Agriculture, Ecosystems and Environment 233 (2016) 7584

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Agriculture, Ecosystems and Environment

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Effects of foliar application of plant growth hormone on methane

emission from tropical rice paddy
Leena Boraha,b , Kushal Kumar Baruaha,*
a
Department of Environmental Science, Tezpur Central University, Tezpur-784028, Assam, India
b
Present address: Department of Environmental Biology and Wildlife Sciences, Cotton College State University, Guwahati-781001, Assam, India

A R T I C L E I N F O A B S T R A C T

Article history:
Received 29 February 2016 Methane (CH4), a major greenhouse gas, is an important agent of global warming and climate change.
Received in revised form 18 July 2016 While rice agriculture is a major source of anthropogenic CH4 emissions, increased production of rice is
Accepted 30 August 2016 essential for ensuring global food security. Mitigating CH4 emissions from rice cultivation with
Available online 5 September 2016 simultaneous increase in grain productivity is a challenging issue. A two-year eld study was conducted
to investigate the effects of foliar application of plant growth hormones on emission reduction of CH4
Keywords: from rice paddies. CH4 emission measurement was done from rice plants treated with plant growth
Methane hormones gibberellic acid (GA3), indole-3-acetic acid (IAA), indole-3-butyric acid (IBA) and kinetin
Plant growth hormones
(KIN) (in 20 mg L!1 concentration) and compared with the emission from untreated/control (CONT)
Stomatal frequency
plants. IAA and KIN applications were found to bring about a reduction in the cumulative CH4 emission
Xylem size
Photosynthate partitioning over control primarily through regulation of leaf growth, stomatal density and xylem vessel size. Foliar
Grain yield application of IAA and KIN enhanced the leaf photosynthetic rate and caused maximum partitioning of
photosynthates to the grains as evident from the higher grain lling ability and grain yield. The IAA and
KIN treatments improved the thousand grain weight and high density grain (%) in the rice plants resulting
in higher grain productivity over untreated plants. It can be concluded that foliar application of indole
acetic acid and kinetin can be an effective measure for regulating methane emission from rice paddies
coupled with increase in economic yield of the most popular crop of this region.
2016 Elsevier B.V. All rights reserved.

1. Introduction Green plants are reported to inuence the production,

Rice (Oryza sativa L.) is the major cereal crop and the staple food
for more than half of the worlds population. Annual rice cropping
area in India is reported to be 44.1 million hectares which is the
largest rice growing area of the world (FAOSTAT, 2013). Rice
production in India needs to be increased for national food
security. On the other hand, rice elds are considered as one of the
major sources for greenhouse gas (GHG) emissions contributing
about 30 and 11% of global agricultural methane (CH4) and nitrous
oxide (N2O) emissions respectively (IPCC, 2007). The global
warming potential (GWP) of CH4 is 34 times higher than that of
CO2 (Myhre et al., 2013). Indian agriculture accounts for
approximately 5% of the global CH4 budget (Wassmann et al.,
2009) with an emission estimate of 4.09 " 1.19 Tg CH4 y!1 from
Indian paddy elds (Gupta et al., 2009).
* Corresponding author.
E-mail addresses: leeborah@gmail.com (L. Borah), kkbaruah14@gmail.com,
kkbaruah@tezu.ernet.in (K.K. Baruah).
consumption, as well as transport of CH4 from the soil to the
atmosphere (Koelbener et al., 2010). Root exudates from plants
play a signicant role in CH4 production from soil organic matter
by determining the C content of the rhizosphere soil (Koelbener
et al., 2010). Rice plants also mediate the transport of CH4 from the
soil to the atmosphere (Nouchi et al., 1990; Aulakh et al., 2000;
Ding et al., 2005; Das and Baruah, 2008a).
Plant growth hormones (gibberellins, auxins, cytokinins) are
widely known to regulate plant growth and developmental
processes such as cell division, cell elongation, protein synthesis
etc. and also determine overall plant architecture (Gray, 2004;
Depuydt and Hardtke, 2011; Durbak et al., 2012). Therefore,
application of plant growth hormones may inuence the CH4
transport capacity of the rice plants by regulating their growth and
development. A two-year eld study was undertaken to investigate
the impact of plant growth hormones on CH4 emissions from a rice
ecosystem planted to cultivar Luit.
Optimization of grain yields have been suggested as an
important measure for substantial CH4 emission reduction from
rice agriculture by several authors (Denier van der Gon et al., 2002;

http://dx.doi.org/10.1016/j.agee.2016.08.033
0167-8809/ 2016 Elsevier B.V. All rights reserved.
76 L. Borah, K.K. Baruah / Agriculture, Ecosystems and Environment 233 (2016) 7584
Das and Baruah, 2008b; Jiang et al., 2013). Plant growth hormones
play crucial role in controlling sink and source size in plants and
thus inuence the grain yield (Ghodrat et al., 2012). Therefore, in
the present study, attempts were also made to determine whether
growth hormone application in rice agriculture can improve the
grain productivity while simultaneously reducing the CH4
emissions.
2. Materials and methods
2.1. Site description
The experiments were conducted inside Tezpur Central
University Campus, Tezpur (26! 410 N, 92! 500 E) situated in the
North Bank Plain Agro-climatic Zone of Assam, India. The region is
subtropical humid having moderately hot-wet summers and dry
cold winters. The soil is characterized by recent and old alluvium
soils of sandy to sandy-loam texture with sand (%) 60.35, silt (%)
14.40, clay (%) 23.50 and slight to moderate acidic soil pH (5.65).
The bulk density of the soil was 1.72 g cm"3 and the percentage of
organic carbon was 0.94. The available nitrogen, phosphorous and
potassium content of the soil were 108.64, 32.72 and 249.18 kg
ha"1 respectively. The meteorological parameters recorded at a
weather station located at the university campus are presented in
Supplementary Fig. 1.
2.2. Field management
Rice was planted to the eld during pre-monsoon season (locally
known as Ahu rice, AprilJuly) of 2012 and 2013. The main eld was
ploughed, puddled thoroughly and leveled properly before trans-
planting of rice. Fertilizers were applied @ 40:20:20 kg N-P2O5-
K2O ha"1 as recommended by the Department of Agriculture,
Assam. One-third of the quantity of N (13.3 kg ha"1 applied as urea)
and the whole quantity of P2O5 (as single super phosphate) and K2O
(as muriate of potash) were applied at the time of nal puddling.
Twenty-ve days old rice seedlings of an improved rice cultivar Luit
were transplanted (2 seedlings hill"1) during the rst week of April
with a spacing of 20 cm between the rows and 15 cm between plant
to plant in plots of size 2 m # 2m = 4 m2. The second 1/3rd
(13.33 kg ha"1) and the third 1/3rd (13.33 kg ha"1) doses of urea
were applied at tillering (28 days after transplanting/28 DAT) and
panicle initiation stages (56 DAT) respectively. The experiment
consisted of ve treatments (section 2.3) with 4 replications each
arranged in a randomized block design.The crop was grown
exclusively under rainfed condition and was properly managed
during the experimental period. Insecticides were sprayed just after
crop establishment (8 DAT) for protection against caseworms which
was well before the rst dose of hormone application (28 DAT).
2.3. Foliar application of plant growth hormones
Four different plant growth hormones viz. gibberellic acid (GA),
indole acetic acid (IAA), indole butyric acid (IBA) and kinetin (KIN)
were selected for this experiment. Hormone solutions were
prepared by rst dissolving 20 mg of GA, IAA and IBA in few
drops of ethanol and 20 mg of KIN in 1N NaOH since they were
insoluble in water. Then the nal volume was made up to 1000 ml
with distilled water in order to get the desired concentration
(20 mg L"1). One set of rice plants was sprayed with distilled water
without any hormone and this treatment was taken as control
(CONT). Thus the total number of treatments was ve, viz.
T1 = CONT, T2 = GA, T3 = IAA, T4 = IBA, T5 = KIN. These solutions
were sprayed on the rice leaves at tillering and panicle initiation
stage of the crop with a back-pack sprayer system (Ghodrat et al.,
2012). Maximum care was taken during spraying to ensure that no
run-off reaches the soil from the leaves which could possibly affect
soil bacterial activity. In one 2 m # 2m = 4 m2 plot, there were 130
numbers of rice plants and 500 ml of 20 mg L"1 solution per plot
was sprayed on the rice leaves.
2.4. CH4 sampling and measurement
CH4 ux from both treated and untreated rice plants was
recorded from 0 DAT at weekly interval up to 21 days after harvest
(DAH) using the static chamber technique described by Buendia
et al. (1997). Chambers of 50 cm length, 30 cm breadth and 70 cm
height made of 6 mm thick acrylic sheets were used for gas
collection. Chambers with height ranging from 90 to 120 cm were
used during the later crop growing period to accommodate the
increasing plant height. Rectangular U shaped aluminum channels
(50 cm # 30 cm) were used to accommodate the chamber. The
aluminum channels were inserted into the soil to a depth of 15 cm
well ahead of gas sample collection. Each aluminum channel
enclosed six plants of rice. A battery-operated fan was xed inside
the chamber to homogenize the inside air. The top of the chamber
was tted with two self-sealing rubber septa. A thermometer was
inserted through one of these septa to record the chamber
temperature. Gas samples were drawn from the chambers using a
50 ml airtight syringe tted with a three-way stop-cock and a ne
needle inserted through the other self-sealing rubber septum. The
samples were drawn at xed interval of 0, 15, 30 and 45 min at
0900 h and again at 1400 h from the day of transplanting (0 DAT)
onwards at weekly interval. The samples were brought to the
laboratory immediately after collection and CH4 concentration in
the gas samples were analyzed using a gas chromatograph (Varian,
CP-3800 GC, USA) equipped with a ame ionization detector (FID)
and a chromopack capillary column. Column, injector and detector
temperatures were maintained at 50 ! C, 90 ! C and 150 ! C,
respectively. The gas chromatographic system was calibrated
periodically with a standard obtained from National Physical
Laboratory, New Delhi, India. Nitrogen (N2) (99.999% pure) was
used as a carrier gas whereas hydrogen (H2) and zero air were used
for ignition of the ame in the FID. CH4 uxes (mg CH4 m"2 h"1)
were calculated from the temporal increase in the gas concentra-
tion inside the box using the equation of Parashar et al. (1996) and
the average of morning and evening uxes were considered as the
ux value for the day. Cumulative CH4 emission for the entire crop
growth period was computed by the method given by Naser et al.
(2007) by using the following formula:
n"1
Cu mulative CH4 emission S Ri # Di
i1
Where Ri is the mean gas emission, Di is the number of days in the
sampling interval and n is the number of sampling times.
Cumulative CH4 emission was expressed in g m"2.
2.5. Analysis of soil parameters
Prior to rice cultivation soil samples were collected randomly
from different locations of the experimental plot from a depth of 0
15 cm and soil samples were analyzed for the basic soil
physicochemical properties (Section 2.1) by following the methods
described in Borah and Baruah (2016).
After transplanting of rice, soil samples were collected from the
rice grown plots at weekly interval on the day of CH4 gas sampling
with the help of a soil core from 015 cm soil depth and analyzed
for soil moisture content (SMC) following the method of Page et al.
(1982). Soil organic carbon was analyzed in a TOC analyzer (Multi
N/C 2100S with HT 1300 module, Analytik Jena, Germany) by the
method of Awale et al. (2013).
 L. Borah, K.K. Baruah / Agriculture, Ecosystems and Environment 233 (2016) 7584
2.6. Analysis of plant morphological and physiological parameters
Plant variables such as leaf area index and tiller number were
recorded at weekly interval on the day of CH4 sampling. Total leaf
area of plants within 1 m2 area of the planted plots were estimated
with the help of portable laser leaf area meter (CID, Model CI-203,
USA) at weekly interval (on the day of CH4 sampling) from four
replications. The leaf area index (LAI) was expressed as the total
leaf area per unit ground area (m2). The results presented are the
mean of four observations.
Five plants were selected at random from each plot and the
number of tillers were counted from the four replications
(n = 5 ! 4 = 20) on the day of CH4 sampling.
Leaf photosynthesis, transpiration and stomatal conductance
were recorded at weekly interval with the help of an infrared gas
analyzer (LI-6400 Portable photosynthesis system, LI-COR, Lincoln,
NE, USA). The middle portion of a fully expanded leaf from the top
of a selected plant was taken for measurement of photosynthesis
and transpiration during pre-owering stage. Three randomly
selected plants from each replication (total 3 replications) were
taken for this measurement. After panicle initiation stage (56 DAT
onwards) photosynthesis and transpiration rate was measured in
the fully expanded ag leaf. Results presented are the mean of total
nine readings for each observation.
For plant biomass determination, four randomly selected plants
of rice were uprooted carefully from three replications of each
treatment (n = 4 ! 3 = 12) at weekly interval (on the day of CH4
sampling) and were brought to the laboratory. The soil adhered to
the roots were washed carefully by placing the roots on a ne sieve
to prevent the washing away of root fragments. After that, different
parts of the plants viz. leaf blade, leaf sheath, culm, root (at
vegetative stage) and panicle (after panicle emergence) were
separated with the help of scissors. Different plant parts were dried
separately in an oven at 75 " C until constant weight was obtained
and then the dry weights were recorded in an electronic balance
(Sartorius CP225D, Germany). Plants within 1 m2 of each 4 m2 plot
were kept intact for determination of grain yield at harvest. Plants
for biomass determination were sampled at random from outside
this 1 m2 area.
2.7. Analysis of plant anatomical parameters
Stomatal frequency of ag leaf and size of the xylem vessels of
the node attached to the ag leaf of the rice plants were analyzed at
panicle initiation stage (56 DAT) with the help of scanning electron
microscope (SEM) during the second year of experimentation
(2013). Leaf sections were cut from the middle portion of the fully
expanded ag leaf with a sharp blade and immediately xed in 3%
glutaraldehyde for 24 h. Primary xation was followed by washing
in 0.1 M sodium cacodylate buffer. Post xation was done by
immersing the samples overnight in 1% osmium tetroxide.
Thereafter, the samples were washed again in 0.1 M sodium
cacodylate buffer. Washing was followed by dehydration which
was done by passing the samples through various acetone grades
(30, 50, 70, 80, 90, 95 and 100% 2 changes of 15 min each) at 4 " C.
Dehydrated samples were then dried using the Tetra Methyl Silane
(TMS) method. Samples were immersed in TMS for 510 min (2
changes) at 4 " C and then brought to room temperature (26 " C) for
drying. The samples were then mounted on metal stubs with a
carbon adhesive tape and platinum sputter coating was done by
Auto Fine Coater (JFC-1600; JEOL, Tokyo, Japan) before examining
the samples under a SEM (JEOL Asia PTE Ltd., Singapore/JEOL JSM,
Model 6390 LV). Three random elds from each sample were taken
for the measurement of stomatal frequency (from the adaxial
surface of leaf). The average stomatal frequency was expressed as
number of stomata per mm2 of leaf surface area. Sections of node
77
adjacent to the ag leaf were prepared for analysis of the xylem
size following the same procedure. Three to four randomly selected
vascular bundles were photographed in SEM and from these, 5
individual xylem vessels were selected at random for calculation of
xylem size. Xylem size was calculated in mm according to the scale
visible on the micrograph and expressed as the average internal
diameter of xylem vessels of the node.
2.8. Analysis of yield and yield attributing parameters
Five panicles each from four replications (n = 5 ! 4 = 20) of the
treatments were taken at random for determining yield param-
eters such as panicle length and lled grain (%) at harvest.
Thousand grain weight and grain yield were recorded from four
replications (n = 4) of each treatment following the method of
Baruah et al., 2012.
2.9. Statistical analysis
The statistical software IBM SPSS version 20.0 was used to
calculate the correlation (Pearson correlation) coefcient of CH4
uxes with means of plant and soil variables. Pooled analysis of
data for two years of experimentation was done by using two-way
ANOVA and Duncans multiple range test (DMRT).
3. Results
3.1. Weekly CH4 emission and cumulative CH4 emission from the rice
plants
The quantity of CH4 emitted from the rice plants varied with
respect to the control depending on the hormones applied (Fig. 1).
However, the trend of CH4 emission was similar in hormone
treated as well as control plants with two distinct emission
maxima-one at tillering and the other at panicle initiation stage of
the crop (Fig. 1). CH4 ux ranged from 0.02 to 1.13 mg m#2 h#1
during the experimental period of 2012 and from 0.09 to
0.89 mg m#2 h#1 during 2013 (Fig. 1). Initially the CH4 ux was
low and gradually started to increase with the growth of the rice
plants. The rst high peak of CH4 emission was observed at 35 and
42 DAT during 2012 and 2013 respectively which coincided with
the tillering stage of the crop. After the rst emission maxima,
there was a drop in CH4 ux for a brief period up to 49 DAT.
Thereafter, the emission increased again through 56 DAT and
reached a peak at 63 DAT which marked the panicle initiation (PI)
stage of the crop. During the later part of the cropping period CH4
ux gradually declined and recorded a minimal level after harvest
(Fig. 1).
The cumulative emission of CH4 (Esif), from the rice plants were
computed at the end of the experiment. Statistical variations in
cumulative CH4 emission pooled for 2012 and 2013 were observed
among different treatments (Fig. 2). IAA and KIN treated plants
recorded signicantly lower ($10% lower) cumulative CH4 ux
compared to control indicating a potential role of plant growth
hormones in suppressing CH4 emission from rice paddies (Fig. 2).
On the other hand, GA treated plants recorded higher cumulative
CH4 emission over CONT while IBA did not signicantly alter CH4
emission. The effect of year and hormone treatment on cumulative
CH4 emission was signicant at p < 0.01 level (Fig. 2).
3.2. Association of soil moisture and soil organic carbon with CH4
emission
The soil moisture content of the experimental eld varied from
30-62 and 15-76% during 2012 and 2013 respectively (Supplemen-
tary Fig. 2). The pattern of soil moisture agreed fairly well with the
78 L. Borah, K.K. Baruah / Agriculture, Ecosystems and Environment 233 (2016) 7584

Fig. 1. CH4 emission (mg m!2 h!1) from hormone treated and untreated Ahu rice variety Luit in 2012 and 2013. Vertical bars represent standard error. DAH = Days after harvest.
Arrows represent date of hormone application (tillering stage-28 DAT; panicle initiation stage-56 DAT).

rainfall pattern during the experimental seasons (Supplementary
Fig. 1). The soil moisture content recorded a positive correlation
with CH4 emission during both the years (p < 0.01 in 2012 and
p < 0.05 in 2013) (Table 1).
The soil organic carbon (SOC) content varied from 1.401.69% in
2012 and 1.401.83% in 2013 (Supplementary Fig. 3). The organic
carbon content recorded a good correlation with CH4 ux (p < 0.01
in 2012 and p < 0.05 in 2013) during both the experimental seasons
(Table 1).
3.3. Effect of plant growth hormones on leaf area index (LAI) and tiller
number
The leaf area index (LAI) of the rice plants ranged from 0.12
5.67 and 0.127.47 in 2012 and 2013 respectively. GA application
was found to bring about an increase in LAI (maximum value of
5.67 at 63 DAT) over control (4.79) (Supplementary Table 1). IBA
application slightly increased the LAI (4.82) whereas IAA and KIN
application resulted in reduction of LAI (4.22 and 4.28 respectively)
 L. Borah, K.K. Baruah / Agriculture, Ecosystems and Environment 233 (2016) 7584 79

Fig. 2. (a) Cumulative CH4 emission (g m"2) and (b) grain yield (kg ha"1) of hormone treated and untreated Ahu rice variety Luit pooled for the years 2012 and 2013. Bars
(mean # standard error) tagged with the same letter are not signicantly different at P < 0.05 level by Duncans Multiple Range Test.

over control in the rst year. A similar trend was observed in the
second year. LAI was found to bear a good correlation with CH4
emission (p < 0.05 in 2012 and p < 0.01 in 2013) in our study
(Table 1).
The mean tiller number per hill ranged from !230 over the
crop growing season in both the years. Tiller number in GA, IAA and
IBA treated plants were slightly lower compared to control and KIN
treated plants (Supplementary Table 1). Tiller number of the rice
plants recorded a positive correlation with CH4 emission (Table 1).
3.4. Effect of plant growth hormones on plant biomass
The leaf blade biomass of the hormone treated rice plants
ranged from 1.156.87 g hill"1 in 2012 and 0.119.05 g hill"1 in
2013. Plants treated with GA recorded higher biomass of leaf blade
over control, whereas IBA application marginally increased the leaf
biomass (Supplementary Table 2). On the other hand, IAA and KIN
treated plants resulted in a lower leaf blade biomass over control
(Supplementary Table 2). Leaf blade biomass showed a good
association with CH4 emission in both the years of experimenta-
tion (p < 0.01) (Table 1).
The biomass of the leaf sheath of the different treatments
ranged from 0.3510.41 and 0.429.73 g hill"1 in 2012 and 2013
respectively while the culm biomass ranged from 0.1515.79 and
0.1612.54 g hill"1 in 2012 and 2013 respectively. Biomass of leaf
sheath and culm were not found to be related to CH4 emission in
the present investigation (Table 1).
The root biomass of the hormone treated plants ranged from
0.035.87 g hill"1 in 2012 and 0.055.68 g hill"1 in 2013 GA and IBA
applications were not effective on the root biomass increment in
the rice plants whereas IAA and KIN treated plants recorded lower
root biomass over control (Supplementary Table 2). The root
biomass was closely associated with CH4 emission in our study in
both the years (p < 0.05 in 2012 and p < 0.01 in 2013) (Table 1).

3.5. Effect of plant growth hormones on leaf photosynthesis,

Table 1
Correlation of various soil and plant parameters with CH4 emission during Ahu rice transpiration and stomatal conductance
growing seasons of 2012 and 2013.
Rate of leaf photosynthesis varied from 5.4131.67 and
Parameters Correlation coefcient
6.2237.65 mmol CO2 m"2 s"1 in 2012 and 2013 respectively. Rice
2012 2013 plants have shown differential response to hormone applications
Leaf area index 0.903* 0.984** in terms of photosynthesis rate (Supplementary Fig. 4). IAA and
Tiller number 0.841* 0.710** KIN applications were found to bring about an enhancement in leaf
Leaf blade biomass (g hill"1) 0.971* 0.984**
photosynthesis rate of the rice plants over control, GA and IBA
Leaf sheath biomass (g hill"1) 0.502 0.522
Culm biomass (g hill"1) 0.486 0.506 applications. The rate of photosynthesis in plants treated with
Root biomass (g hill"1) 0.934* 0.929** exogenous GA was slightly lower compared to control. The leaf
Photosynthesis rate (mmol CO2 m"2 s"1) 0.283 0.406 photosynthesis rate was weekly correlated with CH4 emission in
Transpiration rate (mmol H2O m"2 s"1) 0.656* 0.994**
our study (Table 1).
Stomatal conductance (mol H2O m"2 s"1) 0.655* 0.968*
Soil moisture (%) 0.733** 0.750* Rate of leaf transpiration in the hormone treated plants varied
Soil organic carbon (%) 0.678** 0.852* from 0.818.19 mmol H2O m"2 s"1 in 2012 and 0.7211.16 mmol
*
H2O m"2 s"1 in 2013. GA treated plants mostly recorded higher
Signicant at P < 0.05.
** transpiration rate compared to untreated (control) plants
Signicant at P < 0.01.
80 L. Borah, K.K. Baruah / Agriculture, Ecosystems and Environment 233 (2016) 7584
(Supplementary Fig. 4). IAA treated plants recorded lower
transpiration rate over control at the initial and later stages of
the crop but during the mid-cropping period (3556 DAT) the rate
of transpiration was higher in IAA treated plants over control. IBA
and KIN treated plants generally recorded lower transpiration rate
over control. The rate of leaf transpiration was found to be
positively related to weekly CH4 emission from the rice plants in
both the years (Table 1).
Leaf stomatal conductance of the rice plants varied from 0.22
2.61 and 0.052.61 mol H2O m!2 s!1 in 2012 and 2013 respectively.
GA treated plants usually recorded higher stomatal conductance
compared to untreated plants (Supplementary Fig. 4). IBA
application did not appear to have signicant inuence on the
stomatal conductance. IAA and KIN treated plants generally
recorded lower stomatal conductance over control at the initial
stage of plant growth but subsequently the conductance increased
from 49 to 70 DAT. Leaf stomatal conductance recorded a good
correlation with CH4 emission in our study (Table 1).
3.6. Effect of plant growth hormones on stomatal frequency and xylem
vessel size
Scanning electron micrographs of the adaxial surface of the ag
leaf of treated as well as untreated plants are shown in Fig. 3. The
ag leaf stomatal frequency of GA and IBA treated plants was
higher (765.96 and 691.92 respectively) compared to control plants
(666.67) whereas the stomatal frequency of IAA and KIN treated
plants were signicantly lower (476.95 and 539.01 respectively)
over control plants (Table 2). The stomatal frequency was found to
have good correlation with the cumulative CH4 emission (p < 0.05)
from the rice varieties (Table 2).
Scanning electron micrographs of the xylem vessels of the node
are presented in Fig. 3. Differences in xylem vessel size were
observed in the rice plants (Table 2) due to application of
hormones. IAA and KIN application resulted in a smaller xylem size
(21.23 mm) compared to untreated plants (24.92 mm). The xylem
vessel size bears a signicant correlation with cumulative CH4
emission (p < 0.05) recorded from the rice plants (Table 2).
Fig. 3. Scanning electron micrographs of (AE) stomata of adaxial ag leaf surface and (FJ) topmost node of hormone treated and untreated Ahu rice variety Luit. Circled areas
indicate the leaf stomata and arrows indicate xylem vessels of node.
3.7. Yield response of rice plants to exogenous application of plant
growth hormone
The yield (Fig. 2) and yield attributing parameters (Supple-
mentary Table 3) recorded at harvest were pooled for two seasons
(2012 and 2013). Plant growth hormones were found to inuence
the grain lling efciency in rice in terms of lled grain percentage,
IAA being the most effective with 68% lled grain (Supplementary
Table 3). The thousand grain weight in IAA and KIN treated plants
was slightly higher (18.89 g and 18.77 g respectively) over control,
GA and IBA treated plants (Supplementary Table 3). Foliar
application of IAA and KIN signicantly enhanced the grain yield
over control, whereas IBA application did not produce a similar
effect (Fig. 2).
4. Discussion
4.1. Pattern of methane emission
The occurrence of the high emission peak at tillering stage may
be attributed to the increase in CH4 transport capacity of the rice
plants with increasing growth and development of tillers (Aulakh
et al., 2000) and also due to the expansion of leaf blade surface area
for CH4 emission (Neue et al., 1997; Gogoi et al., 2005) (Fig. 1). A
positive relationship of leaf area index (LAI) with CH4 emission was
observed in our study (Table 1) which further lends support to the
above statement. The occurrence of the high peak at the panicle
initiation stage may be attributed to a further enhancement of leaf
surface area with the emergence of ag leaves and also due to the
greater availability of soil organic carbon for CH4 production due to
higher rhizosphere activity and rhizodeposition (Zhan et al., 2011).
Cause of decline after rst peak in CH4 emission in the later
cropping period may be due to the reduction in leaf area, tiller
number and root growth with the onset of crop senescence. Similar
pattern of CH4 emission from rice has been reported by several
authors (Gogoi et al., 2008; Suryavanshi et al., 2013).
 L. Borah, K.K. Baruah / Agriculture, Ecosystems and Environment 233 (2016) 7584
Table 2
Flag leaf stomatal frequency and xylem size of node of Ahu rice variety Luit and their correlation with cumulative CH4 emission. Within a column, values (mean ! standard
error) followed by same letters are not signicantly different from each other at P < 0.05 level by Duncans multiple range test.
Treatment
CONT
GA
IAA
IBA
KIN
Correlation with cumulative CH4 emission
*
Signicant at P < 0.05.
4.2. Soil moisture and organic carbon
Moisture content determines the availability of anaerobic zones
in the soil which are conducive for CH4 production (Kotiaho et al.,
2010; Yu et al., 2013) and this might be the reason for the observed
positive relationship of soil moisture with CH4 emission in our
study. Soil organic carbon acts as the major source of substrates to
methanogenic bacteria for CH4 production (Zhan et al., 2011;
Conrad et al., 2012; Yuan et al., 2014) which might be the reason for
the observed positive relationship of SOC with CH4 emission. The
higher CH4 emission during the tillering and panicle initiation
stages may be attributed to the greater availability of SOC during
these stages (Supplementary Fig. 3).
4.3. Leaf area index and tiller number
Increase in LAI of GA treated rice plants over control
(Supplementary Table 1) may be attributed to a GA induced
elongation of leaf blade observed in the present study. Similar
ndings have been reported by Asthir et al. (2004) in barley.
Exogenous supply of IAA resulted in a reduction of LAI in rice plants
over control (Supplementary Table 1). The role of auxin in
controlling leaf expansion is reported to have shown conicting
results (Keller et al., 2004). Higher level of auxin in intact plants is
reported to inhibit leaf expansion (Keller et al., 2011) which
possibly might have caused inhibition in leaf growth. Slight
reduction of LAI was observed in kinetin treated rice plants over
control (Supplementary Table 1). There are some reports on
inhibition of leaf area development due to foliar application of
cytokinin in soybean (Leite et al., 2003) and in tomato (Papado-
poulos et al., 2006). Therefore, it is evident that the effect of kinetin
on LAI is dependent on hormone concentration as well as plant
species.
Exogenous GA is reported to mask tiller growth by regulating
the levels of endogenous IAA (Liu et al., 2011), which might be the
reason for the observed reduction in tiller number of GA treated
rice plants over control in our study (Supplementary Table 1).
Auxins such as IAA are also reported to reduce tillering in rice (Choi
et al., 2012) and our ndings are in conformity with the above
report.
4.4. Leaf blade and root biomass
GA induced leaf area enhancement in rice as discussed earlier
(section 4.3) might have resulted in higher leaf blade biomass over
control whereas the lower LAI observed in IAA and KIN treated
plants have resulted in reduction in leaf blade biomass in these
plants (Supplementary Table 2).
Exogenously applied GA is reported to have little effect on root
growth in many plants (Tanimoto, 2005) and similar observations
have been made in our study on rice (Supplementary Table 2).
Deceleration of root growth due to exogenous auxin application is
81
Stomatal frequency (mm"2 of leaf) Xylem size (mm)
666.67 ! 37.53bc 24.92 ! 1.43b
765.96 ! 14.18c 26.40 ! 0.70b
476.95 ! 48.78a 21.23 ! 0.63a
691.92 ! 58.98c 24.46 ! 0.21b
539.01 ! 28.37ab 21.23 ! 0.61a
0.888* 0.882*
reported by many authors (Liu et al., 2002; Tanimoto, 2005;
Kukavica et al., 2007) which might be the reason for the lower root
biomass of IAA treated plants in our study. The role of cytokinin in
the inhibition of lateral root initiation in rice and other plants are
reported by many authors (Debi et al., 2005; Laplaze et al., 2007;
Bielach et al., 2012) and are well corroborated with our ndings of
lower root biomass in kinetin treated plants.
4.5. Leaf photosynthesis, transpiration and stomatal conductance
Inhibition of photosynthesis rate due to external application of
GA is reported by Thetford et al. (1995) and Wang et al. (2013) and
similar ndings are reported in the present study (Supplementary
Fig. 4). Our results on enhancement of leaf photosynthesis rate in
rice by IAA are in good agreement with Aldesuquy (2000) who
have reported increased ag leaf photosynthetic activity and
pigment formation during grain lling due to external application
of IAA in wheat. Positive inuence of auxin on chlorophyll content
and net photosynthetic rate was also reported by Ahmad et al.
(2001) in Brassica juncea and our results are well corroborated with
these ndings. Stimulation of photosynthesis by external applica-
tion of cytokinins is reported by Iqbal and Ashraf (2005) in wheat
and Wu et al. (2012) in eggplant and our results on kinetin induced
enhancement in photosynthetic activity are in conformity with the
ndings of above reports.
The enhancement of leaf transpiration rate and stomatal
conductance by exogenous GA might have been brought about
by the regulation of stomatal behavior as suggested by Dodd
(2003) in barley (Supplementary Fig. 4). Stomatal conductance is
also reported to be inuenced by GA application (Yuan and Xu,
2001). Our ndings on the enhancement of transpiration rate and
stomatal conductance due to GA application are in good agreement
with the above reports.
Responses of stomata to auxins and cytokinins are species
specic as reviewed by Pospilov (2003). In the present
investigation, variations in transpiration rate and stomatal
conductance between CONT-IAA-KIN were not consistent through-
out the experimental seasons (Supplementary Fig. 4). This may be
due to uctuations in environmental factors such as CO2
concentration over the canopy, ambient temperature, water
potential of leaf and xylem, soil water potential, leaf-stem
hydraulics and vapor pressure decit (Hubbard, 2001; Tuzet
et al., 2003; Yu et al., 2004; Zhang et al., 2013).
4.6. Stomatal frequency and xylem vessel size
Stomatal behavior (opening and closing), stomatal pattern,
stomatal density etc. are regulated by hormonal and environmen-
tal factors (Dodd, 2003; Casson and Gray, 2008; zyig !it and Can,
2009). Relatively higher rate of leaf area development in GA and
IBA treated plants might have contributed to a greater leaf stomatal
frequency in these plants (Table 2). Conversely, reduced leaf area in
82 L. Borah, K.K. Baruah / Agriculture, Ecosystems and Environment 233 (2016) 7584
IAA and KIN treated plants as discussed earlier may have resulted
in a lower stomatal frequency over control plants (Table 2).
Gibberellins, auxins and cytokinins are widely reported to
control vascular cell differentiation and determination of continu-
ity and pattern of vascular tissue formation in plants (Berleth et al.,
2000; Fukuda, 2004; Aloni et al., 2006; Sorce et al., 2013). The
control of xylem cell differentiation determines the diameter of
xylem vessels (Sorce et al., 2013). In the present investigation, GA
and IBA application on rice did not alter the xylem vessel size
whereas IAA and kinetin application resulted in reduced size of
xylem vessels over untreated plants (Table 2). Therefore, hormonal
control of xylem vessel size is evident in case of IAA and KIN
applications and is in good agreement with the ndings of above
authors.
4.7. Yield attributes and grain yield
GA application brought about an improvement in grain yield
through its stimulatory effect on panicle length and quantity of
lled grains (Fig. 2; Supplementary Table 3). Improvement of grain
yield by GA application is reported by many authors (Ghodrat et al.,
2012; Tiwari et al., 2011) which support our ndings. IAA and
kinetin application signicantly enhanced the grain yield in Ahu
rice through their stimulatory effect on the yield attributing
parameters. Similar ndings are reported by Sadak et al., 2013 and
Abdoli et al., 2013. Enhancement of yield and yield attributing
parameters by kinetin and auxin application are reported by many
authors (Sawan et al., 2000; Papadopoulos et al., 2006; Abdoli
et al., 2013; Javid et al., 2011; Khalil et al., 2006) and our results are
in conformity with these ndings.
4.8. Mitigation of CH4 emission by hormone application
The reduction in cumulative CH4 emission in IAA and KIN
treated plants (Section 3.1) appears to have been brought about by
hormonal control of vegetative growth, physiological processes
and yield development of the rice plants. The overall vegetative
growth in IAA and KIN treated plants in terms of leaf area index,
leaf biomass, tiller number and root biomass was lower over
control plants which might have brought about a reduction in CH4
emission, whereas the converse was true for GA and IBA treated
plants. The association of plant factors like leaf area index, tiller
number etc. with CH4 emission are well documented (Gogoi et al.,
2005; Gogoi et al., 2008; Aulakh et al., 2000) and similar results
have been observed in the present study as well. The lower root
biomass in IAA and kinetin treated plants might have caused lower
root exudation resulting in lesser plant-derived C transfer to the
root zone (Wang and Adachi, 2000), which is considered to be the
possible reason for lower CH4 production.
Less number of stomata per unit area of leaf and smaller xylem
size of the node (Table 2) induced by exogenous IAA and kinetin
application may also have contributed to lower CH4 emission. The
association of leaf stomatal frequency with CH4 emission from rice
is reported by Das and Baruah (2008a) and is in agreement with
the ndings of the present study.
Higher rate of ag leaf photosynthesis in IAA and KIN treated
plants may have brought about efcient translocation of photo-
synthates towards the developing grains, as evident from the
higher grain yield (Fig. 2) and restricted the translocation of
photosynthates towards the roots, as evident from the lower root
biomass (Supplementary Table 2). An inverse relationship between
grain yield and seasonal CH4 emission is reported by Sass and
Cicerone (2002), Denier van der Gon et al. (2002) and Das and
Baruah (2008b) and we also report here a similar relationship.
Lower root biomass may be a possible but not conclusive reason for
low methane emission in IAA and KIN treated plants.
5. Conclusions
Hormones are known to regulate growth and developmental
processes of plants. In the present investigation, hormone
applications in rice were found to have visible impact on the
growth, physiological and anatomical characteristics of the rice
plants. IAA and kinetin applications in rice agriculture were found
to be benecial in terms of lower GHG emission accompanied by
higher grain production. Our experimental ndings provide
evidences in favor of a regulatory role of plant hormones in
growth and development of rice plants which indirectly inuence
their GHG transport capacity. It is concluded that manipulation of
plant growth and developmental processes and optimization of
grain yields by application of plant growth hormones, especially
indole acetic acid (IAA) and kinetin (KIN) in suitable dosage can
provide a promising option for mitigation of CH4 emissions from
rice ecosystem through management of plant mediated methane
transport.
Acknowledgement
Leena Borah is grateful to Department of Science and
Technology, Government of India, for providing nancial support
to carry out the present research work through DST INSPIRE
Fellowship.
Appendix A. Supplementary data
Supplementary data associated with this article can be found, in
the online version, at http://dx.doi.org/10.1016/j.agee.2016.08.033.
References
Abdoli, M., Saeidi, M., Jalali-Honarmand, S., Azhand, M., 2013. The effect of foliar
application of Indole-3-Acetic Acid (IAA) and roles of ear photosynthesis on
grain yield production of two wheat cultivars (Triticum aestivum L.) under post
anthesis water decit. Int. Res. J. Appl. Basic Sci. 4, 14061413.
Ahmad, A., Hayat, S., Fariduddin, Q., Ahmad, I., 2001. Photosynthetic efciency of
plants of Brassica juncea, treated with chlorosubstituted auxins.
Photosynthetica 39, 565568.
Aldesuquy, H.S., 2000. Effect of indol-3yl acetic acid on photosynthetic
characteristics of wheat ag leaf during grain lling. Photosynthetica 38,
135141.
Aloni, R., Aloni, E., Langhans, M., Ullrich, C.I., 2006. Role of cytokinin and auxin in
shaping root architecture: regulating vascular differentiation lateral root
initiation, root apical dominance and root gravitropism. Ann. Bot. 97, 883893.
Asthir, B., Duffus, C.M., Spoor, W., 2004. Correlation of gibberellin-induced growth,
polyamine levels and amine oxidases in epicotyl: root and leaf blade of barley
seedlings. Plant Growth Regul. 42, 193201.
Aulakh, M.S., Bodenbender, J., Wassmann, R., Rennenberg, H., 2000. Methane
transport capacity of rice plants. II. Variations among different rice cultivars and
relationship with morphological characteristics. Nutr. Cycl. Agroecosyst. 58,
367375.
Awale, R., Chatterjee, A., Franzen, D., 2013. Tillage and N-fertilizer inuences on
selected organic carbon fractions in a North Dakota silty clay soil. Soil Tillage
Res. 134, 213222.
Baruah, K.K., Gogoi, B., Borah, L., Gogoi, M., Boruah, R., 2012. Plant
morphophysiological and anatomical factors associated with nitrous oxide ux
from wheat (Triticum aestivum). J. Plant Res. 125, 507516.
Berleth, T., Mattsson, J., Hardtke, C.S., 2000. Vascular continuity and auxin signals.
Trends Plant Sci. Rev. 5, 387393.
Bielach, A., Ducler, C.Q., Marhav, P., Benkov, E., 2012. Genetic approach towards
the identication of auxincytokinin crosstalk components involved in root
development. Phil. Trans. R. Soc. B 367, 14691478.
Borah, L., Baruah, K.K., 2016. Nitrous oxide emission and mitigation from wheat
agriculture: association of physiological and anatomical characteristics of
wheat genotypes. Environ. Sci. Pollut. Res. 23, 709721.
Buendia, L.V., Neue, H.U., Wassmann, R., Lantin, R.S., Javellana, A.M., Yuchang, X.,
Makarim, A.K., Corton, T.M., Charoensilp, N., 1997. Understanding the nature of
methane emission from rice ecosystems as basis of mitigation strategies. Appl.
Energy 56, 433444.
 L. Borah, K.K. Baruah / Agriculture, Ecosystems and Environment 233 (2016) 7584
Casson, S., Gray, J.E., 2008. Inuence of environmental factors on stomatal
development. New Phytol. 178, 923.
Choi, M.S., Koh, E.B., Wu, M.O., Piao, R., Oh, C.S., Koh, H.J., 2012. Tiller formation in
rice is altered by over expression of OsIAGLU gene encoding an IAA-conjugating
enzyme or exogenous treatment of free IAA. J. Plant Biol. 55, 429435.
Conrad, R., Klose, M., Lu, Y., Chidthaisong, A., 2012. Methanogenic pathway and
archaeal communities in three different anoxic soils amended with rice straw
and maize straw. Front. Microbiol. 3 doi:http://dx.doi.org/10.3389/
fmicb.2012.00004.
Das, K., Baruah, K.K., 2008a. Methane emission associated with anatomical and
morphological characteristics of rice (Oryza sativa L.) plant. Physiol. Plant 134,
303312.
Das, K., Baruah, K.K., 2008b. A comparison of growth and photosynthetic
characteristics of two improved rice cultivars on methane emission from
rainfed agroecosystem of north east India. Agric. Ecosyst. Environ. 124, 105113.
Debi, B.R., Taketa, S., Ichii, M., 2005. Cytokinin inhibits lateral root initiation but
stimulates lateral root elongation in rice (Oryza sativa). J. Plant Physiol. 162,
507515.
Denier van der Gon, H.A.C., Kropff, M.J., Van Breemen, N., Wassmann, R., Lantin, R.S.,
Aduna, E., Corton, T.M., Van Laar, H.H., 2002. Optimizing grain yields reduces
methane emission from rice paddy elds. Proc. Natl. Acad. Sci. 99, 1202112024.
Depuydt, S., Hardtke, C.S., 2011. Hormone signalling crosstalk in plant growth
regulation. Curr. Biol. 21, R365R373.
Ding, W.X., Cai, Z.C., Tsuruta, H., 2005. Plant species effects on methane emissions
from freshwater marshes. Atmos. Environ. 39, 31993207.
Dodd, I.C., 2003. Hormonal interactions and stomatal responses. J. Plant Growth
Regul. 22, 3246.
Durbak, A., Yao, H., McSteen, P., 2012. Hormone signaling in plant development.
Curr. Opin. Plant Biol. 15, 9296.
FAOSTAT, 2013. Food and Agriculture Organization of the United Nations. FAOSTAT.
http://faostat.fao.org/default.aspx.
Fukuda, H., 2004. Signals that control plant vascular cell differentiation. Nat. Rev.
Mol. Cell Biol. 5, 379391.
Ghodrat, V., Rousta, M.J., Tadaion, M.S., Karampour, A., 2012. Yield and yield
components of corn (Zea mays L.) in response to foliar application with indole
butyric acid and gibberellic acid. Am.-Eurasian J. Agric. Environ. Sci. 12,
12461251.
Gogoi, N., Baruah, K.K., Gogoi, B., Gupta, P.K., 2005. Methane emission
characteristics and its relation with plant and soil parameters under irrigated
rice ecosystem of North East India. Chemosphere 59, 16771684.
Gogoi, N., Baruah, K.K., Gupta, P.K., 2008. Selection of rice genotypes for lower
methane emission. Agron. Sustain. Dev. 28, 181186.
Gray, W.M., 2004. Hormonal regulation of plant growth and development. PLoS Biol.
2, 12701273.
Gupta, P.K., Gupta, V., Sharma, C., Das, S.N., Purkait, N., Adhya, T.K., Pathak, H.,
Ramesh, R., Baruah, K.K., Venkatratnam, L., Singh, G., Iyer, C.S.P., 2009.
Development of methane emission factors for Indian paddy elds and
estimation of national methane budget. Chemosphere 74, 590598.
Hubbard, R.M., 2001. Stomatal conductance and photosynthesis vary linearly with
plant hydraulic conductance in ponderosa pine. Plant Cell Environ. 24, 113121.
IPCC, 2007. Summary for Policymakers. Climate Change 2007: Synthesis Report of
Fourth Assessment Report of the Intergovernmental Panel on Climate Change.
Cambridge University Press, Cambridge, UK.
Iqbal, M., Ashraf, M., 2005. Presowing seed treatment with cytokinins and its effect
on growth, photosynthetic rate, ionic levels and yield of two wheat cultivars
differing in salt tolerance. J. Integr. Plant Biol. 47, 13151325.
Javid, M.G., Sorooshzadeh, A., Sanavy, S.A.M.M., Allahdadi, I., Moradi, F., 2011. Effects
of the exogenous application of auxin and cytokinin on carbohydrate
accumulation in grains of rice under salt stress. Plant Growth Regul. 65,
305313.
Jiang, Y., Wang, L.L., Yan, X.J., Tian, Y.L., Deng, A.X., Zhang, W.J., 2013. Super rice
cropping will enhance rice yield and reduce CH4 emission: a case study in
Nanjing. China. Rice Sci. 20, 427433.
Keller, C.P., Stahlberg, R., Barkawi, L.S., Cohen, J.D., 2004. Long-term inhibition by
auxin of leaf blade expansion in bean and Arabidopsis. Plant Physiol. 134,
12171226.
Keller, C.P., Grundstad, M.L., Evanhoff, M.A., Keith, J.D., Lentz, D.S., Wagner, S.L.,
Culler, A.H., Cohen, J.D., 2011. Auxin-induced leaf blade expansion in
Arabidopsis requires both wounding and detachment. Plant Signaling Behav. 6
(12), 19972007.
Khalil, S., El-Saeid, H.M., Shalaby, M., 2006. The role of kinetin in ower abscission
and yield of lentil plant. J. Appl. Sci. Res. 2, 587591.
Koelbener, A., Strm, L., Edwards, P.J., Olde Venterink, H., 2010. Plant species from
mesotrophic wetlands cause relatively high methane emissions from peat soil.
Plant Soil 326, 147158.
Kotiaho, M., Fritze, H., Meril, P., Juottonen, H., Leppl, M., Laine, J., Laiho, R., Yrjl,
K., Tuittila, E.S., 2010. Methanogen activity in relation to water table level in two
boreal fens. Biol. Fertil. Soils 46, 567575.
Kukavica, B., Mitrovi! c, A., Mojovi!
c, M., Veljovi!
c-Jovanovi!
c, S., 2007. Effect of indole-
3-acetic acid on pea root growth, peroxidase proles and hydroxyl radical
formation. Arch. Biol. Sci. 59, 319326.
Laplaze, L., Benkova, E., Casimiro, I., Maes, L., Vanneste, S., Swarup, R., Weijers, D.,
Calvo, V., Parizot, B., Herrera-Rodriguez, M.B., Offringa, R., Graham, N., Doumas,
P., Friml, J., Bogusz, D., Beeckman, T., Bennett, M., 2007. Cytokinins act directly
on lateral root founder cells to inhibit root initiation. Plant Cell 19, 38893900.
83
Leite, V.M., Rosolem, C.A., Rodrigues, J.D., 2003. Gibberellin and cytokinin effects on
soybean growth. Sci. Agric. 60, 537541.
Liu, C., Zhu, J., Liu, Z., Li, L., Pan, R., Jin, L., 2002. Exogenous auxin effects on growth
and phenotype of normal and hairy roots of Pueraria lobata (Willd) Ohwi. Plant
Growth Regul. 38, 3743.
Liu, Y., Wang, Q., Ding, Y., Li, G., Xu, J., Wang, S., 2011. Effects of external ABA, GA3 and
NAA on the tiller bud outgrowth of rice is related to changes in endogenous
hormones. Plant Growth Regul. 65, 247254.
Myhre, G., Shindell, D., Bron, F., Collins, W., Fuglestvedt, J., Huang, J., Koch, D.,
Lamarque, J., Lee, D., Mendoza, B., Nakajima, T., Robock, A., Stephens, G.,
Takemura, T., Zhang, H., 2013. Anthropogenic and natural radiative forcing. In:
Stocker, T.F., Qin, D., Plattner, G.-K., Tignor, M., Allen, S.K., Boschung, J., Nauels, A.,
Xia, Y., Bex, V., Midgley, P.M. (Eds.), Climate Change 2013: The Physical Science
Basis, Contribution of Working Group I to the Fifth Assessment Report of the
Intergovernmental Panel on Climate Change. Cambridge University
PressCambridge, United Kingdom and New York, pp. 649740.
Naser, H.M., Nagata, O., Tamura, S., Hatano, R., 2007. Methane emissions from ve
paddy elds with different amounts of rice straw application in central
Hokkaido Japan. Soil Sci. Plant Nutr. 53, 95101.
Neue, H.U., Wassmann, R., Kludze, H.K., Bujun Wang Lantin, R.S., 1997. Factors and
processes controlling methane emissions from rice elds. Nutr. Cycl.
Agroecosyst. 49, 111117.
Nouchi, I., Mariko, S., Aoki, K., 1990. Mechanism of methane transport from the
rhizosphere to the atmosphere through rice plants. Plant Physiol. 94, 5966.
zyig "it, I.I., Can, S., 2009. Effects of some external treated plant growth regulators on
stomatal aperture of cucumber (Cucumis sativus L.). Afr. J. Agric. Res. 4, 628632.
Page, A.L., Miller, R.H., Keeney, D.R., 1982. Methods of Soil Analysis, Part 2. Soil
Science Society America, Madison, WI.
Papadopoulos, A.P., Saha, U., Hao, X., Khosla, S., 2006. Response of rockwool-grown
greenhouse cucumber, tomato, and pepper to kinetin foliar sprays. HortTechnol
16, 493501.
Parashar, D.C., Mitra, A.P., Gupta, P.K., Rai, J., Sharma, R.C., Singh, N., Koul, S., Ray, H.S.,
Das, S.N., Parida, K.M., Rao, S.B., Kanungo, S.P., Ramasami, T., Nair, B.U., Swamy,
M., Singh, G., Gupta, S.K., Singh, A.R., Saikia, B.K., Baruah, A.K.S., Pathak, M.G.,
Iyer, C.S.P., Gopalakrishnan, M., Sane, P.V., Singh, S.N., Banerjee, R., Sethunathan,
N., Adhya, T.K., Rao, V.R., Palit, P., Saha, A.K., Purkait, N., Chaturvedi, G.S., Sen, S.P.,
Sen, M., Sarkar, B., Banik, A., Subbaraya, B.H., Lal, S., Venkatramani, S., Lal, G.,
Chaudhary, A., Sinha, S.K., 1996. Methane budget from paddy elds in India.
Chemosphere 33, 737757.
Pospilov, J., 2003. Participation of phytohormones in the stomatal regulation of
gas exchange during water stress. Biol. Plant. 46, 491506.
Sadak, M.S., Dawood, M.G., Bakry, B.A., El-Karamany, M.F., 2013. Synergistic effect of
indole acetic acid and kinetin on performance: some biochemical constituents
and yield of faba bean plant grown under newly reclaimed sandy soil. World J.
Agric. Sci. 9, 335344.
Sass, R.L., Cicerone, R.J., 2002. Photosynthate allocations in rice plants: food
production or atmospheric methane. Proc. Natl. Acad. Sci. 99, 1199311995.
Sawan, Z.M., Mohamed, A.A., Sakr, R.A., Tarrad, A.M., 2000. Effect of kinetin
concentration and methods of application on seed germination, yield
components, yield and bre properties of the Egyptian cotton (Gossypium
barbadense). Environ. Exp. Bot. 44, 5968.
Sorce, C., Giovannelli, A., Sebastiani, L., Anfodillo, T., 2013. Hormonal signals
involved in the regulation of cambial activity, xylogenesis and vessel patterning
in trees. Plant Cell Rep. 32, 885898.
Suryavanshi, P., Singh, Y.V., Prasanna, R., Bhatia, A., Shivay, Y.S., 2013. Pattern of
methane emission and water productivity under different methods of rice crop
establishment. Paddy Water Environ. 11, 321329.
Tanimoto, E., 2005. Regulation of root growth by plant hormonesroles for auxin
and gibberellin. Crit. Rev. Plant Sci. 24, 249265.
Thetford, M., Warren, S.L., Blazich, F.A., Thomas, J.F., 1995. Response of Forsythia-x
intermedia Spectabilis to uniconazole. II. Leaf and stem anatomy, chlorophyll,
and photosynthesis. J. Am. Soc. Hortic. Sci. 120, 983988.
Tiwari, D.K., Pandey, P., Giri, S.P., Dwivedi, J.L., 2011. Effect of GA3 and other growth
regulators on hybrid rice seed production. Asian J. Plant Sci. 10, 133139.
Tuzet, A., Perrier, A., Leuning, R., 2003. A coupled model of stomatal conductance,
photosynthesis and transpiration. Plant Cell Environ. 26, 10971116.
Wang, B., Adachi, K., 2000. Differences among rice cultivars in root exudation,
methane oxidation, and populations of methanogenic and methanotrophic
bacteria in relation to methane emission. Nutr. Cycl. Agroecosyst. 58, 349356.
Wang, X., Han, F., Yang, M., Yang, P., Shen, S., 2013. Exploring the response of rice
(Oryza sativa) leaf to gibberellins: a proteomic strategy. Rice 6, 17. http://www.
thericejournal.com/content/6/1/17.
Wassmann, R., Hosen, Y., Sumeth, K., 2009. Reducing methane emissions from
irrigated rice. In: Nelson, G.C. (Ed.), Agriculture and Climate Change: An Agenda
for Negotiation in Copenhagen, 2020 Focus. International Food Policy Research
Institute, pp. 78.
Wu, X., Zhu, Z., Li, X., Zha, D., 2012. Effects of cytokinin on photosynthetic gas
exchange: chlorophyll uorescence parameters and antioxidative system in
seedlings of eggplant (Solanum melongena L.) under salinity stress. Acta Physiol.
Plant. 34, 21052114.
Yu, Q., Zhang, Y., Liu, Y., Shi, P., 2004. Simulation of the stomatal conductance of
winter wheat in response to light, temperature and CO2 changes. Ann. Bot. 93,
435441.
Yu, L., Tang, J., Zhang, R., Wu, Q., Gong, M., 2013. Effects of biochar application on soil
methane emission at different soil moisture levels. Biol. Fertil. Soils 49, 119128.
84 L. Borah, K.K. Baruah / Agriculture, Ecosystems and Environment 233 (2016) 7584

Yuan, L., Xu, D.Q., 2001. Stimulation effect of gibberellic acid short-term treatment
on leaf photosynthesis related to the increase in Rubisco content in broad bean
and soybean. Photosynth. Res. 68, 3947.
Yuan, Q., Pump, J., Conrad, R., 2014. Straw application in paddy soil enhances methane
production also from other carbon sources. Biogeosciences 11, 237246.
Zhan, M., Cao, C., Wang, J., Jiang, Y., Cai, M., Yue, L., Shahrear, A., 2011. Dynamics of
methane emission, active soil organic carbon and their relationships in wetland
integrated rice-duck systems in Southern China. Nutr. Cycl. Agroecosyst. 89,
113.
Zhang, Y.J., Meinzer, F.C., Qi, J.H., Goldstein, G., Cao, K.F., 2013. Midday stomatal
conductance is more related to stem rather than leaf water status in subtropical
deciduous and evergreen broadleaf trees. Plant Cell Environ. 36, 149158.