Plant Physiol. Rep.

https://doi.org/10.1007/s40502-020-00508-6

ORIGINAL ARTICLE

Influence of different light intensities on specific leaf weight,

stomatal density photosynthesis and seed yield in soybean
Kanchan Jumrani1 • Virender Singh Bhatia1

Received: 13 May 2018 / Accepted: 3 March 2020

Ó Indian Society for Plant Physiology 2020

Abstract The availability of low light intensity due to Keywords Intercropping
Photosynthesis
Shading

shading is the critical factor in determining soybean yield Solar radiation
Soybean
Yield
in intercropping system. An experiment was conducted to
search genotypes tolerant to low light intensity, which can
be better grown for intercropping system. The objective of Introduction
this study was to determine the influence of different light
regimes (35%, 50%, 75% of normal light and normal light) Soybean is the world’s most important seed legume and it
on physiological traits such as specific leaf weight, stom- contributes significantly to edible oil, protein concentrate
atal density, photosynthesis, total biomass and seed yield of for animal feed, food uses and various industrial products
soybean genotypes. Different shading conditions were (Bhatia and Jumrani 2016; Jumrani et al. 2017, 2018;
created using shading net of 35, 50 and 75% to cut the Jumrani and Bhatia 2018). Intercropping is a farming
natural solar radiation received by plants to identify practice involving two or more crop species growing
genotypes tolerant to low light intensity. One set of plants together. Intercropping used by small farmers primarily to
was also grown under natural conditions without any increase the diversity of their crop and the stability of their
shading net. Significant negative effect of low light inten- annual production through effective use of land and
sity was observed on specific leaf weight, stomatal density resources. Crop variety in intercropping pattern must be
and photosynthesis in soybean genotypes. Among the carefully chosen to minimize competition and enhance the
shading conditions, the average seed yield was maximum efficient use of water, light and nutrients. Success of
(31.5 g/pl) in plants grown under control condition. When intercropping pattern depends on the ability of the second
plants were grown under 35% shading net the seed yield crop to become established under the canopy of the first
was declined by 37% (20.0 g/pl). However, severe reduc- crop.
tion to the extent of 64 and 76% was observed when plants Soybean is commonly grown in intercrop combinations
were grown under shading net of 50% (11.2 g/pl) and 75% of which cereal/legumes are the most common in tropical
(7.7 g/pl), respectively as compared to plants grown under countries. The planting patterns of maize-soybean inter-
control condition. Therefore, the effects of shade on crops cropping systems can induce changes in the light intensity
should be considered when trying to increase the produc- of the soybean canopy (Awal et al. 2006). Seed yield of
tivity of an intercropping system. soybean reduced under the low light because of reduction
in dry matter production (Kakiuchi and Kobata 2004).
Maize yield is not affected by the presence of soybean
while soybean yield is more often reduced under high
maize population. To obtain the maximum seed yield of
& Kanchan Jumrani intercropped soybean under shading effects of adjacent
[email protected]
maize plants, selection of suitable soybean genotype could
1
Division of Plant Physiology, Indian Institute of Soybean play an important role for intercropping system. New
Research, Indore, India approaches to plant breeding are needed for intercropping

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systems (George et al. 2014). Therefore, the main objective ambient PAR of 1000–1200 lmol m-2 s-1, CO2 concen-
of the present research was to observe influence of different tration (390–400 lmol mol-1air CO2) in each treatment
light on physiological traits such as specific leaf weight, between 10:00 and 12:00 h.
stomatal density, photosynthesis, total biomass and seed
yield and to identify the compatible soybean genotype, Stomatal density
which can be better grown to achieve farmer’s benefit
under intercropping conditions using various physiological To measure the stomatal density, the impression approach
traits. was used and is expressed as the number of stomata per
unit leaf area. Leaves were smeared with transparent nail
varnish in the mid-area between the central vein and the
Materials and methods leaf edge for * 20 min. After drying, the thin film was
peeled off from the leaf surface, mounted on a glass slide,
Shading treatments and number of stomata for each filmstrip was counted
under a microscope (LEICA Model DM 2000 LED,
The experiment was conducted at ICAR-Indian Institute of Germany).
Soybean Research, Indore (22.72° N, 75.83° E). Three
structures with their tops and sides covered by mesh and Seed yield and yield attributes
knitted shade cloths were constructed to provide shading
treatments. Shade treatments consisted of 0% shade (con- At maturity, plants from each treatment and genoptype
trol) in which plants were left uncovered in the 100% day were harvested from 6 pots (2 plants/pot) and data on seed
light and 35, 50 and 75% shade. Solar radiation for the crop yield and total biomass was recorded.
duration was recorded using automatic weather station
(Licor Inc., USA) installed at 100 m away from the Statistical analysis
experimental site.
Analysis of variance was carried out for all the data sets
Planting of soybean using SAS statistical software (ver.9.2; SAS Institute, Cary,
NC). The treatment means (main effect as well as inter-
Eleven genotypes were grown in cement pots (45 cm action means) were compared based on least significant
height and 18 cm diameter) filled with farm soil (black differences (LSD) at P B 0.05 using Duncan multiple
clay) and farm yard manure mixed in the ratio of 2:1.sown range test (DMRT).
under varying solar radiations viz. 0, 35, 50 and 75% shade
conditions.
Results and discussion
Data collection and analysis
The intensity of light is essential for growth, morphogen-
Specific leaf weight (SLW) esis, photosynthesis, adaptation and productivity of plants
(Ali et al. 2005; Schneider et al. 2006). Growing crops in
For recording specific leaf weight (SLW), leaf discs intercropping is the main feature of the conventional
(2 cm2) were collected from intact fully expanded third cropping systems in the tropical countries (Li et al. 2009).
leaf from top in three replications (5 leaves/replication) at During the intercropping, light is one of the most important
R5 stage. The specific leaf weight (SLW) was calculated limiting factors to yields because of inter specific compe-
using the following formula: SLW (g/cm2) = LW/LA, tition (Ghanbari et al. 2010). Therefore, the effects of shade
where LW is the leaf dry weight and LA is the leaf area. on crops should be considered when trying to increase the
productivity of an intercropping system. The present study
Rate of photosynthesis was conducted with the aim to identify the soybean
genotypes which can be better grown under intercropping.
To measure the rate of photosynthesis at R5 stage, five
plants from each treatment were selected from different Weather conditions
pots. Gas exchange parameters were measured on an intact
youngest fully expanded third leaf from top in three Eleven soybean genotypes were grown under ambient
replications (5 leaves/replication) using portable photosyn- conditions. Different shading conditions were created using
thesis system (LI-6400 XT, USA). A leaf was fitted into shading net of 35, 50 and 75% shade to cut the natural solar
6 cm2 leaf chamber and measurements were made at radiation received by plants to identify genotypes tolerant

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to low solar radiation. Daily solar radiation was recorded compared to control conditions (Table 2). Soybean geno-
and the cumulative solar radiation was 1027 MJ/m2 of crop types also differed significantly for the stomatal frequency.
growth period (Fig. 1). The average frequency on the abaxial surface was highest
in JS 97-52 (188/mm2) and lowest for JS 20-29 (158/mm2).
Effect of shading on specific leaf weight (SLW) Higher stomatal density and thicker leaves tend to favor a
higher photosynthetic rate (Beemster et al. 2005).
The maximum average SLW was observed in plants grown
under control condition (4.10 g/cm2) and was reduced by Effect of shading on rate of photosynthesis
30, 42 and 58% when the plants were grown under 35, 50
and 75% shading net, respectively (Table 1). Significant Maximum rate of photosynthesis in soybean genotypes
differences among genotypes for SLW were also observed. (30.9 lmol CO2 m-2 s-1) was observed in plants grown
Genotype JS 335 (3.12 g/cm2), JS 95-60 (3.01 g/cm2), EC under control condition which was reduced by 18, 30, 45%
538828 (2.88 g/cm2), RVS 2001-4 (2.88 g/cm2) and JS when the plants were grown under 35, 50 and 75% shading
97-52 (2.87 g/cm2) showed significantly higher average net, respectively as compared to control condition
SLW while genotypes JS 20-69 (2.27 g/cm2) exhibited (Table 3). Genotypes also differed significantly for their
significantly lowest average SLW. Leaf morphological and average rate of photosynthesis. Among genotypes, rate of
anatomical characteristics play important role in the regu- photosynthesis was maximum in JS 93-05 (25.3 lmol
lation of photosynthetic capacity and provide a structural CO2 m-2 s-1), while minimum was observed in JS 20-69
framework for the diffusion of gases (Jiang et al. 2011). (21.6 lmol CO2 m-2 s-1). Light plays a critical role in
For leaf area, it has been reported that shading inhibited plant growth and development and regulate plant photo-
leaf expansion (Kozuka et al. 2005), which consequently synthetic efficiency. The high photosynthetic rate under
resulted in the reduction of the light interception. Thus, full sunlight was in accordance with other studies (Mielke
thinner leaf have disadvantages in terms of photosynthetic and Schaffer 2010).
capacity consequently, supply of photosynthates reduces in
thinner leaves (Tholen et al. 2012). Effect of shading on seed yield

Effect of shading on stomatal density Among the shading conditions, the average seed yield was
maximum (31.5 g/pl) in plants grown under control con-
In the present study shading led to a significant decrease in dition. When plants were grown under 35% shading net the
stomatal density. The average number of stomata on seed yield was marginally declined by 37% (20.0 g/pl).
abaxial surface in leaves of the plants grown under control However, severe reduction to the extent of 64 and 76% was
conditions was 230/mm2 and reduced to 189 (18%), 154 observed when plants were grown under shading net of
(33%) and 128/mm2 (44%) when the plants were grown 50% (11.2 g/pl) and 75% (7.7 g/pl) as compared to plants
under 35, 50 and 75% shading net, respectively as grown under control condition, respectively (Table 4).

Fig. 1 Solar radiation (daily

and cumulative) profile for the
crop season

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Table 1 Effect of shading on

Genotypes Control 35% shade 50% shade 75% shade Mean
specific leaf weight (g/cm2) in
soybean genotypes EC 538828 4.08 2.94 2.57 1.94 2.88bc
EC 456548 3.49 2.69 2.20 1.59 2.49e
JS 95-60 5.03 3.13 2.38 1.51 3.01ab
JS 20-34 4.44 3.10 2.50 1.85 2.97bc
JS 335 4.74 3.24 2.59 1.91 3.12a
JS 93-05 4.51 2.87 2.30 1.73 2.85c
RVS 2001-4 4.08 2.93 2.65 1.87 2.88bc
JS 20-29 3.69 2.52 2.14 1.67 2.51e
JS 97-52 4.00 3.09 2.50 1.89 2.87bc
EC 602288 3.63 2.86 2.37 1.82 2.67d
JS 20-69 3.38 2.28 2.03 1.38 2.27f
a b c d
Mean 4.10 2.88 2.38 1.74 2.77
ANOVA
Shading \ 0.001
Genotype \ 0.001
S9G \ 0.001
Means within a row and column for each main treatment followed by the same letters are not significantly
different (P B 0.05). Values under ANOVA are the probabilities (P values) of the sources of variation

Table 2 Effect of shading on

Genotypes Control 35% shade 50% shade 75% shade Mean
number of stomata (number/
mm2) in soybean genotypes EC 538828 213 192 160 138 176de
EC 456548 223 193 160 139 179cd
JS 95-60 264 203 153 109 182bc
JS 20-34 226 176 148 114 166f
JS 335 242 196 169 138 186ab
JS 93-05 251 200 146 121 180cd
RVS 2001-4 218 179 149 134 170e
JS 20-29 211 170 137 114 158g
JS 97-52 234 200 169 149 188a
EC 602288 228 194 158 140 180cd
JS 20-69 220 172 143 115 163fg
a b c d
Mean 230 189 154 128 175
ANOVA
Shading \ 0.001
Genotype \ 0.001
S9G \ 0.001
Means within a row and column for each main treatment followed by the same letters are not significantly
different (P B 0.05). Values under ANOVA are the probabilities (P values) of the sources of variation

Among genotypes, the average yield was significantly high Effect of shading on total biomass (TBM)
in JS 97-52 (29.3 g/pl), EC 602288 (24.4 g/pl), EC 456548
(22.3 g/pl) and RVS 2001-4 (20.7 g/pl) as compared to Maximum TBM (51.5 g/pl) was observed in plants grown
genotypes such as JS 95-60 (10.9 g/pl) and JS 93-05 (8.2 g/ under control condition, which was reduced by 35, 62, and
pl) which gave lowest seed yield (Fig. 2). Based on percent 75% when the plants were grown under 35, 50 and 75%
reduction in yield with decreasing solar radiation among 11 shading net, respectively as compared to control conditions
genotypes studied JS 97-52 and EC 602288 showed rela- (Table 5). Hence, similar to the seed yield, the magnitude
tively less reduction in yield while maximum reduction in of reduction in biomass was more severe under 50 and 75%
seed yield was observed in JS 95-60. net conditions (Table 5). Among genotypes, the average

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Table 3 Effect of shading on

Genotypes Control 35% shade 50% shade 75% shade Mean
rate of photosynthesis
(lmol CO2 m-2 s-1) in EC 538828 29.5 25.1 23.3 17.2 23.8c
soybean genotypes
EC 456548 29.9 25.5 22.7 17.5 23.9bc
JS 95-60 33.6 26.5 20.2 15.1 23.8c
JS 20-34 30.6 24.8 19.2 16.2 22.7d
JS 335 31.4 25.7 21.2 17.6 24.0bc
JS 93-05 34.7 28.1 21.2 17.0 25.3a
RVS 2001-4 30.8 26.5 21.9 19.4 24.6ab
JS 20-29 29.6 23.1 20.9 16.0 22.4d
JS 97-52 30.1 25.2 23.2 17.6 24.0bc
EC 602288 30.8 26.3 23.7 16.8 24.4bc
JS 20-69 29.0 23.1 19.0 15.3 21.6e
a b c d
Mean 30.9 25.4 21.5 16.9 23.7
ANOVA
Shading \ 0.001
Genotype \ 0.001
S9G \ 0.001
Means within a row and column for each main treatment followed by the same letters are not significantly
different (P B 0.05). Values under ANOVA are the probabilities (P values) of the sources of variation

Table 4 Effect of shading on

Genotypes Control 35% shade 50% shade 75% shade Mean
seed yield (g/plant) in soybean
genotypes EC 538828 22.9 15.4 9.6 6.5 13.6f
EC 456548 39.4 24.9 14.5 10.5 22.3c
JS 95-60 25.1 12.3 4.5 1.9 10.9g
JS 20-34 23.3 12.5 5.7 3.9 11.4g
JS 335 37.4 23.1 11.2 7.8 19.9de
JS 93-05 17.0 9.1 4.2 2.6 8.2h
RVS 2001-4 36.6 22.9 13.4 9.9 20.7d
JS 20-29 25.9 16.7 7.2 5.1 13.7f
JS 97-52 44.1 33.4 23.2 16.6 29.3a
EC 602288 38.4 27.2 19.3 12.7 24.4b
JS 20-69 36.8 22.3 10.8 7.1 19.2e
a b c d
Mean 31.5 20.0 11.2 7.7 17.6
ANOVA
Shading \ 0.001
Genotype \ 0.001
S9G \ 0.001
Means within a row and column for each main treatment followed by the same letters are not significantly
different (P B 0.05). Values under ANOVA are the probabilities (P values) of the sources of variation

TBM was maximum for JS 97-52 (51.1 g/pl) while the intercropping. Decline in the seed yield of soybean, when
minimum TBM was observed in JS 95-60 (17.1 g/pl) and intercropped with maize has been reported (Hayder et al.
JS 20-34 (16.9 g/pl). Seed yield is usually positively cor- 2003). The low yield in intercropped soybean as compared
related with total dry matter accumulation and therefore to monoculture is mainly due to shading resulting in weak
any effect on total dry matter accumulation affects yield. plant growth. The above results indicated that high light
According to previous comparative studies, the total bio- grown plants yielded greater productivity than low light
mass decreased under low light (Zhang et al. 2003). grown plants. Significant variation in yield and other
The availability of low light intensity due to shading is physiological processes were observed among the 11 soy-
the critical factor in determining legume yield in bean genotypes evaluated in the present study. Also,

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Fig. 2 Reduction in seed yield

(%) in different soybean
genotypes grown under 35, 50
and 75% shading conditions as
compared to control

Table 5 Effect of shading on

Genotypes Control 35% shade 50% shade 75% shade Mean
total biomass (g/plant) in
soybean genotypes EC 538828 33.7 22.5 16.0 9.3 20.4h
EC 456548 55.8 37.8 21.9 15.5 32.7d
JS 95-60 35.5 19.5 8.2 5.2 17.1j
JS 20-34 34.9 18.5 8.3 6.1 16.9j
JS 335 45.7 28.6 15.8 9.9 25.0g
JS 93-05 36.4 20.4 11.7 7.2 18.9i
RVS 2001-4 66.9 42.6 26.9 15.9 38.1c
JS 20-29 48.5 30.6 15.2 10.6 26.2f
JS 97-52 77.8 59.4 39.6 27.7 51.1a
EC 602288 77.2 53.9 35.4 24.0 47.6b
JS 20-69 53.9 34.7 15.8 11.0 28.8e
a b c d
Mean 51.5 33.5 19.5 12.9 29.4
ANOVA
Shading \ 0.001
Genotype \ 0.001
S9G \ 0.001
Means within a row and column for each main treatment followed by the same letters are not significantly
different (P B 0.05). Values under ANOVA are the probabilities (P values) of the sources of variation

shading and genotype interaction was also significant for Conclusion

most of the parameters studied, which indicated that the
response of these genotypes to shading differed consider- New approaches to plant breeding are needed for inter-
ably. The adverse effect of shading on physiological cropping systems. Selection of suitable shade-tolerant
parameters such as specific weight, stomatal density, total soybean varieties can be used as a direct and efficient
biomass and photosynthesis was relatively less in geno- approach to relieve the effect of shade resulting from the
types, which showed less reduction in yield as compared to maize canopy in intercropping. To obtain the maximum
highly sensitive genotypes. yield of the intercrop soybean under low light stress,
selection of suitable soybean cultivar plays an important
role for intercropping systems. The cultivars may respond
to shading stress differently in terms of growth and yield.
Therefore, the genotype JS 97-52 had a higher potential
yield advantage in inter-cropping systems in which low

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light intensity is a major limiting factor on seed yield. performance and high-light tolerance in sorghum. Plant Phys-
Thus, using these physiological traits as selection criteria, iology, 155, 1416–1424.
Jumrani, K., & Bhatia, V. S. (2018). Impact of combined stress of
soybean genotypes better grown for intercropping can be high temperature and water deficit on growth and seed yield of
identified which require more studies to confirm the find- soybean. Physiology and Molecular biology of Plants, 24,
ings of the study. 37–50.
Jumrani, K., Bhatia, V. S., & Pandey, G. P. (2017). Impact of elevated
Acknowledgements The authors acknowledge funding for this work temperatures on specific leaf weight, stomatal density, photo-
from Indian Council of Agricultural Research. synthesis and chlorophyll fluorescence in soybean. Photosyn-
thesis Research, 131, 333–350.
Jumrani, K., Bhatia, V. S., & Pandey, G. P. (2018). Screening
soybean genotypes for high temperature tolerance by in vitro
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