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1
Institute of Soil & Environmental Sciences, 2Department of Plant Breeding & Genetics,
University of Agriculture, Faisalabad, Pakistan
Corresponding authors’e-mail: wazir1604@yahoo.com1, .qurbanalisaim@yahoo.com2

#
Seed dormancy is key factor for late germination of seeds. Therefore, it is common practice to use
various techniques to break down dormancy of dormant seeds for the improvement of
germination rate. Exogenous ethylene is known to stimulate germination of dormant and non-
dormant seeds if applied in right concentration as excessive ethylene delays or inhibits
germination. In order to assess germination variations for ethylene in to
exogenously applied ethylene released from calcium carbide, five cross pollinated genotypes
were screened at the seedling stage by growing them under the influence of 10, 20, 30 and 40 mg
petri-dish-1 calcium carbide in Sanyo Incubator MIR 253 in disposable plastic petri-dishes with
rubber stopper at one side and sealed air tightly with silicone. A treatment containing calcium
sulfate equivalent to the amount of calcium in CaC2 was added in control was also maintained
during study for comparison. Findings were that CaC2 @ 40 mg petri-dish-1 inhibited germination
of cucumber seeds but lower rates (10, 20 and 30 mg petri-dish-1) stimulated germination In short,
summarized results exploited that there was 72% increase in germination percentage, 98%
increase in shoot weight, 13% increase in shoot due to addition of CaC2 compared to control.
/ ' 0 $ calcium carbide germination, tomato, root-to-shoot ratio, seedling vigor
$ "
A seed is programmed to survive after being dispersed from the mother plant until the
establishment of a photosynthetically competent seedling. Seed dormancy prevents germination
during periods unfavourable to seedling growth. Primary dormancy develops during seed
formation on the mother plant and can be relieved by dry storage or by imbibition at certain
temperatures over a particular time period. The breaking of dormancy is characterized by changes
in several physiological parameters that affect the subsequent germination responsel3, 14, 28
.
Ethylene is known to stimulate the germination6, 24
and break the dormancy of many kinds of
29, 28
seeds . In most species, exogenous ethylene or ethephon (an ethylene releasing compound)
stimulates the germination of dormant and non dormant seeds 18, 16, 9, 8. Ethylene at concentration
of 0.1 to 200 µL is usually sufficient to stimulate seed germination of many species. Ethylene or
ethephon removes primary dormancy in peanut17. The response of ethylene may depend on time
of application12, 27
. Ethylene can promote germination of non-dormant seeds at non-optimal
conditions. The inhibition of lettuce2 and chickpea11 seed germination due to supra-optimal
temperate could be alleviated by ethylene. Thus, in many species, the inhibition of seed
germination due to dormancy or stress conditions can be completely or partially reversed by
ethylene or ethephon. But the level of exogenous ethylene is a limiting factor in the germination
of these seeds. After germination effect of ethylene starts on root growth which may be especially
influential in fashioning the root system34, 25, 28, 21, 33, 4.
Calcium carbide (CaC2) has been now well known to influence wide range of growth and
development processes of plant1,26,31 from germination (seed dormancy and germination
processes, 28,.21,33,4 to maturity due to bifacit action of gases released from it 13,14,20. The two gases
released from calcium carbide are acetylene (C2H2) and ethylene (C2H4)31 but ethylene being
potent plant hormone23 to stimulate early and enhanced root growth32, 31
and fruit set19. These
gases have shown great impact in improving the quality and production of almost all types of
crops23, 15, 5, 20, 28, 21, 33, 4.
In the present manuscript, effect of calcium carbide released gas, ethylene on seed germination
and rate of root and shoot growth in tomato genotypes under controlled conditions was
investigated by using different rates of calcium carbide.
" $ + $
The germination of tomato ( L.) seeds under the influence of CaC2 was
investigated with the help of a lab experiment in Soil Fertility & Plant Nutrition Lab, Institute of
Soil & Environmental Sciences, University of Agriculture, Faisalabad (Pakistan) during 2009-10.
A Completely Randomized Design (CRD) with four replications was followed during this
experiment. Disposable plastic petri dishes of 80cc (cm3) in volume with covers were used.
Before starting the experiment, a hole of 4 mm diameter was made near edge in the cover of these
petri dishes and then plugged this hole with the help of a rubber stopper and sealed with silicone.
These holes were later on use to inject water. Analytical grade calcium carbide (27% a.i. CaC2,
Ningxia National Chemical Group Co. Ltd. China) in powdered form from Sigma was used for
all the experiments.
Seeds of tomato were washed with 70% ethanol to remove any fungus/bacteria present on seed
coat. Then seeds were washed 5 times with distilled water and stored at 4°C till used for
experiments. Three autoclaved Whatman No. I filter papers were used in each petri dish. 1st filter
paper was placed at the bottom of petri dish and required dose of CaC2 was spread over the paper.
Second filter paper was placed over it and 5 pretreated seeds were spread on it. Third filter paper
was placed above the seeds to check their movement or floating of seeds. For the treatments
without calcium carbide, three filter papers were used as above and de-ionized water was used to
supply moisture in all treatment. During the study, four levels of CaC2 (3, 6, 9, 12 and 15 mg
petri-dish-1) were placed at the middle of first filter paper so that gases released from CaC2 would
uniformly diffuse to all directions. A treatment containing calcium sulphate salt (CaSO4)
equivalent to calcium in was also run as control for comparison. Five genotypes of tomato i.e. Rio
Grandi +1, Tenzila, Romor Ser-France, Top Gord and F-1 3560-S were used during study.
Each petri dish was closed with cover having rubber septum and rapped and sealed with the help
of Parafilm tape (Parafilm “M” Laboratory Film, Pechiney Plastic Packing, Chicago, IL 60631).
De-ionized water (7 mL) was injected to each petri dish by using a 12 mL disposable syringe.
Then these petri dishes were placed in a Sanyo Incubator MIR 253 under the following
experimental conditions: 24+2oC temperature and 12/12 hours day and night i.e. light/dark
period. Germination was counted after every 24 hours interval. At completion of seed
germination, data about root and shoot weights and length was taken after 120 hours growth
period in an incubator and analyzed by using standard statistical methods30.
At completion of seed germination, data about root and shoot weights and length was taken after
120 hours growth period in an incubator and analyzed by using standard statistical methods30.
1
This research was operated under controlled conditions. Outcomes disclose that calcium carbide
not only hastened germination rate of seeds but also impelled root and shoot growth of five
genotypes of tomato i.e. Rio Grandi+1, Tenzila, Romor Ser-France, F-1 3560-S and Top Gord.
Scrutiny on planting materials was switched on just when there was commence of germination in
seeds in any treatment. Indication of seed germination set off to become visible after 48 h in some
petri dishes and germination process was finalized at 120 h. All the genotypes of tomato unveiled
high but variable response to calcium carbide application at different rates.
Data belonging to germination percentage of five tomato genotypes (Tables 1a, 1b & 1c) exposed
existence of significant differences among tomato genotypes for seed germination due to the
effect of rate of application of calcium carbide, time of incubation, genotype and their interaction.
Data also illuminated that CaC2 cut down germination period by causing early breakage of
dormancy which escorted to higher germination percentage in CaC2 treated seeds. Calcium
carbide treated seeds got under the way to germinate after 48 h compared to those seeds without
calcium carbide i.e. control where seed germination was set up at 72 h. After 72 h, there were
only 11 % seeds germinated in the control treatment whereas at this time interval 22-67 %
germination was perceived in seeds with supplanted with calcium carbide (Table 1a & Fig. 1).
Germination percentage was multiplied with raising level of calcium carbide (Tables 1a & 1c).
End results also pointed out that germination rate picked up the pace with raise in rate of CaC2
applied after 72 h of incubation growth period in all the tomato genotypes except Top Gord.
Assessment among germination percent at different rates of CaC2 at 72 h sign post that highest
germination (67.0 %) was observed in the treatments having 15 mg CaC2 and it was statistically
chased by the treatment of 12 mg CaC2 with 69.6 % germination. Response of all tomato
genotypes to added CaC2 was statistically at par at 72 h of growth period (Table 1b). But after 96
h of growth period, Tenzila responded totally differently by showing highest germination (78 %)
at 15 mg CaC2 (Table 1c). This genotype also paraded the highest germination (51.9 %) at 3 mg
CaC2 treatment which was statistically at par to that at 6 and 9 mg CaC2. Least germination was
detected in the control. Germination of genotype Tenzila went on increasing afterwards with the
increase in rate of CaC2 application. It reached to 70.6 % at 12 mg CaC2 and then to a maximum
value of 78 % at 15 mg CaC2 at 96 h growth periods. Comparison of seed germination at 120 h
revealed that minimum germination percentage was noted in control which augmented to 80.3 %
due to supplement dose of CaC2. Statistical comparison of effect of various rates of calcium
carbide and five genotypes of tomato (Figures 1) bring to light that Rio Grandi +1 gave 100 %
germination at 12 and 15 mg CaC2 whereas F-1 3560-S showed off 100 % germination at 15 mg
CaC2. Genotype, Rio Grandi+1 showed excellent results regarding germination among all the
tomato genotypes used in the study.
2 " #"$ $ + 3 0+ 3 '
A lot of variations were discerned for root length under the impact of varying rates of calcium
carbide. Minimum mean root length (30 mm plant-1) was noted in control which amplified to 41 -
88 % due to CaC2. Minimum root length (22 mm plant-1) was shown by genotype Romor Ser-
France in control while maximum (62 mm plant-1) was shown by F-1 3560-S at 15 mg CaC2
among all CaC2 treatments and genotypes. Rio Grandi +1 and Tenzila closely followed F-1 3560-
S and had root length 58 mm plant-1 at 15 mg CaC2 (Table 2).
Similar response of tomato genotype was observed for shoot length (Table 3). Addition of
calcium carbide resulted in 55 to 113 % increase in mean shoot length over mean shoot length in
control. Statistical comparison among five tomato genotypes and average across calcium carbide
levels revealed that Rio Grandi +1 exhibited the largest shoot length.
2 " #"$ + " 3 '
Results regarding root-to-shoot ratio on length basis (Table 4) indicated that all CaC2 applied
rates except 3 mg CaC2 (1.96) showed lower mean root-to-shoot ratio than control (1.93). This
ratio ranged from 1.62 to 2.28 at control and increased from 1.67 to 2.41 at 3 mg CaC2. This ratio
was then decreased with increasing levels of CaC2 and ranged from 1.51 to 2.15 1.38 to 2.13,
1.62 to 2.02 and 1.49 to 2.05 at 6, 9, 12 and 15 mg CaC2, respectively. Minimum root-to-shoot
ratio (1.38) was recorded in the genotype Tenzila in the treatment where 9 mg CaC2 was applied.
However, maximum root-to-shoot ratio was observed in genotype Romor Ser-France at 3 mg
CaC2. These results indicate that root-to-shoot ratio increased up to 3 mg CaC2 level and
afterwards it tended to decrease with increase in level of CaC2.
4" "
Finalized conclusions suggest that there was a positive correlation to a critical concentration
between calcium carbide and seed germination. Calcium carbide had pronounced influence on
germination7, 10, 13
. Application of calcium carbide stimulated root growth28 and root-to-shoot
ratio28 due to classical triple response of ethylene 7, 10, 13, 14, 21
. . Siddiq 33
and Anwar .4
also reported similar results in agronomic and vegetable crops. Exogenous application of small
amounts of C2H45 in the rhizosphere physiologically is active in influencing the growth22 and
development1 of tomato plants22, 14, 28. However, its higher amount can retard growth rather than
to stimulatel3. Conclusively, slow and consistent release/conversion of C2H4 from C2H222 released
from CaC2 is pre-requisite to obtain improved quality produce of vegetables20, 28, 21,33, 4.
5 0 $3 We are thankful to Higher Education Commission (HEC) Pakistan for
financial assistance during smooth and sound running of the study.
1
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, N. Claassen, H. Flessa, W.B. Frommer, H. Goldbach, H.W. Olfs and V. Romheld Effect
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& # Comparative effect of different rates of calcium carbide on germination percentage of tomato
genotypes from 72 h to 120 h of growth period in incubator.
&" 1 & 3 '
" 6 ! !
Control 11.0 gh* 11.0 gh 11.0 gh 0h 11.0 gh
3 mg 33.0 e-g 44.3 d-f 33.0 e-g 33.0 e-g 22.0 f-h
6 mg 44.3 d-f 44.3 d-f 44.3 d-f 44.3 d-f 44.3 d-f %7
72 h 9 mg 55.7 c-e 33.0 e-g 44.3 d-f 55.7 c-e 55.7 c -e 8 9%
12 mg 55.7 c -e 55.7 c -e 55.7 c -e 67.0 b-d 45.7 d-f 8 29
15 mg 67.0 b-
67.0 b-d 55.7 c -e 33.0 e -g 67.0 b-d : 8 4 9
d
Control 11.0 gh 44.3 d-f 11.0 gh 11.0 gh 11.0 gh : :
3 mg 33.0 e-g 55.7 c -e 33.0 e -g 33.0 e -g 33.0 e -g : 7
6 mg 55.7 c -e 55.7 c -e 55.7 c -e 55.7 c -e 55.7 c -e : 29
9 mg 55.7 c -e 55.7 c -e 55.7 c -e 55.7 c -e 55.7 c -e : 29
96 h
12 mg 67.0 b-
67.0 b-d 67.0 b-d 67.0 b-d 67.0 b-d : ; 4
d
15 mg 67.0 b-
78.0 a -c 67.0 b-d 67.0 b-d 67.0 b-d 8 ;!
d
Control 36.7 ef 44.3 d-f 22.0 f-h 22.0 f-h 33.0 e-g
3 mg 67.0 b-d 55.7 c -e 44.3 d-f 44.3 d-f 44.3 d-f 2 %
6 mg 81.7 ab 55.7 c -e 55.7 c -e 55.7 c -e 55.7 c -e 8 ! 2
120 h
9 mg 96.3 a 67.0 b-d 67.0 b-d 78.0 a -c 67.0 b-d : ;
12 mg 100.0 a 89.0 ab 84.7 ab 89.0 ab 67.0 b-d 8
15 mg 100.0 a 89.0 ab 95.7 a 67.0 b-d 100.0 a 8
:: ; ; ;
*Values sharing same letter(s) in a column do not differ significantly at P=0.05 according to LSD test
V-1 = Rio Grandi +1, V-2 = Tenzila, V-3 = Romor Ser-France, V-4 = Top Gord, V-5 = F-1 3560-S
& # # Summarized table on comparative response of five tomato genotypes for seed germination (%) at
different interval of growth period
&" " 6 & 3 '
<+ = Rio Grandi +1 Tenzila Romor Ser- Top Gord F-1
France 3560-S
72 44.4 c* 42.6 c 40.7 c 38.8 c 40.9 c !
96 48.2 c 59.4 b 48.2 c 48.2 c 48.2 c ;
120 80.3 a 66.8 b 61.6 b 59.3 b 61.2 b
:: ; ; ;
*Values sharing same letter(s) in a column do not differ significantly at P=0.05 according to LSD test

& # Summarized table on comparative response of five tomato genotypes for seed germination (%) at
different levels of calcium carbide
& 3 '
&

Control 19.6 hi* 33.2 gh 14.7i 11.0 i 8.3 i 8 4

3 mg CaC2 44.3 fg 51.9 ef 36.8 g 36.8 g 33.1 gh !
6 mg CaC2 60.6 b-e 51.9ef 51.9ef 51.9ef 51.9ef ;
9 mg CaC2 69.2 a -d 51.9ef 55.7d-f 63.1b-e 59.4c -e 88;
12 mg CaC2 74.2 ab 70.6a -c 69.1a -d 74.3ab 59.9c -e 8
15 mg CaC2 78.0 a 78.0a 72.8a -c 55.7d-f 78.0a :
:: ; ; ;
*Values sharing same letter(s) in a column do not differ significantly at P=0.05 according to LSD test
V-1 = Rio Grandi +1, V-2 = Tenzila, V-3 = Romor Ser-France, V-4 = Top Gord, V-5 = F-1 3560-S
& # Comparative effect of different rates of calcium carbide on root length (mm) of tomato genotypes
after 120 h growth period in incubator
1 ! ! < 3> " $" + =
& 3 '
! 8
Rio Grandi +1 33 m* 44 g 52 ef 52 f 55 b-d 58 b 8 ;
Tenzila 33 m 43 gh 43 gh 42 hi 51 f 58 b !
Romor Ser-France 22 o 36 k-m 38 i-k 39 ij 39 ij 52 f ::2
Top Gord 28 n 37 i-l 38 i-k 37 j-l 56 b-d 52 f 4
F-1 3560-S 34 lm 52 f 55 b-d 54 d-f 57 bc 62 a
2 4 ! ! ;
*Values sharing same letter(s) in a column do not differ significantly at P=0.05 according to LSD test
V-1 = Rio Grandi +1, V-2 = Tenzila, V-3 = Romor Ser-France, V-4 = Top Gord, V-5 = F-1 3560-S
& # Comparative effect of different rates of calcium carbide on shoot length (mm) of tomato genotypes
after 120 h growth period
1 ! ! < 3> " $" + =
& 3 '
! 8
Rio Grandi +1 18.3 j-l* 27.3 ef 28.3 e 28.6 e 33 cd 38.3 ab 8
Tenzila 16.6 lm 27.3 ef 25.3 e-h 28.3 e 28.3 e 39.3 a : ;
Romor Ser-France 13.6 mn 20.6 i-l 21.6 g-j 23.6 f-i 25.3 e-h 25.6 e-g :!
Top Gord 12.3 n 20.6 i-l 21.3 h-k 23 g-i 25.6 e-g 28.6 e 8!
F-1 3560-S 17.3k-m 25.3 e-h 25.6 e-g 29 de 29 de 34.3 bc ;
4 ! ! ;! ;
*Values sharing same letter(s) in a column do not differ significantly at P=0.05 according to LSD test
V-1 = Rio Grandi +1, V-2 = Tenzila, V-3 = Romor Ser-France, V-4 = Top Gord, V-5 = F-1 3560-S

& # Comparative effect of different rates of calcium carbide on root-to-shoot ratio of tomato genotypes
on length basis after 120 h growth period in incubator
1 ! ! < 3> " $" + =
& 3 '
! 8
Rio Grandi +1 1.80 jk* 1.67 e-k 1.52 i-k 1.90 c-i 1.82 d-j 1.82 d-j : ;
Tenzila 1.99 e-k 1.53 h-k 1.71 e -k 1.38 k 1.62 g-k 1.49 j k ;
Romor Ser-France 1.62 g-k 2.41 a 2.15 a -d 1.73 e -k 1.91 c -h 1.63 f -k 8
Top Gord 2.28 ab 2.25 a -c 1.51 j k 1.95 b-g 2.01 b-f 1.60 g-k 8
F-1 3560-S 1.96 b-f 1.92 c-g 1.82 d-j 2.13 a-d 2.02 b-f 2.05 a-e 8
8 ; 8 : ! ;! ; : !
*Values sharing same letter(s) in a column do not differ significantly at P=0.05 according to LSD test
V-1 = Rio Grandi +1, V-2 = Tenzila, V-3 = Romor Ser-France, V-4 = Top Gord, V-5 = F-1 3560-S
 Values sharing same letter(s) in a column do not differ significantly at P=0.05 according to
LSD test.
V-1 = Rio Grandi +1, V-2 = Tenzila, V-3 = Romor Ser-France, V-4 = Top Gord, V-5 = F-1
3560-S

9"3 Effect of different levels of calcium carbide on germination of five tomato genotypes
after 120 h growth period in incubator.