H. I. MALKAWI, N. A. AL-QURAAN AND W. M.

OWAI S
Department of Biological Sciences, Yarmouk University, Irbid, Jordan
Department of Biotechnology and Genetic Engineering, Jordan University of Science and Technology,
Irbid, Jordan
Presented by:
INTRODUCTION
Chlorsulfuron [2-chloro-N-[(4-methoxy-6-methyl-1.3.5-
triazin-2-yL) amino carbonyl benzene sulfonamide]
compound (trade name : Glean) (Ray 1986) belongs to the
sulfonylurea class of herbicides and has been used widely
in weed control (Ray 1984)
Sulfonylurea herbicides inhibit the synthesis of the three
essential amino acids : isoleucine, leucine and valine
(Umbarger 1978), by inhibiting the activity of the rst
enzyme acetolactate synthase (ALS) or acetohydroxy acid
synthase (AHAS) in the synthetic pathway of these
branched chain amino acids (Ray 1984)
Sensitivity of nontolerant crops is like wise due to the
lack of metabolic detoxication of the herbicide in
plants. Many farm crops such as barley (Hordeum
vulgare L.), wheat (Triticum aestivum L.), sugar beet
(Beta vulgaris L.), corn (Zea mays L.) and lentil (Lens
culinaris Medik.)
Resistance to ALS-inhibiting herbicides has been
developed for many crop plants (Kuch & Bright 1981 ;
Newhouse et al. 1992 ; Joel et al. 1995). Such
resistance has been introduced by somatic cell selec-
tion (Anderson & Georgeson 1989), mutation breed-
ing, genetic engineering and interspecic
hybridization (Joel et al. 1995).
MATERIALS AND METHODS
Plant material
The two lentil cultivars used in this study were grown in
Jordan and were provided by the National Center for
Agricultural Research and Technology Transfer (NCARTT)

irradiation material
Gamma-irradiation was carried out at room temperature
in a cobalt-60 gamma cell (Ministry of Energy and Mineral
Resources, Amman-Jordan) of 5000 C activity and at a
dose rate of 57.511 Gray/min.


Irradiation prosedure
Seeds (1000 seeds/treatment) were sub-
jected to gamma-irradiation with the
following doses (4 treatments) :
1. 0 Gray (for 0 min),
2. 90 Gray (for 1.16 min),
3. 100 Gray (for 1.27 min) and
4. 110 Gray (for 1.37 min).
The treated seeds M1 (90, 100 and 110 Gray) and the
control non-treated seeds (at 0 Gray) were grown in the
eld at Marrow Agricultural Station, Irbid, Jordan in a
randomized complete block design with two
replications for each treatment for each cultivar.
Plant treatment with herbicide
when the plants of the treated seeds (M1)
reached about 10 cm in height, chlorsulfuron
(Glean,98 %, DPX, W4189-Dupont, USA) was
applied at 3 g a.i./ha to all treatments using a
CO2-pressurized sprayer of 5-litre capacity.
Surfactant (Tween-20)

RESULTS
Eect of gamma-irradiation treatment
(taken after47 days of seed planting)
The germination percentge of Jordan 1 was 98 % at 0 Gray,
6.2 % at 90 Gray, 5.7 % at 100 Gray and 3.4 % at 110 Gray
while the percentage of germination of Jordan 2 was 95 %
at 0 Gray, 5.1 % at 90 Gray, 4.3 % at 100 Gray and 1.7% at
110 Gray.
Also the average plant height (cm) after complete
germination was reduced as the radiation dose increased,
being for Jordan 1 : 21 cm at 0 Gray, 12.5 cm at 90 Gray, 11.8
cm at 100 Gray and 10 cm at 110 Gray. For Jordan 2, the
average plant height ranged from 18 cm at 0 Gray, 11.8 cm
at 90 Gray, 9.5 cm at 100 Gray and 9 cm at 110 Gray

Response of lentil cultivars to chlorsulfuron

Chlorsulfuron herbicide was spread over the green,
healthy and mature plants (after 7 weeks of germi-
nation). In both lentil cultivars, the colour of the
sensitive plants leaves changed from green to yellow to
brown after the rst week of herbicide application and
died after the second week of herbicide application
when compared with the healthy green tolerant plants
(M1) (data not shown).

Response of lentil cultivars to chlorsulfuron


Response of lentil cultivars to chlorsulfuron


Response of lentil cultivars to chlorsulfuron

Response of lentil cultivars to chlorsulfuron
Result of Purication of ALS enzyme from lentil
plants








CONCLUSION
Especially at the 90 Gray treatment, The decrease in
sensitivity levels of the ALS enzyme to inhibition by
chlorsulfuron herbicide might be explained by the
inheritance of an alteration of the gene(s) coding for
the enzyme, that could lead to overproduction of the
ALS in the plants, via the increase of the expression
level of the ALS gene(s),

Semarang, 22 Mei 2014
ALS (asetolaktat sintase) merupakan enzim yang terbentuk saat awal
pembentukan rantai cabang asam amino (valin, isolisin dan lisin)
Chlorosuforon dapat:
Menghambat sintesis DNA tanaman akibat produksi asam amino
terganggu
Berfungsi baik pada tanaman rumput maupun dikotil
Merupakan herbisida paling aman (ALS tidak terdapat pada sel
hewan)
Contoh herbisida: sulfonylureas (SUs), imidazolinones (IMIs),
triazolopyrimidines (TPs), dll


menghambat kerja dari enzim acetolactate synthase (ALS) dan
acetohydroxy synthase (AHAS) dengan menghambat
perubahan dari ketoglutarate menjadi 2-acetohydroxybutyrate
dan piruvat menjadi 2-acetolactate sehingga mengakibatkan
rantai cabang-cabang asam amino valine, leucine, dan
isoleucine tidak dihasilkan. Tanpa adanya asam amino yang
penting ini, maka protein tidak dapat terbentuk dan tanaman
mengalami kematian (Ross and Childs, 2010).
Tanaman
lentil (Lens
culinaris
Medik.)
radiasi
ti
d
ak
Merubah (mutasi)
susuanan DNA
DNA tetap
Menambahkan
chlorsulfuron
Menambahkan
chlorsulfuron
Enzim
acetolactate
synthase
terhambat
drastis
Enzim
acetolactate
synthase masih
dapat disintesis
lentil (Lens
culinaris Medik.)
rentan terhadap
chlorosurforon
lentil (Lens
culinaris Medik.)
toleran dari
chlorosurforon
Biosintesis leusin melibatkan 8 langkah konversi, yaitu konversi
piruvat menjadi a- acetolactate, isomer a-acetolactate, a--
dihydroxyisovalerate, a-ketoisovalerat, aisopropylmalate, -
isopropylmalate, a-ketoisocaproate, dan akhrinya menjadi leucine.
Proses biosintesis ini dikatalisis oleh enzim acetolactate synthase,
acetohydroxyacid isomeroreductase, acetohydroxy acid
isomeroreductase, dihydroxy acid dehydratase, aisopropylmalate
synthase, isopropylmalate isomerase, dihydrogenase, dan leucine
aminotransferase secara berurutan
Biosintesis leusin