Modulation of embryo-forming capacity in culture through the expression of Brassica genes involved in the regulation of the shoot apical meristem.
Modulation of embryo-forming capacity in culture through the expression of Brassica genes involved in the regulation of the shoot apical meristem.
复制标题
DOI：
10.1093/jxb/erq222
复制
发表时间：
2010-09
影响因子：
6.9
通讯作者：
Stasolla C
中科院分区：
文献类型：
--
作者：
Elhiti M;Tahir M;Gulden RH;Khamiss K;Stasolla C
拟南芥中的体细胞胚胎发生是通过在含有 2,4-D 的诱导培养基上培养弯曲子叶胚胎 14 天，然后转移到无激素的发育培养基上来实现的。从甘蓝 (Bo)、白菜 (Br) 和甘蓝型油菜 (Bn) 中分离出与拟南芥 SHOOTMERISTEMLESS (STM)、CLAVATA 1 (CLV1) 和 ZWILLE (ZLL) 同源的几个基因，并在拟南芥中异位表达，以评估它们对体细胞胚胎发生的影响。 BoSTM、BrSTM 和 BnSTM 的异位表达增加了体细胞胚胎的数量，而在过表达 BnCLV1 的品系中观察到不同的效果，其中体细胞胚胎的形成受到严重抑制。 BnZLL 的引入对拟南芥体细胞胚胎发生没有任何影响。在过表达芸苔属 STM 的品系中观察到胚胎形成能力的增加与诱导信号 2,4-D 的较低需求以及界定胚胎发生细胞形成的 WUSCHEL (WUS) 的较高表达有关。这与 35S::BnCLV1 系形成对比，35S::BnCLV1 系表现出对外源 2,4-D 的最高需求和减少的 WUS 表达。进行微阵列研究以监测拟南芥体细胞胚胎发生过程中野生型 (WT) 系和 BoSTM 过表达系之间转录水平的整体变化，这表明体细胞胚胎产量的提高最为显着。 BoSTM 的引入影响了许多涉及激素感知和信号传导的基因的表达，以及编码 DNA 甲基转移酶和谷胱甘肽代谢酶的基因。为证实一些微阵列结果而进行的药理学实验表明，诱导阶段 DNA 的整体低甲基化以及随后发育阶段谷胱甘肽池向氧化状态的转变促进了拟南芥体细胞胚胎发生。这两个事件都发生在 35S::BoSTM 系中，但不在 WT 系中。甘蓝 STM 表达的改变也对欧洲油菜小孢子衍生的胚胎发生产生深远影响。过表达 BnSTM 的品系中小孢子衍生胚的产量增加，而下调 BnSTM 的反义品系中小孢子衍生胚的产量显着降低。
Somatic embryogenesis in Arabidopsis is achieved by culturing bending-cotyledon embryos on a 2,4-D-containing induction medium for 14 d followed by a transfer on to a hormone-free development medium. Several genes orthologous to Arabidopsis SHOOTMERISTEMLESS (STM), CLAVATA 1 (CLV1), and ZWILLE (ZLL) were isolated from Brassica oleracea (Bo), B. rapa (Br), and B. napus (Bn), and ectopically expressed in Arabidopsis to assess their effects on somatic embryogenesis. Ectopic expression of BoSTM, BrSTM, and BnSTM increased the number of somatic embryos, whereas a different effect was observed in lines overexpressing BnCLV1 in which somatic embryo formation was severely repressed. The introduction of BnZLL did not have any effects on Arabidopsis somatic embryogenesis. The increased embryo-forming capacity observed in lines overexpressing Brassica STM was associated with a lower requirement for the inductive signal 2,4-D, and a higher expression of WUSCHEL (WUS) which demarcates the formation of embryogenic cells. This was in contrast to the 35S::BnCLV1 lines which showed the highest requirement for exogenous 2,4-D and a reduced WUS expression. Microarray studies were conducted to monitor global changes in transcript levels during Arabidopsis somatic embryogenesis between the wild-type (WT) line and a BoSTM-overexpressing line, which showed the most pronounced enhancement of somatic embryo yield. The introduction of BoSTM affected the expression of many genes involved in hormone perception and signalling, as well as genes encoding DNA methyltransferases and enzymes of glutathione metabolism. Pharmacological experiments performed to confirm some of the microarray results showed that Arabidopsis somatic embryogenesis is encouraged by a global hypomethylation of the DNA during the induction phase and by a switch of the glutathione pool towards an oxidized state during the subsequent development phase. Both events occurred in the 35S::BoSTM line, but not in the WT line. Altered expression of Brassica STM also had profound effects on B. napus microspore-derived embryogenesis. The yield of microspore-derived embryos increased in lines overexpressing BnSTM and significantly decreased in antisense lines down-regulating BnSTM.
登录
查看更多内容
影响因子：
6.2
作者：
Bassuner, Burgund M.;Lam, Russell;Yeung, Edward C.
通讯作者：
Yeung, Edward C.
影响因子：
5.4
作者：
LOSCHIAVO, F;PITTO, L;TERZI, M
通讯作者：
TERZI, M
影响因子：
7.2
作者：
CHAUDHURY, AM;LETHAM, S;DENNIS, ES
通讯作者：
DENNIS, ES
DOI：
10.1016/s0044-328x(82)80040-8
发表时间：
1982-01-01
期刊：
ZEITSCHRIFT FUR PFLANZENPHYSIOLOGIE
影响因子：
--
作者：
LICHTER, R
通讯作者：
LICHTER, R
DOI：
10.1111/j.2517-6161.1995.tb02031.x
发表时间：
1995-01-01
影响因子：
5.8
作者：
BENJAMINI, Y;HOCHBERG, Y
通讯作者：
HOCHBERG, Y