Journal of Experimental Botany, Vol. 28, No. 103, pp.

484-492, April 1977

Some Primary Features of Salt Tolerance in Senna

(Cassia acutifolia)
ALI T. AYOUB
Hudeiba Research Station, P.O. Box 31, Ed-Damer, Sudan

Received 1 March 1976

Downloaded from http://jxb.oxfordjournals.org/ at University of York on August 19, 2014

ABSTRACT
Studies on the germination of Cassia acutifolia (senna) seeds indicated that they were highly
tolerant of salinities up to 160 mmhos cm"1, and 50 per cent reduction in germination occurred
at about 20-0 mmhos cm"1. Air temperature had a significant effect on senna germination at
substrate salinities between 10-0 and 25-0 mmhos cm"1.
Seedling growth was more sensitive to salinity and alkalinity than the germination stage.
Young plant survival and total pod yield were significantly reduced in soil salinities higher than
11-0 minhoa cm"1, and this was more pronounced when plants were irrigated at short intervals
•with saline waters. The sensitivity of senna to higher levels of salinities was correlated with the
higher rate of chloride accumulation in the tissue resulting in specific chloride injury. However,
the possibility of sodium injury cannot be excluded.

INTRODUCTION
In a previous paper (Ayoub, 1975), the importance of senna (Cassia acutifolia) as a
promising irrigated cash crop was put forward. The crop requires high temperatures
and an abundance of sunlight (Gupta, 1971), and these conditions prevail in the
more arid parts of the Sudan where soil salinity and alkalinity problems may be
important. A study was, therefore, undertaken on the effects of salt levels, salt
types, and soil alkalinity on senna germination, vegetative growth, yield, and
mineral composition. Some environmental factors which may effect salt tolerance
by senna, such as watering interval and air temperature, were also included in
these studies. The present paper reports some primary features of these studies.

MATERIALS AND METHODS

Germination studies
Germ Lnation experiments were carried out i n 10 -cm-diameter Petri dishes in a single randomized
design with 11 levels of NaCl solutions (0-40-0 mmhos cm"1) replicated four times. In each
sterilized Petri dish, 10 healthy surface-disinfected (HgCla) senna seeds were placed on a filter
paper bed, covered with another sheet of filter paper, and soaked with 10 ml of the specified
solution. This volume was sufficient to saturate the filter papers and to partially immerse the
seeds. The Petri dishes were covered and placed in the dark in an incubator maintained at
30 ± 2 °C. The germination coimt was taken every 24 h.
Interaction of temperature and salinity on germination
The germination tests were carried out in Petri dishes as described above. Salt levels used
were 0,50, 8-3, 16-3, 24-5, and 33-6 mmhos cm"1 NaCl at 25 °C and each test was replicated four
 Ayoub—Salt Tolerance in Senna 485
times. Replicates were incubated at 12-15, 28-30, 32—35, and 40-45 °C and germination counts
were made on the 7th day.

ESP tolerance studies

Hudeiba Research Station Farm heavy clay soil (Vertisols) was air-dried, crushed, and sieved
through 2 mm mesh. Soils of different exchangeable sodium percentage (ESP) levels were pro-
duced following the method of Chang and Dregne (1955). Final ESP levels obtained were
approximately 5, 15, 24, 35, 45, and 55.
These soils were again air-dried, sieved, and 5'5 kg of each soil placed in 21-cm-diameter
plastic pots and randomized on a ventilated greenhouse bench. Each soil test was replicated
five times. Ten seeds of senna were sown in each pot and irrigated when necessary with low-
salt tap water (0-20 mmhos).
Germination counts were recorded and after a period of 30 d plants were harvested for dry
matter determinations.
Effect of salt types and levels on plant growth

Downloaded from http://jxb.oxfordjournals.org/ at University of York on August 19, 2014

About 5 • 5 kg of washed sandy loam soil were put in 60 plastic pots (21 cm diameter) with drain-
age holes and 10 seeds of senna were sown in each pot. For a period of 2 weeks the pots were
irrigated alternately with tap water and Hoagland nutrient solution after which the seedlings
were thinned to four plants per pot and 45 pots of uniformly growing plants were selected for
salt treatments.
Equal concentrations of NaCl, CaCla, Na2SO4, and an equal mixture of NaCl and CaCl2
were compared at the levels shown in Table 2. Saline waters were applied gradually to allow
for osmotic adjustment until the final specified salt level was maintained. The plants were
harvested after 8 weeks of growth in salt treatments and the fresh weight of the new growth
and its mineral contents were determined. Observations such as plant survival and leaf shedding
were also taken.
On dry ashed plant material Na and K were determined by a flame photometer, Ca and Mg
by titrating with EDTA, and Cl using the AgNOs method (Chapman and Pratt, 1961). Soil
salinity (EC) was measured following the U.S. Salinity Laboratory method (Richards, 1954).
All experiments reported above were repeated and comparable results were obtained.
Frequency of irrigation with NaCl on senna growth
Seeds of senna were planted in metallic lysimeters with drainage holes at the bottom and
containing about 130 kg of river silt. The lysimeters used were oil drums 65 cm high and 60 cm
in diameter and painted with asphalt. The experimental design was a factorial combination of
five concentrations of NaCl (0-2, 2-5, 5-0, 10-0, and 20-0 mmhos cm"1) and two intervals of
watering (every 7 or 14 d with the salt solution). These lysimeters, with three replicates, were
arranged randomly in the open air where daily temperatures ranged between 25 and 39 °C.
They were watered with tap water whenever necessary until the salt treatments were applied.
Just before applying salt treatments the plants were thinned to eight per lysimeter.
Salt was applied after 4 weeks from sowing in increments of 2-5 mmhos NaCl in the irrigation
water every other day to allow for osmotic adjustment until the plants were receiving solutions
of assigned concentration. Salt concentrations in the soils at harvest are shown in Table 3 as
mmhos cm"1 of saturated extract. The volumes of solutions added were sufficient to allow
copious drainage and thorough flushing out of the root medium.
Observations concerning plant survival and development were recorded. Samples of dead
plants were also taken throughout the duration of the experiment. Pod yields were recorded
periodically and at the termination of the experiment yield components and some plant charac-
ters were determined. Leaf samples were also taken for Na and Cl analysis.

RESULTS
Effect and NaCl levels on seed germination
Senna seed germination never exceeded 75 per cent and under field conditions the
maximum germination obtained was usually less than 60 per cent. This low germina-
tion percentage is mainly due to the presence of 'hard' seeds which do not imbibe
486 Ayoub—Salt Tolerance in Senna

100-

90

80-

— 70-

| 6 0 -

I 50-
8

Downloaded from http://jxb.oxfordjournals.org/ at University of York on August 19, 2014

I 40

30-

20-

10

0
4 5 6
Days from sowing
FIG. 1. Senna seed germination at different NaCl levels. The numbers on the graph are NaCl
solution conductivities in mmhos cm"1 at 25 °C.

water even after 10 d soaking. Laboratory tests have indicated that these seeds
have an impermeable seed coat. The data shown in Fig. 1, presenting germination
percentage in relation to different salt levels of the substrate, are therefore based on
germinable seeds.
Control seeds completed germination on the 7th day. Seed germination was
satisfactory up to a salt level of about 16-0 mmhos cm"1 NaCl. Between salt levels
of 20-0 and 24-0 mmhos cm"1 germination was reduced 50 per cent and beyond the
salt level of 30-0 mmhos cm"1 germination was almost inhibited.

Interaction of salinity and temperature on senna germination

Fig. 2 shows the effect of salinity at different ah" temperatures on senna germina-
tion. Substrate salinity, ah- temperature, and their interaction highly significantly
affected senna germination. Up to a salinity of 6-0 mmhos cm"1 there was no differ-
ence between temperatures from 28 to 45 °C hi germination percentage. Air tem-
perature had its greatest effect on germination at salinities between 10-0 and 25-0
mmhoa cm"1. Fifty per cent reduction in germination occurred at salinities of about
10-0, 170, and 22-0 mmhos cm"1 when air temperatures were 40-45,32-35, and 28-
30 °C, respectively. Senna did not germinate at temperatures below 15 °C.

Effect of ESP levels on senna germination and seedling groioth

The data ha Table 1 shows the effect of different ESP levels on senna seed germina-
tion and seedling growth. Seed germination was satisfactory up to an ESP level of
 Ayovb—Salt Tolerance in Senna 487

Downloaded from http://jxb.oxfordjournals.org/ at University of York on August 19, 2014

o-o 5-0 10-0 15-0 20-0 25-0 300 350
NsCI levd (mmhos cm"1)
F i o . 2. Effect of NaCl levels a t different air t e m p e r a t u r e s on senna seed germination: -O,
28-30 °C; • • , 32-35 °C; A A, 40-45 °C.

35. Beyond that level some seeds germinated, but most of the seedlings died shortly
after that. Seedlings growing in ESP levels of 25 or more were stunted and had
bleached-yellowish (chlorotic) secondary leaves. The effect of ESP was, there-
fore, more pronounced at the seedling growth stage than during seed germination.

TABLE 1. Effect of exchangeable sodium percentage (ESP) on senna seed germination

and seedling growth
ESP Relative Relative
germination (%) growth (%)

5 100-0 100-0
15 96-3 601
25 88-9 45-8
35 88-9 32-8
45 48-2 25-0
55 33-3 100

Effect of salt types and salt levels

Plant growth, survival, and mineral composition from the different treatments as
obtained at the end of the 8-week period of growth are shown in Table 2.
Plants receiving additions of C&GI2 were affected more markedly than those re-
ceiving the corresponding levels of NaCl while plants grown in Na2SO4 media showed
less changes in growth than those grown in NaCl.
 ownloaded from http://jxb.oxfordjournals.org/ at University of York on August 19, 2014
00
00

TABLE 2. Effect of equal conductances of different salt types on senna growth, survival, and mineral composition
Sftlt treatments Relative Plant Mineral contents (mBq per 100 g dry wt.)
growth survival
Type mmhos cm"1 ITIM Na Mg Ca Cl

No salt 0-2 1-4 100 100 4-6 73-9 60-9 69-9 4-9
NaCl 2-5 22-0 96 100 4-4 67-5 44-4 87-3 18-2
50 43-9 66 88 6-9 60-9 51-0 95-8 34-2
100 87-8 29 49 19-9 57-0 41-9 103-8 106-6
20-0 175-6 17 19 38-0 42-7 38-7 124-8 121-2

CaCla 2-5 14-6 100 100 4-5 64-5 105-3 87-3 190
5-0
10-0
20-0
29-3
58-0
117-2
41
23
12
86
54
14
7-1
31
8-2
54-5
52-4
53-2
116-0
120-0
100-3
108-3
112-3
143-7
04-0
94-0
186-0
r
Na 2 SO 4 2-5 16-5 79 94 12-3 64-7 105-3 05-9 7-5
5-0 33-0 89 97 14-1 63-4 83-9 60-4 7-7
10-0 66-0 61 85 13-6 65-2 60-0 91-3
20-0 132-0 23 25 16-7 58-1 53-5 87-3 9-5

CaCla+NaCl 10-0 73-2 46 64 19-9 49-9 46-1 128-7 106-6

20-0 146-4 20 25 30-1 52-4 69-9 141-2 107-2
 Ayoub—Salt Tolerance in Senna 489
There was a progressive reduction in growth with increasing salinity of the sub-
strate. This effect was more apparent in the plants exposed to chloride salinity.
Fifty per cent reductions in growth were found at 4-0 mmhos cm"1 CaCl2, at 7-0
mmhos cm"1 NaCl, and at about 13-0 mmhos cm"1 Na2 SO4. Data on leaf shedding
and plant survival confirmed the results reported for plant growth.

Mineral composition of shoots

Mineral contents of plant shoots as mEq per 100 g dry wt. are also shown in
Table 2. The control plants contained very little Na and Cl, but their K contents
were high. Increasing NaCl in the substrate increased Cl and Na and decreased K
contents of shoots, Cl being accumulated at much higher rates. Sodium accumulated
in larger amounts in the shoots from NaCl treatments than from Na2S(>4, and Cl

Downloaded from http://jxb.oxfordjournals.org/ at University of York on August 19, 2014

TABLE 3. Effect of irrigation frequency with different NaCl levels on senna yield
and yield components
Irrigation interval NaCl levels (mmhos cm- 1 at 25 CC) of saturated soil Watering
extract effect

0-9 7-5 10-1 16-1 27-8

Pod yield (g/pot)
7d 47-0 52-0 16-1 1-4 1-5 23-6
14 d 18-2 24-3 11-6 5-3 5-6 130
Salt effect 32-6 38-2 12-9 3-3 3-6
S.e. means salt effect = 5-42, watering effect = 3-44.
No. of pods/pot
7d 661-3 691-3 228-3 25-3 32-0 327-7
14 d 254-7 330-0 176-7 81-3 93-3 187-2
Salt effect 458-0 510-7 202-5 53-3 62-7
S.e. means salt effect = 67-20, watering effect = 42-49.
Final plant survival/pot
7d 8-0 8-0 4-7 0-3 0-3 4-3
14 d 7-7 8-0 8-0 2-7 0-7 5-4
Salt effect 7-9 8-0 6-4 1-5 0-5
S.e. means salt effect = 0-86, watering effect = 0-54.
Wt/dry pod (mg)
7d 73-3 71-0 69-3 690 47-0 65-9
14 d 68-0 72-7 68-3 650 60-3 66-ft
Salt effect 70-7 71-9 68-8 67-0 53-7
S.e. means salt effect = 5-07, watering effect = 3-21.
Dry wt. plant (g)
7d 13-4 111 31 11 1-4 60
14 d 11-4 7-5 3-5 1-7 1-8 5-2
Salt effect 12-4 9-3 3-3 1-4 1-6
S.e. means salt effect = 1-85, watering effect = 1-17.
Plant height (cm)
7d 72-7 60-7 33-0 25-0 250 43-3
14 d 59-7 52-7 40-7 28-3 26-0 41-6
Salt effect 66-2 56-7 36-9 26-7 25-5
S.e. means salt effect = 4-55, watering effect = 2-86.
490 Ayoub—Salt Tolerance in Senna
accumulated in greater amounts from CaCl2 treatments than from NaCl. Increasing
substrate content of CaCl2 increased shoot Cl, Ca, and Mg contents with slight de-
crease in K contents.

Effect of frequency of irrigation ivith salt water on senna yield and yield components
Results of senna pod yield and yield components from different NaCl levels are
shown in Table 3.
' Pod yield was significantly affected by salt levels, frequency of irrigation, and
their interaction. Watering every 7 d with lower-salt waters resulted in appreciably
higher yields than watering every 14 d. Contrary to this, frequent irrigation with
high-salt waters injured plants and resulted in lower pod yields than irrigation
every 14 d with the same water.
The number of pods per pot in the various treatments were comparable to pod

Downloaded from http://jxb.oxfordjournals.org/ at University of York on August 19, 2014

yield while increasing salt level resulted in a small but significant (P < 0-05)
depression in pod weight.
Plant survival was significantly reduced by salt levels higher than 10-0 mmhos
cm" 1 in the soil. This effect was more noticeable at the shorter watering interval
than at the longer watering interval, indicating that the effect of NaCl salinity on
senna was not solely an osmotic phenomenon. Injured plants contained considerable
amounts of Cl which was closely related to the degree of visible injury.
Pod yield was highly correlated with pod number (r = 0-9947), less with plant
survival (r = 0-7221), and least with pod weight (r = 0-3818).

Dry vxight per plant and plant Tieight

Significant reductions in dry matter yields and plant heights occurred at soil
salinities of 10-0 mmhos cm""1 and above. When soil salinity was raised from 7-5

6-0-
• Logy =4-9889-00129*
r - -0-895 /><0001
5-0-
ve pod yield

•
3 3-0-

2
eo
3 2-0-

l-0<

ft-0
55 110 165 220 275
Leaf bUde O (mEq/100 g dry wt.)
F I G . 3. Relation between senna leaf chloride content and log per cent relative pod yield.
 Ayoub—Salt Tolerance in Senna 491
1
to 10-1 mmhos cm" reductions of 65 and 35 per cent were found in dry matter and
plant height, respectively. Increasing the watering interval from 7 to 14 d reduced
both parameters but not significantly.
The relationship between pod yield and Cl contents of senna leaves is demon-
strated in Fig. 3. Significant yield reduction occurred when the Cl content of the
blade was above 28 mEq per 100 g dry wt. Fifty per cent yield reduction occurred
at Cl content of about 62 mEq, and at Cl content of greater than 110 mEq pod yield
was very low. The Na contents also varied with NaCl levels, but these variations
seemed to be of minor magnitude.

DISCUSSION
Senna was shown to be fairly tolerant of salinity at the germination stage. A 50

Downloaded from http://jxb.oxfordjournals.org/ at University of York on August 19, 2014

per cent reduction in germination was reached at salt levels between 20-0 and 24-0
mmhos cm"1. By way of contrast, a 50 per cent reduction in germination was found
at 13-0 for groundnuts (Shalhevet, Reiniger, and Shimshi, 1969), 14-0 for tomatoes
(van Hoorn, 1970), and 21-0-30-5 mmhos cm"1 for rice (Pearson, Ayers, and Eber-
hard, 1966). The relatively higher tolerance of senna to salinity during germina-
tion is not surprising since it is a crop originating from regions with a dry climate
and where salinity problems usually exist. The optimum temperature at which
senna could germinate satisfactorily at high salinity levels was found to be between
28 and 30 °C. The significant interaction of soil salinity and air temperature re-
ported in this study might be important in determining optimum sowing dates on
saline soils. Senna seeds germinated in soils high in exchangeable sodium (ESP of
35), but most of the seedlings died shortly after. The chlorotic symptoms appearing
in seedlings growing in high ESP levels could be due to the toxic effects of bicarbonate
ions resulting in high pH values.
Salt type effects were considered important in senna since Na2SC>4 effects were
generally less detrimental to the plant growth than were those of CaCl2 or NaCl
of the same salinity. Plants treated with chloride salts were damaged to a greater
extent indicating the more deleterious effects of chloride ion. With chloride salinity,
CaCl2 was more toxic than NaCl. It would appear that the high chloride concentra-
tion in the plant tissue produces the drastic decrease in plant growth. It is possible
that the high sodium levels in plants found in some treatments could be due to
chloride injury to membranes giving rise to passive sodium uptake.
Senna very rapidly absorbed chloride from the root media and accumulated toxic
amounts of it in the leaves. Visible chloride injury symptoms occurred at foliar
chloride concentration of about 42—70 mEq per 100 g dry wt., plant growth was
retarded at about 85 mEq, and plant survival was highly impaired at chloride levels
of more than 140 mEq. In its sensitivity to chloride injury, senna behaves in a like
manner to avocado, citrus, and grapes but contrary to most plant species which are
more sensitive to sulphate salts (Mengel, 1973).
For good pod yields of senna, NaCl salinity should not exceed about 8-0 mmhos
cm"1 in the soil, but satisfactory yields may be obtained up to 15-0 mmhos cm"1,
similar to safnower and wheat (Bernstein, 1970).
492 Ayoub—Salt Tolerance in Senna
More frequent irrigation with saline water induced, earlier and more severe injury
than less frequent irrigation. Bernstein and Francois (1975) found that more fre-
quent sprinkling with saline waters resulted in greater osmotic shock than less
frequent watering. Moreover, it was noted in this study that water use by the crop
was reduced greatly under high salinity. It is, therefore, inadvisable to give extra
water to senna under saline conditions.
In conclusion, the following points can be mentioned. Senna is moderately toler-
ant to salinity. It seems to be more salt tolerant at germination and later growth
stages, the seedling stage being the most sensitive. Avoiding adverse environmental
conditions like high air temperature (Francois and Goodin, 1972), low relative
humidity (O'Leary, 1975), and bad cultural practices (Sandoval and Benz, 1973)
at that critical stage may improve the salinity tolerance by senna and its survival
in the field.

Downloaded from http://jxb.oxfordjournals.org/ at University of York on August 19, 2014

ACKNOWLEDGEMENTS
The author is grateful to the Director General, A.R.C, Sudan for permission to
publish this paper and to Dr. R. H. Nieman of U.S. Salinity Laboratory, Riverside,
for his valuable comments and encouragement.

LITERATURE CITED
AYOTTB, ALI T., 1975. J. exp. Bot. 26, 891-96.
BERNSTEIN, L., 1970. Agric. Inform. Bull. No. 283, U.S. Dept. Agrio.
and FRANCOIS, L. E., 1975. Agron. J. 67, 185-90.
CHANG, C. E., and DBEGNE, H. E., 1955. Proc. Soil Set. Soc. Am. 19, 29-35.
CHAPMAN, H. D., and PRATT, P. F., 1961. In Methods of analysis for soils, plants and waters.
Univ. Calif., Riverside, Div. Agrio. Sei.
FBANCOIS, L. E., and GOODIN, J. R., 1972. Agron. J. 64, 272-73.
GUPTA, R., 1971. Indian Fmg. 21, 29-32.
MENGEL, K., 1973. First FAO/SIDA Seminar for plant scientists from Africa and Near East,
Cairo, Egypt, 1-20 September, 1973.
O'LEARY, J. W., 1975. PL Soil, 42, 717-21.
PEARSON, J. A., AYERS, A. D., and EBEBHABD, D. L., 1966. Soil. Sri. 38, 777-80.
RICHARDS, L. A. (Ed.), 1954. Diagnosis and improvement of saline and alkali soils. U.S. Dept.
Agric. Handbook No. 60.
SANDOVAL, F. M., and BENZ, L. C, 1973. Soil Sci., 166, 100-5.
SHALHEVET, J., REINIGER, P., and SHXHSHI, D. 1969. Agron, J. 61, 384-7.
VAN HOOBN, J. W., FAO/UNDP/TA seminar on methods of amelioration of saline and water-
logged soils, Beghdad, Iraq, 5-14 December 1970.