test the
tolerance to water stress of the callus derived from these two grape cvs.
and from their shoots that cultured in vitro. The calli and shoots were
cultured separately in the culture media supplemented with Poly Ethylene
Glycol (PEG) with molecular weight 8000 gm/Mole with concentrations
0, 5, 10, 15 and 20 % for four weeks. Callus and shoot growth and their
content of proline, protein and sugar and enzymatic activity of peroxidase
and catalase were examined as indicators for the determination of the
effect of water stress.
The results of these experiments can be summarized as following:
1. The results of the experiment of callus induction showed the
difference between the two grape cultivars in their ability for callus
induction where Amber Queen had higher ability than Cardinal in
the percentage of response for callus induction and fresh callus
weight (67.50% and 164.58 mg versus 58.06% and 143.92 mg
respectively).
In addition, there was a difference regarding the type of
explant used for callus induction where the nodes were better than
leaves with
a higher significance in the percentage of the
response and callus fresh weight (70.38% and 209.24 mg for nodes
versus 55.72% and 99.25 mg in leaves). Moreover, the volume of
callus derived from nodes was bigger than that derived from
leaves.

It was also found, from the results of this experiments, the

better medium for callus induction of both grape cultivars was that
used for node culture which consist of MS medium supplemented
with 1mg / L BA and 2 mg / L NAA; while there was different
responses in the other callus induction media according to the
different concentrations of NAA, BA and 2,4-D.
2. Results of the second experiment of grape callus tolerance to water
stress demonstrated that supplementation of PEG to the media for
callus induction led to significant decrease in the percentage of
relative growth rate of callus, its dry weight and soluble proteins
where the decreases were in the proportion to the increases in the
concentration of PEG. Whereas, there was an increase in soluble
sugar content, proline, and enzymatic activity of both peroxidase
and catalase.
However, there was no significant effect of grape cultivars
on relative growth of callus and the percentage of dry matter in it.
On the other hand, Amber Queen cv. had more callus content of
total protein, soluble sugar, proline and enzymatic activity of
peroxidase and catalase than that of Cardinal cv.
3. Results of the third experiment of grape shoots tolerance to water
stress showed a significant effect of PEG in some growth
characteristics of shoots in tissue culture for both studied grape
cvs. with increase PEG concentration in the culture medium. There
was a significant decrease in the percentage of number and length
of the shoots and the percentage of their fresh and dry weights,
leaves number and the shoots contents of soluble proteins; while
there was an increase in shoots contents of soluble sugar, proline
and the enzymatic activity of peroxidase and catalase with the
increase PEG concentrations.
In addition, the results showed a differences between grape
cvs. in the stress response that resulted from the presence of PEG
in culture medium, where Amber Queen cv. had a significantly
higher percentage of number and length of shoots, percentage of
fresh and dry weight, leaves number, total protein, soluble sugar,

proline and higher enzymatic activity of peroxidase and catalase

than that of Cardinal cv.
From the results of this study, it can be concluded that it is possible
to use the characteristics of callus and shoot growth in the evaluation of
the tolerance of grape cvs. to water stress in vitro.
Moreover, due to the behavior of Amber Queen cv. under water
stress, depending on its studied characteristics in vitro with PEG, it can be
concluded that Amber Queen cv. is more tolerant to water stress than
Cardinal cv.

..
2011

( ) (2,4-D BA
+NAA) Amber QueenCardinal

PEG)(Poly ethylene glycol 8888

(85080508)% 4


:

-0

Amber Queen

Cardinal

%6775164.58 %58760143.92

Cardinal

(78708%209.24 ) (55% 99.25 )


MS

0 BA+0

NAA

NAABA2,4-D

0-
PEG

Amber Queen

Cardinal

-0
PEG





PEG
PEG

Amber Queen

Cardinal

Amber Queen

PEG

Cardinal