Lyu, Qinghao, Jiang, Bo, He, Pengfei, Bai, Shuang, Sun, Xiyu, Liu, Jinling, and Li, Lingxu
Backgroud: Echinochloa curs-galli (barnyardgrass) is the most prominent weed in rice fields, and its herbicide resistance seriously threatens rice yields. Penoxsulam failed to control an E. crus-galli var. zelayensis population (R) in field. However, the resistance level, cross-resistance pattern and resistance mechanism is not characterized.The whole plant method was employed to determine the resistance level to penoxsulam and the cross-resistance pattern in R. Enzyme activity, gene cloning and sequencing, quantative real-time quantitative PCR (qRT-PCR) were conducted to determine the target-site resistance mechanisms. Metabolism enzyme inhibitors pretreatment, RNA sequencing (RNA-Seq) and qRT-PCR validation were performed to identify the penoxsulam-resistance related metabolism genes.Here we identified an E. crus-galli var. zelayensis population (R) highly resistant to acetolactate synthase (ALS) inhibitor penoxsulam, cross-resistant to bispyribac-sodium and pyribenzoxim, and multiple-resistant to auxin mimics florpyrauxifen, but susceptible to acetyl coenzyme A carboxylase (ACCase) cyhalofop-butyl and the 4-hydroxyphenylpyruvate dioxygenase (HPPD) inhibitor benzobicyclon. Molecular investigation indicated that the penoxsulam resistance was unrelated to amino acid substitution or overexpression of the target enzyme. The metabolism inhibitors were employed to investigate the non-target site resistance mechanism consequently. P450s inhibitor malathion or GSTs inhibitor 4-chloro-7-nitro-2,1,3-benzoxadiazole (NBD-Cl) both significantly decreased the resistance level to penoxsulam in population R. RNA-seq and qRT-PCR validation indicated that 15 P450s and 5 glutathione S-transferase (GST) genes were upregulated by penoxsulam in R plants.We present an E. crus-galli var. zelayensis population with resistance to penoxsulam and multiple-resistance to auxin mimics. Enhanced herbicide metabolism via P450s and GSTs played a vital role in penoxsulam resistance.Methods: Echinochloa curs-galli (barnyardgrass) is the most prominent weed in rice fields, and its herbicide resistance seriously threatens rice yields. Penoxsulam failed to control an E. crus-galli var. zelayensis population (R) in field. However, the resistance level, cross-resistance pattern and resistance mechanism is not characterized.The whole plant method was employed to determine the resistance level to penoxsulam and the cross-resistance pattern in R. Enzyme activity, gene cloning and sequencing, quantative real-time quantitative PCR (qRT-PCR) were conducted to determine the target-site resistance mechanisms. Metabolism enzyme inhibitors pretreatment, RNA sequencing (RNA-Seq) and qRT-PCR validation were performed to identify the penoxsulam-resistance related metabolism genes.Here we identified an E. crus-galli var. zelayensis population (R) highly resistant to acetolactate synthase (ALS) inhibitor penoxsulam, cross-resistant to bispyribac-sodium and pyribenzoxim, and multiple-resistant to auxin mimics florpyrauxifen, but susceptible to acetyl coenzyme A carboxylase (ACCase) cyhalofop-butyl and the 4-hydroxyphenylpyruvate dioxygenase (HPPD) inhibitor benzobicyclon. Molecular investigation indicated that the penoxsulam resistance was unrelated to amino acid substitution or overexpression of the target enzyme. The metabolism inhibitors were employed to investigate the non-target site resistance mechanism consequently. P450s inhibitor malathion or GSTs inhibitor 4-chloro-7-nitro-2,1,3-benzoxadiazole (NBD-Cl) both significantly decreased the resistance level to penoxsulam in population R. RNA-seq and qRT-PCR validation indicated that 15 P450s and 5 glutathione S-transferase (GST) genes were upregulated by penoxsulam in R plants.We present an E. crus-galli var. zelayensis population with resistance to penoxsulam and multiple-resistance to auxin mimics. Enhanced herbicide metabolism via P450s and GSTs played a vital role in penoxsulam resistance.Results: Echinochloa curs-galli (barnyardgrass) is the most prominent weed in rice fields, and its herbicide resistance seriously threatens rice yields. Penoxsulam failed to control an E. crus-galli var. zelayensis population (R) in field. However, the resistance level, cross-resistance pattern and resistance mechanism is not characterized.The whole plant method was employed to determine the resistance level to penoxsulam and the cross-resistance pattern in R. Enzyme activity, gene cloning and sequencing, quantative real-time quantitative PCR (qRT-PCR) were conducted to determine the target-site resistance mechanisms. Metabolism enzyme inhibitors pretreatment, RNA sequencing (RNA-Seq) and qRT-PCR validation were performed to identify the penoxsulam-resistance related metabolism genes.Here we identified an E. crus-galli var. zelayensis population (R) highly resistant to acetolactate synthase (ALS) inhibitor penoxsulam, cross-resistant to bispyribac-sodium and pyribenzoxim, and multiple-resistant to auxin mimics florpyrauxifen, but susceptible to acetyl coenzyme A carboxylase (ACCase) cyhalofop-butyl and the 4-hydroxyphenylpyruvate dioxygenase (HPPD) inhibitor benzobicyclon. Molecular investigation indicated that the penoxsulam resistance was unrelated to amino acid substitution or overexpression of the target enzyme. The metabolism inhibitors were employed to investigate the non-target site resistance mechanism consequently. P450s inhibitor malathion or GSTs inhibitor 4-chloro-7-nitro-2,1,3-benzoxadiazole (NBD-Cl) both significantly decreased the resistance level to penoxsulam in population R. RNA-seq and qRT-PCR validation indicated that 15 P450s and 5 glutathione S-transferase (GST) genes were upregulated by penoxsulam in R plants.We present an E. crus-galli var. zelayensis population with resistance to penoxsulam and multiple-resistance to auxin mimics. Enhanced herbicide metabolism via P450s and GSTs played a vital role in penoxsulam resistance.Conclusion: Echinochloa curs-galli (barnyardgrass) is the most prominent weed in rice fields, and its herbicide resistance seriously threatens rice yields. Penoxsulam failed to control an E. crus-galli var. zelayensis population (R) in field. However, the resistance level, cross-resistance pattern and resistance mechanism is not characterized.The whole plant method was employed to determine the resistance level to penoxsulam and the cross-resistance pattern in R. Enzyme activity, gene cloning and sequencing, quantative real-time quantitative PCR (qRT-PCR) were conducted to determine the target-site resistance mechanisms. Metabolism enzyme inhibitors pretreatment, RNA sequencing (RNA-Seq) and qRT-PCR validation were performed to identify the penoxsulam-resistance related metabolism genes.Here we identified an E. crus-galli var. zelayensis population (R) highly resistant to acetolactate synthase (ALS) inhibitor penoxsulam, cross-resistant to bispyribac-sodium and pyribenzoxim, and multiple-resistant to auxin mimics florpyrauxifen, but susceptible to acetyl coenzyme A carboxylase (ACCase) cyhalofop-butyl and the 4-hydroxyphenylpyruvate dioxygenase (HPPD) inhibitor benzobicyclon. Molecular investigation indicated that the penoxsulam resistance was unrelated to amino acid substitution or overexpression of the target enzyme. The metabolism inhibitors were employed to investigate the non-target site resistance mechanism consequently. P450s inhibitor malathion or GSTs inhibitor 4-chloro-7-nitro-2,1,3-benzoxadiazole (NBD-Cl) both significantly decreased the resistance level to penoxsulam in population R. RNA-seq and qRT-PCR validation indicated that 15 P450s and 5 glutathione S-transferase (GST) genes were upregulated by penoxsulam in R plants.We present an E. crus-galli var. zelayensis population with resistance to penoxsulam and multiple-resistance to auxin mimics. Enhanced herbicide metabolism via P450s and GSTs played a vital role in penoxsulam resistance. [ABSTRACT FROM AUTHOR]