Scientia Agricultura Sinica ›› 2023, Vol. 56 ›› Issue (14): 2713-2723.doi: 10.3864/j.issn.0578-1752.2023.14.007

• PLANT PROTECTION • Previous Articles     Next Articles

Resistance Mechanism of Barnyard Grass (Echinochloa crus-galli) to Penoxsulam and Screening Herbicides for Its Control in Rice Fields

ZHANG LiYa1(), LI Qi1(), SHI ShanShan1, MA YuMeng1, LIU YaQi1, ZHAO ChaoWei1, WANG HeRu2, CAO HaiQun1, LIAO Min1, ZHAO Ning1()   

  1. 1 School of Plant Protection, Anhui Agricultural University/Anhui Province Key Laboratory of Crop Integrated Pest Management, Hefei 230036
    2 Tianchang Agricultural Science and Technology Center Plant Protection Station, Tianchang 239300, Anhui
  • Received:2023-04-11 Accepted:2023-05-14 Online:2023-07-16 Published:2023-07-21
  • Contact: ZHAO Ning

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Abstract:

【Objective】Barnyard grass (Echinochloa crus-galli) is one of the main malignant weeds in rice fields in China. Acetolactate synthase (ALS) inhibitors, such as penoxsulam, are the main herbicides for controlling E. crus-galli in rice fields. Previously, our research team identified a suspected penoxsulam-resistant E. crus-galli population, AHTC-01, in the main rice production area of Tianchang City, Anhui Province, China. The objective of this study is to clarify its resistance levels to major herbicides, investigate the possible resistance molecular mechanisms, and to provide a theoretical basis for the effective control of resistant E. crus-galli and the delay of further development of herbicide resistance.【Method】Using the greenhouse potting method, the resistance levels of AHTC-01 to penoxsulam and its resistance patterns to different herbicides were determined at the whole-plant level. The target resistance molecular mechanism was explored through target gene sequencing and real-time quantitative PCR (RT-qPCR) analysis.【Result】Compared with the susceptible E. crus-galli population AHFY-01, the suspected resistant population AHTC-01 had developed high-level resistance to penoxsulam, with a resistance index (RI) of 620. Analysis of the target-site based resistance mechanisms showed that the ALS2 gene copy in the AHTC-01 population had a mutation from tryptophan (Trp) to leucine (Leu) at codon position 574, with a population mutation frequency of 100%. At 12 h after penoxsulam treatment, the relative expression level of ALS in the resistant E. crus-galli population AHTC-01 was 2.26 times of that in the susceptible E. crus-galli population AHFY-01. AHTC-01 also exhibited varying levels of cross-resistance to three other ALS inhibitors, bispyribac sodium, pyribenzoxim, and imazamox, with RIs of 8.24, 13.36, and 20.36, respectively. However, it remained susceptible to other herbicides with different modes of action (MOAs), including the acetyl-CoA carboxylase (ACCase) inhibitors cyhalofop-butyl, fenoxaprop-P-ethyl, and clethodim, the 4-hydroxyphenylpyruvate dioxygenase (HPPD) inhibitor tripyrasulfone, and the synthetic auxin mimic florpyrauxifen-benzyl.【Conclusion】Mutation of the ALS2 at amino acid position 574 and ALS overexpression are one of the main reasons for the resistance of E. crus-galli population AHTC-01 to penoxsulam, which also confers cross-resistance to different ALS inhibitors. In actual agricultural production, effective control of this type of resistant E. crus-galli can still be achieved by rotating the use of other herbicides with different MOAs.

Key words: Echinochloa crus-galli, penoxsulam, acetolactate synthase (ALS), gene mutation, gene overexpression

Table 1

Detailed information of the herbicide treatments applied in the whole-plant dose-response experiments"

除草剂
Herbicide		 系列剂量 Series of doses (g a.i.·hm-2)
抗性种群AHTC-01
Resistant population AHTC-01		 敏感种群AHFY-01
Susceptible population AHFY-01
五氟磺草胺Penoxsulam 0, 60, 120, 240, 480, 960, 1920 0, 0.04, 0.12, 0.37, 1.11, 3.33, 10
双草醚Bispyribac-sodium 0, 45, 90, 180, 360, 720, 1440 0, 5.60, 11.30, 22.50, 45, 90, 180
嘧啶肟草醚Pyribenzoxim 0, 37.50, 75, 150, 300, 600, 1200 0, 4.70, 9.40, 18.80, 37.50, 75, 150
甲氧咪草烟Imazamox 0, 24, 48, 96, 192, 384, 768 0, 3, 6, 12, 24, 48, 96
氰氟草酯Cyhalofop-butyl 0, 0.49, 1.48, 4.44, 13.33, 40, 120 0, 0.49, 1.48, 4.44, 13.33, 40, 120
精噁唑禾草灵Fenoxaprop-P-ethyl 0, 0.10, 0.50, 2.50, 12.40, 62.10, 310.50 0, 0.02, 0.10, 0.50, 2.50, 12.40, 62.10
烯草酮Clethodim 0, 0.03, 0.17, 0.86, 4.32, 21.60, 108 0, 0.03, 0.17, 0.86, 4.32, 21.60, 108
三唑磺草酮Tripyrasulfone 0, 0.04, 0.22, 1.08, 5.40, 27, 135 0, 0.04, 0.22, 1.08, 5.40, 27, 135
氯氟吡啶酯Florpyrauxifen-benzyl 0, 0.15, 0.44, 1.33, 4, 12, 36 0, 0.15, 0.44, 1.33, 4, 12, 36

Table 2

Resistance level of different E. crus-galli populations to penoxsulam"

种群
Population		 整株剂量响应曲线参数 Parameters for whole-plant dose-response curves GR50
(g a.i.·hm-2)		 抗性倍数
Resistance index
C D b R2
AHTC-01 26.90 (9.20) 93.25 (16.53) 5.72 (6.23) 0.91 155.00 (53.80) 620
AHFY-01 32.89 (3.82) 102.17 (7.65) 1.89 (0.67) 0.99 0.25 (0.06) -

Fig. 1

Trp574 codons of susceptible (AHFY-01, left) and resistant (AHTC-01, right) populations of E. crus-galli ALS2"

Table 3

Alignment of ALS2 copies cloned from the susceptible and resistant populations of E. crus-galli"

物种/种群
Species/Population		 各氨基酸的位置和碱基及相应氨基酸序列
The amino acid position, relative sequence of nucleotide and derived amino acid
122 197 205 376 377 574 653 654
拟南芥
A. thaliana		 GCA CCT GCT GAT CGT TGG AGT GGT
Ala Pro Ala Asp Arg Trp Ser Gly
AHFY-01 GCC CCC GCC GAT CGT TGG AGC GGT
Ala Pro Ala Asp Arg Trp Ser Gly
AHTC-01 GCC CCC GCC GAT CGT TTG AGC GGT
Ala Pro Ala Asp Arg Leu Ser Gly

Fig. 2

Relative expression ratio of ALS in the resistant relative to susceptible population of E. crus-galli before and after penoxsulam treatment"

Fig. 3

Dose-response curves for relative fresh weights of the resistant (AHTC-01) and susceptible (AHFY-01) plants of E. crus-galli treated with different herbicides"

Table 4

Resistance levels of the resistant E. crus-galli population AHTC-01 to other herbicides"

除草剂
Herbicide		 GR50 (g a.i.·hm-2) 抗性倍数
Resistance index
AHTC-01 AHFY-01
双草醚Bispyribac-sodium 190.09 (8.30) 23.07 (2.56) 8.24
嘧啶肟草醚Pyribenzoxim 207.05 (45.02) 15.50 (6.68) 13.36
甲氧咪草烟Imazamox 197.32 (55.85) 9.69 (2.18) 20.36
氰氟草酯Cyhalofop-butyl 7.79 (0.45) 5.74 (0.35) 1.37
精噁唑禾草灵Fenoxaprop-P-ethyl 3.44 (0.27) 2.06 (0.30) 1.67
烯草酮Clethodim 4.75 (2.20) 3.27 (1.55) 1.45
三唑磺草酮Tripyrasulfone 5.27 (1.80) 5.04 (2.58) 1.05
氯氟吡啶酯Florpyrauxifen-benzyl 1.04 (0.12) 1.31 (0.49) 0.80
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