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1.
感性、耐性菜豆从萌芽到24d的EPSP合成酶比活性是不同的,但变化规律相似。出苗时比活性较高,10d时降至最低点,随后上升,16d时达到最高点,然后缓慢下降;两种菜豆在生长的同一天酶比活性差异小,最大倍数仅为1.1;用草甘膦处理两种三出复叶时的菜豆1d后酶比活性上升15%～25%。实验测定了经4步纯化,比活性达6219.4nmol· min-1· mg-1蛋白以上的两种菜豆EPSP合成酶的一些酶学性质。PEP是酶的最适底物,感性、耐性 Km(PEP)分别为3.5和7.1μmol· L-1,两者相差2倍,亲和力Ki(草甘膦)分别是6.2和 16.7μmol· L-1,两者相差2.7倍。两种菜豆对草甘膦耐性不同的酶学机理在于耐性菜豆的EPSP合成酶与草甘膦的亲和力小,与酶底物亲和力大;而感性菜豆EPSP酶则与草甘膦的亲和力大,与其底物亲和力小。 相似文献
2.
用耐性菜豆根尖为材料，以λgt10为载体获得了4.8×105个重组噬菌体的cDNA文库。以植物EPSP合成酶基因保守序列合成二段探针，经噬菌体原位杂交筛选出了阳性克隆，并进行了酶切鉴定。测定 cDNA 序列长度为2024个核苷酸，其中编码长1569个核苷酸，共编码523个氨基酸和一个终止密码子，与已发表的其它植物成熟EPSP合成酶氨基酸序列相比具有很高的同源性（≥84.8%），但进入叶绿体运输肽的氨基酸同源性低。以耐性菜豆EPSP合成酶的cDNA序列为模板，用RT-PCR方法扩增感性菜豆EPSP合成酶cDNA片段，并克隆于pUC18质粒上，经测定序列并比较发现:感性菜豆EPSP合成酶cDNA核苷酸序列1737位碱基为G，而耐性的为C，从而表达出的513位氨基酸残基感性的为E，耐性的为Q，表明两种菜豆EPSP合成酶cDNA核苷酸序列一位点的差异是它们对草甘膦耐性不同的原因。 相似文献
3.
EPSP 合成酶的特性及新抑制剂的研究进展 总被引：4，自引：0，他引：4
概述了EPSP 合成酶的生理作用。高等植物前体EPSP 合成酶进入叶绿体后, 经剪切而成成熟EPSP 合成酶并定位于叶绿体内质膜上; EPSP 合成酶的三维结构上存在三个活性区域, 改变活性区域的氨基酸残基可对草甘膦产生抗性, 同时表明了酶与草甘膦的结合位点; 不同生物的EPSP 合成酶有高的同源性。EPSP 合成酶催化底物反应机理为: PEP 首先与酶形成过渡态, 而后和S3P 形成缩酮四面体, 最后生成EPSP。草甘膦与EPSP 合成酶、EPSP形成三元复合物而阻断了EPSP 合成酶的催化作用; 以EPSP 和S3P ·glypho sate 为分子模型, 设计合成的化合物对EPSP 合成酶的活性有抑制作用。 相似文献
4.
EPSP合成酶 ( 5 -烯醇丙酮酸莽草酸 - 3-磷酸合酶 )是除草剂靶标酶之一 [1 ] ,也是抗草甘膦转基因作物的关键性酶 [2 ] ,它催化一分子的莽草酸 - 3-磷酸 ( S3P)和烯醇式丙酮酸 ( PEP)成EPSP,从而导致芳香族氨基酸——色氨酸、酪氨酸、苯丙氨酸的生物合成。 EPSP合成酶存在于细菌、真菌和植物中 ,但由于其性质的不稳定性、不均匀性和低丰度 ,造成分离纯化困难 [3] 。作者以菜豆 ( Phaselusvulgaris L .)幼苗为原料 ,经快速纯化 (整个纯化过程少于 1 .5 h)获得的EPSP合成酶产品能用于除草剂的筛选工作 ,为以 EPSP合成酶为靶标的生… 相似文献
5.
<正>1草甘膦抗性现状及面临的其他问题草甘膦为内吸传导型慢性广谱灭生性除草剂,主要抑制植物物体内烯醇丙酮基莽草素磷酸合成酶(EPSP),从而抑制莽草素向苯丙氨酸、酪氨酸及色氨酸的转化,使蛋白质的合成受到干扰,从而导致植物死亡。由于草甘膦优异的杀草活性、广泛的杀草谱、较低的土壤残留、较长的控草时间,加上抗除草剂转基因作物的广泛种植,使其成为全球销量第一的除草剂品种。然而由于长时间大量单一连续使用草甘膦,杂草的抗性问题已经非常突出。到目前已经公布了有31种 相似文献
6.
草甘膦与硫酸铵混合后处理空心莲子草,测定其对植株的抑制作用和体内草甘膦的吸收与传导量。结果表明,加入硫酸铵(1.20 g/L)后草甘膦(300 mg/L)对地下根茎抑制率比对照提高了12.2个百分点。植株经硫酸铵喷雾处理后,地下茎和根系中14C-草甘膦含量分别是对照处理的1.39和1.86倍。药液中水的硬度达到342.0 mg/L时明显降低草甘膦的除草活性,硫酸铵浓度达到12.0 g/L则可基本消除水的硬度对草甘膦除草活性的影响。 相似文献
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从2004年开始,延边地区稻田大量发生抗磺酰脲类除草剂苄嘧磺隆的抗药性生态型雨久花。为解析其抗药性突变机制,采用离体法测定了抗、感性雨久花乙酰乳酸合成酶活性。结果表明,抑制抗、感性雨久花乙酰乳酸合成酶活性50%的苄嘧磺隆剂量为253.44×10-7mol/L和2.80×10-7mol/L;抑制抗、感性雨久花乙酰乳酸合成酶活性50%的吡嘧磺隆剂量为1802.15×10-7mol/L和20.85×10-7mol/L。抗药性雨久花对苄嘧磺隆和吡嘧磺隆的抗性系数(RI50/SI50)值分别为90.6和86.5,并存在交互抗药性。确认抗药性突变系由其乙酰乳酸合成酶对磺酰脲类除草剂苄嘧磺隆、吡嘧磺隆反应钝化所致。 相似文献
8.
辣椒碱对小菜蛾体内谷胱甘肽S-转移酶和Na+ -K+ -ATP酶活性的影响 总被引：3，自引：0，他引：3
采用常规生化方法测定了辣椒碱对小菜蛾体内谷胱甘肽S-转移酶和Na+ -K+ -ATP酶活性的影响。结果表明,经不同浓度辣椒碱处理12 h后,小菜蛾体内谷胱甘肽S-转移酶活力均显著低于空白对照,在100.56~141.53 μmol ·L-1 ·mg-1Pro ·min-1 之间;而Na+ -K+ -ATP酶活力均显著高于空白对照,在12.43~20.36 μmol ·mg-1Pro ·min-1之间。说明辣椒碱能抑制小菜蛾体内谷胱甘肽S-转移酶的活性,而对Na+ -K+ -ATP酶活性则表现为促进作用。 相似文献
9.
为采用生物技术防控草地贪夜蛾Spodoptera frugiperda的扩散为害,对草地贪夜蛾5龄和6龄幼虫注射浓度为1×109CFU/mL的大肠杆菌Escherichia coli菌液,并以注射等量磷酸盐缓冲液(phosphate buffer solution,PBS)和未做任何处理(CK)为对照,24 h后测定幼虫体重、集结数和酚氧化酶(phenoloxidase,PO)活性。结果显示,注射大肠杆菌菌液24 h后,草地贪夜蛾5龄和6龄幼虫体重均受到抑制,其体重分别为0.170 g和0.411 g,均显著低于CK的0.181 g和0.484 g;注射大肠杆菌菌液24 h后,草地贪夜蛾5龄和6龄幼虫集结数分别为135.0、338.4个索引集结数(the indexed nodules,INs),前者极显著低于后者,且均显著高于CK和PBS处理,分别为0.4、10.2个INs和0.3、10.9个INs;注射大肠杆菌菌液24 h后,草地贪夜蛾5龄幼虫PO活性为0.156 ABS·min-1·mg-1,显著高于CK和PBS处理,分别为0.046 ABS·min-1·mg-1和0.066 ABS·min-1·mg-1,但草地贪夜蛾6龄幼虫的PO活性为0.050 ABS·min-1·mg-1,显著低于CK和PBS处理,分别为0.066 ABS·min-1·mg-1和0.069 ABS·min-1·mg-1,且草地贪夜蛾5龄幼虫的PO活性显著高于6龄幼虫的PO活性。表明细菌侵染后草地贪夜蛾不同高龄幼虫的免疫应激反应存在差异,而这种差异可能受幼虫生长发育及细胞免疫和体液免疫功能之间权衡现象的影响。 相似文献
10.
利用异源表达于酵母细胞中的小麦细胞色素P450cDNA(CYP71C6v1)研究了磺酰脲类除草剂绿磺隆、醚苯磺隆的代谢作用。结果表明,代谢产物5-羟基绿磺隆和5-羟基醚苯磺隆能够抑制乙酰乳酸合成酶(ALS酶)活性,且代谢产物与母体化合物绿磺隆、醚苯磺隆抑制ALS酶活性的IC50值差异小,但是代谢产物在茎叶喷雾小麦和菜豆时,均未表现出活性。绿磺隆及其代谢产物抑制小麦ALS酶活性的IC50值分别为7.1×10-9和7.9×10-9mol/L,抑制菜豆ALS酶活性的IC50分别为3.6×10-9和4.1×10-9mol/L;醚苯磺隆及其代谢产物抑制小麦ALS酶活性的IC50分别为4.6×10-9和5.3×10-9mol/L,抑制菜豆ALS酶活性的IC50分别为4.7×10-9和4.9×10-9mol/L。结果表明,在磺酰脲类分子苯环5位上进行结构改造,有可能得到高活性的化合物。 相似文献
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12.
Glyphosate resistance in common ragweed (
A
mbrosia artemisiifolia L.) from Mississippi,USA 
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下载免费PDF全文 Glyphosate is one of the most commonly used broad‐spectrum herbicides over the last 40 years. Due to the widespread adoption of glyphosate‐resistant (GR) crop technology, especially corn, cotton and soybean, several weed species have evolved resistance to this herbicide. Research was conducted to confirm and characterize the magnitude and mechanism of glyphosate resistance in two GR common ragweed ( A mbrosia artemisiifolia L.) biotypes from Mississippi, USA. A glyphosate‐susceptible (GS) biotype was included for comparison. The effective glyphosate dose to reduce the growth of the treated plants by 50% for the GR1, GR2 and GS biotypes was 0.58, 0.46 and 0.11 kg ae ha?1, respectively, indicating that the level of resistance was five and fourfold that of the GS biotype for GR1 and GR2, respectively. Studies using 14 C‐glyphosate have not indicated any difference in its absorption between the biotypes, but the GR1 and GR2 biotypes translocated more 14 C‐glyphosate, compared to the GS biotype. This difference in translocation within resistant biotypes is unique. There was no amino acid substitution at codon 106 that was detected by the 5‐enolpyruvylshikimate‐3‐phosphate synthase gene sequence analysis of the resistant and susceptible biotypes. Therefore, the mechanism of resistance to glyphosate in common ragweed biotypes from Mississippi is not related to a target site mutation or reduced absorption and/or translocation of glyphosate. 相似文献
13.
Debrah F Lorraine-Colwill Tim R Hawkes Patricia H Williams Simon AJ Warner Peter B Sutton Stephen B Powles Christopher Preston 《Pest management science》1999,55(4):489-491
Annual ryegrass (Lolium rigidum) is a widespread and important weed of Australia and populations of this weed have developed resistance to most major herbicides, including glyphosate. The possible mechanisms of resistance have been examined in one glyphosate-resistant Lolium population. No major differences were observed between resistant and susceptible biotypes in respect of (i) the target enzyme (EPSP synthase), (ii) DAHP synthase, the first enzyme of the target (shikimate) pathway, (iii) absorption of glyphosate, or (iv) translocation. Following treatment with glyphosate, there was greater accumulation of shikimate (derived from shikimate-3-Pi) in susceptible than in resistant plants. In addition, the resistant population exhibited cross-resistance to 2-hydroxy-3-(1,2,4-triazol-1-yl)propyl phosphonate, a herbicide which, although structurally similar to glyphosate, acts at an unrelated target site. On the basis of these observations we speculate that movement of glyphosate to its site of action in the plastid is involved in the resistance mechanism. © 1999 Society of Chemical Industry 相似文献
14.
Three cell lines of groundnut (Arachis hypogaea L), an important oilseed legume, were selected on glyphosate using in-vitro culture techniques. The cell lines isolated through single as well as stepwise selection procedures showed c 20-fold increase in glyphosate tolerance as compared to the unselected control cell line. Studies on the biochemical mechanism of glyphosate tolerance in these cell lines showed a significant increase in the total extractable activity of the target enzyme, 5-enolpyruvyl shikimate-3-phosphate (EPSP) synthase (EC 2.5.1.19), which was further confirmed with immunological data. The over-expressed EPSP synthase activity was, however, subject to inhibition by glyphosate in vitro. Two other key regulated enzymes of the shikimic acid pathway, 3-deoxy-D -arabino heptulosonate 7-phosphate (DAHP) synthase (EC 4.1.2.15) and chorismate mutase (CM) (EC 5.4.99.5) did not show any change in specific activity in the selected cell lines. The enhanced activity of EPSP synthase in the tolerant cell lines was found to be stably inherited in the absence of selection pressure. © 1999 Society of Chemical Industry 相似文献
15.
Gene polymorphisms in glyphosate-resistant and -susceptible biotypes of Eleusine indica from Malaysia 总被引：3，自引：0，他引：3
Summary Resistant (R) and susceptible (S) biotypes of Eleusine indica were collected from four areas, namely Chaah, Lenggeng, Bidor and Temerloh, in Malaysia. Restriction fragment length polymorphism (RFLP) and polymerase chain reaction (PCR)-RFLP analyses using Sph I restriction enzyme were able to differentiate the R biotype from the S biotype by showing R-specific and S-specific polymorphisms in E. indica from three of the areas, with the exception of Temerloh where no polymorphisms were detected. The different DNA profiles for the R biotypes obtained indicate that Sph I is not a useful diagnostic marker. The DNA polymorphisms detected in the 5-enolpyruvylshikimate 3-phosphate (EPSP) synthase gene suggest that there are different mutation events leading to development of resistance to glyphosate. Partial sequencing of the EPSP synthase gene confirmed different mutations occurring with substitution of proline with serine or threonine at amino acid 106 for the R biotype in Chaah, Bidor and Temerloh. 相似文献
16.
麦田抗性生物型荠菜对苯磺隆的抗性机制研究 总被引：4，自引：1，他引：3
为明确抗性生物型荠菜对苯磺隆的抗性机制,分别测定了苯磺隆对抗性和敏感生物型荠菜体内乙酰乳酸合成酶(ALS)、谷胱甘肽-S-转移酶(GSTs)、超氧化物歧化酶(SOD)、过氧化物酶(POD)和过氧化氢酶(CAT)的影响。结果表明:离体条件下,抗性生物型荠菜体内ALS对苯磺隆的敏感性明显降低,苯磺隆对荠菜抗性和敏感生物型ALS的抑制中浓度(I50)分别为0.722 8和0.052 1 μmol/L,抗性与敏感生物型I50的比值为13.87;活体条件下,施用苯磺隆后,抗性和敏感生物型荠菜ALS活性均受到一定程度的抑制,但抗性生物型ALS活性受到抑制后能逐渐恢复,而敏感生物型则不能恢复;经苯磺隆处理后,抗性生物型GSTs相对活力明显高于敏感生物型,而抗性和敏感生物型体内POD、SOD和CAT相对活力无明显差异。研究表明,抗性生物型荠菜体内ALS对苯磺隆敏感性降低是其抗药性产生的原因之一,而GSTs对苯磺隆代谢能力的差异也可能与荠菜对苯磺隆的抗性有关。 相似文献