BIOLOGIA P L A N T A R U M (PRAHA)

1 (1):9--15, 1959

Application of Humus Substances to Overground

Organs of Plants

ZDEN~K S L A D K ~ a n d VLADIlYI[R T~CHf

Laboratory for Plant Physiology and Anatomy, Natural Science Faculty, Brno University

Received January 16, 1958

Souhrn

N6kte~i auto~i se domnivajl, ~ie humusov6 1Atky nepflsobi na rostlinu p~imo,

n~br~ zvyhuji pouze rozvoj ko~enov~ a pddnl mikroflory, jeji~ pflsobeni je pak
vlastni p~i6inou lep~iho rflstu rostlin. Abyehom mohli studovat p~izniv:~ vliv humu-
sov~ch 1Atek na rdst a metabolismus rostliny, vylouSili jsme jejich pflsobeni na
pfldni a ko~enovou mikrofloru tim, ~e jsme jo pods rostlin6 post~ikem na list.
SouSasn6 n&m ~1o o to, zjistit mo~nost vyu~iti p~izniv6ho pflsobeni humusovs}ch
l&tek p~i post~iku rostlin v praxi.
Na z~klad6 p~edb~n:~ch pokusfl byla pro dalai pr&ci vybr~na rostlina Begonia
semperflorens LINK et OTTO, kter~ citliv6 reagovala ha humusov~ ls K post~iku
bylo pou~iito dlalysovan6ho extraktu zahradni pfldy, obsahujici hum&t sodnp
s p~im6si frakce fulvokyselin v celkov~ koncentraci 300 rag/1. Jedrlotliv~ dAvky,
aplikovan6 dvakr&t t~dn6 v dob6 hlavni rflstov~ raze, byly 2 ml.
Bylo zjiw ~e humusov6 1Atky aplikovan6 na list pflsobi prodlu~ovAnl
rostlinn~ch orghnfi i zv~t~enl jejich s v ~ i v~hy a su~iny. Je zv:~-~en i obsah chloro-
fylu a mno~stvi vyd~chan~ho kysli6niku uhli6it~ho. KoneSnp efekt zhvisi na dob~,
kdy byl post~ik proveden, i na pou~ita koncentraci. Lze mit za to, ~e tyto d6inky
jsou v~sledkern bezprost~edniho pflsobenl humusov~ch 1Atek a nikoliv pflsobeni
stimulovan~ pfidni 5i ko~enov6 mikroflory nebo zlep~en~ch fysiks resp.
fysikhln~ chemick~'ch vlastnosti pfidniho prost~edi.

Summary
The authors studied the effect of the application of humus substances to the
leaves of Begoniasemperflorens LINK et OTTO. For spraying they used a dialysed
extract of garden soil containing sodium humate with admixture of fulvic acid
in total concentration of 300 mg/1. Doses of 2 ml. were applied twice a week
during the period of the main growth phase.
I t was found t h a t humus substances applied to the leaves cause a leng-
thening of plant organs and increase of their fresh and dry weights. The chloro-
phyll content and the amount of expired carbon dioxide are increased. The
final result depends on the time when spraying was carried out and on the
concentration used. I t can be assumed t h a t these effects are the result of direct
influence by the humus substances and are not due to the action of stimulated
soil or root micro flora nor to improvement of the physieochemical character
of the soil environment.
 ]0 Z. S L A D K ~ a n d V. T I C H ~

Introduction

The favourable influence of humus and humus substances on the growth

and development of plants has been demonstrated in the past b y many authors
and is t o d a y more or less generally recognised (extensive literature on this
subject was collected b y PRAT 1955). However, the way this influence is
exerted, b y what paths the humus substances impinge on the flow of meta-
bolic processes or biochemical reactions taking place in the plant organism
remained undecided. The fact that humus substances give rise to striking
changes in such fundamental vital processes as respiration, glycide conversions,
reception of mineral ions, cell division or growth, led to the acceptance as
more or less self-evident, of the supposition that these substances enter into
organs and cells and there directly influence the metabolic processes. Never-
theless, some authors hold the view (e. g. CHIZ~EVSKu and DIKUSAR 1955)
that humus substances do not act in this direct manner on the plant, but
that they merely stimulate the development of root and soil micro flora, the
influence of which is then the actual cause of improved plant growth, or
that they simply influence the physical and physicochemical conditions ob-
taining in the soil. It is indisputable that humus substances act both on the
soil microflora and on some important qualities of the soil. It is therefore
necessary, if we wish to study the direct influence of these substances on
plants, to eliminate both these factors of the external environment. This
requirement can be partly met b y using sterile water or sand cultures.
In our work we have chosen another method of eliminating the influence
of soil environment and its microflora, that is the application of humus
substances to the leaves b y spraying. At the same time we wished to deter-
mine whether it would be possible to employ the stimulatory qualities of
humus substances b y spraying plants, as this could be of considerable signifi-
cance in market gardening and in agriculture.

Material and Methods

W e carried o u t m a n y p r e l i m i n a r y e x p e r i m e n t s , w h i c h s h o w e d t h a t it is n e c e s s a r y a b o v e
all to choose s u i t a b l e e x p e r i m e n t a l m a t e r i a l , a s c e r t a i n t h e o p t i m a l c o n c e n t r a t i o n of t h e prepar-
a t i o n to be u s e d a n d t h e m o s t f a v o u r a b l e t i m e for a p p l i c a t i o n w i t h r e g a r d to t h e g r o w t h a n d
d e v e l o p m e n t of t h e p l a n t . As a r e s u l t of t h e s e e x p e r i m e n t s we chose for f u r t h e r w o r k t h e p l a n t
Begonia semperflgrens LINK et OTTO, w h i c h r e a c t e d s e n s i t i v e l y to h u m u s s u b s t a n c e s . R e s u l t s
o b t a i n e d in e x p e r i m e n t s w i t h t h i s p l a n t are t h e s u b j e c t of t h i s paper.
Begonia semperflgrens w a s g r o w n f r o m seed in a glass h o u s e to t h e s t a g e of two large assimil-
a t i o n leaves. T h e seedlings were t h e n t r a n s p l a n t e d to flower pots, one set b e i n g 11 cm. in d i a m e t e r
w i t h a c o n t e n t of 800 g. of g a r d e n soil a n d t h e o t h e r 9 era. in d i a m e t e r w i t h a c o n t e n t of 400 g.
of g a r d e n soil. F r o m a larger n u m b e r of t h e s e t r a n s p l a n t e d seedlings 25 pairs c o r r e s p o n d i n g in
a p p e a r a n c e a n d g r o w t h were selected f r o m t h o s e in t h e large flower p o t s a n d 15 p a i r s f r o m t h e
s m a l l pots. I n b o t h g r o u p s one h a l f w a s s p r a y e d w i t h h u m u s s u b s t a n c e s a n d t h e o t h e r half,
s e r v i n g as control, w i t h distilled water. T h e p l a n t s were k e p t as before in t h e glass h o u s e . A h a n d
s p r a y e r w a s u s e d , w h i c h f o r m e d m i s t y drops. I n t h i s w a y t h e s o l u t i o n w a s d i s t r i b u t e d e v e n l y o n
t h e leaves, s t e m s a n d flowers to f o r m a fine, e v e n coating. Care w a s t a k e n t h a t t h e s o l u t i o n s h o u l d
n o t drip o n to t h e soil in t h e flower pots. T h e p l a n t s were s p r a y e d in t h i s w a y twice a w e e k d u r i n g
J u n e a n d J u l y , i. e. d u r i n g t h e period of g r e a t e s t g r o w t h u p to t h e flowering a n d r e n e w e d slowing
of growth.
 AP1)LICATIOM O F H U M U S S U B S T A N C E S 11

The dose for one s p r a y i n g of one p l a n t w a s a b o u t 2 ml. of solution of h u m u s substances, con-

centration 300mg.]l. The solution was p r e p a r e d as follows: dried garden soil w a s e x t r a c t e d cold
w i t h 96% ethanol in order to s e p a r a t e the alcoholic fraction. After d r y i n g it w a s decalcified w i t h
l % I-IC1 a n d following t h o r o u g h rinsing w i t h distilled w a t e r e x t r a c t e d w i t h 0.3 N N a O H . The
solution was t h e n filtered, c o n c e n t r a t e d b y freezing out a n d dialysed for 70 hours. Finally the
concentrate was diluted to 300 rag. of h u m u s substance in one litre of solution, the final p H of
which w a s 6.5. The composition corresponded to s o d i u m h u m a t e w i t h a d m i x t u r e of fulvie acid
a n d o t h e r s u b s t a n c e s of this fraction. F o r the period during which we used it the solution wa~
kept in a refrigerator at a t e m p e r a t u r e of - - 1 to - - 5 ~ C.
The differences between the experimental p l a n t s and the controls were evaluated b y m e a s u r -
ing the lengths of stems a n d roots, d e t e r m i n i n g fresh and d r y weights of o v e r g r o u n d p a r t s
a n d roots a n d finally by m e a s u r i n g the length a n d b r e a d t h of the blades of leaves at each level.
The values o b t a i n e d were averaged a n d the differences in relation to the controls ascertained.
The chlorophyll c o n t e n t was established b y the colorimetric m e t h o d according to GODNEV
(1952) a n d the a m o u n t of expired c a r b o n dioxide was d e t e r m i n e d b y a b s o r p t i o n in b a r i u m h y d r o -
xide.

Results

The experiments showed that spraying of Begonia semperflgrens with a sol-

ution of humus substances affects growth favourably. This was apparent
from the length of stems and roots and the fresh and dry weights of these
organs. The values measured and the differences as compared with the control
are given in table 1 and photographically recorded on figs. 1 and 2. Humus
substances affect both the length and breadth of leaf blades. This is graphic-
ally illustrated b y fig. 4. Spraying with humus substances also influenced
the chl~ophyll content and the production of carbon dioxide b y leaves in
the dark as can be clearly seen from table 2. Overdosing with humus substances
through spraying has an inhibitory effect. Growth is slowed up, leaves are
wrinkled, branching of the stems occurs. The generM appearance of such a plant
is shown in fig. 3.

Diseussion

The results of experiments with Begonia semperflorens show that humus

substances affect the whole plant organism even when they are applied to
the leaves. At the same time the results of this action do not in general differ
from the effects caused b y humus substances when present in the soil. There
was a considerable increase in the fresh and dry weights of the overground
parts and roots and this increase was percentually greater in the ease of plants
in the small flower pots than in those in the large pots. The dry weights
of stems and roots of plants in the large pots remained on the whole within
the limits of variability of biological material, but showed significant increases
in the case of plants in the small pots (table 1). It is necessary, however, to
point out that changes in the length and weight of the root system are un-
doubtedly distorted to a certain extent b y the procedure of washing out traces
of earth. Since the root system is very delicate even our great care in prepar-
ation could not ensure against the possibility of causing changes in weight.
For this reason we regard the changes in the root system recorded in table
1 as preliminary results which will have to be verified in water culture. Both
12 Z. S L A D K ~ r a n d V. T I C H Y

T a b l e 1. D i m e n s i o n s a n d w e i g h t s of o r g a n s of Begonia semperflorens t h e o v e r g r o u n d p a r t s of
w h i c h were s p r a y e d w i t h solution of h u m u s s u b s t a n c e s of t~otal c o n c e n t r a t i o n 300 mg./1. Control
p l a n t s were s p r a y e d w i t h w a t e r

Difference
Plants sprayed with in ~o of Signifi-
Control p l a n t s
h u m u s solution control cance t

P l a n t s g r o w n in large p o t s (25 p l a n t s )

H e i g h t in cm. 27.1 -4- 3 . 1.22 33-9 3 . 1.66 25"1"* 3-30

F r e s h w e i g h t in g. 29.8 -4- 3 . 1.21 37.3 3 . 1.73 25"1"* 3.55
D r y w e i g h t in g. 1.42 -4- 3 . 0 . 1 5 1.48 i 3 9 0.48 4.1t 0.12

L e n g t h in cm. 17.7 3 . 0 . 8 2 17-1 3 . 0 . 5 5 --2.6? 0.60

F r e s h w e i g h t in g. 1.62 i 3 . 0 . 0 8 1.84 3 . 0 - 0 6 14.2" 2.20
D r y w e i g h t in g. 0.28 3 . 0 - 0 9 0.26 3 . 0 - 1 0 --8.4? 0.15
I
P l a n t s g r o w n in s m a l l p o t s (15 p l a n t s )

H e i g h t in cm. 16"5 3 . 0 " 5 8 21-5 3 . 0 ' 5 6 30"5** 6.17

F r e s h w e i g h t in g. 12"0 3 . 0 - 5 2 16"8 3 . 0 ' 6 9 40-1"* 5.58
D r y w e i g h t in g. 0'49 3 . 0'06 0'72 3 . 0'03 47"8** 3-43

L e n g t h in cm. 14"5 3 . 0 ' 8 0 19'4 ~ 3 . 1"13 33.8** 3.55

F r e s h w e i g h t in g. 0-63 3 . 0"04 0"92 3 . 0'08 46-2** 3-22
D r y w e i g h t in g. 0"10 3 . 0"09 0'16 3 . 0-07 60.8? 0.54

N o t e : Difference t48 tzs

** h i g h l y significant (P < 0.01) > 2.69 > 2.76
* significant (P < 0.05) ) 2.01 > 2.05
Jr n o t significant

T a b l e 2. C h l o r o p h y l l c o n t e n t of leaves a n d o u t p u t of c a r b o n dioxide of leaves of Begonia semper-

florens, t h e o v e r g r o u n d p a r t s of w h i c h were s p r a y e d w i t h a solution of h u m u s s u b s t a n c e s of
t o t a l c o n c e n t r a t i o n 300 mg./1. Control p l a n t s were s p r a y e d w i t h w a t e r

Plants spray-
Control Difference in
Pots ed w i t h h u -
plants ~ of c o n t r o l
m u s solution

C h l o r o p h y l l c o n t e n t in mg. in l0 g. large 9.850 10.966 11.4

of fresh w e i g h t of leaves. small 7.780 9.706 24"8

A m o u n t of CO 2 in m g . e x p i r e d b y large 5.3 6.9 30.0

15 g. of f r e s h w e i g h t of leaves. small 5.9 6.9 16"5
 Z. S L A D K ~ and V. TICHS~

APPLICATION OF HUMUS SUBSTANCES

Fig. 1. Influence of h u m u s s u b s t a n c e s applied to t h e leaves of Begonia semperflorens in larger

flower pots. Control o n t h e right, p l a n t s p r a y e d w i t h h u m u s o n t h e left.

Fig. 2. Fig. 3.
Fig. 2. I n f l u e n c e of h u m u s s u b s t a n c e s a p p l i e d to t h e leaves of Begonia semperflorens in s m a l l e r
flower pots. Control o n t h e right, p l a n t s p r a y e d w i t h h u m u s on t h e left.
F~g. 3. I n h i b i t o r y effect of h i g h c o n c e n t r a t i o n s a n d large doses of h u m u s s u b s t a n c e s a p p l i e d
to t h e leaves of Bego~ia semperflorens in larger flower pots. L e a v e s are d e f o r m e d , side
s t e m s develop.
 APPLICATION OF HUMUS SUBSTANCES 13

in the large and the small pots stem lengths were increased due to spraying.
Increase in length of stems was not the same in all parts: the lower internodes
showed greater increases t h a n those formed at a later stage of the plant's
growth. Similarly the size of leaf blades (length and breadth) does not in-
crease evenly over the surface but, as shown in fig. 4, with a maximum in
areas 4, 5, 6 and 7 of the leaf. From
this it m a y be concluded t h a t the 100
effect of humus substances applied
to the plant on the leaves is most
/," ',
intense in the growth of those organs, ' ~ ' , I
'h
which at the time of application are
in the early stages of tissue develop-
ment. With the slowing down of the
growth of a plant organ its sensitiv-
ity to the action of humus sub- 50
stances also decreases. The special
sensitivity of meristems is also
evident from the reaction to exces-
sive doses of humus substances (fig. 3). p--
The more frequent occurrence of
side stems in this case indicates t h a t
the inhibitory effect of the growing
points on the development of axillary 0 2 3 4 5 6 Z 8 9 10 11
buds is influenced and limited.
A further remarkable fact is the
difference in the effect of humus sub- Fig. 4. I n f l u e n c e of s p r a y i n g w i t h h u n m s s u b -
s t a n c e s on lengl~h ( u n b r o k e n line) a n d b r e a d t h
stances on plants in large and small ( d o t t e d line) of leaves a t different levels of Be,
flower pots. This phenomenon can gonia semperflsrens (large pots). C u r v e 1 , 2 -
Mso be explained in our experiment m e a s u r e m e n t s of s p r a y e d leaves, c u r v e 3.4 ---
by the fact t h a t humus substances m e a s u r e m e n t s of leaves o f control. C u r v e 5 -
differences in b r e a d t h b e t w e e n s p r a y e d leaves
improve the utilization of those ele- a n d control, c u r v e 6 - - differences in l e n g t h , x:
ments of nutrition which are avail- order of leaves o n s t e m f r o m t h e oldest (1) to
able to the plant in minimal a- t h e y o u n g e s t (I1L y: m e a s u r e m e n t in ram.
mounts. Therefore the effect of spray-
ing is greater in the smaller pots, where the source of nutrient is more limited.
The increase in total dry weight of sprayed plants also indicates an increase
in intensity of photosynthetic assimilation, t h a t is a more powerful assimi-
latory apparatus. This is confirmed by the colorimetric determination of
chlorophyll content: its value increased for sprayed plants in large pots by
11.4%, in small pots by 24.80/0. Thus in this case too humus substances act
through spraying in the same way as when they are taken in by the roots
(for example NIKLEWSKI and WOJCIECHOWSKI 1947, WOJCIECHOWSKI 1948,
KHRISTEVA 1951).
Besides more intense growth we also observed an increase in intensity of
respiration (output of carbon dioxide in the dark). The increase for sprayed
plants in large pots was 30% and 16.5% for those in small pots. This effect,
too, has already been observed in cases where humus substances were given
to plants as a part of the root nutrition (e. g. KHRISTEVA 1951).
14 Z. S L A D K ~ and V. TICH~r

In conclusion we may say that humus substances applied to the leaves

cause lengthening of the plant organs and increase in their fresh and dry
weights. Chlorophyll content and the amount of expired carbon dioxide are
also increased. The final effect depends on the period of spraying and on the
concentration used. In view of the fact that these effects do not differ from
the effects caused by humus substances given as a part of root nutrition.
we may assume that they are the result of direct action of humus substances
and not of the action of stimulated soil or root mieroflora, nor to the improve-
ment of physical or physico-chemical character of the soil en~/ironment.
A prerequisite for the practical utilization of humus sprays is, however,
the extension of experiments to a greater number of plant species, the determin-
ation of their sensitivity, suitable concentrations of the solutions and the
most suitable time of application.

References

CttIZHEVSKI, M. ~., DIKUSAR, M. M.: Rol gumusa i mikroorganizmov v kornevom pitauii

vysshikh rasteny v usloviyakh vodnikh i peschanikh kultur. (The role of humus and micro-
organisms in t h e r o o t n u t r i t i o n o f higher plants in water and sand cultures.) - - Izv. Timiryaz.
s-kh. Akad. Moskva 2 : 173--192, 1955.
GODfrEY, T. N.: Stroenie chlorofilla i metodi ego kolichestvennogo opredelenya. (The structure
o f chlorophyll and methods o f its quantitative determination.) - - Minsk 1952.
KH~IST~VA, L. A.: Rol guminovoy kisloti v pitanii rasteny i guminovie udobreniya. (The role
o f humic acids in plant nutrition and humic fertilizers.) - - Tr. pochv. Inst. ira. Dokuchacva
3 8 : 108--184, 1951.
NIKLEWSKI, B., WOJCIECHOWS~I, J.: Wplyw kwas6w pr6chnicznych na rozwSj ro~lin. (The
influence of humic acids on the development of plants.) - - A c t a Soc. Bet. Pol. 18 : 63--90, 1947.
PRAT, S.: Literatura a pozng~mky o biologick~m vlivu humolitfl (hlavni~ kapuclnfl). (Literature
and notes on the biological influence of humolites.) - - Cyklostyl. Praha 1955.
~roJc~EcI~OWSXi, J.: Por6wnawcze badania nad wptywem ielaza i zwi~zk6w pr6chnicznych na
ro~liny. (Comparative research on the influence of iron and humic compounds on plants.) ---
Rozpr. Pol. Ak. Umiej. 73, dz. B: 26--28, 1948.

Address: Dr ZDEN]~K SLADK~ and :Dr VLADIMiR TICH~2, Laboratory for P l a n t Physiology and
Anatomy, Natural Science Faculty, Brno University, Brno K o t l ~ s k ~ 2.
 APPLICATION OF HUMUS SUBSTANCES 15

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