Journal of Integrative Agriculture 2015, 14(5): 919–925

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RESEARCH ARTICLE

Characterization of glyphosate-resistant goosegrass (Eleusine

indica) populations in China

CHEN Jing-chao*, HUANG Hong-juan*, WEI Shou-hui, ZHANG Chao-xian, HUANG Zhao-feng

Key Laboratory of Weed and Rodent Biology and Management, Institute of Plant Protection, Chinese Academy of Agricultural
Sciences, Beijing 100193, P.R.China

Abstract
Goosegrass is a worst grass weed in orchards and turf. The increased use of glyphosate for goosegrass control has led to
the occurrence of many resistant populations. Although glyphosate has been used to control weeds for the past 30 years
in China, few reports are available on glyphosate-resistant (GR) googegrass. In this study, we determined the GR level
of 14 goosegrass populations from Chengdu and Guangzhou, China. Glyphosate only controlled 3.1 and 25.0% of the
populations SL5 and SL1, respectively, at the dose of 1 680 g acid equivalent (ae) ha–1 at 14 days after treatment (DAT).
In contrast, the susceptible population (XD1) was completely (100%) controlled. The resistant index (RI) of SL5 and SL1
were 5.1 and 4.5, and the RI for SL2, SL3 and ZC1 were 4.2, 3.2 and 2.6, respectively. The RI for other populations was
range from 1.8 to 2.5. Under the dose of glyphosate at 1 640 g ae ha–1 at 10 DAT, shikimate accumulation in susceptible
population XD1 was 17.6 and 16.4 times higher than SL5 and SL1, respectively. And the chlorophyll content in the plant
leaf of populations SL1, SL2 and SL5 were decreased slightly ranging from 22.6 to 28.0. These results confirmed that the
SL1, SL2, SL3, ZC1 and SL5 populations had evolved moderate resistance to glyphosate. This is the first report for the
GR goosegrass populations confirmed in Chengdu, China.

Keywords: goosegrass, glyphosate, resistant, turf

et al. 1977). Each goosegrass plant can produce 140 000

1. Introduction
Goosegrass is an all-season prolific grass that is widely
distributed in the tropics, particularly in Asia, Africa, South
America, and the southern parts of North America (Holm
Received 4 August, 2014 Accepted 17 November, 2014
CHEN Jing-chao, E-mail: jcchenws@126.com;
Correspondence ZHANG Chao-xian, Tel: +86-10-62815937,
Fax: +86-10-62810289, E-mail: cxzhang@wssc.org.cn
*
These authors contributed equally to this study.
© 2015, CAAS. All rights reserved. Published by Elsevier Ltd.
doi: 10.1016/S2095-3119(14)60910-2
seeds, it is considered one of the five “world’s worst weeds”
(Chin and Raja 1979). In orchard and vegetable farm, the
goosegrass can compete for the nutrient with the crops. It
is also problematic for the golf and sports turf. The roadside
goosegrass in the turf farm can spread the seeds to the turf.
Infestations of the weeds during growth season affect the
turfgrass quality (Busey 2004).
Glyphosate is used extensively in orchards, non-till crop
production and none crop areas worldwide for post-emer-
gence control of annual and perennial broadleaf, grass and
sedge weeds since it was on the market (Wilcut and Askew
1999; Woodburn 2000; Duke and Powles 2008). Studies
have shown that glyphosate can control more than 96%
goosegrass at the dose of 840 g acid equivalent (ae) ha–1
920 CHEN Jing-chao et al. Journal of Integrative Agriculture 2015, 14(5): 919–925

(Corbett et al. 2004; Clewis et al. 2006; Thomas et al. 2006).
The increased use of glyphosate for weed control has
led to the selection of resistant weed populations. The first
glyphosate-resistant (GR) weed, rigid ryegrass (Lolium
rigidum), was reported in 1996 (Pratley et al. 1996). Since
then, 31 GR weed species including goosegrass (Eleusine
indica (L.) Gaertn.) have been reported (Tran et al. 1999).
GR goosegrass was first found in Malaysia, and subsequent-
ly, it has been reported in Taiwan of China (Yuan et al. 2005),
Philippines (Kaundun et al. 2008), Tennessee (Mueller et al.
2011) and Mississippi (Molin et al. 2013), and the resistant
rate was evolved to 12 (Lim and Jeremy 2000).
Glyphosate has been applied in China (mainly in
orchards) for 30 years. With the development of no-till
farming methods and the directed spray between crops,
the demand of glyphosate in China is growing. Studies
have shown that the glyphosate control efficiency is dif-
ferent for goosegrass in orchards and vegetable farms
in Guangdong Province, China (Yang et al. 2012). The
turf managers in Sichuan Province, China, used gly-
phosate to control the roadside weeds >3 times each
year. However, herbicides used for goosegrass control
are limited. In Guangzhou City, Guangdong Province,
glyphosate is a favored herbicide to control the weeds
in orchard and the roadside of the vegetable farm. After
years of glyphosate selection pressure, it became difficult
to control the goosegrass at the dose of 840–1 200 g
ae ha–1. The objective of this research was to determine the
sensitivity of some goosegrass populations collected from
those regions to verify the occurrence of GR goosegrass.
2. Results
2.1. Whole-plant bioassay
The responses of 12 suspected GR goosegrass populations
to glyphosate were different (Table 1). The fresh weight was
decreased with the increases of glyphosate doses. The
shoot growth of sensitive populations XD1 and BY3 was
completely inhibited by glyphosate, whereas the growth of
SL1, SL2 and SL5 was slightly reduced. At 14 days after
treatment (DAT), the death rate of XD1 and BY3 were all
100% at the dose of 1 680 g ae ha–1 (Fig. 1). In contrast,
the survival rates of both SL1 and SL2 populations were
over 70%, and SL5 was 97% at this dose. The populations
of SL6, WJ1 and PX1 were more sensitive than SL1. The
survival rate of WJ1 was 3.3% at the dose of 1 680 g ae
ha–1, while the population SL1 was 75%.
The GR50 (herbicide doses required to reduce 50% of
the test population’s growth) value calculated from the
fresh weight were similar for SL1 and SL5; both can reach
1 262.5 g ae ha–1. Compared to the sensitive population
XD1, the resistant index (RI) of the most resistance popu-
lation SL5 was 5.1. The RIs of the most populations from
Shuangliu County were more than 3.0, whereas, the RIs of
the populations in Guangzhou were around 2.3.
2.2. Chlorophyll content and shikimate assay
The chlorophyll content in the plant leaf of BY3, SL3, WJ1
and XD1 populations decreased rapidly after treatment,

Table 1 Parameter estimates (±SE) of regression equations for shoot fresh weight for the goosegrass populations
Population1) b2) C3) D4) GR505) R2 P RI6)
ZC1 2.2±0.46 0.69±0.31 6.2±0.44 892.6 (706.7–1 078.6) 0.95 <0.0001 2.6
BY1 2.4±0.38 0.07±0.24 7.1±0.37 778.6 (659.8–897.5) 0.97 <0.0001 2.2
BY2 3.3±0.97 0.16±0.20 4.6±0.32 790.6 (653.9–926.8) 0.93 <0.0001 2.3
HD1 2.9±0.61 0.28±0.36 9.7±0.56 819.2 (694.9–943.6) 0.96 <0.0001 2.4
BY3 2.1±0.30 0.40±0.20 10.1±0.50 346.2 (283.6–408.8) 0.96 <0.0001 –
SL1 4.7±0.39 0.32±0.16 10.4±0.22 1 262.5 (1 186.7–1 338.4) 0.99 <0.0001 4.5
SL2 4.0±0.88 0.30±0.16 3.8±0.22 1 168.7 (966.6–1370.8) 0.93 <0.0001 4.2
SL3 3.8±0.91 1.60±0.38 11.2±0.57 894.9 (791.0–998.9) 0.95 <0.0001 3.2
SL4 2.9±0.28 0.10±0.05 10.4±0.29 685.2 (646.5–724.0) 0.99 <0.0001 2.5
SL5 5.9±1.90 0.93±0.38 11.0±0.49 1 431.3 (1 241.9–1 620.6) 0.96 <0.0001 5.1
SL6 2.2±0.34 1.11±0.40 15.1±0.82 537.8 (443.2–632.3) 0.97 <0.0001 1.9
PX1 6.7±0.58 0.15±0.13 11.1±0.21 576.6 (539.6–613.5) 0.99 <0.0001 2.1
WJ1 7.5±3.20 0.26±0.04 11.1±0.42 504.8 (416.0–593.7) 0.98 <0.0001 1.8
XD1 2.2±0.14 -0.10±0.13 13.7±0.26 279.4 (261.5–297.3) 0.96 <0.0001 –
1)
BY3 was as the susceptible population for the Guangdong populations; XD1 was as the susceptible population for the Sichuan
populations. The same as below.
2)
b, the slope of the line at GR50.
3)
C, the lower limit.
4)
D, the upper limit.
5)
GR50, herbicide doses required to reduce 50% of the test population's growth.
6)
RI, resistance index.
–, the susceptible populations GR50 divided by itself.
 CHEN Jing-chao et al. Journal of Integrative Agriculture 2015, 14(5): 919–925 921

120

a ab a
100 abc
bcd
d cd
80
de d
Death rate (%)

de
60 ef

f
40 f

20
g

0
ZC1 BY1 BY2 HD1 BY3 SL1 SL2 SL3 SL4 SL5 SL6 PX1 WJ1 XD1
Population

Fig. 1 The death rates of 14 goosegrass populations at 14 days after treatment (DAT) at glyphosate dose of 1 680 g acid equivalent
(ae) ha–1. Vertical bars represent standard errors of the mean. Different lowercase letters mean significant difference by Duncan’s
new multiple range test (P<0.05).

whereas, it decreased slightly in the SL1, SL2 and SL5
populations, this consistent with the results of fresh weight.
The populations SL1 and SL2 had similar chlorophyll con-
tent in the plant leaf which can reach to 22.6 at the dose of
1 680 g ae ha–1 at 10 DAT, while the chlorophyll content in the
XD1 population was 1.5 (Table 2). The chlorophyll contents
in the populations ZC1, BY2 and HD1 were lower than that in
the control range from 16.4 to 16.9 at the dose of 1 680 g ae
ha–1 at 10 DAT, although the SPAD (soil and plant analyzer
development) value was higher than the population XD1.
Shikimate concentration increased slightly in populations
SL1, SL2 and SL5 (from 188.3 to 201.2 µg g–1, at 10 DAT).
However, shikimate was strongly accumulated in XD1, BY3,
PX1, and WJ1 populations and moderately accumulated in
other populations. Shikimate concentration in the plants
of population XD1 at 10 DAT was 3 228.2 µg g–1, which
was 17.6 and 16.4 times higher than that in SL5 and SL1
populations, respectively (Table 3).
3. Discussion
3.1. Whole-plant bioassay
Glyphosate completely (100%) controlled XD1 population at

Table 2 Chlorophyll content (mean±SE) in the leaves of different goosegrass populations following the application of the glyphosate
at 1 680 g acid equivalent (ae) ha–1 at different days after treatment (DAT)
Population 2 DAT 4 DAT 6 DAT 8 DAT 10 DAT
ZC1 26.0±2.7 c 25.9±1.3 bcd 23.8±2.0 b 19.9±2.8 b 16.7±0.2 c
BY1 27.7±0.3 abc 21.1±2.4 de 16.7±1.4 cd 5.6±2.1 cd 5.7±2.4 de
BY2 28.2±0.5 abc 23.9±0.8 cde 26.7±0.9 ab 22.0±1.7 ab 16.4±1.0 c
HD1 28.8±0.6 abc 27.9±0.6 abc 19.5±1.8 c 18.3±0.8 b 14.9±0.7 c
BY3 14.9±1.4 e 5.3±1.5 h 2.4±0.6 e 3.9±0.6 cd 0.3±0.3 f
SL1 28.7±2.4 abc 30.2±0.9 ab 26.8±0.3 ab 22.2±1.4 ab 22.6±3.1 b
SL2 30.4±0.6 ab 24.1±2.4 cde 25.8±2.8 ab 23.1±2.1 ab 24.8±1.7 ab
SL3 27.8±0.2 abc 19.8±2.6 e 14.1±2.8 d 7.0±1.6 c 4.4±0.8 def
SL4 29.0±0.8 abc 29.2±1.2 ab 19.0±0.9 c 17.9±2.1 b 12.0±0.7 c
SL5 31.8±0.3 a 32.3±1.0 a 30.2±1.0 a 26.9±1.1 a 28.0±2.6 a
SL6 28.1±0.3 abc 22.0±1.9 de 13.9±2.8 d 6.8±2.8 c 6.0±1.6 d
PX1 13.5±1.4 e 7.2±1.8 g 6.3±1.2 e 6.0±0.8 cd 0.9±0.6 e
WJ1 20.1±1.7 d 12.2±1.1 f 12.9±1.1 d 6.7±1.0 c 1.4±1.1 def
XD1 27.4±0.3 bc 11.1±0.6 fg 6.2±1.0 e 1.1±0.3 d 1.5±0.5 def
Different lowercase letters in the same column meant significant difference by Duncan’s new multiple range test (P<0.05). The same as
below.
922 CHEN Jing-chao et al. Journal of Integrative Agriculture 2015, 14(5): 919–925

Table 3 Shikimate accumulation (mean±SE) in shoots of different goosegrass populations following the application of the glyphosate
at 1 680 g ae ha–1 at different DAT
Populations 2 DAT 4 DAT 6 DAT 8 DAT 10 DAT
ZC1 185.9±3.8 d 353.6±39.0 e 386.8±10.6 e 241.0±3.9 gh 320.5±9.7 h
BY1 158.1±0.9 d 255.3±18.9 fg 266.5±13.0 efg 367.5±8.1 fgh 858.1±8.1 e
BY2 150.8±2.3 d 238.1±11.7 fg 252.6±12.1 efg 273.9±11.2 i 333.3±16.1 h
HD1 229.7±41.5 cd 245.0±37.1 fg 373.1±35.4 ef 480.2±58.6 f 553.2±40.2 fg
BY3 361.8±7.0 a 1 207.9±62.4 c 1 735.9±144.5 b 3 787.2±124.7 b 3 787.2±124.7 b
SL1 196.7±23.1 d 238.6±15.4 fg 223.1±13.0 g 410.1±35.9 fgh 196.3±41.1 i
SL2 220.3±19.8 cd 201.3±21.8 g 188.3±18.7 g 428.6±18.9 fg 201.2±20.1 i
SL3 150.0±9.3 d 233.9±8.0 fg 247.8±38.0 fg 350.4±6.3 fgh 520.1±6.7 g
SL4 430.1±33.5 b 581.3±28.5 d 646.2±34.9 d 893.3±63.6 e 658.6±51.2 f
SL5 167.9±3.6 d 167.9±4.6 g 195.7±3.7 g 155.1±2.9 j 183.3±7.8 i
SL6 183.5±3.9 d 234.2±2.9 fg 294.0±8.3 efg 345.3±11.9 fgh 496.2±1.3 g
PX1 278.6±7.6 c 314.3±4.3 ef 1 034.6±29.0 c 1 026.9±26.9 d 1 026.9±26.9 d
WJ1 696.8±6.5 a 1 105.1±31.7 b 1 043.6±23.7 c 1 717.9±31.1 c 2 045.6±62.0 c
XD1 1 309.6±49.4 b 2 149.6±71.9 a 3 155.1±143.2 a 3 228.2±148.1 a 3 228.2±148.1 a

the dose of 1 680 g ae ha–1, which is consistent with the stud-
ies conducted in cotton and soybean (Corbett et al. 2004;
Clewis et al. 2006; Thomas et al. 2006). Previous studies
showed that 45% of GR population found in Tennessee
could be controlled by 840 g ae ha–1 glyphosate (Mueller
et al. 2011), in our research, glyphosate at the dose of 1 680
g ae ha–1 had no significant effect on SL5 population, and
only controlled 25% for SL1 population.
According to the dose-response assay, the SL1, SL2,
SL3, SL5 and ZC1 populations were moderately resistant to
glyphosate. The RI of population SL5 was 5.1, which was
similar to the population found in Malaysia (Baerson et al.
2002) and more resistant than the GR populations in Guang-
zhou City. The strong resistant population was also found
in USA with a RI of 12 (Mueller et al. 2011; Heap 2014). A
3–10-fold levels of glyphosate resistance has been reported
in other plant species including Conyza canadensis (Dinelli
et al. 2008), Lolium multiliorum (Perez-Jones et al. 2007),
and Lolium rigidum (Wakelin et al. 2004). Dose-response
studies indicated that the populations in Chengdu were
more tolerance to glyphosate than those in Guangzhou.
To date, most of the GR weeds were found under a high
glyphosate selection pressure. In Chengdu and Guang-
zhou, the goosegrass is a terrible grass in the roadside of
the vegetable and turf farm. The glyphosate is an effective
and economic herbicide to control them. The frequently
using glyphosate and limiting other herbicide to control the
goosegrass is one of the reason for the resistance evolved.
3.2. Chlorophyll content and shikimate assay
Chlorosis, necrosis and atrophy, of the plant, developed
several days after application of glyphosate (Ruiter et al.
1999). These effects were correlated to the dose and days of
glyphosate treatment. Yanniccari et al. (2012) reported that
the chlorophyll content in the glyphosate-resistant Lolium
perenne L. was not altered. We showed that the chlorophyll
content in the populations SL1, SL2, SL4 and SL5 was sig-
nificantly higher than that in the sensitive population XD1
at 10 DAT. And this result also found in ZC1, BY2 and HD1
populations in Guangzhou City.
Accumulation of shikimate in glyphosate-treated plants
indicates that the herbicide affects the activity of 5-enolpy-
ruvylshikimate-3-phosphate synthase (EPSPS) (Mueller
et al. 2003). Many GR weeds had lower accumulation of
shikimate than the susceptive populations (Perez-Jones
et al. 2004, 2005; Ribeiro et al. 2008). Our results showed
that the shikimate in the four populations from Guangzhou
was below 860 µg g–1, and it was lower than 660 µg g–1
in six populations from Chengdu. Mueller et al. (2011)
found that shikimate in the plants of glyphosate-resistant
goosegrass was below 850 µg g–1 while shikimate in the
sensitive populations could reach 3 500 µg g–1 at 6 DAT with
a dose of 840 g ae ha–1. High accumulation of shikimate in
the XD1 population indicated susceptibility to glyphosate,
while low accumulation of shikimate in the other popula-
tions indicated resistance to glyphosate. Glyphosate has
been applied in China (mainly in orchards) for 30 years.
Glyphosate resistant Conyza canadensis has been found
in Southern China (Song et al. 2011). The diversity of the
farming system in China leads to the spread of seeds with
glyphosate resistance. Glyphosate should be combined
with herbicides exerting different modes of action to reduce
selection pressure for additional resistant populations. The
knowledge obtained in this study will provide basis for GR
weed control in China.
4. Conclusion
The dose-response assay, chlorophyll content and the
 CHEN Jing-chao et al. Journal of Integrative Agriculture 2015, 14(5): 919–925 923

shikimate assay indicated that moderate resistance to 5.2. Greenhouse experiment

glyphosate were present in the populations of SL1, SL2,
SL3 and SL5 in turf farm of Chengdu City, China, and ZC1
in the vegetable farm in Guangzhou City, China. The RI of
SL5 can reach to 5.1, and the RI for the other populations
was range from 1.8 to 2.5.
5. Materials and methods
5.1. Plant culture
Seeds of putative GR goosegrass populations were
collected during 2010–2014 from different plantations in
Chengdu, Sichuan Province and Guangzhou, Guangdong
Province, China (Table 4). These sites had been intensive-
ly treated with glyphosate for more than three times per
year at the dose of 840 g ae ha–1. Two known susceptible
goosegrass populations (XD1 and BY3) were included as
control obtained from the field in Chengdu and Guangzhou,
respectively.
Seeds from all the populations were sowed in plastic pots
(250 mL) containing a 1:1 (v:v) mixture of peat (organic fer-
tilizer, Kai Yin LLC, Beijing) and loam. The plastic pots were
kept in the greenhouse ((27/20±2)°C day/night temperature)
with a 14-h photoperiod. Daily watering was applied to
maintain good plant growth. Each pot has eight plants.
Seven doses (0, 210, 420, 840, 1 680, 3 360 and 6 720 g ae
ha–1) of glyphosate (Roundup, isopropylamine salt of gly-
phosate, 410 g ae L–1, Monsanto Company, St. Louis, MO)
were applied with a sprayer mounted with a flat-fan nozzles
calibrated to deliver 280 L ha–1 of spray volume when the
plants reached five-leaf stage. 14 days after treatment, plant
survival was evaluated by recording the number of controlled
plants on every replicate. Then, the shoot biomass in each
pot was harvested and the fresh weight was measured with
four replicates.
5.3. Chlorophyll content and shikimate bioassay
Seedlings were grown as previously described. The five-leaf
stage plant treated with the glyphosate at a dose of 1 680 g
ae ha–1. Chlorophyll content was recorded with a portable
chlorophyll meter (SPAD-502, Minolta, Japan) every 48 h
until 10 DAT. The value of chlorophyll per plant was the
average of three measurements taken on the middle 3rd of
the penultimate expanded leaf (the 3rd leaf) with 30 repli-
cates per treatment. After detecting the chlorophyll content,
tissue samples from four populations of goosegrass were
collected by clipping the tissues above the soil surface. The
plant tissue was finely ground in liquid nitrogen by mortar

Table 4 The information of the goosegrass seeds for this study

Collect data
Location Habitat Latitude and longitude
(mon/year)
Zengcheng, Guangzhou City, Guangdong (ZC1) Vegetable farm 23°09´29.47´´N 1/2014
113°48´19.76´´E
Baiyun, Guangzhou City, Guangdong (BY1) Vegetable farm 23°12´28.57´´N 1/2014
113°12´04.73´´E
Baiyun, Guangzhou City, Guangdong (BY2) Vegetable farm 23°12´36.03´´N 1/2014
113°12´06.50´´E
Huadu, Guangzhou City, Guangdong (HD1) Vegetable farm No data 9/2010
Baiyun, Guangzhou City, Guangdong (BY3) Wasteland 23°12´37.34´´N 11/2013
113°12´02.14´´E
Dongsheng, Shuangliu County, Chengdu City, Sichuan Roadside of the turf 30°32´57´´N 11/2012
(SL1) 103°55´55´´E
Huangshui, Shuangliu County, Chengdu City, Sichuan Roadside of the turf 30°31´13´´N 11/2012
(SL2) 103°50´25´´E
Dongsheng, Shuangliu County, Chengdu City, Sichuan Roadside of the turf 30°32´48.7´´N 11/2013
(SL3) 103°54´53.2´´E
Jinqiao, Shuangliu County, Chengdu City, Sichuan (SL4) Roadside of the turf 30°32´14´´N 11/2012
103°51´13´´E
Dongsheng, Shuangliu County, Chengdu City, Sichuan Roadside of the turf 30°32´48.7´´N 11/2013
(SL5) 103°54´53.2´´E
Dongsheng, Shuangliu County, Chengdu City, Sichuan Roadside of the turf 30°32´48.7´´N 11/2013
(SL6) 103°54´53.2´´E
Youai, Pi County, Chengdu City, Sichuan (PX1) Nursery 30°50´39.43´´N 11/2013
103°48´51.22´´E
Wenjiang, Chengdu City, Sichuan (WJ1) Nursery No data 11/2013
Xindu, Chengdu City, Sichuan (XD1) Wasteland 30°46´54´´N 11/2012
104°12´39´´E
924 CHEN Jing-chao et al. Journal of Integrative Agriculture 2015, 14(5): 919–925
and pestle. After grinding, the tissue was weighted quickly.
Approximately 0.5 g (depending upon the amount of shi-
kimate present in the extract) was placed into 2-mL screw-
cap polypropylene centrifuge tubes and 1 mL 0.25 mol L–1
HCl was added. The centrifuge tubes were vortexed for 5 s
to ensure that the tissue was in solution. The extract was
centrifuged at 25 000×g for 15 min at 4°C and 0.2 mL super-
natant was collected to a 10-mL screw-cap polypropylene
centrifuge tubes for the shikimate assay.
Shikimate was determined according to the previously
described method with some modifications (Gaitonde and
Gordon 1958). Briefly, the supernatant (0.2 mL) was mixed
with 2 mL periodic acid (1%) to oxidize shikimate at 25°C.
After 3 h, the sample was mixed with 2 mL 1 mol L–1 NaOH,
followed by addition of 1.2 mL of 0.1 mol L–1 glycine. The
optical density at 380 nm was measured immediately after
thorough mixing.
A shikimate standard curve was developed by adding
known amount of shikimate (Sigma-Aldrich, St. Louis, MO;
99% purity) to the tubes. The shikimate was diluted from
10 to 250 µg mL–1 using 0.25 mol L–1 HCl.
5.4. Statistical analysis
The experiments were arranged in a completely randomized
design and repeated twice. Log-logistic regression analy-
sis was conducted to evaluate the data of fresh biomass
using the following model proposed previously (Seefeldt
et al. 1995).
Y=C+(D–C)/[1+(X/GR50)b]
In this equation, Y represents the fresh or dry weight at
the herbicide dose of X; C is the lower limit for Y; D is the
upper limit for Y; GR50 is the glyphosate rate that results in
a 50% reduction in fresh weight; b is the slope of the curve
around GR50. Pseudo R2 values were calculated to assess
the goodness of fit for the appropriate equation. The re-
gression analyses were conducted using Sigma Plot 12.0
(Systat software, San Jose, CA). The level of resistance
was determined by calculating the ratio of the GR50 of the
resistant population to the GR50 of the susceptible popula-
tion. The shikimate concentration data, chlorophyll content
and survival rate were analyzed using SPSS 13.0 (SPSS,
Chicago, USA). Duncan’s multiple range test was used to
assess the effects of the days after application.
Acknowledgements
This research was funded by the Special Fund for Agro-Sci-
entific Research in the Public Interest, China (201303022).
References
Baerson S R, Rodriguez D J, Tran M, Feng Y, Biest N A, Dill
G M. 2002. Glyphosate-resistant goosegrass. Identification
of a mutation in the target enzyme 5-enolpyruvylshikimate-
3-phosphate synthase. Plant Physiology, 129, 1265–1275.
Busey P. 2004. Goosegrass (Eleusine indica) control with
foramsulfuron in bermudagrass (Cynodon spp.) turf. Weed
Technology, 18, 634–640.
Chin H F, Raja H. 1979. Ecology and physiology of Eleusine
indica seeds. In: Proceedings of the 17th Asia-Pacific Weed
Science Society Conference. Sydney. pp. 313–315.
Clewis S B, Wilcut J W, Porterfield D. 2006. Weed management
with S-metolachlor and glyphosate mixtures in glyphosate-
resistant strip- and conventional-tillage cotton (Gossypium
hirsutum L.). Weed Technology, 20, 232–241.
Corbett J L, Askew S D, Thomas W E, Wilcut J W. 2004. Weed
efficacy evaluations of bromoxynil, glufosinate, glyphosate
and pyrithiobac and sulfosate. Weed Technololgy, 18,
443–453.
Dinelli G, Marotti I, Bonetti A, Catizone P, Urbano J M, Barnes
J. 2008. Physiological and molecular basis of glyphosate
resistance in Conyza bonariensis biotypes from Spain.
Weed Research, 48, 257–265.
Duke S O, Powles S B. 2008. Glyphosate: a once-in-a-century
herbicide. Pest Management Science, 64, 319–325.
Gaitonde M K, Gordon M W. 1958. A microchemical method for
the detection and determination of shikimic acid. Journal of
Biological Chemistry, 230, 1043–1050.
Heap I. 2014. The international survey of herbicide resistant
weeds. [2014-10-22]. http://www.weedscience.com
Holm L G, Pluncknett D L, Pancho J V, Herberger J P. 1977.
Distribution and Biology. In: The World’s Worst Weeds. The
University Press of Hawaii, Honolulu. pp. 47–53.
Kaundun S S, Zelaya I A, Dale R P, Lycett A J, Carter P,
Sharples K R, McIndoe E. 2008. Importance of the P106S
target-site mutation in conferring resistance to glyphosate
in a goosegrass (Eleusine indica) population from the
Philippines. Weed Science, 56, 637–646.
Lim J L, Jeremy N. 2000. A first report of glyphosate-resistant
goosegrass (Eleusine indica (L.) Gaertn) in Malaysia. Pest
Management Science, 56, 336–339.
Molin W T, Wright A A, Nandula V K. 2013. Glyphosate-resistant
goosegrass from Mississippi. Agronomy, 3, 474–487.
Mueller T C, Barnett K A, Brosnan J T, Steckel L E. 2011.
Glyphosate-resistant goosegrass (Eleusine indica)
confirmed in Tennessee. Weed Science, 59, 562–566.
Mueller T C, Massey J M, Hayes C L, Main C L, Stewart C N.
2003. Shikimate accumulates in both glyphosate-sensitive
and glyphosate-resistant horseweed (Conyza Canadensis
L. Cronq.). Journal of Agriculture and Food Chemistry, 51,
680–684.
Perez-Jones A, Park K W, Colquhoun J, Mallory-Smith C,
Shaner D. 2005. Identification of glyphosate-resistant italian
ryegrass (Lolium multiflorum) in Oregon. Weed Science,
53, 775–779.
Perez-Jones A, Park K W, Polge N, Colquhoun J, Mallory-Smith
C A. 2007. Investigating the mechanisms of glyphosate
resistance in Lolium multiflorum. Planta, 226, 395–404.
Pratley J E, Baines P, Eberbach P, Incerti M, Broster J. 1996.
 CHEN Jing-chao et al. Journal of Integrative Agriculture 2015, 14(5): 919–925
Glyphosate resistance in annual ryegrass. In: Proceedings
of the 11th Annual Conference of the Grassland Society of
New South Wales. The Grassl Society of NSW, Wagga.
p. 122.
Ribeiro D N, Gil D, Cruz-Hipolito H E, Ruiz-Santaella J P,
Christoffoleti P J, Vidal R A, de Prado R. 2008. Rapid assays
for detection of glyphosate-resistant Lolium spp. Journal of
Plant Diseases and Protection, 21, 95–99.
Ruiter H, Meinen E, Ribot S A, Uffing A J M. 1999. A new
approach for monitoring the influence of adjuvants on
glyphosate transfer from drop deposit to sink leaf. In:
Proceedings of the International Symposium on Crop
Protection. Belgium. pp. 679–685.
Seefeldt S S, Jensen J E, Fuerst E P. 1995. Log-logistic analysis
of herbicide dose-response relationship. Weed Technology,
9, 218–227.
Song X L, Wu J J, Zhang H J, Qiang S. 2011. Occurrence
of glyphosate-resistant horseweed (Conyza canadensis)
population in China. Agricultural Sciences in China, 10,
1049–1055.
Thomas W E, Britton T T, Clewis S B, Askew S D, Wilcut J W.
2006. Glyphosate-resistant cotton (Gossypium hirsutum)
response and weed management with trifloxysulfuron,
glyphosate, prometryn and MSMA. Weed Technology,
925
20, 6–13.
Tran M, Baerson S, Brinker R. 1999. Characterization of
glyphosate resistant Eleusine indica biotypes from Malaysia.
In: Proceedings of the 17th Asia-Pacific Weed Science
Society Conference. Bangkok. pp. 527–536.
Wakelin A M, Lorraine-Colwill D F, Preston C. 2004. Glyphosate
resistance in four different populations of Lolium rigidum
is associated with reduced translocation of glyphosate to
meristematic zones. Weed Research, 44, 453–459.
Wilcut J W, Askew S D. 1999. In: Ruberson J R, Marcel D,
eds., Handbook of Pest Management. CRC Press, Boca
Raton. pp. 627–661.
Woodburn A T. 2000. Glyphosate: production, pricing and use
worldwide. Pest Management Science, 56, 309–312.
Yang C H, Tian X S, Feng L, Yue M F. 2012. Resistance of
Eleusine indica to glyphosate. Scientia Agriculture Sinica,
45, 2093–2098. (in Chinese)
Yanniccari M, Istilart C, Giménez D O, Castro A M. 2012.
Glyphosate resistance in perennial ryegrass (Lolium
perenne L.) from Argentina. Crop Protection, 32, 12–16.
Yuan C I, Hsieh Y C, Chiang M Y. 2005. Glyphosate-resistant
goosegrass (Eleusine indica (L.) Gaertn.) in Taiwan: Dose-
response and enzyme activity. Plant Protection Bulletin, 47,
143–154. (in Chinese)
(Managing editor ZHANG Juan)