Centre for Environmental Risk Assessment and Remediation, University of South Australia, Building X, Mawson Lakes Campus, Mawson Lakes, SA 5095, Australia.
Anal Bioanal Chem. 2012 Apr;402(10):3287-98. doi: 10.1007/s00216-011-5624-9. Epub 2011 Dec 27.
Functional characterisation of the genes regulating metal(loid) homeostasis in plants is a major focus for phytoremediation, crop biofortification and food security research. Recent advances in X-ray focussing optics and fluorescence detection have greatly improved the potential to use synchrotron techniques in plant science research. With use of methods such as micro X-ray fluorescence mapping, micro computed tomography and micro X-ray absorption near edge spectroscopy, metal(loids) can be imaged in vivo in hydrated plant tissues at submicron resolution, and laterally resolved metal(loid) speciation can also be determined under physiologically relevant conditions. This article focuses on the benefits of combining molecular biology and synchrotron-based techniques. By using molecular techniques to probe the location of gene expression and protein production in combination with laterally resolved synchrotron techniques, one can effectively and efficiently assign functional information to specific genes. A review of the state of the art in this field is presented, together with examples as to how synchrotron-based methods can be combined with molecular techniques to facilitate functional characterisation of genes in planta. The article concludes with a summary of the technical challenges still remaining for synchrotron-based hard X-ray plant science research, particularly those relating to subcellular level research.
研究植物中调控金属(类)内稳态的基因的功能特征是植物修复、作物生物强化和食品安全研究的主要重点。X 射线聚焦光学和荧光检测的最新进展极大地提高了同步加速器技术在植物科学研究中的应用潜力。利用微 X 射线荧光映射、微计算机断层扫描和微 X 射线吸收近边光谱学等方法,可以在水合植物组织中以亚微米分辨率对金属(类)进行体内成像,并且还可以在生理相关条件下确定横向分辨的金属(类)形态。本文重点介绍了将分子生物学和基于同步加速器的技术相结合的好处。通过使用分子技术探测基因表达和蛋白质产生的位置,并结合横向分辨的同步加速器技术,可以有效地将功能信息分配给特定的基因。本文介绍了该领域的最新进展,并举例说明了如何将基于同步加速器的方法与分子技术相结合,以促进植物中基因的功能特征分析。文章最后总结了基于同步加速器的硬 X 射线植物科学研究中仍然存在的技术挑战,特别是与亚细胞水平研究相关的挑战。