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重力持续信号 1(GPS1)揭示了参与向重力性的新型细胞色素 P450。

Gravity Persistent Signal 1 (GPS1) reveals novel cytochrome P450s involved in gravitropism.

机构信息

Department of Environmental and Plant Biology, 317 Porter Hall, Ohio University, Athens, Ohio 45701, USA.

出版信息

Am J Bot. 2013 Jan;100(1):183-93. doi: 10.3732/ajb.1200436.

Abstract

PREMISE

Gravity is an important environmental factor that affects growth and development of plants. In response to changes in gravity, directional growth occurs along the major axes and lateral branches of both shoots and roots. The gravity persistent signal (gps) mutants of Arabidopsis thaliana were previously identified as having an altered response to gravity when reoriented relative to the gravity vector in the cold, with the gps1 mutant exhibiting a complete loss of tropic response under these conditions.

METHODS

Thermal asymmetric interlaced (TAIL) PCR was used to identify the gene defective in gps1. Gene expression data, molecular modeling and computational substrate dockings, quantitative RT-PCR analyses, reporter gene fusions, and physiological analyses of knockout mutants were used to characterize the genes identified.

RESULTS

Cloning of the gene defective in gps1 and genetic complementation revealed that GPS1 encodes CYP705A22, a cytochrome P450 monooxygenase (P450). CYP705A5, a closely related family member, was identified as expressed specifically in roots in response to gravistimulation, and a mutation affecting its expression resulted in a delayed gravity response, increased flavonol levels, and decreased basipetal auxin transport. Molecular modeling coupled with in silico substrate docking and diphenylboric acid 2-aminoethyl ester (DBPA) staining indicated that these P450s are involved in biosynthesis of flavonoids potentially involved in auxin transport.

CONCLUSION

The characterization of two novel P450s (CYP705A22 and CYP705A5) and their role in the gravity response has offered new insights into the regulation of the genetic and physiological controls of plant gravitropism.

摘要

前提

重力是影响植物生长和发育的重要环境因素。植物对重力的响应是通过沿茎和根的主轴和侧枝的定向生长来体现的。先前已经鉴定出拟南芥的重力持续信号(GPS)突变体在相对于重力矢量重新定向时对重力的响应发生改变,gps1 突变体在这些条件下表现出完全丧失向性反应。

方法

使用热不对称交错(TAIL)PCR 来鉴定 gps1 中缺陷的基因。使用基因表达数据、分子建模和计算底物对接、定量 RT-PCR 分析、报告基因融合以及敲除突变体的生理分析来表征鉴定出的基因。

结果

克隆 gps1 中缺陷的基因并进行遗传互补表明,GPS1 编码细胞色素 P450 单加氧酶(P450)CYP705A22。鉴定出密切相关的家族成员 CYP705A5,其在根中特异性表达以响应重刺激,并且影响其表达的突变导致重力反应延迟、类黄酮水平增加和向基生长素运输减少。分子建模结合计算机模拟底物对接和二苯硼酸 2-氨基乙酯(DBPA)染色表明,这些 P450 参与类黄酮的生物合成,类黄酮可能参与生长素运输。

结论

对两种新型 P450(CYP705A22 和 CYP705A5)的特性及其在重力响应中的作用的研究,为植物向重力性的遗传和生理控制的调节提供了新的见解。

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