种康院士2015年Cell文章Ca²⁺流实验介绍

期刊：Journalof integrative plant biology（JIPB）
主题：OsCIPK7改变激酶活性调控水稻耐寒机制（钙信号)
标题：OsCIPK7point-mutation leads to conformation and kinase-activity change for sensingcold response
影响因子：3.129
检测指标：Ca²⁺
Ca²⁺流实验方法：两周龄的水稻幼苗、低温瞬时胁迫，检测距离根尖200um。
Ca²⁺流实验测试液成份： 0.1 mM KCl, 0.1 mM CaCl2, 0.1 mM MgCl2, 0.5 mM NaCl, 0.3 mM MES, 0.2 mM Na2SO4, pH6.0（参考种康老师Cell文献信息）
作者：中科院植物所 种康
 

英文摘要

Calcineurin B-like interacting proteinkinases (CIPKs) play important roles via environmental stress. However, less isknown how to sense the stress in molecular structure conformation level.

Here, an OsCIPK7 mutant via TILLINGprocedure with a point mutation in the kinase domain showed increased chillingtolerance, which could be potentially used in the molecular breeding.

We found that this point mutation ofOsCIPK7 led to a conformational change in the activation loop of the kinasedomain, subsequently with an increase of protein kinase activity, thusconferred an increased tolerance to chilling stress.
 

中文摘要（谷歌机翻译）

钙调神经磷酸酶B样相互作用蛋白激酶（CIPKs）通过环境压力发挥重要作用。 然而，人们对如何感知分子结构构象水平的应力知之甚少。

在此，通过TILLING程序在激酶结构域中具有点突变的OsCIPK7突变体显示出增加的耐寒性，其可能潜在地用于分子育种。

我们发现OsCIPK7的这一点突变导致激酶结构域的激活环的构象变化，随后随着蛋白激酶活性的增加，因此赋予对低温胁迫的增加的耐受性。
 

NMT measurements show dynamic change of extracellular Ca²⁺ influx in live roots of various genetic backgrounds responding to cold shock. The blue background represents the duration of cold treatment. The number was the cold treatment time on the abscissa. The minus numbers were the time before the treatment.

NMT瞬时处理实验流速数据分析思路

 

文章链接：http://www.jipb.net/EN/10.1111/jipb.12800