15DB01水稻对几种除草剂的抗性及其相关抗性基因的克隆
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摘要
以AHAS为靶标的除草剂是目前生产中应用较为广泛的除草剂,包括咪唑啉酮、嘧啶水杨酸、磺酰脲、三唑嘧啶以及磺酰胺等不同化合物类型.在前期工作中筛选到一株咪唑啉酮类除草剂"普施特"的抗性水稻"选一".本文报道了"选一"后代(15DB01)对我国农业生产中较为常见的6种以AHAS为靶标的除草剂的抗性实验结果和相关抗性基因的鉴定结果.研究结果表明,15DB01对咪唑啉酮类甲氧咪草烟、两种嘧啶水杨酸类双草醚和嘧啶肟草醚、两种磺酰脲类甲基二磺隆和烟嘧磺隆都有不同程度的抗性,但对甲氧咪草烟的抗性显著大于比对其他几种除草剂的抗性.实验中并未发现15DB01水稻与对照13lb05在对砜嘧磺隆的抗性上存在明显差异."选一"中AHAS基因的克隆结果显示,基因序列1880位点的G突变成了A,导致编码的氨基酸由丝氨酸变成了天冬酰胺.这些结果对于"选一"抗除草剂种质的利用和水稻生产中除草剂的选择有重要参考价值.
Herbicides targeting AHAS are widely used in crop production, including imidazolinone, pyrimidinylthio-benzoates, sulfonylurea, triazolopyrimidines and sulfonamide. In our previous work, we screened a resistant rice strain of imidazolinone herbicide,named as " Xuan 1". This paper reports the results of resistance test and identification of related resistance genes in posterity 15 DB01 of Xuan 1 to six targeting AHAS herbicides, which are commonly used in agricultural production in China. The results showed that 15 DB01 had different degrees of resistance to imidazolinone-based imazamox, two pyrimidinylthio-benzoates based bispyribac-sodium and pyribenzoxim, two sulfonylurea-based Mesosulfuron-methyl and nicosulfuron, but its resistance to imazamox was significantly greater than that of other herbicides. No significant difference was found between 15 DB01 rice and 13 lb05 control in resistance to sulfuron-methyl.The cloning of AHAS gene in " Xuan 1" showed that G mutated to A at site 1880 of gene sequence, resulting in the transformation of the encoded amino acid from serine to asparagine. These results have important reference value for the utilization of herbicide-resistant germplasm and the selection of herbicides in rice production.
Herbicides targeting AHAS are widely used in crop production, including imidazolinone, pyrimidinylthio-benzoates, sulfonylurea, triazolopyrimidines and sulfonamide. In our previous work, we screened a resistant rice strain of imidazolinone herbicide,named as " Xuan 1". This paper reports the results of resistance test and identification of related resistance genes in posterity 15 DB01 of Xuan 1 to six targeting AHAS herbicides, which are commonly used in agricultural production in China. The results showed that 15 DB01 had different degrees of resistance to imidazolinone-based imazamox, two pyrimidinylthio-benzoates based bispyribac-sodium and pyribenzoxim, two sulfonylurea-based Mesosulfuron-methyl and nicosulfuron, but its resistance to imazamox was significantly greater than that of other herbicides. No significant difference was found between 15 DB01 rice and 13 lb05 control in resistance to sulfuron-methyl.The cloning of AHAS gene in " Xuan 1" showed that G mutated to A at site 1880 of gene sequence, resulting in the transformation of the encoded amino acid from serine to asparagine. These results have important reference value for the utilization of herbicide-resistant germplasm and the selection of herbicides in rice production.
引文
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5徐志军,徐建欣,杨洁.水稻免耕直播秸秆覆盖条件下稻田杂草的发生规律和防治策略[J].作物研究,2018,32(2):121-126.
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11陈涛,张善磊,赵凌,等.ALS抑制剂类除草剂抗性水稻功能标记的开发与验证[J].中国水稻科学,2018,32(2):137-145.
12张保龙,王金彦,陈天子,等.粳稻的ALS突变型基因及其蛋白和应用:中国,CN201710120061.5[P].2017-03-02.
13 Okuzaki A,Shimizu T,Kaku K,et al.A novel mutated acetolactate synthase gene conferring specific resistance to pyrimidinyl carboxy herbicides in rice[J].Plant Mol Biol,2007,64:219-224.
14 Webster E P,Masson J A.Acetolactate synthase-inhibiting herbicides on imidazolinone-tolerant rice[J].Weed Science,2001,49(5):652-657.
15陈竹锋,王承旭,柳威,等.水稻抗除草剂蛋白及其在植物育种中的应用:中国,CN201210037789.9[P].2013-04-17.
16 Tan S,Evans R R,Dahmer M L,et al.Imidazolinone-tolerant crops:history[J].Current Status and Future,2005,61(3):246-257.
2 Goulart I C G R,Matzenbacher F O,Merotto A J R.Differential germination pattern of rice cultivars resistant to imidazolinone herbicides carrying different acetolactate synthase gene mutations[J].Weed Res,2012,52:224-232.
3刘兴林,孙涛,付声姣,等.水稻田除草剂的应用及杂草抗药性现状[J].西北农林科技大学学报:自然科学版,2015,43(7):115-126.
4吴文革,季雅岚,习敏,等.杂草稻的生物学特性及利用研究进展[J].安徽农业科学,2018,46(17):30-33.
5徐志军,徐建欣,杨洁.水稻免耕直播秸秆覆盖条件下稻田杂草的发生规律和防治策略[J].作物研究,2018,32(2):121-126.
6陈宇博,赖朝晖,许燎原,等.直播早稻杂草发生规律研究[J].农学学报,2015,5(8):27-30.
7张云月,卢宗志,李洪鑫,等.杂草对乙酰乳酸合成酶抑制剂类除草剂抗药性的研究进展[J].杂草科学,2013,31(2):1-5.
8 Green J M.Current state of herbicides in herbicide-resistant crops[J].Pest Manag Sci,2014,70:1 351-1 357.
9 Duggleby R G,McCourt J A,Guddat L W.Structure and mechanism of inhibition of plant acetohydroxyacid synthase[J].Plant Physiol Biochem,2008,46:309-324.
10 Yu Q,Powles S B.Resistance to AHAS inhibitor herbicides:current understanding[J].Pest Manag Sci,2014,70:1 340-1 350.
11陈涛,张善磊,赵凌,等.ALS抑制剂类除草剂抗性水稻功能标记的开发与验证[J].中国水稻科学,2018,32(2):137-145.
12张保龙,王金彦,陈天子,等.粳稻的ALS突变型基因及其蛋白和应用:中国,CN201710120061.5[P].2017-03-02.
13 Okuzaki A,Shimizu T,Kaku K,et al.A novel mutated acetolactate synthase gene conferring specific resistance to pyrimidinyl carboxy herbicides in rice[J].Plant Mol Biol,2007,64:219-224.
14 Webster E P,Masson J A.Acetolactate synthase-inhibiting herbicides on imidazolinone-tolerant rice[J].Weed Science,2001,49(5):652-657.
15陈竹锋,王承旭,柳威,等.水稻抗除草剂蛋白及其在植物育种中的应用:中国,CN201210037789.9[P].2013-04-17.
16 Tan S,Evans R R,Dahmer M L,et al.Imidazolinone-tolerant crops:history[J].Current Status and Future,2005,61(3):246-257.