| Electro-fusion of Gametes and Subsequent Culture of Zygotes in Rice
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Author:
Date:
2016-12-20
[Abstract] Electro-fusion system with isolated gametes has been utilized to dissect fertilization-induced events in angiosperms, such as egg activation, zygote development and early embryogenesis, since the female gametophytes of plants are deeply embedded within ovaries. In this protocol, procedures for isolation of rice gametes, electro-fusion of gametes, and culture of the produced zygotes are described.
[摘要] 已经利用具有孤立配子的电融合系统来解剖被子植物中受精诱导的事件，例如卵活化，合子发育和早期胚胎发生，因为植物的雌配子体深深嵌入卵巢内。在该方案中，描述了分离水稻配子，配子电融合和产生的受精卵培养的程序。
背景 被子植物的受精和后续事件，如胚胎发生和胚乳发育，发生在深入嵌入卵母细胞的胚囊中（Nawaschin，1898; Guignard，1899; Russell，1992; Raghavan，2003）。因此，分离的配子已被用于体外受精（IVF）系统，以观察和分析受精和后处理过程（Wang等人，2006年）。用于被子植物的IVF系统包括三种基本微技术的组合：（i）男性和女性配子的分离和选择; （ii）配对对和（iii）单细胞培养物的融合（Kranz，1999）。已经在广泛的植物物种中建立了分离活的配子的程序，包括单子叶植物和双子叶植物（综述于Kranz，1999和Okamoto，2011）。分离的配子可以电融合（Kranz等人，1991; Uchiumi等人，2006和2007）或化学地使用钙（Faure等人，1994; Kranz和Lörz，1994; Khalequzzaman和Haq，2005），聚乙二醇（Sun等，1995; ...
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| Purification of Flagellin from Acidovorax avenae and Analysis of Plant Immune Responses Induced by the Purified Flagellin
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Author:
Date:
2016-08-20
[Abstract] Plants sense potential pathogens by recognizing conserved pathogen-associated molecular patterns (PAMPs) that cause PAMP-triggered immunity (PTI) including the generation of reactive oxygen species, callose deposition, and expression of several PTI-related genes. Acidovorax avenae is a Gam-negative bacterium that causes a seedling disease characterized by the deposition of brown stripes on the sheaths of infected plants. We previously reported that flagellin isolated from the rice avirulent A. avenae N1141 strain induces PTI, while flagellin isolated from the rice virulent A. avenae K1 strain does not induce PTI. To examine the molecular mechanism of specific PTI induction by N1141 flagellin, highly purified flagellin from N1141 or K1 strains is required. Here, ...
[摘要] 植物通过识别引起PAMP触发的免疫（PTI）的保守的病原体相关分子模式（PAMP）来感测潜在的病原体，包括活性氧的产生，胼lose质沉积和几个PTI相关基因的表达。 酸性弧菌是一种革兰氏阴性细菌，其导致幼苗疾病，其特征在于在感染的植物的鞘上沉积棕色条纹。 我们以前报告了从无毒力的大米中分离的鞭毛蛋白。 avenae N1141菌株诱导PTI，而从水稻毒力分离的鞭毛蛋白。 avenae K1株不诱导PTI。 为了检查N1141鞭毛蛋白的特异性PTI诱导的分子机制，需要来自N1141或K1菌株的高度纯化的鞭毛蛋白。 在这里，我们描述了一种高质量的纯化方法。 avenae鞭毛蛋白，并用于PTI诱导研究。
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| Arabidopsis Metabolome Analysis Using Infusion ESI FT-ICR/MS
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Author:
Date:
2015-05-05
[Abstract] We made the method for Arabidopsis metabolome analysis based on direct-infusion Fourier transform ion cyclotron resonance mass spectrometry (FT-ICR/MS) (IonSpec). This method was sufficiently applied to metabolic phenotyping of Arabidopsis. This method is simple in that after homogenizing samples, powdered samples are dissolved in extraction solvents (acetone and methanol) to 20% fresh weight/volume. Extracted sample solutions are dried and dissolved in 50% (v/v) acetonitrile. Mass analysis using FT-ICR/MS (IonSpec) is performed in positive and negative ionization operation modes. Mass spectra are acquired over the 100-1,000 m/z range and accumulated to improve the S/N ratio.
[摘要] 我们基于直接输注傅立叶变换离子回旋共振质谱（FT-ICR/MS）（IonSpec）制备了拟南芥分析的方法。 该方法充分应用于拟南芥的代谢表型。 该方法的简单之处在于，在均质化样品后，将粉末样品溶解在提取溶剂（丙酮和甲醇）中至20％鲜重/体积。 将提取的样品溶液干燥并溶解在50％（v/v）乙腈中。 使用FT-ICR/MS（IonSpec）的质量分析在正和负电离操作模式中进行。 质谱在100-1,000m/z范围内获得并累积以提高S/N比。
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