J. Hon. Sci. 
Vol. 1 (1): 19-23, 2006 

Response of grape rootstocks to soil moisture stress 

J. Satisha, G S. Prakash', G S. R. Murti^ and K. K. Upreti^ 
National Research Centre for Grapes 

P.B. # 3, Manjari Farm 
Solapur Road, Pune-412 307, India 
E-mail: j.satisha@nrcgrapes.res.in 

ABSTRACT 

Studies on root and shoot morphology, endogenous hormones and water use efficiency in five grape rootstocks 
namely Dogridge, 1613 C, Salt Creek, St. George and Vitis champinii clone (VC Clone) at three levels of moisture 
stress viz., no stress (100% irrigation), 50% stress (50% irrigation) and 100% stress (without irrigation) for 14 
days revealed that Dogridge and Salt Creek rootstocks maintained the highest ratios of root to shoot length and 
root to shoot dry weight as compared to other rootstocks. Water use efficiency increased with increased soil 
moisture stress and was the highest in Dogridge and Salt Creek. The abscisic acid content in the leaves of Dogridge 
was maximum at 50% stress followed by that of Salt Creek. Similarly the cytokinin content (both t-ZR and 
DHZR) was minimum in Dogridge and Salt Creek at 50% stress while it was maximum in 1613 C and St.George. 
The root to shoot length ratio was positively correlated with ABA content under moisture stress conditions. The 
higher levels of abscisic acid content in Dogridge and Salt Creek under soil moisture stress suggested their better 
drought tolerance capacity through a reduction of stomatal conductance and increased water use efficiency. 

Key words: ABA, cytokinins, grape rootstocks, root to shoot length ratio, root to shoot dry weight ratio, water use 
efficiency 

INTRODUCTION 

In India, use of rootstocks in viticulture is gaining 
importance as grape is cultivated predominantly under arid 
and semi arid regions. Rootstocks are known for their deep 
root system to exploit water from deeper layers of soil and 
also have selective absorption capacity of mineral elements 
to restrict toxic elements in saline soils. Among the several 
adaptive strategies, improving water use efficiency is 
perhaps the most relevant mechanism in drought tolerance 
(Lincoln and Eduardo, 2002). Another important adaptation 
to moisture stress is that root growth is generally less 
inhibited than shoot growth. This characteristic is thought 
to be adaptive, wherein roots continue to explore soil for 
water while inhibition of shoot growth together with 
stomatal closure restricts transpiration. Abscisic acid (ABA) 
is mostly synthesized in root system and translocated into 
leaves during moisture stress to close stomata and maintain 
relative water content in the plant system. The ability to 
synthesize ABA is specific to variety. Rootstocks are known 
to influence the physiological and biochemical processes 
of scions after budding or grafting (Hartmann and Kester, 
1976). This investigation aims to study the levels of 
endogenous ABA and cytokinin levels in the leaves of 

commonly employed rootstocks, their root and shoot growth 
patterns under varying levels of soil moisture stress with 
the ultimate objective of identifying the drought tolerant 
rootstocks in terms of increased water use efficiency by 
production of more ABA and increased root growth. 

MATERIAL AND METHODS 

Experiments were conducted at the research farm 
of Indian Institute of Horticultural Research, Hessaraghatta, 
Bangalore, India during October-May, 2002-03. The 
experimental site is situated at 14°N latitude and 77°E 
latitude. It is in the elevated plain at an altitude of 863 m 
above the mean sea level. The climate is mild and slightly 
humid, maximum and minimum temperatures were between 
30-34.5 °C and 18-21 "C respectively; the relative humidity 
was 65-70% in the morning hour (8 A.M) and 30-35% 
during afternoon (1 RM), while the evaporation rate ranged 
between 6.25 to 8.3 mm during the experimental period. 

Rooted cuttings of five grape rootstocks namely; 
Dogridge, 1613 C, Salt Creek, St. George and V C clone 
from the nursery beds were transplanted to 14" cement pots 
containing 20 kg potting mixture of red sandy loam soil, 
farm yard manure and sand (2:1:1). The water holding 

'Division of Fruit Crops, Indian Institute of Horticultural Research, Bangalore-560 089, India 
^Division of Plant Physiology and Biochemistry, Indian Institute of Horticultural Research, Bangalore-560 089, India 

mailto:j.satisha@nrcgrapes.res.in


Satisha et al 

capacity of the potting mixture was 30%. The plants, when 
attained the age of six months, were subjected to two levels 
of moisture stress viz., 50% stress and 100% stress and a 
check was maintained with 100% irrigation (no stress). The 
period of stress was 14 days. Plants were irrigated manually 
with 1.5 1 of water for control (no stress), 0.75 1 for 50% 
stress and 100% stress treatment received no irrigation of 
water. The experiment was designed in factorial randomized 
design with three replications consisting of two factors viz., 
irrigation levels and grape rootstocks. Each replication 
consisted of 10 vines. 

Observations were recorded on gas exchange 
parameters using portable photosynthesis system (Model 
6200, LiCor, USA). Water use efficiency (WUE) was 
derived from photosynthesis rate and transpiration rate. 
Fully developed immature leaves were collected in icebox 
and washed in distilled water; 10 g leaf tissue was ground 
in 80% methanol and filtered using Whatman filter paper. 
Residue was incubated overnight with 80% methanol, 
filtered, filtrates were pooled and dried using rotary flash 
evaporator at 35 °C under vacuum and the residue was 
dissolved in distilled water. The water extract was 
partitioned thrice against di-ethyl ether after adjusting pH 
to 3.0 with 0.1 N HCl. The ether fraction was dried in the 
flash evaporator at 35 "C and the residue was dissolved in 
5ml tris buffer (20 mM, pH 7.4) for ABA esfimation. 
Aqueous phase was partitioned with water saturated n-
butanol at pH of 8.0. The butanol fraction was dried under 
reduced pressure at 35 °C in the flash evaporator and the 
residue was dissolved in 5ml tris buffer (20 mM, pH 7.4) 
for estimation of cytokinins like, zeatin riboside (ZR) and 
dihydro zeatin riboside (DHZR). ELISA technique was 
used to quantify ABA (Weiler, 1982) and Cytokinins (Barthe 
and Stewart, 1985) employing laboratory raised polyclonal 
antibodies. The quantity of hormones was expressed as ng/ 
g fresh weight of the tissue. 

Statistical analysis was performed following 
Gomez and Gomez (1984). 

RESULTS AND DISCUSSION 

Gas exchange parameters 

Photosynthetic rate in all the rootstocks reduced 
from fully irrigated control to 50 % stress. None of the 
rootstocks could survive for 14 days without irrigation 
(100% stress). Dogridge rootstock maintained higher 
photosynthesis on 14* day of stress at 50 % stress. Drastic 
reduction in transpiration rate was recorded on 14* day of 
stress cycle in all the rootstocks. Dogridge and Salt Creek 

maintained lower transpiration rate among the rootstocks. 
Water use efficiency increased from control to 50% stress 
in all the rootstocks, highest being in Dogridge. Similar 
results were also reported by Allweldt and Ruht (1982) in 
grapevines with marginal reduction in photosynthesis and 
greater reduction in transpiration under moisture stress 
conditions. Prakash et al (2001) also observed reduction in 
photosynthesis and transpiration rate of grape rootstocks 
with increased soil moisture stress. Maintenance of high 
photosynthetic rate and decreased transpiration rate under 
50% moisture stress could have contributed for the 
increased WUE in the rootstocks, Dogridge and Salt Creek. 

Endogenous Hormones 

Significant differences in hormonal content of 
rootstocks at all stress levels were observed. ABA content 
of all the rootstocks increased with increased soil moisture 
stress. At 50% moisture stress, Dogridge recorded maximum 
ABA content followed by Salt Creek, while it was least in 
1613 C and St. George. Simultaneously, there was a 
reduction in t-ZR content in all the rootstocks with increased 
moisture stress. But DHAR content decreased in all the 
rootstocks except in Dogridge and St. George. Among the 
rootstocks, 1613 C had the highest cytokinin content at 50% 
stress, while it was least in Dogridge and Salt Creek. 
Reynolds and Naylor (1994) and Schultz (1998) also 
reported such increases in ABA: cytokinin ratio and 
decreased stomatal conductance when grape vine Pinot Noir 
and Reisling were subjected to moisture stress. The 
variation in ABA levels among the rootstocks may be a 
varietal behaviour as suggested by Regina and Carbonneau, 
(1997). In spite of the high ABA content in Dogridge leaves, 
there was higher photosynthesis and lower transpiration 
rate, which might be due to relatively higher stomatal 
conductance in this rootstock under moistures stress 
revealing its drought tolerance. 

Morphological parameters 

As the soil moisture stress increased, root length 
increased among grape rootstocks with a concurrent 
reduction in total shoot length. The total root length was 
maximum in Dogridge followed by Salt Creek and both 
recorded lowest shoot length. The root to shoot length ratio 
was maximum in Dogridge (3.50) and was minimum in St. 
George (1.37). Similarly, there was an increase in total root 
dry matter and decrease in shoot dry mass with increased 
soil moisture stress. The root to shoot dry weight ratio was 
maximum in Dogridge (2.01) followed by Salt Creek (1.38) 
and was least in St. George (1.10). Increase in ABA content 
of drying roots combined with availability of water drawn 

J. Hon. Sci. 
Vol. 1(1): 19-23, 2006 

20 



Response of grape rootstocks to soil moisture stress 

Table 1. Influence of moisture stress on gas exchange parameters in grape rootstoclis 

Rootstock Rate of photosynthesis 
(H mol CO / m^ /sec) 

Transpiration rate 
(H mol HO / m^ /sec) 

Stomatal Conductance 
(H mol CO^/ m- /sec) 

Water use efficiency 
{\i mol CO, / m mol Ĥ O) 

Oday 14'" day Oday H'" day Oday U'" day Oday H-'-day 

Dogridge 
1613 C 
Salt Creek 
St. George 
VC clone 

13.70 
8.33 

11.20 
11.10 
12.03 

SI 
8.30 
5.70 
6.80 

10.60 
7.03 

S2 
9.36 
4,83 
9.23 
7.20 
8.40 

S3 
* 
* 
* 
* 
* 

9.06 
7.80 
9.33 
9.43 
9.56 

SI 
9.76 
9.46 
9.76 

10.33 
10.06 

S2 
7.06 
8.33 
7.63 
9.40 
7.93 

S3 
* 
* 
* 
* 
* 

0.82 
0.38 
0.62 
0.62 
0.72 

SI 
0.53 
0.44 
0.58 
0.52 
0.44 

S2 
0.35 
0.36 
0.39 
0.38 
0.39 

S3 
* 
* 
* 
* 
* 

1.50 
1.05 
1.19 
1.16 
1.25 

SI 
0.84 
0.60 
0.69 
1.02 
0.69 

S2 
1.31 
0.58 
1.21 
0.76 
1.06 

S3 

R S RxS R S RxS R S 
SEM± 1.184 0.324 0.251 0.262 0.420 0.113 0.087 0.196 0.099 0.016 0.013 
CD at 5 % NS 0.937 0.726 1.624 NS 0.327 0.253 0.567 NS NS 0.037 

Table 2. Influence of soil moisture stress on endogenous hormonal content in grape rootstocks 

Rootstock Abscisic acid) Zeatin riboside 
(ng /g tissue)) (ng / g tissue) 

Oday H'^day Oday 14'" day 
SI S2 S3 SI S2 S3 

R x S 
0.029 

NS 

Oday 

R S R x S 
0.09 0.040 0.0310.070 
NS 0.118 0.0910.204 

Dihydrozeatin riboside 
(ng / g tissue) 

/ 14'" day 
SI S2 S3 

Dogridge 
1613 C 
Salt Creek 
St. George 
VC clone 

73.80 
17.37 
33.41 
22.20 
13.68 

66.37 
19.35 
32.46 
12.17 
22.46 

163.8 
25.92 
78.05 
31.38 
34.42 

41.24 
63.74 
51.28 
52.98 
75.21 

43.41 
63.49 
52.18 
71.16 
53.43 

32.85 
47.12 
34.32 
30.92 
35.16 

30.22 
40.98 
27.04 
31.81 
51.50 

33.56 
41.92 
27.85 
52.97 
31.61 

26.60 
44.42 
29.70 
30.25 
39.62 

R RxS R RxS R RxS 
SEM± 
CD at 5 % 

1.908 
6.01 

2.005 
5.914 

1.268 
3.740 

2.835 
8.363 

1.361 
4.288 

1.096 
3.235 

0.693 
2.046 

1.551 
4.575 

1.294 
4.079 

1.176 
3.469 

14.975 
2.193 

33.488 
4.905 

Table 3. Influence of soil moisture stress on plant morphological characters on M* day of stress cycle in grape genotypes 

Rootstock 

Dogridge 
1613 C 
Salt Creek 
St. George 
VC clone 

SEm± 
CD at 5% 

S 1 
356.33 
189.00 
252.66 
277.66 
201.66 

V 
23.68 
14.976 

Total Root Length 
(cm) 
S 2 

647.66 
190.00 
420.00 
322.66 
323.33 

S 
69.84 

44.175 

S 3 
* 
* 
* 
* 
* 

VxS 
95.52 

60.248 

S 1 
330.00 
170.66 
277.00 
325.00 
266.66 

V 
8.600 
5.439 

Total Shoot Length 
(cm) 
S 2 

183.66 
125 
191.00 
235.33 
162.66 

S 
25.366 
16.043 

S 3 
* 
* 
* 
* 
* 

VxS 
34.596 
21.880 

Root to shoot length 

SI 
1.09 
1.17 
0.91 
0.85 
0.75 
V 

0.130 
0.384 

S2 
3.50 
1.52 
2.19 
1.37 
1.99 
S 

0.082 
0.243 

ratio 

S3 
* 
* 
* 
* 
* 

VxS 
0.184 
0.543 

Table 4. Influence of soil moisture stress on plant morphological characters on 14"' day of stress cycle in grape genotypes 

Rootstock 

Dogridge 
1613 C 
Salt Creek 
St. George 
VC clone 

SEm± 
CD at 5% 

S 1 
26.75 
28.45 
22.91 
25.31 
32.20 

R 
2.528 

NS 

Total root dry weight 

(g) 
S 2 

47.89 
33.12 
40.03 
27.91 
30.85 

S 
1.599 
4.717 

S 3 
* 
* 
* 
* 
* 

RxS 
3.576 
10.548 

S 1 
38.64 
28.92 
31.38 
28.00 
29.93 

R 
1.469 
4.415 

Total shoot dry 

(g) 
S 2 

24.19 
28.59 
29.15 
25.13 
25.34 

S 
0.946 
2.792 

weight 

S 3 
* 
* 
* 
* 
* 

RxS 
2.116 
6.244 

SI 
0.69 
0.74 
0.79 
0.98 
1.09 
V 

0.129 
0.382 

Root to shoot dry 
weight ratio 

S2 
2.01 
1.21 
1.38 
1.10 
1.22 
S 

0.081 
0.241 

S3 
* 
* 
* 
* 
* 
VxS 
0.183 
0.540 

S 1: Control (100% irrigation) S 2: 50% stress (50% irrigation) S 3: 100% stress (0% irrigation); *: Plants died and the observations were not 
recorded 

/ Hon. Sci. 
Vol. 1 (1): 19-23, 2006 

21 



Satisha et al 

from the wetter roots have an impact on root growth as 
revealed by Dry et al (2000), where partial root drying 
treatment recorded higher root length than fully irrigated 
treatment. Sharp (1996) also suggested that ABA can 
maintain root growth under conditions of low soil moisture 
which results in inhibition of shoot growth. The increased 
root elongation in Dogridge and Salt Creek under moisture 
stress may be due to ABA mediated regulation of osmotic 
adjustment in root tips, wall loosening enzymes and 
restriction of ethylene biosynthesis as hypothesized by 
Sharp et al (1994). The reduction in shoot length of 
rootstocks at 50% moisture stress may be due to reduced 
cytokinin available from the roots, suggesting its important 
role in stomatal conductance. This was confirmed by Stoll 
et al (2000) where, spraying grapevines exposed to water 
deficit, with cytokinin restored stomatal conductance to 
values close to that of fully irrigated vines. Though ABA 
content of 1613 C and St. George increased marginally, 
there was a reduction in stomatal conductance. This might 
be due to accumulation of compounds similar to that of 
ABA that could not be detected by assay system used for 
ABA. Parry et al (1988) reported increased accumulation 
of t-xanthoxin in water stress mutants of tomato at a similar 
rate as ABA in wild mutants of tomato. Similarly Netting 
et al (1997) observed an increased accumulation of 
precursors of ABA biosynthesis pathway in ABA deficient 
mutant of tomato. 

The ABA content was positively correlated (r = 
0.89) with root to shoot length ratio (Fig. I) and negatively 
correlated (r = -0.69) with transpiration rate (Fig. 2). 

The increased WUE, high root to shoot length ratio 
and root to shoot dry weight ratio in response to increased 
ABA content and reduced cytokinin content under soil 

4.5 

4 

5 

3 

5 

2 

5 

1 i 
5 

S3 
C 
« 2 
"5 o 
.c 
in 

o 1 
o o 
a. 

r =0.89 

F i g l 

0 50 100 150 

ABA (ng/g tissue) 

Relationship between ABA and root to shoot length ratio 

200 

l | 
I?" 
II 

12 

10 

8 -

6 

4 

2 

0 

r = -0.69 

50 100 150 200 

ABA (ng/g tissue) 

Fig 2. Relationship between ABA and transpiration rate 

moisture stress conditions in Dogridge and Salt Creek 
rootstocks suggests their better drought tolerance capacity 
than 1613 C and St. George rootstocks. 

ACKNOWLEDGEMENTS 

The authors are thankful to Mr. Jayaram and Mr. 
Nageshwara Rao for their technical assistance during this 
investigation. 

REFERENCES 

Allweldt, G and Ruhl, E. 1982. Investigation on gas 
exchange in grape vine: Influence of extended soil 
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Barthe, GA. and Stewart, I. 1985. Enzyme immunoassay 
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Dry, RR. Loveys, B.R. and During, H. 2000. Partial drying 
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Gomez, A.K. and Gomez, A.A. 1984. Statistical procedure 
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Hartmann, H. and Kester, D.E. 1976. Plant propagation: 
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J. Hon. Sci. 
Vol. 1 (1); 19-23, 2006 

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Response of grape rootstocks to soil moisture stress 

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(MS Received 29 June, 2006 Revised 14 July, 2006) 

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Vol, 1 (1): 19-23,2006 

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