Republic of Iraq

Ministry of Higher Education and Scientific Research

University of Kufa
College of Agriculture
Department of Horticulture and Landscape

In Vitro production of Vincristine and Vinblastine

from Catharanthus rosues(L.) G.Don callus
cultures under salt stress

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
Ihsan Jaly Ozbeeb Al-Hujamy
Supervisors
Assist. Prof. Muslim A. A. Abdulhussein
2010

Abstract
Number of experiments were conducted at Plant Tissue Culture
Laboratory / Horticulture Department / Agriculture College / Kufa
University during 2008-2009 to study the effects of NaCl on
Catharanthus roseus cv. rosea callus growth, which initiated from in
vitro multiple shoots, and its total indole alkaloids, vincristine and
vinblastine contents.
This study was conducted with three stages. First stage was to
study the effect of Benzyladenin (BA) in MS medium at ( 0, 0.1, 0.2 or 0.3
mg/l ) concentrations on axillary bud burst percentage. The results
showed that 0.3 mg/l BA gave the highest percentage( 100 % ) . For
Shoot multiplication, Benzyladenin (BA) at concentration of ( 0, 1, 2, 3)
mg/l and Naphthaleneacetic acid (NAA) at ( 0, 0.1, 0.2 or 0.3 mg/l) were
added to MS medium alone or with combination . Result showed that
addition of 2 mg/l BA + 0.1 mg/l NAA gave the highest shoot number ,
shoot length and fresh and dry weights as compared with other
treatments..
The second Stage included studying the efficiency of some growth
regulators (2,4-D ; 2,4-D and BA , or NAA and BA) on the initiation and
growth of callus from axillary bud and leaf of invitro multiple shoots .
The results indicated that axillary bud

gaved the best result as

compared with the leaf, and the best medium for callus initiation and
growth was MS medium with 0.5 mg NAA and 1 mg BA or0.5 mg NAA
and 2 mg BA .

The third stage was conducted to study the response of callus growth
in MS medium which supplied with ( 0, 25 , 50 or 75 mM ) NaCl for 30
days and its productivity of

total indole alkaloids, vincristine and

vinblastine. The results showed significant differences in content of

callus in total indole alkaloids, vincristine and vinblastine which cultured
under different NaCl salt concentrations in MS medium. The results olso
showed that NaCl gave a significant increase in these parameters as
compared with the control ( with out NaCl ). The highest amount of total
indole alkaloids, vincristine and vinblastine (81.3
81.3 mg, 21.5 g and 11..
g / gm dry weight ) respectivily was found in the callus cultured in MS
medium that supplied with 50 mM NaCl as compared with the lowest
values( .1..mg , .121g and 81.1g / gm dry weight ) in the control.

Catharanthus roseus
(L.) G. Don

( )

...
50.0

/ / 2009 - 2008

rosea NaCl

Catharanthus roseus

BA

MS

)3 % .00

BA ) 8 5 . 0 /

0.1 BA /

3 NAA

2,4-D

MS 2 /

NAA ) 018 015 01. 0 /

) 0 4 018 015 01. 0 /

03. /

NAA

BA

)/(BA

3 012 / ) .10 + NAA

0.5 / ) 2 + NAA / ) BA

Vincristine

Vinblastine

Vincristine

21.5

.121

81.1

11..

.1..
.

MS 20

81.3

Vincristine

Vinblastine

80

3Vinblastine

NaCl

.2 20 52 0) NaCl