Identification and analysis of MKK and MPK gene families in canola (Brassica napus L.).
Identification and analysis of MKK and MPK gene families in canola (Brassica napus L.).
复制标题
双低油菜 MKK 和 MPK 基因家族的鉴定与分析
DOI：
10.1186/1471-2164-14-392
复制
发表时间：
2013-06-11
期刊：
影响因子：
4.4
通讯作者：
Jiang YQ
中科院分区：
文献类型：
--
作者：
Liang W;Yang B;Yu BJ;Zhou Z;Li C;Jia M;Sun Y;Zhang Y;Wu F;Zhang H;Wang B;Deyholos MK;Jiang YQ
背景真核细胞有丝分裂原激活蛋白激酶（MAPK/MPK）信号级联反应通过三种类型的可逆磷酸化激酶转导和放大环境信号以激活防御基因表达。油菜（油菜，Brassica napus）是温带地区的主要作物。MAPK和MAPK激酶（MAPKK/MKK）的鉴定和表征的油菜将有助于阐明它们在响应非生物和生物stress.ResultsWe描述的7 MKK（BnaMKK）和12 MPK（BnaMPK）的成员从油菜的鉴定和分析的作用。序列比对和系统发育分析的预测氨基酸序列的BnaMKKs和BnaMPKs分为四个不同的群体。我们还研究了亚细胞定位的四个和两个成员的BnaMKK和BnaMPK基因家族，分别使用绿色荧光蛋白（GFP），发现GFP信号在细胞核和细胞质。此外，我们通过酵母双杂交（Y2 H）分析BnaMKK和BnaMPKs之间的相互作用，以及BnaMPK和BnaWRKYs之间的相互作用，确定了几个有趣的相互作用对。我们定义了包括BnaMKK 9-BnaMPK 1/2-BnaWRKY 53、BnaMKK 2/4/5-BnaMPK 3/6-BnaWRKY 20/26和BnaMKK 9-BnaMPK 5/9/19/20的连续信令模块。其中，有几种相互作用以前没有在任何物种中描述过。通过双分子荧光互补（BiFC）测定在体内验证所选相互作用。通过实时荧光定量RT-PCR分析了油菜MKK和MPK基因对病原真菌、激素和非生物胁迫处理的转录响应，并鉴定了几个对水杨酸（SA）、草酸（OA）、核盘菌（Sclerotinia scleroecuminescence）或其他胁迫条件有响应的BnaMKK和BnaMPK基因。在油菜和拟南芥中的假定的直系同源物的表达模式的比较表明，转录表达模式一般是保守的，与一些差异暗示sub-functionalization.ConclusionsWe确定了七MKK和12 MPK基因从油菜，并研究其系统发育关系，转录表达模式，亚细胞定位，和蛋白质-蛋白质相互作用。并不是所有的表达模式和相互作用的油菜和拟南芥之间的保守，突出了从模式物种中推断作物的局限性。本文提供的数据首次系统描述了油菜中的MKK-MPK-WRKY信号模块，将进一步提高我们对防御反应的总体理解，并为未来的作物改良提供基础。
BackgroundEukaryotic mitogen-activated protein kinase (MAPK/MPK) signaling cascades transduce and amplify environmental signals via three types of reversibly phosphorylated kinases to activate defense gene expression. Canola (oilseed rape,Brassica napus) is a major crop in temperate regions. Identification and characterization of MAPK and MAPK kinases (MAPKK/MKK) of canola will help to elucidate their role in responses to abiotic and biotic stresses.ResultsWe describe the identification and analysis of sevenMKK(BnaMKK) and 12MPK(BnaMPK) members from canola. Sequence alignments and phylogenetic analyses of the predicted amino acid sequences of BnaMKKs and BnaMPKs classified them into four different groups. We also examined the subcellular localization of four and two members of BnaMKK and BnaMPK gene families, respectively, using green fluorescent protein (GFP) and, found GFP signals in both nuclei and cytoplasm. Furthermore, we identified several interesting interaction pairs through yeast two-hybrid (Y2H) analysis of interactions between BnaMKKs and BnaMPKs, as well as BnaMPK and BnaWRKYs. We defined contiguous signaling modules including BnaMKK9-BnaMPK1/2-BnaWRKY53, BnaMKK2/4/5-BnaMPK3/6-BnaWRKY20/26 and BnaMKK9-BnaMPK5/9/19/20. Of these, several interactions had not been previously described in any species. Selected interactions were validatedin vivoby a bimolecular fluorescence complementation (BiFC) assay. Transcriptional responses of a subset of canola MKK and MPK genes to stimuli including fungal pathogens, hormones and abiotic stress treatments were analyzed through real-time RT-PCR and we identified a few ofBnaMKKsandBnaMPKsresponding to salicylic acid (SA), oxalic acid (OA),Sclerotinia sclerotiorumor other stress conditions. Comparisons of expression patterns of putative orthologs in canola and Arabidopsis showed that transcript expression patterns were generally conserved, with some differences suggestive of sub-functionalization.ConclusionsWe identified sevenMKKand 12MPKgenes from canola and examined their phylogenetic relationships, transcript expression patterns, subcellular localization, and protein-protein interactions. Not all expression patterns and interactions were conserved between canola and Arabidopsis, highlighting the limitations of drawing inferences about crops from model species. The data presented here provide the first systematic description of MKK-MPK-WRKY signaling modules in canola and will further improve our understanding of defense responses in general and provide a basis for future crop improvement.
登录
查看更多内容
影响因子：
11.6
作者：
Ishihama, Nobuaki;Yamada, Reiko;Yoshioka, Hirofumi
通讯作者：
Yoshioka, Hirofumi
影响因子：
7.4
作者：
Holley, SR;Yalamanchili, RD;Stratmann, JW
通讯作者：
Stratmann, JW
影响因子：
11.6
作者：
Cessna, SG;Sears, VE;Low, PS
通讯作者：
Low, PS
影响因子：
64.8
作者：
Asai, T;Tena, G;Sheen, J
通讯作者：
Sheen, J
影响因子：
11.6
作者：
Doczi, Robert;Brader, Gunter;Hirt, Heribert
通讯作者：
Hirt, Heribert