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Reduced Absorption of Glyphosate and Decreased Translocation of Dicamba Contribute to Poor Control of Kochia ( Kochia scoparia ) at High Temperature


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Background: Plant growth temperature is one of the important factors that can influence postemergent herbicide efficacy and impact weed control. Control of kochia (Kochia scoparia), a major broadleaf weed throughout the North American Great Plains, often is unsatisfactory when either glyphosate or dicamba are applied on hot summer days. We tested effects of plant growth temperature on glyphosate and dicamba phytotoxicity on two Kansas kochia populations (P1 and P2) grown under the following three day/night (d/n) temperature regimes: T1, 17.5/7.5 °C; T2, 25/15 °C; and T3, 32.5/22.5 °C. Results: Visual injury and above-ground dry biomass data from herbicide dose response experiments indicated greater susceptibility to both glyphosate and dicamba when kochia was grown under the two cooler temperature regimes, i.e. T1 and T2. At T1, the ED50 of P1 and P2 kochia were 39 and 36 g · ha(-1) of glyphosate and 52 and 105 g · ha (-1) of dicamba, respectively. In comparison, at T3 the ED50 increased to 173 and 186 g · ha(-1) for glyphosate and 106 and 410 g · ha(-1) for dicamba, respectively, for P1 and P2. We also investigated the physiological basis of decreased glyphosate and dicamba efficacy under elevated temperatures. Kochia absorbed more glyphosate at T1 and T2 compared to T3. Conversely, there was more dicamba translocated towards meristems at T1 and T2, compared to T3. Conclusion: Reduced efficacy of dicamba or glyphosate to control kochia under elevated temperatures can be attributed to decreased absorption and translocation of glyphosate and dicamba, respectively. Therefore, it is recommended to apply glyphosate or dicamba when the temperature is low (e.g. d/n temperature at 25/15 °C) and seedlings are small (less than 12 cm) to maximize kochia control.
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Research Article
Received: 2 June 2016 Revised: 28 September 2016 Accepted article published: 21 October 2016 Published online in Wiley Online Library:
( DOI 10.1002/ps.4463
Reduced absorption of glyphosate and
decreased translocation of dicamba contribute
to poor control of kochia (Kochia scoparia)at
high temperature
Junjun Ou,aPhillip W Stahlmanband Mithila Jugulama*
BACKGROUND: Plant growth temperature is one of the important factors that can influence postemergent herbicide efficacy
and impact weed control. Control of kochia (Kochia scoparia), a major broadleaf weed throughout the North American Great
Plains, often is unsatisfactory when either glyphosate or dicamba are applied on hot summer days. We tested effects of plant
growth temperature on glyphosate and dicamba phytotoxicity on two Kansas kochia populations (P1 and P2) grown under the
following three day/night (d/n) temperature regimes: T1, 17.5/7.5C; T2, 25/15C; and T3, 32.5/22.5C.
RESULTS: Visual injury and above-ground dry biomass data from herbicide dose– response experiments indicated greater
susceptibility to both glyphosate and dicamba when kochia was grown under the two cooler temperature regimes, i.e. T1 and
T2. At T1, the ED50 of P1 and P2 kochia were 39 and 36 g ha1of glyphosate and 52 and 105 g ha1of dicamba, respectively. In
comparison, at T3 the ED50 increased to 173 and 186 g ha1for glyphosate and 106 and 410 g ha1for dicamba, respectively,
for P1 and P2. We also investigated the physiological basis of decreased glyphosate and dicamba efficacy under elevated
temperatures. Kochia absorbed more glyphosateat T1 and T2 compared to T3. Conversely, there wasmore dicamba translocated
towards meristems at T1 and T2, compared to T3.
CONCLUSION: Reduced efficacy of dicamba or glyphosate to control kochia under elevated temperatures can be attributed
to decreased absorption and translocation of glyphosate and dicamba, respectively. Therefore, it is recommended to apply
glyphosate or dicamba when the temperature is low (e.g. d/n temperature at 25/15C) and seedlings are small (less than 12
cm) to maximize kochia control.
© 2016 Society of Chemical Industry
Keywords: glyphosate; dicamba; growth temperature; kochia
Kochia is one of the most troublesome annual C4 broadleaf weeds
in croplands in the Great Plains of North America.1Kochia can
emerge early in spring (early March in Kansas2) before most other
spring and summer annual weeds and spring-sown crops and can
grow rapidly under cool as well as warm temperatures.1,2Due
to its aggressive growth habit, kochia can cause huge yield loss
in grain crops.1,3In addition, mature plants of kochia accumu-
late saponins, alkaloids, oxalates, and nitrates, which are toxic to
domestic animals.4More than 30 kochia populations across the
USA have been reported to have evolved resistance to one or more
herbicide modes of action.5Yet, herbicide application is still one of
the most effective methods to manage kochia in croplands. Weed
resistance to herbicide sites of action is evolving at a rapid rate
while no new herbicide modes of action have been developed in
more than two decades.6Thus, more efficient use of existing her-
bicides is vital to maintain their effectiveness in the future.
Poor control of kochia in western Kansas has been observed
numerous times when glyphosate [N-(phosphonomethyl)-
glycine] or dicamba (3,6-dichloro-o-anisic acid) was applied in hot
weather (P.W. Stahlman, Personal communication). Incomplete
control of kochia can accelerate the evolution of glyphosate
or dicamba resistance, since long-term or constant exposure
to a low/ineffective concentration of a specific herbicide can
significantly contribute to the evolution of resistance in weeds.7,8
Several studies have found that the efficacy of commonly used
herbicides such as glyphosate, glufosinate, and mesotrione can
be affected by temperature. Increased temperature has altered
the efficacy of glyphosate on wild oat (Avena fatua),9liverseed
grass (Urochloa panicoides),9velvetleaf (Abutilon theophrasti),10
and awnless barnyardgrass (Echinochloa colona).11 However, only
a few studies have investigated the underlying mechanism of
Correspondence to: M Jugulam, Department of Agronomy, Kansas State Uni-
versity, 2004 Throckmorton Plant Sciences Center, 1721 Claflin Road, Manhat-
tan, KS 66506, USA. Email:
aDepartment of Agronomy, Kansas State University, 2004 Throckmorton Plant
Sciences Center, 1712 Claflin Road, Manhattan, KS, USA
bAgricultural Research Center-Hays,Kansas State Universit y,1232 240th Avenue,
Hays, KS, USA
Pest Manag Sci (2016) © 2016 Society of Chemical Industry J Ou, PW Stahlman, M Jugulam
altered glyphosate efficacy under different temperature regimes.
For instance, Jordan11 reported glyphosate controlled bermuda-
grass (Cynodon dactylon) better at high than low temperature
because more glyphosate was absorbed and translocated out of
the treated leaves. Similarly, Coupland12 found elevated basipetal
translocation enhanced glyphosate activity at high temperature
in couch grass (Elymus repens). However, in quackgrass (Agropy-
ron repens), Devine et al.13 concluded altered efficacy of glyphosate
at different temperatures was not due to differential absorption
or translocation of the herbicide. Similarly, Friesen and Dew14
reported phytotoxicity of dicamba on tartary buckwheat (Fagopy-
rum tataricum) was not affected when temperature was increased.
This study was conducted based on the hypothesis that the tem-
perature can alter absorption and/or translocation of glyphosate
or dicamba, thereby affecting kochia control. The objectives of this
study were to: (a) evaluate the differential efficacy of glyphosate or
dicamba at varying temperatures on kochia control and (b) inves-
tigate the mechanisms underlying the differential efficacy of these
herbicides on kochia control.
2.1 Plant materials and growth conditions
Kochia seed was collected from field sites in Pratt County15 (Pop-
ulation 1, P1) and Riley County (Population 2, P2), Kansas, in
2012. Because of the short seed longevity of kochia, 5 10 plants
from each population annually were grown together in isolation
from other kochia and mature seed bulked and stored in dark
at 4C. Seeds from P1 and P2 produced in 2014 were used to
conduct glyphosate and dicamba dose–response experiments in
growth chambers under different temperature regimes (described
in detail in section 2.1). However, only P1 was used to conduct
glyphosate and dicamba absorption and translocation experi-
ments at different temperatures.
In 2015, kochia seed of P1 and P2 were germinated in small
trays (25 ×15 ×2.5 cm) filled with commercial potting mixture
(Pro-Mix Potting-Mix; Premier Tech Horticulture, Mississauga,
Ontario, Canada). Individual seedlings 2– 3 cm tall were trans-
planted into plastic pots (6.5 ×6.5 ×9 cm) in a greenhouse on the
campus of Kansas State University in Manhattan. The following
greenhouse conditions were maintained: 25/20C (day/night, d/n)
temperatures, 60 ±10% relative humidity, and 15/9 h day/night
photoperiod supplemented with 120 μmol m2s1illumination
provided with sodium vapor lamps. One week after transplanting,
healthy kochia plants (5 cm tall) were transferred to growth
chambers that were maintained at different d/n temperatures: T1:
17.5/7.5C; T2: 25/15C; and T3: 32.5/22.5C. Light in all growth
chambers was provided by incandescent and fluorescent bulbs
delivering 750 μmol m2s1photon flux (15/9 h, d/n) at plant
canopy level. Due to the unavailability of settings for constant
vapor pressure deficit, all the growth chambers were set to main-
tain 60 ±10% relative humidity throughout the experiment. Plants
were watered daily.
2.2 Glyphosate and dicamba dose–response experiment
2.2.1 Glyphosate and dicamba treatment
Kochia plants were treated with glyphosate (Roundup Weather-
Max; Monsanto Co., St. Louis, MO, USA) at dosages of 0, 26.3, 52.5,
105, 210, 420, 840, and 1680 g ha1with 2.5% (w/v) ammonium
sulfate (AMS) or dicamba (Clarity; BASF Corp., Florham Park, NJ,
USA) without AMS at dosages of 0, 17.5, 35, 70, 140, 280, 560, and
1120 g ha1when the plants were 10– 12 cm tall. Herbicides were
mixed in water and applied using a bench-type sprayer (Research
Track Sprayer; De Vries Manufacturing, Hollandale, MN, USA)
equipped with a single moving flat-fan nozzle tip (80015LP TeeJet
tip; Spraying Systems Co., Wheaton, IL, USA) delivering 187 L ha1
at 222 kPa in a single pass at 3.21 km h1. Following treatment,
plants were returned to corresponding growth chambers within
30 min after treatment.
2.2.2 Visual injury and biomass measurement
Glyphosate- and dicamba-induced injury was rated based on com-
posite visual estimations of growth inhibition, curling, necrosis,
and plant vigor on a scale of 0 (no effect) to 100 (plant death). Visual
injury ratings were taken at 1, 2, 3, and 4 weeks after treatment
(WAT). At 4 WAT, plant stems were cut at soil level and individual
plants were placed in separate paper sacks. After oven drying at
60C for 72 h, plants were weighed once more to calculate dry
2.3 Absorption and translocation experiments
Results of the dose–response experiments showed that the
two kochia populations, P1 and P2, responded similarly to
glyphosate and dicamba at each temperature regime. There-
fore, the glyphosate or dicamba absorption and translocation
experiments were conducted using only one population, i.e. P1.
Prior to conducting the absorption and translocation exper-
iments, we tested whether absorption or translocation of
14C-glyphosate or 14 C-dicamba in kochia would be affected
by spraying plants with formulated products of either herbicide
before 14C-herbicide treatment using the method described by
Perez-Jones et al.16 Briefly, on six 10–12 cm tall kochia seedlings,
two newly expanded leaves were marked and wrapped with small
pieces of aluminium foil, then the plants were sprayed with formu-
lated product of 840 g ha1of glyphosate or 560 g ha1of dicamba
using the methods described in section 2.1. After 30 min, when
the herbicide droplets dried, the aluminium foil was removed.
Likewise, another set of six untreated kochia seedlings of the same
size were selected and two newly expanded leaves were marked
on these plants as well. On both sets of kochia, the absorption
and translocation of 14C-glyphosate or 14 C-dicamba were tested
under T2 using the method described in detail in sections 3.1
and 3.2. Results (data not shown) showed that neither absorption
nor translocation of 14C-dicamba or 14 C-glyphosate was affected
by spraying the plants with formulated herbicide. Hence, the
absorption and translocation experiments using 14C-glyphosate
or 14C-dicamba reported here were not sprayed with formulated
Additionally, preliminarily testing of 14C-glyphosate or
14C-dicamba translocation in kochia grown at T2 revealed that less
than 0.5% of 14C-glyphosate and only 1.3% of 14 C-dicamba was
translocated to roots at 72 h after treatments (HAT). At the same
time, 88– 95% and 92 96% of 14C-dicamba and 14 C-glyphosate,
respectively, was recovered from the aboveground parts of kochia.
Hence, the translocation of 14C-glyphosate or 14 C-dicamba to plant
roots was not measured in subsequent experiments.
2.3.1 Absorption and translocation of glyphosate
One milliliter of 14C-glyphosate working solution with 0.33
kBq μL1of radioactivity was prepared by mixing 93.6 μLof
phosphonomethyl-14C-glyphosate water solution (3.7 kBq μL1,
specific activity: 2.04 kBq μg1, PerkinElmer, Inc., Boston, MA, USA), © 2016 Society of Chemical Industry Pest Manag Sci (2016)
Effect of growth temperature on glyphosate and dicamba efficacy in Kochia
9.2 μL of Roundup Weathermax herbicide (Monsanto Co.), 73.5
μL of ammonium sulfate (AMS) aqueous solution (34%, w/v) and
823.7 μL of water, which was equivalent to 840 g of glyphosate in
a carrier volume of 187 L water with 2.5% (w/v) of AMS.
Kochia seedlings (10–12 cm tall) grown under three tempera-
ture regimes (as described above) were used. On the upper sur-
face of two newly expanded leaves, 10 μLof14C-glyphosate work-
ing solution (5 μL per leaf) was applied using Wiretrol®(10 μL;
Drummond Scientific Co., Broomall, PA, USA). After 30 min, plants
were returned to growth chambers. Plantswere har vestedat 24, 48
and 72 HAT and separated into treated leaf (TL), tissue above the
treated leaf (ATL), and tissue below the treated leaf (BTL). Treated
leaves were washed twice with 5 mL wash solution [10% (v/v)
ethanol aqueous solution with 0.5% of Tween-20] in 20-mL scin-
tillation vials for 1 min. After adding 15 mL Ecolite-(R) (MP Biomed-
icals, LLC, Santa Ana, CA, USA), radioactivity in leaf rinsate was mea-
sured by using liquid scintillation spectrometry (LSS; Tricarb 2100
TR Liquid Scintillation Analyzer, Packard Instrument Co., Meriden,
CT, USA). Plant sections were dried at 60C for 72 h and radioactiv-
ity in each plant part was quantified by LSS after combusting for 3
min with a biological oxidizer (OX-501; RJ Harvey Instrument, New
Yor k , NY, U SA).
2.3.2 Absorption and translocation of dicamba
The methods of 14C-dicamba application and sample collection
were the same as described above for the 14C-glyphosate exper-
iment, except that the 1 mL of 14C-dicamba working solution was
obtained by mixing 29.3 μL of dicamba-(ring-UL-14C) ethanol solu-
tion (11.4 kBq μL1, specific activity: 2.87 kBq μg1, BASF Corp.), 6.4
μL of Clarity herbicide (BASF Corp.) and 964.3 μLofwater,which
was equal to 560 g of dicamba in a carrier volume of 187 L.
2.3.3 Data analysis
The data from absorption and translocation experiments of both
herbicides was converted into percentages for further analysis
using the following equations:
percentage recovery =Rrinsate +RATL +RTL +RBTL
×100 (1)
percentage absorption =Rapplied Rrinsate
×100 (2)
percentage translocation =100 RTL
Rapplied Rrinsate ×100
percentage in ATL =RATL
Rapplied Rrinsate
×100 (4)
percentage in TL =RTL
Rapplied Rrinsate
×100 (5)
percentage in BTL =RBTL
Rapplied Rrinsate
×100 (6)
In Eqns 1–6, Rrinsate is the radioactivity recovered in leaf rinsate;
Rapplied is total amount of radioactivity applied on the plant; RATL
is the radioactivity recovered in tissue above the treated leaf (ATL);
RTL is the radioactivity recovered in the treated leaf (TL); and RBTL is
the radioactivity recovered in tissue below the treated leaf (BTL).
2.4 Experimental design and statistical analysis
Split-plot experimental design was used for all experiments. In the
glyphosate and dicamba dose–response experiments, tempera-
ture and herbicide doses were main and subplots, respectively. In
absorption and translocation of 14C-dicamba and 14 C-glyphosate
experiments, temperature and harvesting time were the main and
subplot, respectively. At least four replicates of each dose were
included in both studies and all the experiments were repeated
twice in time, and the growth chambers were rotated to avoid
In the whole-plant dose–response experiments, treatments
were arranged in a factorial combination of three levels of growth
temperatures (T1, T2, and T3) and different herbicide doses.
There was no interaction between experimental runs and treat-
ments; hence, data from the two dose– response experiments
were pooled for each population prior to analysis. Using the drc
package17 in R (v.3.2.1, R Foundation for Statistical Computing,
Vienna, Austria), visual injury and dry biomass were subjected to
non-linear regression analysis using four parameter log-logistic
1+exp blog (x)log I50 (7)
In Eqn 7, Yrefers to the percentage of control or untreated, Cis
the lower limit, Dis the upper limit, bis the slope, and I50 is the
dose required for 50% response of plant injury or biomass reduc-
tion. This model was used to estimate ED50 (effective dose for
50% control of kochia) and GR50 (effective dose for 50% biomass
reduction) values from the visual injury and dry biomass of kochia,
For experiments involving absorption and translocation, treat-
ments were arranged in a factorial combination of three levels of
growth temperatures (T1, T2, and T3) as main factors, and four lev-
els of measurement time (12, 24, 48, and 72 h) as simple factors.
There was no interaction between experimental runs and treat-
ments; hence, data from the two experiments were combined and
analyzed by fitting to an asymptotic regression, rectangular hyper-
bolic or linear model using the method developed by Kniss et al.19
based on drc17 and qpcR20 packages in R program. Furthermore,
the bias-corrected Akaike information criteria (AICc) of these three
models were compared and the rectangular hyperbolic model
[Eqn 8] with the lowest AICc value19 was chosen for analyzing
glyphosate or dicamba absorption data. However, none of these
three regression models could be used to analyze glyphosate or
dicamba translocation data. Therefore, all translocation data were
analyzed using two-way ANOVA (P<0.05) in Prism 6 (GraphPad
Software, Inc., La Jolla, CA, USA):
absorption =Amax ×t
90)×t90 +t(8)
In Eqn 8, Amax is the upper limit (maximum) for absorption of
herbicide, tis the time, Absorption is the percentage of absorbed
herbicide at time t,andt90 refers to the time required to achieve
90% of the maximum absorption.
3.1 Dose–response of glyphosate
At 4 WAT, ED50 values for glyphosate on P1 kochia at T1 and T2 were
39 and 68 g ha1[Table 1; Fig. 1(a)], respectively. However, the GR50
values for glyphosate at T1 and T2 were 34 and 42 g ha1for this
Pest Manag Sci (2016) © 2016 Society of Chemical Industry J Ou, PW Stahlman, M Jugulam
Table 1. Glyphosate and dicamba dose– response analysis of kochia visual injury and dry biomass under three different temperatures at 4 weeks
after treatment*
Parameter estimate (dry biomass)
Kochia population
(site of collection)
Tem pe ra tu re
(day/night, C) ED50 (g ·ha1)GR
50 (g ·ha1)bC(g) D(g)
Glyphosate P1(Pratt County, KS) 17.5/7.5 39 (2.4)a34 (5.2)a4.34 (1.78) 0.04 (0.01) 0.22 (0.02)a
25/15 68 (4.4)b42 (11)a2.10 (1.18) 0.06 (0.08) 0.95 (0.11)b
32.5/22.5 173 (10)c171 (55)b2.90 (2.28) 0.03 (0.20) 1.27 (0.19)bc
P2(Riley County, KS) 17.5/7.5 36 (2.2)a46 (1.2)a4.32 (0.55) 0.05 (0.01) 0.55 (0.01)a
25/15 68 (6.3)b67 (1.8)b3.40 (0.22) 0.11 (0.01) 1.24 (0.02)ab
32.5/22.5 186 (9.8)c187 (8.3)c3.49 (0.43) 0.08 (0.03) 1.48 (0.23)b
Dicamba P1(Pratt County, KS) 17.5/7.5 52 (2.4)a21 (15)a1.96 (3.06) 0.07 (0.06) 0.60 (0.07)a
25/15 54 (3.4)a26 (16)a1.46 (1.94) 0.08 (0.16) 1.28 (0.04)b
32.5/22.5 106 (6.5)b73 (19)b3.99 (5.16) 0.06 (0.26) 1.46 (0.07)c
P2(Riley County, KS) 17.5/7.5 105 (9.7)a46 (15)a0.48 (0.08) 0.24 (0.12) 0.59 (0.05)a
25/15 167 (34)a114 (35)a0.68 (0.37) 0.43 (0.11) 0.95 (0.06)b
32.5/22.5 410 (36)b225 (6.3)b2.76 (0.16) 0.01 (0.02) 1.51 (0.02)c
*Values (mean ±standard error) followed by different letters are significantly (P<0.05) different in each column for each population.
ED50 values were calculated using visual injury data.
The four parameters log-logistic model was used for estimation [see Eqn 7, for details].
population [Table 1; Fig. 1(b)]. Differences between T1 and T2 were
significant (P<0.05) for ED50 but not GR50. However, when d/n
temperature was increased to T3, both the ED50 and GR50 increased
significantly (P<0.05) to 173 and 171 g ha1, respectively in P1
kochia. The results of glyphosate dose– response on P2 kochia
population [Table 1; Fig. 1(c and d)] showed similar tendency of
growth temperature effects on glyphosate efficacy as described
above for P1 kochia. ED50 values for glyphosate on P2 were 36, 68
and 176 g ha1at T1, T2, and T3, respectively, whereas the GR50
were estimated as 46, 67, and 187 g ha1, respectively. Both ED50
and GR50 of glyphosate on P2 increased significantly as growth
temperature increased. When the GR50 values were estimated in
the four parameters log-logistic model using the raw data of dry
biomass, the estimates for other parameters were also generated
for glyphosate and listed in Table 1. The estimation of Dvalues
(the upper limit, which represents the dry biomass accumulation
of untreated samples) of P1 and P2 were significantly different
at T1 and T3 (Table 1). In general, the untreated kochia plants
grown under cooler temperature (T1) produced three times more
biomass than at high temperature [T3; Fig. 1(b and d) and Fig. 2(b
and d)].
3.2 Dose–response of dicamba
At 4 WAT, both P1 and P2 kochia showed similar response to
dicamba when grown at different temperatures. The ED50 [Table 1,
Fig. 2(a)] of dicamba for P1 kochia was 52, 54, and 106 g ha1
at T1, T2, and T3, respectively. On the basis of dry biomass, GR50
[Table 1, Fig. 2(b)] of dicamba for P1 kochia was 21, 26, and 73
1at T1, T2, and T3, respectively. Likewise, ED50 of 105, 167,
and 410 g ha1and GR50 of 46, 114 and 225 g ha1at T1, T2,
and T3 [Table 1, Fig. 2(c and d)], respectively, are estimated for
P2 kochia. The efficacy of dicamba on both P1 and P2 decreased
when temperature was increased from T2 to T3, but not from T1
to T2. Also, estimation of the four parameters for dicamba using
raw dry biomass data was also determined and listed in Table 1.
The dry biomass accumulation of untreated samples (Dvalues)
was significantly different among the three temperature regimes,
which indicates temperature has siginificant effect on growth of
3.3 Absorption and translocation of glyphosate
Analysis of the data of 14C-glyphosate absorption/translocation
(Table 2) indicates the upper limit of absorption of 14C-glyphosate
(Amax) as 71, 70, and 41% at T1, T2, and T3, respectively. When
the Amax at different temperatures was compared, significantly
less 14C-glyphosate was absorbed by kochia at T3 than at T1 or
T2. Similarly, analysis of the data by regression model also sug-
gest the time required to achieve 90% of the maximum absorp-
tion (t90, Table 2) as 188, 144, and 313 h for T1, T2, and T3,
respectively, but the comparison of t90 at different temperatures
showed the time differences were not significant among the three
temperature regimes. Interestingly, regardless of the amount of
14C-glyphosate absorbed, there was no significant difference in the
percent of 14C-glyphosate translocated (Fig. 3b) either to ATL or
BTL of kochia grown under any of the temperature regimes tested
(Fig. s 3d to 3e). Overall, absorption of 14C-glyphosate was signif-
icantly reduced when kochia was grown under T3 (Fig. 3a). How-
ever, translocation of 14C-glyphosate in kochia appeared not to
be influenced by alterations in temperature (Fig. 3b). Therefore,
reduced absorption of glyphosate may contribute to the lack of
control of kochia grown under high temperature.
3.4 Absorption and translocation of dicamba
Similar to absorption of glyphosate, the upper limit of dicamba
absorption (Amax) and time required to achieve 90% of the max-
imum absorption (t90) were generated using regression analysis,
and the results are listed in Table 2. The data suggest Amax of 99,
98, and 100%, and t90 of 57, 36, and 48 h for T1, T2, and T3, respec-
tively. However, in contrast to glyphosate, the data of the Amax
and t90 of dicamba was not significantly affected by temperature.
While absorption of dicamba increased with time, translocation
out of the TL also increased [Fig. 4(b)], regardless of temperature.
Translocation of 14C-dicamba at 12 and 72 HAT increased from 26
to 47% and 20 to 58% at T1 and T2, respectively [Fig. 4(b)]. In © 2016 Society of Chemical Industry Pest Manag Sci (2016)
Effect of growth temperature on glyphosate and dicamba efficacy in Kochia
(a) (b)
(c) (d)
Figure 1. Whole-plant glyphosate dose– response of kochia at different temperatures as measured by (a) visual injury (P1), (b) dry biomass (P1), (c) visual
injury (P2), and (d) dry biomass (P2) at 4 WAT.
contrast, at 72 HAT translocation of 14C-dicamba increased from
only 6.9% to 21% in kochia grown at T3. This means 20– 30% more
14C-dicamba was retained in the TL [Fig. 4(c)] of kochia grown
at T3, than in kochia grown at T1 or T2. More importantly, at 12
HAT, 16.5% and 16% of 14C-dicamba was translocated to ATL at
T1 and T2, respectively, but only 3.2% moved towards meristems
in kochia grown at T3 [Fig. 4(d)]. Conversely, there was no dif-
ference (P>0.05) in the amount of 14 C-dicamba translocated to
BTL [Fig. 4(e)] in kochia grown at any of the temperature regimes
tested. Thus, the poor control of kochia grown under high temper-
ature may be attributed to decreased translocation of dicamba to
above treated leaves.
In western Kansas, kochia emerges early- to mid-March and con-
tinues into April2when d/n temperatures are normally about
17.5/7.5C.21 Thereafter, kochia emergence slows down but some
seeds can still emerge throughout the growing season. After the
major flush of emergence in March to April, kochia starts to grow
and accumulates biomass when the d/n temperatures increase
to 25/15C.21 Post-application of glyphosate or dicamba to con-
trol kochia is normally done in mid- to late-June after crop emer-
gence or in July for post-wheat harvest applications when the d/n
temperatures are soaring to 32.5/22.5C or higher. This validates
selection of these three d/n temperature regimes in this study.
In the dose–response experiments, we found the efficacy of
glyphosate decreased significantly when the d/n temperatures
were increased from 25/15C to 32.5/22.5C. Similar results were
observed for GR50 of glyphosate for P2 kochia [Fig. 1(d)], except
the GR50 of glyphosate at T1 and T2 on P1 kochia were not sig-
nificantly different whereas the ED50 of glyphosate on P1 kochia,
and both the ED50 and GR50 of glyphosate on P2 kochia were
significantly different. These results clearly indicate that plant
growth temperature had substantial impact on the efficacy of
glyphosate in controlling kochia. Additionally, the nonsignificant
estimation of Cvalues (data not shown) in the four parameter
log-logistic model indicates that kochia (both P1 and P2) accumu-
lated different amounts of dry biomass at all temperatures tested
in response to the high rates (lethal rates or higher) of glyphosate
or dicamba. This difference in biomass accumulation within each
population can be attributed to the inherent genetic variability,
which is expected among field populations of kochia. In contrast,
in response to any rate of glyphosate or dicamba applied, (except
for P1 at T1), the estimation of dry biomass accumulation of
untreated samples (Dvalues) of both P1 and P2 (Table 2) was
significant at all temperature regimes. Specifically, there was
significantly higher (about two times) biomass accumulation at T3
Pest Manag Sci (2016) © 2016 Society of Chemical Industry J Ou, PW Stahlman, M Jugulam
(a) (b)
(c) (d)
Figure 2. Whole-plant dicamba dose– response of kochia at different temperatures as measured by (a) visual injury (P1), (b) dry biomass (P1), (c) visual
injury (P2), and (d) dry biomass (P2) at 4 WAT.
than at T1, for P1 and P2 kochia [Fig. 1(b and d) and Fig. 2(b and
d)], which clearly suggests that kochia growth was substantially
affected by temperature. The difference in biomass accumuling
of kochia at different temperatures may influence the absorption
or translocation of herbicides. In general, larger plants are more
tolerant to herbicides than the smaller plants. The decreased
efficacy of dicamba or glyphosate on kochia grown under high
temepratures, possibly because of dilution effect that caused by
rapid growth and high biomass accumulation.22
It is known that even with the addition of surfactants, relatively
low amounts of applied glyphosate is absorbed by leaves23 com-
pared to other systemic herbicides such as dicamba. Our data
also show less than 60% of glyphosate absorbed by kochia at 72
HAT [Fig. 3(a)]. More importantly, plants typically develop thick,
lipophilic cuticles to prevent water loss at high temperature.24,25
Therefore, when grown under high temperatures (T3) kochia may
develop thicker cuticle, which may have contributed to reduced
absorption of glyphosate even when the herbicide was formu-
lated with surfactants.26 As we observed in our glyphosate dose
response experiments, efficacy of glyphosate was decreased at
high temperatures, which is highly interrelated with our absorp-
tion and translocation data. We conclude the decreased efficacy
of glyphosate on kochia at high growth temperature was due to
decreased absorption of this herbicide.
In the dicamba experiment, GR50 and ED50 dosages for P2 kochia
plants were three and four times higher, respectively, compared
to GR50 and ED50 dosages for P1 kochia plants (Table 1), indicat-
ing greater tolerance to dicamba in P2 kochia. Yet, the increase
in d/n temperature from 25/15C to 32.5/22.5C reduced the
efficacy of dicamba on both P1 and P2 kochia. Based on the
dose–response results it is evident that efficacy of dicamba on
kochia control did not differ when plants were grown under tem-
perature regimes of 17.5/7.5C or 25/15C; however, efficacy was
significantly decreased when they were exposed to 32.5/22.5C. In
the physiological mechanism study, no difference was found in the
amount of 14C-dicamba absorbed by kochia grown at the temper-
atures tested in this experiment. However, less 14C-dicamba was
translocated to ATL in kochia grown at T3 than at T1 or T2, while
the amount of 14C-dicamba translocated to BTL was not affected
by temperature. Reduced dicamba efficacy on kochia grown at T3
compared to T1 or T2 (Fig. 2), likely was because of reducedtranslo -
cation of dicamba to actively growing meristems at T3 [Fig. 4(b)].
Dicamba is a systematic herbicide and must be translocated to
the meristems27 to obtain satisfactory weed control. Therefore, the
lack of kochia control with dicamba treatment at high temper-
ature (i.e. T3) can be attributed to reduced translocation of this
herbicide. © 2016 Society of Chemical Industry Pest Manag Sci (2016)
Effect of growth temperature on glyphosate and dicamba efficacy in Kochia
Hours after treatment (h)
0 20406080100
24 48 7212
Time (h) after treatment
Translocation of 14C compounds
(% of absorbed)
24 48 7212
Time (h) after treatment
14C compounds in TL
(% of absorbed)
24 48 7212
Time (h) after treatment
14C compounds in ATL
(% of absorbed)
24 48 7212
Time (h) after treatment
14C compounds in BTL
(% of absorbed)
Absorption of 14C glyphosate
(% of applied)
Figure 3. (a) 14C-glyphosate absorption, (b) translocation, (c) retained in treated leaf, (d) translocation to above treated-leaf, and (e) below treated-leaf
at three different temperatures. (*P-value <0.05, **P-value <0.01, ***P-value <0.001, which indicate the levels of significance within each time points at
different temperatures; error bars represent standard error).
Table 2. Regression parameter estimates of glyphosate absorp-
tion in kochia at different temperatures using rectangular hyperbolic
model *
Parameter estimate
Tem pe ra tu re
(day/night, C) Amax t90
Glyphosate 17.5/7.5 70.58 (5.77)a188.03 (44.23)a
25/15 70.22 (4.32)a144.55 (29.79)a
32.5/22.5 41.28 (9.05)b313.67 (155.89)a
Dicamba 17.5/7.5 98.66 (3.38)a57.19 (11.73)a
25/15 97.78 (3.13)a35.62 (8.86)a
32.5/22.5 100.0 (3.05)a47.92 (9.46)a
*See Eqn 8 for the equation of the rectangular hyperbolic model.
Values with different superscript letters are significantly (P<0.05)
different in each column for each herbicide.
Dicamba absorbed into plant cells can be trapped in phospho-
lipid vesicles due to a hydrophobic interaction between the non-
polar portion of dicamba molecule and the hydrocarbons present
in the phospholipid vesicles.28 Since dicamba is predominantly
translocated via symplast,29 it is prone to becoming trapped in
phospholipid vesicles. It is also known that increased temperature
can enhance the strength of hydrophobic interactions of organic
molecules.30 Therefore, in this study, when dicamba was applied
on kochia grown under higher temperature, though the absorp-
tion of dicamba was not affected [Fig. 4(a)], it is possible that
dicamba may have attached to phospholipid vesicles in leaf cells,
resulting in lack of movement of this molecule from the site of
absorption. Additional study is needed to test this hypothesis.
Furthermore, dicamba is volatile and increased temperature can
also accelerate the volatilization of dicamba, regardless of the type
of dicamba formulation used.31 Under field conditions, vapor or
spray drift of dicamba can cause severe crop damage on soybean,3
tomatoes,32 and corn,33 especially on hot days. Therefore, applying
dicamba during periods of high temperature not only reduces
kochia control but also increases the risk of off-target crop injury.
Dicamba is an auxinic herbicide and sensitive plants show severe
injury symptoms (e.g. epinasty, meristem inhibition, etc.) when
treated or exposed to low doses34 of off-target drift. However,
dicamba kills susceptible plants slowly. Some of the plants treated
with higher than field recommended doses of dicamba in this
experiment, although injured severely, still had green tissues at
4 WAT. As a result, it is easy to underestimate dicamba injury
symptoms. This can explain the variation in values obtained for
ED50 when compared to GR50 at each temperature regime.
Although the mechanisms responsible for the reduced efficacy of
dicamba or glyphosate may differ, our results clearly show that
kochia is less sensitive to both these herbicides when grown under
higher temperatures, especially at 32.5C. This research provides
evidence to support the anecdotal observations made in the
field regarding reduced efficacy of herbicides such as dicamba or
glyphosate at high temperature. Therefore, to maximize efficacy of
glyphosate and dicamba on kochia and minimize the chances of
losing these effective tools for controlling kochia, it will be critical
to take action and apply glyphosate or dicamba early in the season
after the main flush of kochia emergence when the temperature is
low (e.g. day/night temperature at 25/15C, or even lower) and the
kochia seedlings are small (less than 12 cm).
Pest Manag Sci (2016) © 2016 Society of Chemical Industry J Ou, PW Stahlman, M Jugulam
Hours after treatment (h)
0 20406080100
Absorption of 14C dicamba
(% of applied)
24 48 7212
Time (h) after treatment
Trans location of 14C compounds
(% of abs orbed)
*** *** ***
24 48 7212
Time (h) after treatment
14C compounds in TL
(% of abs orbed)
24 48 7212
Time (h) after treatment
14C compounds in ATL
(% of absorbed)
*** **
24 48 7212
Time (h) after treatment
14C compounds in BTL
(% of absorbed)
Figure 4. (a) 14C-dicamba absorption, (b) translocation, (c) retained in treated leaf, (d) translocation to above treated-leaf, and (e) below treated-leaf atthree
different temperatures. (**P-value <0.01, ***P-value <0.001, which indicate the levels of significance within each time points at different temperatures;
error bars represent standard error).
Thanks to Dr Aruna Varanasi for comments to improve this
manuscript. A graduate student assistantship to J. Ou from BASF
Corp. is highly appreciated. This study is contribution 16-267-J
from the Kansas Agricultural Experiment Station, Manhattan,
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... In contrast to the TSR mechanisms, the glyphosate resistance can be conferred by multiple non-target-site based (NTSR) mechanisms. For example, reduced glyphosate foliar uptake and/or translocation in GR Sorghum halepense and L. rigidum, vacuolar sequestration and enhanced metabolism by an aldo-keto reductase in GR Echinochloa colona [14][15][16][17][18][19]. ...
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The Dongting and Poyang Lakes are the important rice growing areas, and the Bohai Rim and Loess Plateau are the main producing areas of apples in China, where glyphosate has been used continuously to control weeds including Eleusine. indica for many years. In this study, the resistance levels and target-site based resistance (TSR) mechanisms to glyphosate in E. indica populations, which were collected from above areas were investigated. A total of 35 out of 50 (70%) E. indica populations have evolved resistance to glyphosate with resistance index (RI) of 2.01~10.43. The glyphosate-resistant (GR) E. indica accumulated less shikimic acid than glyphosate-susceptible (GS) populations, when treated by 1.0 mg/L, 10 mg/L or 100 mg/L glyphosate. There was no mutation at Thr102 and Pro106 in 5-enolpyruvate shikimate-3-phosphate synthase (EPSPS), which endowed glyphosate resistance in E. indica and other weed species. A Pro-381-Leu was found in EPSPS in GR populations. In contrast, the expression level of EPSPS gene was highly correlated with glyphosate resistance in E. indica with a Pearson coefficient of 0.73. These indicate that the glyphosate resistance in aforementioned E. indica populations was mainly caused by the overexpression of EPSPS, not by amino acid mutation in EPSPS.
... Higher absorption and more intense translocation of 2,4-D in common ragweed (Ambrosia artemisiifolia) and giant ragweed (Ambrosia trifida) were recorded under high temperature compared to low temperature (Ganie et al. 2017). Similar to these findings, Ou et al. (2018) reported a reduced glyphosate and dicamba efficacy to control Kochia scoparia when temperature was increased. ...
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Recently, there has been growing concern over the potential impact of CO2 concentration and temperature on herbicide efficacy. The aim of the study was to examine the influence of single elevated CO2 (400 vs. 800 ppm) and elevated CO2 in combination with temperature (21 °C vs. 25 °C) on the effects of auxin herbicide 4-chloro-2-methylphenoxyacetic acid (MCPA) (0.5–2 × field recommended rate) to wild mustard (Sinapis arvensis L.) grown in mixed-culture with spring barley (Hordeum vulgare L.). MCPA had a detrimental effect on aboveground and belowground biomass, content of chlorophylls, enzymatic and non-enzymatic antioxidants and induced oxidative stress. The significant decline in photosynthetic rate, stomatal conductance and transpiration with MCPA dose was detected. Elevated CO2 reinforced MCPA efficacy on S. arvensis: sharper decline in biomass, photosynthetic rate and antioxidant enzymes and more pronounced lipid peroxidation were detected. Under elevated CO2 and temperature, MCPA efficacy to control S. arvensis dropped due to herbicide dilution because of increased root:shoot ratio, higher activity of antioxidants and less pronounced oxidative damage. Reinforced MCPA impact on weeds under elevated CO2 resulted in higher H. vulgare biomass, while decreased MCPA efficacy under elevated CO2 and temperature reduced H. vulgare biomass.
... However, there is an increasing number of cases of glyphosate failure in regions where summer fallow weed species occur, threatening the long-term viability of glyphosate in these regions (Peltzer et al. 2009; Thornby and Walker 2009;Walker et al. 2011). Reduced efficacy of glyphosate to control summer fallow weeds can be attributed to decreased absorption and translocation of herbicide due to morpho-physiological changes under elevated temperatures (Ou et al. 2018), referred to as temperature-induced herbicide tolerance. Previous studies have reported the differential effect of elevated temperature on the efficacy of glyphosate in controlling awnless barnyard grass [Echinochloa colona (L.) Link.], wild oats (Avena fatua L.), and liverseed grass (Urochloa panicoides (P.) Beauv.] in the NGR (Tanpipat et al. 1997;Adkins et al. 1998). ...
A temperature-controlled glasshouse study was conducted to evaluate the influence of elevated temperature (eT – 34/24 ± 2°C) on the growth and glyphosate susceptibility of windmill grass (Chloris truncata R.Br.), common sowthistle (Sonchus oleraceus L.), and flaxleaf fleabane [Conyza bonariensis (L.) Cronquist]; and to determine the morpho-physiological factors involved in differential glyphosate tolerance under eT. Results showed that elevation of temperature from ambient temperature (aT – 28/20 ± 2°C) to 34/24 ± 2°C increased growth and biomass production of C. truncata. In contrast, eT suppressed growth of S. oleraceus and C. bonariensis, resulting in fewer, thicker, and smaller leaves with reduced stomatal conductivity and less total plant biomass. In terms of herbicide susceptibility, the responses to glyphosate under different temperature regimes were species- and rate-specific. Slight variations in glyphosate susceptibility were observed when sprayed at sub-lethal rates at eT. Under eT, C. truncata, S. oleraceus, and C. bonariensis required 1.5, 2.0-, and 1.6-times higher glyphosate rates, respectively, to suppress biomass by 50% compared with plants grown at aT. Depending upon the species and glyphosate rate, differences in leaf characteristics (i.e. leaf chlorophyll content, leaf area/thickness, and stomatal conductance) could have promoted or delayed glyphosate activity under eT over the period, especially at sub-lethal rates. Overall, the glyphosate efficacy was unaffected since herbicide within the recommended rates completely controlled all tested weed species under both temperatures.
... Different studies have reported that glyphosate is more efficient when applied at 6 am (Mohr et al. 2007), and temperatures near to 25 °C (Degreeff et al. 2018). Applications made under temperatures below 20°C provided lower uptake and translocation, therefore lower herbicide efficiency (Nguyen et al. 2016;Ou et al. 2018). ...
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Weed management with herbicides requires favorable environmental conditions, that maximize efficiency, such as soil humidity and timing of application. The aim of this study was to assess the effect of the application timing of bentazon and glyphosate herbicides on the control and activity of antioxidant enzymes in Ipomoea grandifolia, under different conditions of soil water availability. Two experiments, one for each herbicide (bentazon and glyphosate), were conducted in a factorial design with four replicates. The first factor was the two rates of each herbicide (504 and 720 g i.a. ha-1 of bentazon and 651 and 911.4 g i.a. ha-1 of glyphosate). The second factor was the six application times (1 am; 5 am; 9 am; 1 pm; 5 pm and 9 pm). The third factor, soil water content (100% and 50% of field capacity). Plus, two controls without herbicide application. At 21 days after application (DAA) of bentazon and 28 DAA of glyphosate, the fresh mass of the aboveground plants was measured. In addition, the activities of the enzymes superoxide dismutase (SOD), catalase (CAT) and peroxidase (POX) were determined. For plants under water restriction, an increase of approximately 20% in fresh mass was observed compared in plants without water restriction, indicating lower control efficiency under water stress condition. For bentazon, at 1 pm has been observed the lowest herbicide efficiency, the other times were the most efficient, and did not differ. For glyphosate, the application at 9 am was the most efficient, while at 1 am provided the worst control efficiency. Higher CAT and SOD activities after bentazon application were observed at 1 pm. Among the three enzymes evaluated, SOD presented the highest activity after glyphosate application. Generally, the times of the day with the highest peak of enzymatic activity were distinct between with and without water restriction.
... 74 In fact, NTR mechanisms such as decreased translocation have already been reported in other dicamba or 2,4-D resistant weeds. [75][76][77][78] Future research could focus on using the newly available kochia genome to identify the genetic basis for compensatory evolution. 79 For example, identifying specific secondary mutations or genes that are involved in the co-selected weediness traits. ...
Full-text available
Background Lack of fitness cost have been reported for multiple herbicide resistance traits, but the underlying evolutionary mechanisms are not well understood. Compensatory evolution that ameliorates resistance costs, has been documented in bacteria and insects but rarely studied in weeds. Dicamba resistant IAA16 (G73N) mutated kochia was previously found to have high fecundity in the absence of competition, regardless of significant vegetative growth defects. To understand if costs of dicamba resistance can be compensated through traits promoting reproductive success in kochia, we thoroughly characterized the reproductive growth and development of different G73N kochia biotypes. Flowering phenology, seed production and reproductive allocation were quantified through greenhouse studies, floral (stigma‐anthers distance) and seed morphology, as well as resulting mating and seed dispersal systems were studied through time‐course microcopy images. Results G73N covaried with multiple phenological, morphological and ecological traits that improve reproductive fitness: 1) 16‐60% higher reproductive allocation; 2) longer reproduction phase through early flowering (2‐7d); 3) smaller stigma‐anthers separation (up to 60% reduction of herkogamy and dichogamy) that can potentially promote selfing and reproductive assurance; 4) “winged” seeds with 30‐70% longer sepals that facilitate long‐distance seed dispersal. Conclusion The current study demonstrates that costs of herbicide resistance can be ameliorated through coevolution of other fitness penalty alleviating traits. As illustrated in a hypothetical model, the evolution of herbicide resistance is an ongoing fitness maximization process, which posts challenges to contain the spread of resistance.
... account for some variance on the magnitude of resistance. 37,38 Therefore, we believe that visual injury combined with mortality data, as reported in the current study, is a better proxy with which to characterize resistance to synthetic auxin herbicides (Fig. S3). ...
Full-text available
Background: Precise quantification of the fitness cost of synthetic auxins resistance has been impeded by lack of knowledge for the genetic basis of resistance in weeds. Recent elucidation of a resistance endowing IAA16 mutation (G73N) in a key weed species kochia (Bassia scoparia), allows detailed characterization of the contribution of resistance alleles to weed fitness, both in the presence and absence of herbicides. Different G73N genotypes from a segregating resistant parental line (9425) were characterized for cross resistance to dicamba, 2,4-D and fluroxypyr, and changes on stem/leaf morphology and plant architecture. Plant competitiveness and dominance of the fitness effects was quantified through measuring biomass and seed production of three F2 lines in two runs of glasshouse replacement series studies. Results: G73N confers robust resistance to dicamba but only moderate to weak resistance to 2,4-D and fluroxypyr. G73N mutant plants displayed significant vegetative growth defects: 1) being 30-50% shorter with a more tumbling style plant architecture; 2) had thicker and more ovate (versus lanceolate and linear) leaf blades with lower photosynthesis efficiency, and 40-60% smaller stems with less developed vascular bundle systems. F2 mutant plants had impaired plant competitiveness, which produced up to 90% less biomass and seeds in the replacement series study. The pleiotropic effects of G73N was mostly semi-dominant (0.5) and fluctuated with the environments and traits measured. Conclusion: G73N is associated with significant vegetative growth defects and reduced competitiveness in synthetic auxin resistant kochia. Management practices should target resistant kochia's high vulnerability to competition to effectively contain the spread of resistance. This article is protected by copyright. All rights reserved.
Shading interferes with the weed's biology, which can change their sensitivity to post-emergence herbicides. The objective was to evaluate the control of Merremia cissoides with glyphosate in full sunlight and shade conditions in two plant growth stages (30 and 73 days after sowing (DAS)). At 30 and 73 DAS, treatments were established in a 2 × 5 and 2 × 6 factorial scheme, respectively. In both experiments, the growth environments constituted the first factor, and the glyphosate doses the second factor. Shading promoted 50 and 40% reductions in glyphosate doses at 30 and 73 DAS, respectively. At 73 DAS, M. cissoides is 177.77 and 131.48% more tolerant to glyphosate than 30 DAS in shading and full sunlight, respectively. Due to the increase in glyphosate tolerance as the plant grows, the management of M. cissoides should be carried out until the stage of six fully expanded leaves. Increasing glyphosate doses reduced the quantum yield of photosystem II and electron transport rate (ETR) in both growth environments, with ETR data showing a high negative correlation with the control. The doses reductions promoted by shading and glyphosate application in the initial growth stage of M. cissoides reduces costs and the negative environmental impacts of this herbicide use.
Control of glyphosate-resistant (GR) junglerice is quite a challenging task in eastern Australia. There is limited information on the efficacy and reliability of alternate herbicides for GR populations of junglerice, especially when targeting large plants and when temperatures are high. A series of experiments were conducted to confirm the level of glyphosate resistance in three populations of junglerice and to evaluate the efficacy of alternate herbicides for the control of GR-junglerice populations. The lethal dose (LD 50 ) of glyphosate required to kill 50% plants of B17/7, B17/34, and B17/35 populations was found to be 298, 2260, and 1715 g ae ha ⁻¹ , respectively, suggesting populations B17/34 and B17/35 were highly resistant to glyphosate. Glyphosate efficacy was reduced at high-temperature (35/25 C day/night) compared with low-temperature conditions (25/15 C day/night), suggesting that control of susceptible populations may also be reduced if glyphosate is sprayed under hot conditions. PRE herbicides dimethenamid (1000 g ai ha ⁻¹ ) and pendimethalin (1500 g ai ha ⁻¹ ) provided 100% control of GR populations (B17/34 and 17/35). POST herbicides, such as clethodim (60 or 90 g ai ha ⁻¹ ), glufosinate (750 g ai ha ⁻¹ ), haloxyfop (52 or 78 g ai ha ⁻¹ ), and paraquat (400 or 600 g ai ha ⁻¹ ), applied at the 4-leaf stage provided 100% control of GR populations. For larger junglerice plants (8-leaf stage), POST applications of paraquat (400 or 600 g ai ha ⁻¹ ) provided greater weed control than clethodim, glufosinate, and haloxyfop. A mixture of either glufosinate or haloxyfop with glyphosate provided poor control of GR-junglerice populations compared with application of glufosinate or haloxyfop applied alone. Efficacy of glufosinate and haloxyfop for the control of GR populations reduced when applied in the sequential spray after glyphosate application. This study identified alternative herbicide options for GR-junglerice populations that can be used in herbicide rotation programs for sustainable weed management.
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Conyza sp. was the first glyphosate-resistant weed in Brazil’s soybean crop. Synthetic auxin herbicides followed by paraquat has improved the hairy fleabane control, and time of day of herbicide application can affect the control efficacy. There are no studies reporting the effects of the application time of synthetic auxins in tank-mixture with paraquat, or in sequential application with paraquat at two growth stages of hairy fleabane, applied at different times of the day. The herbicides were sprayed during the morning and night. 2,4-D applied alone was more effective applied during the day, while dicamba efficiency was higher when applied at night in the rosette stage. The mixture of 2,4-D and paraquat was more efficient when applied during the night. Tall hairy fleabane were more effectively controlled by dicamba + paraquat as well as any synthetic auxin followed by paraquat. When herbicides were applied at night, efficiency was slightly higher. Keywords: Conyza bonariensis; 2,4-D; dicamba; sequential; night application; morning application
The impacts of weeds in cropping systems are diverse and costly. Direct expenditure on control and biosecurity measures costs society billions each year. Even with such heavy investment in prevention and control, weeds continue to reduce the quality and quantity of agricultural produce and represent a significant threat to global food production. The challenge of managing weeds in cropping systems is rendered increasingly complex given the diverse and unpredictable impacts of climate change on both weeds and crops. Atmospheric CO2, temperature and precipitation are key drivers of plant growth, and weeds, like all other plant species, will need to respond to climate change in order to survive. Weed species are by their very nature survivors, able to relocate, acclimate or adapt to changing environmental conditions, with genetic diversity that could confer a natural competitive advantage over crop species. Conversely, modern crops are the result of extensive and highly sophisticated breeding to improve their genetic potential to survive in challenging conditions, including herbicide application, limited soil moisture and high temperatures. Moreover, agricultural weeds evolve in highly managed environments, and management intervention through crop selection, crop planting strategies and weed control measures may exert stronger selection pressures on weed species relative to climate change. It is, however, reasonable to assert that evolution driven by management pressures could occur simultaneously to climate-driven adaptation. For this reason, even given the rapid advancement of increasingly sophisticated weed control technology, weed management now and in the future should be guided a sound understanding of evolutionary biology.
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Timing of weed emergence and seed persistence in the soil influence the ability to implement timely and effective control practices. Emergence patterns and seed persistence of kochia populations were monitored in 2010 and 2011 at sites in Kansas, Colorado, Wyoming, Nebraska, and South Dakota. Weekly observations of emergence were initiated in March and continued until no new emergence occurred. Seed was harvested from each site, placed into 100-seed mesh packets, and buried at depths of 0, 2.5, and 10 cm in fall of 2010 and 2011. Packets were exhumed at 6-mo intervals over 2 yr. Viability of exhumed seeds was evaluated. Nonlinear mixed-effects Weibull models were fit to cumulative emergence (%) across growing degree days (GDD) and to viable seed (%) across burial time to describe their fixed and random effects across site-years. Final emergence densities varied among site-years and ranged from as few as 4 to almost 380,000 seedlings m ⁻² . Across 11 site-years in Kansas, cumulative GDD needed for 10% emergence were 168, while across 6 site-years in Wyoming and Nebraska, only 90 GDD were needed; on the calendar, this date shifted from early to late March. The majority (>95%) of kochia seed did not persist for more than 2 yr. Remaining seed viability was generally >80% when seeds were exhumed within 6 mo after burial in March, and declined to <5% by October of the first year after burial. Burial did not appear to increase or decrease seed viability over time but placed seed in a position from which seedling emergence would not be possible. High seedling emergence that occurs very early in the spring emphasizes the need for fall or early spring PRE weed control such as tillage, herbicides, and cover crops, while continued emergence into midsummer emphasizes the need for extended periods of kochia management.
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Background: Overreliance on glyphosate as a single tool for weed management in agricultural systems in Brazil has selected glyphosate-resistant populations of tall windmill grass (Chloris elata Bisch.). Results: Two C. elata populations, one glyphosate-resistant (GR) and one glyphosate-susceptible (GS), were studied in detail for a dose-response experiment and for resistance mechanism. The dose causing 50% reduction in dry weight (GR50 ) for GR was 620 g a.e. ha(-1) and 114 g ha(-1) for GS, resulting in an R/S ratio of 5.4. GS had significantly higher maximum (14) C-glyphosate absorption (Amax ) into the treated leaf (51.3%) than GR (39.5%), a difference of 11.8% in maximum absorption. GR also retained more (14) C-Glyphosate in the treated leaf (74%) than GS (51%), and GR translocated less glyphosate (27%) to other plant parts (stems, roots, and root exudation) than GS (36%). There were no mutations at the Pro106 codon in the gene encoding 5-enolpyruvylshikimate-3-phosphate synthase (EPSPS). There was no difference in EPSPS genomic copy number or EPSPS transcription between populations GS and GR. Conclusion: Based on these data, reduced glyphosate absorption and increased glyphosate retention in the treated leaf contribute to glyphosate resistance in this C. elata population from Brazil.
The translocation of 2-methoxy-3,6-dichlorobenzoic acid (dicamba-7- ¹⁴ C) in purple nutsedge ( Cyperus rotundus L.) at different stages of development and under various light regimes was studied using autoradiography and liquid scintillation counting techniques. The fate of the herbicide was investigated by means of thin-layer chromatography. Dicamba was slowly but appreciably translocated in purple nutsedge following application to the leaves or roots. Foliarly applied dicamba movement proceeded both acropetally and basipetally, and the herbicide became widely distributed throughout the aerial parts of the plant and accumulated in the regions of meristematic activity. Dicamba was barely detectable in the underground organs, although it was excreted into the culture medium and passed through the rhizomes and tubers into daughter plants. Root-applied dicamba distribution was general throughout the plant except within the tubers and the tips of the leaves. Dicamba was barely detectable within the tubers, although it was present in quantity at or near the surface. Dicamba accumulation was evident at the tips of the leaves. Translocation was enhanced under low light conditions. Distribution of dicamba was profoundly affected by the stage of development of the plant. Greater translocation and broader distribution of the herbicide occurred in the vegetative stage of development. After flowering, movement out of the treated leaf decreased. Studies indicated that the herbicide was not degraded by the plant during the 10 days following treatment.
Background: When used at effective doses, weed resistance to auxinic herbicides has been slow to evolve when compared to other modes of action. Here we report the evolutionary response of a herbicide-susceptible population of wild radish (Raphanus raphanistrum L.) and confirm that sub-lethal doses of 2,4-D amine can lead to the rapid evolution of 2,4-D resistance and cross resistance to acetolactate synthase (ALS) inhibiting herbicides. Results: Following four generations of 2,4-D selection, the progeny of a herbicide-susceptible wild radish population evolved 2,4-D resistance, increasing the LD50 from 16 g ha(-1) to 138 g ha(-1) . Along with 2,4-D resistance, cross resistance to the ALS-inhibiting herbicides metosulam (4.0-fold), and chlorsulfuron (4.5-fold) was evident. Pre-treatment of the 2,4-D-selected population with the cytochrome P450 inhibitor, malathion, restored chlorsulfuron to full efficacy, indicating that cross resistance to chlorsulfuron was likely due to P450 catalysed enhanced rates of herbicide metabolism. Conclusion: This study is the first to confirm the rapid evolution of auxinic herbicide resistance through the use of low doses of 2,4-D and serves as a reminder that 2,4-D must always be used at highly effective doses. With the introduction of transgenic auxinic herbicide resistant crops in the America's there will be a marked increase in auxinic herbicide use and therefore the risk of resistance evolution. Auxinic herbicides should be used only at effective doses and with diversity if resistance is to remain a minimal issue.
Factors influencing dicamba drift, especially vapor drift, were examined in field and growth chamber studies. In field experiments, potted soybeans [Glycine max (L.) Merr.]. exposed to vapors arising from corn ( Zea mays L.) foliarly treated with the sodium (Na), dimethylamine (DMA), diethanolamine (DEOA), or N -tallow- N,N ¹ , N ¹ -trimethyl-1,3-diaminopropane (TA) salts of dicamba (3,6-dichloro- o -anisic acid), developed dicamba injury symptoms. Dicamba volatilization from treated corn was detected with soybeans for 3 days after the application. Dicamba vapors caused symptoms on soybeans placed up to 60m downwind of the treated corn. When vapor and/or spray drift caused soybean terminal bud kill, yields were reduced. In growth chamber studies, dicamba volatility effects on soybeans could be reduced by lowering the temperature or increasing the relative humidity. Rainfall of 1mm or more on treated corn ended dicamba volatilization. The dicamba volatilization was greater from corn and soybean leaves than from velvetleaf ( Abutilon theophrasti Medic.) leaves and blotter paper. The volatilization of dicamba formulations varied in growth chamber comparisons with the acid being most volatile and the inorganic salts being the least volatile. However, under field conditions, use of less volatile formulations did not eliminate dicamba symptoms on soybeans. The volatile component of the commercial DMA salt of dicamba was identified by gas chromatography-mass spectrometry as free dicamba acid.
Greenhouse studies indicated that 3,6-dichloro- o -anisic acid (dicamba) or its metabolic derivative was strongly accumulated in meristematic tissues of Tartary buckwheat ( Fagopyrum tataricum (L.) Gaertn.) and wild mustard ( Sinapis arvensis L.) following both foliar and root uptake. In barley ( Hordeum vulgare L.) and wheat ( Triticum vulgare L.), it was distributed throughout the plants. Detoxification of dicamba occurred in all four species though not at equal rates, and a common major metabolite was identified chromatographically as 5-hydroxy-3,6-dichloro- o -anisic acid. A minor metabolite, 3,6-dichlorosalicylic acid, was found in barley and wheat but not in Tartary buckwheat or wild mustard. The four species tolerated dicamba treatment in the order of wheat, barley, wild mustard, and Tartary buckwheat. This ranking corresponds with the ability of the plants to detoxify dicamba and is inversely related to the extent of dicamba absorption and translocation in them.
Glyphosate [ N -(phosphonomethyl)glycine] toxicity to bermudagrass [ Cynodon dactylon (L.) Pers.] increased significantly with each rate increase from 0.14 to 1.12 kg/ha. Under greenhouse conditions approximately 50% bermudagrass control was obtained at 0.56 kg/ha glyphosate. Visible toxicity and fresh wt of treated plants and regrowth of plants clipped at the soil surface 24 h after treatment were used as indices for penetration and translocation of glyphosate. Visible injury to bermudagrass with 0.56 kg/ha glyphosate was greater at 100% than at 40% relative humidity (RH) at both 22 and 32 C. Fresh wt data indicated that 0.56 kg/ha glyphosate was more toxic at 32 C than at 22 C at 40% RH, but no difference was observed at 100% RH. Less than 10% of the applied ¹⁴ C-glyphosate penetrated the treated bermudagrass leaf at 22 C and 40% RH; whereas, more than 70% penetrated the treated leaf at 32 C and 100% RH. Five to six times more ¹⁴ C-label was translocated into the plant at 100% than at 40% RH. Significantly more ¹⁴ C-label translocated out of the treated leaf and into the plant at 32 C than at 22 C at 40% RH but no significant increase was observed at 100% RH.
Field research was conducted to evaluate the response of soybean to various herbicides applied at rates to simulate drift damage. Dicamba, glyphosate, glufosinate, and the sulfonylurea herbicides CGA-152005, primisulfuron, nicosulfuron, rimsulfuron plus thifensulfuron, and CGA-152005 plus primisulfuron were applied to soybean at the two to three trifoliolate leaf stage in 1997 and 1998 at 1/100, 1/33, 1/10, and 1/3 of the recommended use rates. The order of yield reduction after herbicide treatment was CGA-152005 > dicamba > CGA-152005 plus primisulfuron > rimsulfuron plus thifensulfuron > primisulfuron. Soybean yields were not reduced by glyphosate, glufosinate, and nicosulfuron. Applications of all herbicides at rates higher than 1/33 of the use rate caused injury symptoms within 30 d after treatment. However, soybean plants had partially or fully recovered by the end of the growing season. Therefore, early-season injury symptoms from herbicide drift are not reliable indicators for soybean yield reduction.
Absorption, translocation, and distribution of ¹⁴ C-glyphosate [ N -(phosphonomethyl)glycine] were examined in quackgrass [ Agropyron repens (L.) Beauv.] plants growing at 10/8, 15/12, and 21/18 C day/night temperatures at 300 μE·m ⁻² ·s ⁻¹ . Absorption of ¹⁴ C-glyphosate followed similar trends in all environments, apart from an initial delay at the highest temperature. Approximately 67% of the applied ¹⁴ C-glyphosate was absorbed after 120 h. Glyphosate translocation to the rhizomes was initially slower in plants growing at 10/8 C than at the higher temperatures, but after 24 h continued at a rate similar to that observed at the higher temperatures. CO 2 exchange rates (CER) were only slightly influenced by temperature between 5 and 25 C, which may explain the lack of temperature effect on the rate of glyphosate translocation. Approximately 47% of the applied ¹⁴ C-glyphosate was recovered in the rhizomes and associated roots 120 h after application in all environments. Glyphosate accumulated predominantly in new rhizomes under all growing conditions.
Dose-response studies are an important tool in weed science. The use of such studies has become especially prevalent following the widespread development of herbicide resistant weeds. In the past, analyses of dose-response studies have utilized various types of transformations and equations which can be validated with several statistical techniques. Most dose-response analysis methods 1) do not accurately describe data at the extremes of doses and 2) do not provide a proper statistical test for the difference(s) between two or more dose-response curves. Consequently, results of dose-response studies are analyzed and reported in a great variety of ways, and comparison of results among various researchers is not possible. The objective of this paper is to review the principles involved in dose-response research and explain the log-logistic analysis of herbicide dose-response relationships. In this paper the log-logistic model is illustrated using a nonlinear computer analysis of experimental data. The log-logistic model is an appropriate method for analyzing most dose-response studies. This model has been used widely and successfully in weed science for many years in Europe. The log-logistic model possesses several clear advantages over other analysis methods and the authors suggest that it should be widely adopted as a standard herbicide dose-response analysis method.