EFFECT OF SAND CULTURE AND SODIUM CHLORIDE ON GROWTH PHYSICAL STRUCTURE

AND ORGANIC ACID METABOLISM IN "BRYOPHYLLUM PINNATUM"

Author(s): S. M. KARMARKAR and G. V. JOSHI
Source: Plant and Soil, Vol. 30, No. 1 (February 1969), pp. 41-48
Published by: Springer
Stable URL: https://www.jstor.org/stable/42949523
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Plant and Soil XXX, no. 1 February 1969

EFFECT OF SAND CULTURE AND

SODIUM CHLORIDE ON GROWTH PHYSICAL
STRUCTURE AND ORGANIC ACID METABOLISM
IN BRYOPHYLLUM PINNATUM

by S. M. KARMARKAR* and G. V. JOSHI**

INTRODUCTION

Chlorophyllous parts of succulent plants are characteri

special adaptive feature by means of which they accumu
amounts of malic acid in dark and utilise it as a source of carbon
dioxide in light. This diurnal rhythm in acid fluctuation is known as
Crassulacean Acid Metabolism 10 : (CAM), of which Bryophyllum
pinnatum is a classical example. It is well known that due to CO2-
fixation oxaloacetate is produced which is then converted by malic
dehydrogenase to malic acid. From the same pool of oxaloacetate
aspartate also can be synthesised.
Many times B. pinnatum is grown in sand cultures to study CAM.
It is obvious that sand will be a different medium than soil and is
likely to affect growth and structure of the plant. The present in-
vestigation was undertaken to study effect of the sand medium on
growth and physical structure in B. pinnatum . As CAM is a promi-
nent feature of the Crassulacean succulent the effect of the rooting
medium on CAM was also investigated to give indication of the
effect of subtratum on the metabolism.
CAM is also affected by presence of NaÇl in the nutrient medium.
It has been reported by Joshi et al.6 that sodium chloride concentra-
tions have a profound effect on dark CO2 fixation where chloride
ions stimulate transaminases due to which more aminoacids are

* Department of Botany, R. J. College, Bombay 77, India.

♦♦ Department of Botany, Shivaji University, Kolhapur, India.

- 41 -

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42 S. M. KARMARKAR AND G. V. JOSHI

synthesised rather than organic acids. This observation was further

confirmed by Webb and Burley 14 while working on salt marsh
halophytes. Hiatt and Evans 4 have demonstrated that the activ
ty of isolated malic dehydrogenase is considerably altered due to th
presence of chlorides in the medium. These results clearly indicate
that NaCl can have a profound effect on the patterns of acid fo
mation during dark CCVfixation. It was felt that the study of the
effect of subtratum and NaCl on physical structure, growth, an
organic acid metabolism in Bryophyllum pinnatum would add to th
existing knowledge of plant nutrition. With this view in min
Bryophyllum plants were grown in sand culture and subjected t
increasing concentrations of NaCl in the nutrient medium. Th
results of these experiments are discussed in this communication.

EXPERIMENTAL

Effect of sand culture

Young plantlets of approximately the same size and age of Bry
pinnatum were selected from a patch of plants reared in the Coll
The plants were raised in glass jars which were perforated at the b
of jars had acid free sand while others had a garden soil. Both the
equal illumination. The culture jars were flushed with nutrient
Hoagland and Arnon 5. The solution was prepared in a double glass-
distilled water by using A.R. grade reagents. Flushing of the nutrient was
carried out at four hour intervals. The sampling and harvesting of plants was
carried out after 15 weeks of growth in sand culture. Before harvesting the
diurnal variations in titratable acid number (TAN) in the leaves was deter-
mined according to the method of Thomas and Bee vers. 11 The leaves
were further analysed for their physical properties by the method described
by Linacre 8.

Effect of sodium chloride

The experimental procedure was the same as followed for the sand culture.
In an exploratory experiment plants were raised in a nutrient containing
NaCl. The latter was added to make up a series 0.1, 0.2, 0.3, 0.4, 0.5, 0.6, 0.7,
0.8, 0.9, and 1 M NaCl solutions. This experiment showed that in concentra-
tions above 0.1 M NaCl plantlets could not survive and hence in further ex-
periments lower concentrations ranging from 0.0 2M, 0.04, 0.06, 0.08 and
0.1 M NaCl were employed. The technique employed for supplying higher
concentrations of NaCl was the same as that of Black *. For the first week
the plants were flushed only with the basic nutrient solutions till they were
well established in the changed environment. The plants required for higher
NaCl concentrations were moved up the series at four day intervals. After

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 EFFECT OF SAND AND NaCl ON GROWTH OF BRYOPHYLLUM 43

the final concentrations were reached, plants were allowed to grow for a
further period of eight weeks. The plants were analysed after a period of
fifteen weeks. The experiments were carried out in the month of February
when maximum and minimum temperatures were 35°C and 22°C respectively.
The relative humidity was 70%. The experiments were repeated several
times and the results of a single set of experiments are presented in the tables.
Other experiments showed similar type of results.

RESULTS AND DISCUSSION

The results of the effect of sand subtratum on growth

and CAM are shown in Table 1 while 2 records effect of NaCl.

Effect of sand subtratum

The results clearly indicate that the rooting medium has a definite
effect on growth, structure and organic acid metabolism in the
Crassulacean succulent. The roots under sand culture are shorter in
lenght and profusely branched. The weight of the root system was
2.62 as compared to 1 .68 of that grow iň soil. However, in soil the

TABLE 1

Effect of sand culture on physical structure and crassulacean acid metabolism

leaves of Bryophyttum pinnatum
Leaf from Leaf from
soil culture sand culture

No. of stornata
Upper surface 77 68
Lower surface 91 69

Size * of stornata

Upper surface L 44.2 41.7

B 21.4 32.5
Lower surface L 49.4 42.7
B 37.8 31.1
Mass of leaf, M

Area *♦ of leaf, A

M/A

Thickness, d

Density, 20 M/ Ad

TèA .N

7 p.m

7 a.m

% increase

* L = Length. B = Breadth.
** Area is that of both surfaces.

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44 S. M. KARMARKAR AND G. V. JOSHI

root system reaches to a length of 22.7 cm while in sand it could

only upto 12.2 cm. The profusely branched root system in san
affords a large area for absorption.
The effect of rooting medium is very clear on the shoot of
pinnatum. The plant in sand is slightly higher as it reaches to
height of 16.9 cm as compared to 15.8 cm of that of soil medium.

Fig. 1. Effect of rooting medium on leaf production of Bryophyllum pinna-

tum. Left : Soil. Right : Sand.

sand B. pinnatum produces leaves which are large in size and more
in number (Table 1, Fig. 1). However, thickness as well as densi
of the leaves remains same. There is a striking difference in t
number and size of stornata seen on upper and lower surfaces. Und
soil conditions the number of stornata are much more than those

produced under sand conditions. The leaves from the plants grown
on soil develop longer and broader stornata. However, breadth of the
stornata from upper surface in plants with sand subtratum is more.
Inspite of this minor difference it is possible that numerous, large
and broad stornata under soil condition can provide more CO2 for
dark carboxylation. This results in more acid accumulation and

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 #

EFFECT OF SAND AND NaCl ON GROWTH OF BRYOPHYLLUM 45

subsequently more percentage increase in T.A.N, in the plants

grown on soil. The shoot/root ratio of the plant grown in garden soil
is 0.69 while that from the sand is 1 .37. In terms of growth sand is a
better a medium for B. pinnatum but due to better development of
stomatal aperture in soil substratum more acid accumulation takes
place.

Effect of NaCl on growth

The role played by chloride ions in growth metabolism is increasing-
ly understood by the works of Lip m an and Gre en way 3. In buck
wheat and peas Lip m an found that chloride ions stimulate growth
when supplied with lower concentrations. However when present in
higher concentrations these ions reduce growth in Hordeum vulgare
was the observations of Greenway. Eaton 2 also found that high
amount of chlorides inhibit growth. Our observations are also on
similar lines as growth is inhibited in B. pinnatum. Even a concen-
tration of 0.02 M NaCl in the nutrient retards growth (Fig. 2). It was
found that control plants reached a height of 22.5 cm which progres-
sively decreased to 10.8 cm when 0 AM NaCl was present in the
nutrient solution.

Fig. 2. Effect of sodium chloride on growth of Bryophyllum pinnatum.

NaCl-concentrations, from left to right: Control, 0.02, 0.04, 0.06, 0.08, 0 AM.

The presence of sodium chloride in nutrient solution has a consi-

derable effect on physical structure of the leaves. It was found that
the thickness of the leaves increases upto a concentration of 0.04 M

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 46 S. M. KARMARKAR AND G. V. JOSHI

NaCl and then decreases gradually at the higher concentrations. Th

weight of the leaves increases with increase in concentration of NaCl
This takes place till the external concentration is 0.0AM and her
after it follows on irregular pattern. At the concentration of 0.1 M
NaCl the weight is less than in control there by indicating toxicity
has set in plants.
The size of the leaf also shows an increase till 0.04 M NaCl and
afterwards follows an irregular pattern. There is increase in weight
with increase in NaCl in the medium except at 0.1 M NaCl where it
decreases. The increase in weight is not due to water absorption but
due to increase in organic matter. This indicates that presence of
NaCl in the medium stimulates synthesis of certain organic com-
pounds. It is obvious that an increases in concentration of NaCl
results in production of thicker, larger and heavier leaves. Van
Eijk12 as well as Walter13 observed this effect while studying
problem of halophytes. Walter found that the succulent effect of
leaves was due to Cl-ions and not by SC>4-ions. He explained that
sulphates cause dehydration of proteins and hence produce thin and
flacid leaves. In order to confirm this observation B. pinnatum
plants were cultured for fifteen weeks in increasing concentrations
of sulphates. It was found that the leaves did not show a healthy
development. They were thin and flacid. Our investigations reveal
that chloride ions and not sulphate ions cause production of thick
and succulent leaves in B. pinnatum
Presence of NaCl in the nutrient medium also affects CAM in В .
pinnatum . From Table 2 it is obvious that there is a stimulation of
CAk by NaCl when the concentration in the outside environment is
0.04 M. Beyond this there is an inhibition which increases with
TABLE 2

Effect of NaCl on physical properties and T.A.N, in leaves of Bryophyllum

Den- T.A.N., me acid 100 g
NaCl, Mois" °rganic Ash Mass of Area of м/д Thick- 2Q fresh tissu

(%) (%) (%) I (S) M <mm> (g/cm3)

molarity (%) 7"er (%) (%) le*7 (S) M (g/cm») n,ess'd <mm> M/Ad 7 p.m. 7 a.m.
Õ 9Ï3 Ï27 IÃ2 0.626 22Л 0.0278 Õ53 TÕ5 2X1 18.85 830
0.02 94.0 4.82 1.18 0.755 27.1 0.0278 0.59 0.94 1.36 13.11 960
0.04 92.3 6.47 1.32 0.937 31.8 0.0294 0.67 0.88 1.4 22.68 1620
0.06 91.6 7.29 1.02 0.817 31.5 0.0259 0.61 0.85 2.73 13.74 503
0.08 91.9 7.92 2.08 0.903 37.3 0.0242 0.6 0.81 2.57 12.12 472
0.10 98.6 8.45 1.92 0.563 22.8 0.0246 0.62 0.80 2.57 10.53 140

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 EFFECT OF SAND AND NaCl ON GROWTH OF BRYOPHYLLUM 47

further rise in NaCl-concentration in the external medium. When

the plants were supplied with C14 it was found 7 that increase in
organic acid production is due to stimulation of rate of C02-fixation.
This increased carboxylation is responsible for more production of
organic acids which results in more titrable acidity.

SUMMARY

Bryophyllum pinnatum, a classical example of Crassulacean Acid Meta

lism, was grown in sand and soils as well as in increasing concentration
chloride and sulphate ions to test the effect of rooting medium and salt
growth, physical structure of leaves and on organic acids. Plants grown in
medium show fewer and smaller stornata, a shorter and profusely branc
root system and also a decrease in organic acid metabolism, as compare
with plants grown in garden soil. This indicates that the subtratum play
important role in controlling growth, physical structure of leaves and C
sulacean Acid Metabolism. The presence of chlorides tends to produ
thicker leaves which have more organic matter and less moisture. Incr
in NaCl-concentration retards growth and also affects CAM. In plants tr
with 0.04 M NaCl there is a stimulation in organic acid synthesis which
results in higher TAN values. Another interesting observation is that th
development of a succulent habit is due to effect of chlorides and not
sulphate or sodium ions.

Received August 3, 1967

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