NO, ROS, and cell death associated with caspase-like activity increase in stress-induced microspore embryogenesis of barley.
NO, ROS, and cell death associated with caspase-like activity increase in stress-induced microspore embryogenesis of barley.
复制标题
DOI：
10.1093/jxb/err400
复制
发表时间：
2012-03
影响因子：
6.9
通讯作者：
Testillano PS
中科院分区：
文献类型：
--
作者：
Rodríguez-Serrano M;Bárány I;Prem D;Coronado MJ;Risueño MC;Testillano PS
在特定的胁迫处理（冷、饥饿）下，离体小孢子可被诱导偏离其配子体发育，而转向胚胎发生，在短时间内形成单倍体胚和纯合育种系。诱导应激产生活性氧（ROS）和一氧化氮（NO），它们是介导细胞反应和细胞死亡的信号分子，改变了胚性小孢子反应，因此改变了该过程的效率。这项工作分析了细胞死亡，半胱天冬酶3样活性，ROS和NO的生产（使用荧光探针和共聚焦分析）诱导应力后，大麦小孢子培养物和胚性悬浮培养物，作为一个体外系统，允许容易处理的比较。在小孢子和悬浮培养物中，应激处理后半胱天冬酶3样活性和细胞死亡增加，而在非诱导的小孢子和悬浮培养物中ROS增加。用半胱天冬酶3抑制剂DEVD-CHO处理培养物显著降低了细胞死亡百分比。应激处理的胚性悬浮培养物表现出高NO信号和细胞死亡，而在对照悬浮培养物中用S-亚硝基谷胱甘肽（NO供体）处理导致甚至更高的细胞死亡。相比之下，在小孢子培养物中，在应激后检测到NO的产生，并且在4天的小孢子培养物中，在伴随细胞分裂启动的胚性小孢子中检测到NO的产生。随后的处理应力处理的小孢子培养物与ROS和NO清除剂导致在早期阶段的细胞死亡减少，但后来他们产生了延迟胚胎发育以及在小孢子中的胚胎发生的百分比减少。结果表明，在非诱导小孢子和胚性悬浮培养物中，ROS的增加参与了胁迫诱导的细胞程序性死亡的早期发生;而NO在两种离体培养体系中，在胁迫后发挥双重作用，一种参与了胚性悬浮培养物中的细胞程序性死亡，另一种参与了重编程小孢子中导致胚胎发生的细胞分裂的启动。
Under specific stress treatments (cold, starvation), in vitro microspores can be induced to deviate from their gametophytic development and switch to embryogenesis, forming haploid embryos and homozygous breeding lines in a short period of time. The inductive stress produces reactive oxygen species (ROS) and nitric oxide (NO), signalling molecules mediating cellular responses, and cell death, modifying the embryogenic microspore response and therefore, the efficiency of the process. This work analysed cell death, caspase 3-like activity, and ROS and NO production (using fluorescence probes and confocal analysis) after inductive stress in barley microspore cultures and embryogenic suspension cultures, as an in vitro system which permitted easy handling for comparison. There was an increase in caspase 3-like activity and cell death after stress treatment in microspore and suspension cultures, while ROS increased in non-induced microspores and suspension cultures. Treatments of the cultures with a caspase 3 inhibitor, DEVD-CHO, significantly reduced the cell death percentages. Stress-treated embryogenic suspension cultures exhibited high NO signals and cell death, while treatment with S-nitrosoglutathione (NO donor) in control suspension cultures resulted in even higher cell death. In contrast, in microspore cultures, NO production was detected after stress, and, in the case of 4-day microspore cultures, in embryogenic microspores accompanying the initiation of cell divisions. Subsequent treatments of stress-treated microspore cultures with ROS and NO scavengers resulted in a decreasing cell death during the early stages, but later they produced a delay in embryo development as well as a decrease in the percentage of embryogenesis in microspores. Results showed that the ROS increase was involved in the stress-induced programmed cell death occurring at early stages in both non-induced microspores and embryogenic suspension cultures; whereas NO played a dual role after stress in the two in vitro systems, one involved in programmed cell death in embryogenic suspension cultures and the other in the initiation of cell division leading to embryogenesis in reprogrammed microspores.
登录
查看更多内容
影响因子：
6.9
作者：
Bárány I;Fadón B;Risueño MC;Testillano PS
通讯作者：
Testillano PS
影响因子：
7.4
作者：
de Pinto, MC;Tommasi, F;De Gara, L
通讯作者：
De Gara, L
影响因子：
19
作者：
Green, D;Kroemer, G
通讯作者：
Kroemer, G
影响因子：
2.9
作者：
Fortes, Ana M.;Costa, Joana;Pais, Maria S.
通讯作者：
Pais, Maria S.
影响因子：
7.2
作者：
Clarke, A;Desikan, R;Neill, SJ
通讯作者：
Neill, SJ