中文摘要：本发明公开一种基于叶绿素荧光成像的转基因玉米草甘膦耐受性表型的检测方法，包括：(1)制备实验样本；(2)获取受草甘膦胁迫不同天数的转基因玉米和非转基因玉米植株冠层叶片的莽草酸含量；(3)通过叶绿素荧光成像系统获取受草甘膦胁迫不同天数的转基因玉米和非转基因玉米植株冠层的叶绿素荧光参数；(4)建立PLSR模型，剔除异常样本，经K‑Means选择建模集和预测集；(5)利用主成分分析选择对草甘膦胁迫响应最敏感的叶绿素荧光参数；(6)基于选择的叶绿素荧光参数和转基因玉米植株实际的莽草酸含量建立PLSR回归分析模型，建模集和预测集决定系数为R2＝0.75，R2＝0.63。本发明鉴别精度较高、操作简单，可实现转基因玉米草甘膦耐受性表型的快速无损检测。
外文摘要：NOVELTY - Method for detecting glyphosate tolerance phenotype of transgenic corn involves planting transgenic corn and non-transgenic corn, spraying glyphosate solution and water, determining shikimic acid content in leaves of corn, using chlorophyll fluorescence imaging system, performing principal component analysis, selecting e.g. modeling set, establishing transgenic corn shikimic partial least squares regression (PLSR) analysis model, and inputting the chlorophyll fluorescence parameter of sample prediction set in the PLSR regression analysis model.
USE - The method is useful for detecting glyphosate tolerance phenotype of transgenic corn.
ADVANTAGE - The method is easy, and rapid and enables to detect glyphosate tolerance phenotype of transgenic corn with high precision.
DETAILED DESCRIPTION - Method for detecting glyphosate tolerance phenotype of transgenic corn involves (a) planting transgenic corn and non-transgenic corn, spraying 1080 g/ha glyphosate solution on portion of the transgenic corn and non-transgenic corn as experimental group, and then spraying water on transgenic corn and non-transgenic corn as control group, (b) determining shikimic acid content in leaves of transgenic corn and non-transgenic corn using standard curve of shikimic acid and UV spectrophotometry, (c) using chlorophyll fluorescence imaging system to obtain the fluorescence parameters of the canopy of transgenic corn and non-transgenic corn with different days of glyphosate stress, (d) selecting corn which is more sensitive to the response to glyphosate stress by performing principal component analysis, (e) selecting modeling set and prediction set by k-means method in the experimental group and control group, (f) using chlorophyll fluorescence parameters from the samples in the modeling set as input, and shikimic acid content as output, and establishing transgenic corn shikimic partial least squares regression (PLSR) analysis model, and (g) in! putting the chlorophyll fluorescence parameter of sample prediction set in the transgenic corn shikimic acid PLSR regression analysis model to obtain shikimic acid content of the sample.
主权项：一种基于叶绿素荧光成像的转基因玉米草甘膦耐受性表型的检测方法，其特征在于，包括以下步骤：(1)：种植同等数量的转基因玉米和非转基因玉米，在玉米生长过程中，对一部分数量的转基因玉米和非转基因玉米喷施浓度为1080g a.e.ha-1的草甘膦溶液作为实验组，对剩余的转基因玉米和非转基因玉米喷施等量的水作为对照组；(2)通过制定莽草酸标准曲线结合紫外分光光度法，获得受草甘膦胁迫不同天数的转基因玉米和非转基因玉米的植株冠层叶片的莽草酸含量；(3)应用叶绿素荧光成像系统，获取受草甘膦胁迫不同天数的转基因玉米和非转基因玉米的植株冠层的荧光参数；(4)采用主成分分析，选择对草甘膦胁迫响应最敏感的叶绿素荧光参数；(5)通过k-means方法在实验组和对照组内选择建模集和预测集；(6)以建模集内样本选择的叶绿素荧光参数作为输入，莽草酸含量作为输出，建立转基因玉米莽草酸PLSR回归分析模型：(7)将预测集内样本的叶绿素荧光参数输入所述转基因玉米莽草酸PLSR回归分析模型，得到对样本的莽草酸含量。
申请号：CN201710295204.6
专利号：CN106896077-A
公开/公告号：CN106896077A
申请/专利权人：浙江大学;
发明/设计人：冯旭萍; 何勇; 刘小丹
分类号：G01N21/33; G01N21/64;
主分类号：G01N21/33
法律状态：实质审查
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