Black spot disease in poplar is a disease of the leaf caused by fungus. The major pathogen is Marssonina brunnea f. sp. multigermtubi. To date, little is known about the molecular mechanism of poplar （M. brunnea） in...Black spot disease in poplar is a disease of the leaf caused by fungus. The major pathogen is Marssonina brunnea f. sp. multigermtubi. To date, little is known about the molecular mechanism of poplar （M. brunnea） interaction. In order to identify the proteins related to disease resistance and understand its molecular basis, the clone ＂NL895＂ （P. euramericana CL＂NL895＂）, which is highly resistant to M. brunnea f. sp. multigermtubi, was used in this study. We used two-dimensional gel electrophoresis （2-DE） and mass spectrometry （MS） to identify the proteins in poplar leaves that were differentially expressed in response to black spot disease pathogen, M. brunnea f. sp. multigermtubi. Proteins extracted from poplar leaves at 0, 12, 24, 48, and 72 h after pathogen-inoculation were separated by 2-DE, About 500 reproducible protein spots were detected, of which 40 protein spots displayed differential expression in levels and were subjected to Matrix assisted laser desorption/ionization time of flight mass spectrometry （MALDI-TOF MS） followed by database searching. According to the function, the identified proteins were sorted into five categories, that is, protein synthesis, metabolism, defense response and unclassified proteins.展开更多
为准确检测大豆过敏原,筛查验证表征5类主要大豆过敏原蛋白的特征肽段,建立基于超高效液相色谱-串联质谱技术的大豆主要过敏原检测方法。优化样品总蛋白提取方法,将大豆蛋白经测序级胰蛋白酶酶解后,利用超高效液相色谱-四极杆-飞行时间...为准确检测大豆过敏原,筛查验证表征5类主要大豆过敏原蛋白的特征肽段,建立基于超高效液相色谱-串联质谱技术的大豆主要过敏原检测方法。优化样品总蛋白提取方法,将大豆蛋白经测序级胰蛋白酶酶解后,利用超高效液相色谱-四极杆-飞行时间质谱对酶解产物进行分析,结合Uniprot数据库完成计算机检索,针对性地鉴定24种大豆样品的11种主要过敏原(大豆球蛋白的G1、G2、G3、G4和G5亚基、β-伴大豆球蛋白的α’、α和β亚基、Gly m Bd 30K、Gly m Bd 28K及Kunitz型胰蛋白酶抑制剂)的多肽片段,并从中挖掘用以表征各过敏原的肽段,最终得到29条响应值高、重复性好的适于质谱检测的特征肽段;进一步利用高效液相色谱-三重四极杆质谱多反应监测模式对这些特征肽段进行验证,发现这些肽段的特异性、灵敏度和稳定性均符合质谱检测要求,可为食品中大豆过敏原的全面、准确检测提供一定的理论基础和技术支持。展开更多
目的建立超高效液相色谱串联四级杆飞行时间质谱法快速检测草乌中毒者血清中乌头类生物碱及其代谢产物。方法血样经离心后,取0.2 m L血清用甲醇定容至2.0 mL,涡旋混匀过滤膜后,对草乌中毒者血样中14种乌头类生物碱及其代谢产物进行定性...目的建立超高效液相色谱串联四级杆飞行时间质谱法快速检测草乌中毒者血清中乌头类生物碱及其代谢产物。方法血样经离心后,取0.2 m L血清用甲醇定容至2.0 mL,涡旋混匀过滤膜后,对草乌中毒者血样中14种乌头类生物碱及其代谢产物进行定性定量检测。结果14种乌头类生物碱及其代谢产物在1.0~100ng/m L的浓度范围内线性关系良好,相关系数均大于0.99,加标回收率为80.2%~川.9%,相对标准偏差为0.5〜3.3%。11个草乌中毒者血清中含有苯甲酰新乌头原碱,含量约为2.11-2.38 mg/kg,部分血清样品中还检测到滇乌头碱、印乌头碱,含量分别为3.16〜3.30mg/kg、1.09~1.15mg/kg。结论该方法灵敏,准确,适用于血清中乌头类化合物的检测。展开更多
目的建立超高效液相色谱-飞行时间质谱法测定蔬菜中辛硫磷农药残留的分析方法。方法蔬菜样品经乙腈提取,45℃水浴氮吹至近干,丙酮定容,经超高效液相色谱-飞行时间质谱(ultra performance liquid chromatography/time of flight mass spe...目的建立超高效液相色谱-飞行时间质谱法测定蔬菜中辛硫磷农药残留的分析方法。方法蔬菜样品经乙腈提取,45℃水浴氮吹至近干,丙酮定容,经超高效液相色谱-飞行时间质谱(ultra performance liquid chromatography/time of flight mass spectrometry(UPLC-TOF MS)检测。结果辛硫磷的保留时间为13.34 min,在1~400μg/L范围内线性良好,相关系数r大于0.999,方法检出限为1μg/L。将该方法用于西葫芦、白萝卜、小白菜、西红柿、葱等常见蔬菜的辛硫磷残留检测,结果表明辛硫磷的添加回收率在87.0%~99.9%之间,相对标准偏差为5.14%~8.30%。结论该方法操作简单、准确高效,可以用于蔬菜中辛硫磷残留的日常检测。展开更多
基金This work was supported by the National Natural Science Foundation of China （No. 30230300）.
文摘Black spot disease in poplar is a disease of the leaf caused by fungus. The major pathogen is Marssonina brunnea f. sp. multigermtubi. To date, little is known about the molecular mechanism of poplar （M. brunnea） interaction. In order to identify the proteins related to disease resistance and understand its molecular basis, the clone ＂NL895＂ （P. euramericana CL＂NL895＂）, which is highly resistant to M. brunnea f. sp. multigermtubi, was used in this study. We used two-dimensional gel electrophoresis （2-DE） and mass spectrometry （MS） to identify the proteins in poplar leaves that were differentially expressed in response to black spot disease pathogen, M. brunnea f. sp. multigermtubi. Proteins extracted from poplar leaves at 0, 12, 24, 48, and 72 h after pathogen-inoculation were separated by 2-DE, About 500 reproducible protein spots were detected, of which 40 protein spots displayed differential expression in levels and were subjected to Matrix assisted laser desorption/ionization time of flight mass spectrometry （MALDI-TOF MS） followed by database searching. According to the function, the identified proteins were sorted into five categories, that is, protein synthesis, metabolism, defense response and unclassified proteins.
文摘为准确检测大豆过敏原,筛查验证表征5类主要大豆过敏原蛋白的特征肽段,建立基于超高效液相色谱-串联质谱技术的大豆主要过敏原检测方法。优化样品总蛋白提取方法,将大豆蛋白经测序级胰蛋白酶酶解后,利用超高效液相色谱-四极杆-飞行时间质谱对酶解产物进行分析,结合Uniprot数据库完成计算机检索,针对性地鉴定24种大豆样品的11种主要过敏原(大豆球蛋白的G1、G2、G3、G4和G5亚基、β-伴大豆球蛋白的α’、α和β亚基、Gly m Bd 30K、Gly m Bd 28K及Kunitz型胰蛋白酶抑制剂)的多肽片段,并从中挖掘用以表征各过敏原的肽段,最终得到29条响应值高、重复性好的适于质谱检测的特征肽段;进一步利用高效液相色谱-三重四极杆质谱多反应监测模式对这些特征肽段进行验证,发现这些肽段的特异性、灵敏度和稳定性均符合质谱检测要求,可为食品中大豆过敏原的全面、准确检测提供一定的理论基础和技术支持。
文摘目的建立超高效液相色谱-飞行时间质谱法测定蔬菜中辛硫磷农药残留的分析方法。方法蔬菜样品经乙腈提取,45℃水浴氮吹至近干,丙酮定容,经超高效液相色谱-飞行时间质谱(ultra performance liquid chromatography/time of flight mass spectrometry(UPLC-TOF MS)检测。结果辛硫磷的保留时间为13.34 min,在1~400μg/L范围内线性良好,相关系数r大于0.999,方法检出限为1μg/L。将该方法用于西葫芦、白萝卜、小白菜、西红柿、葱等常见蔬菜的辛硫磷残留检测,结果表明辛硫磷的添加回收率在87.0%~99.9%之间,相对标准偏差为5.14%~8.30%。结论该方法操作简单、准确高效,可以用于蔬菜中辛硫磷残留的日常检测。
