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Comparison of Pollen Grain Treatments Without Mechanical Fracturation Prior to Protein Quantification
Journal of Insect Science ( IF 2.1 ) Pub Date : 2021-06-18 , DOI: 10.1093/jisesa/ieab043
Lila R Westreich 1 , Patrick C Tobin 1
Affiliation  

Protein and amino acids in pollen are important nutritional components for larval development in several insect species, especially in Apoidea. The Bradford assay is a widely used method to measure relative protein content of pollen, which can shed light on pollen quality and consequences to fitness. Prior to using the Bradford assay, protein must be released from pollen grains, often using a mixture of chemical and mechanical fracturation methods. In this study, we tested the efficacy of protein extraction without using mechanical fracturation. We used pollen collected by the solitary bee Osmia lignaria Say to compare two known buffers associated with pollen protein analysis: phosphate-buffered saline and sodium hydroxide, and deionized water, and with different pollen weights from which we quantified protein using the Bradford assay. While all buffers and deionized water were useful in releasing protein from pollen grains collected by O. lignaria, the use of sodium hydroxide resulted in significantly higher protein quantification across all pollen weights. This methodological study can inform future studies of pollen nutrition in pollen-foraging species.

中文翻译:

在蛋白质定量之前没有机械破碎的花粉粒处理的比较

花粉中的蛋白质和氨基酸是几种昆虫物种幼虫发育的重要营养成分,特别是在蚜虫中。布拉德福德法是一种广泛使用的测量花粉相对蛋白质含量的方法,它可以揭示花粉质量和对健康的影响。在使用 Bradford 测定之前,必须从花粉粒中释放蛋白质,通常使用化学和机械断裂方法的混合物。在这项研究中,我们在不使用机械破碎的情况下测试了蛋白质提取的功效。我们使用由独居蜜蜂 Osmia lignaria Say 收集的花粉来比较与花粉蛋白分析相关的两种已知缓冲液:磷酸盐缓冲盐水和氢氧化钠,以及去离子水,并使用不同的花粉重量,我们使用 Bradford 测定法对蛋白质进行量化。虽然所有缓冲液和去离子水都可用于从 O. lignaria 收集的花粉粒中释放蛋白质,但使用氢氧化钠可显着提高所有花粉重量的蛋白质定量。这项方法学研究可以为花粉觅食物种中花粉营养的未来研究提供信息。
更新日期:2021-06-18
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