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首页> 外文期刊>Journal of Experimental Botany >Identification of AHK2-and AHK3-like cytokinin receptors in Brassica napus reveals two subfamilies of AHK2 orthologues
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Identification of AHK2-and AHK3-like cytokinin receptors in Brassica napus reveals two subfamilies of AHK2 orthologues

机译：甘蓝型油菜中AHK2和AHK3样细胞素受体的鉴定揭示了AHK2矫形器的两个亚属

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摘要

Cytokinin (CK) signalling is known to play key roles in the regulation of plant growth and development, crop yields, and tolerance to both abiotic stress and pathogen defences, but the mechanisms involved are poorly characterized in dicotyledonous crops. Here the identification and functional characterization of sensor histidine kinases homologous to Arabidopsis CK receptors AHK2 and AHK3 in winter oilseed rape are presented. Five CHASE-containing His kinases were identified in Brassica napus var. Tapidor (BnCHK1-BnCHK5) by heterologous hybridization of its genomic library with gene-specific probes from Arabidopsis. The identified bacterial artificial chromosome (BAC) clones were fingerprinted and representative clones in five distinct groups were sequenced. Using a bioinformatic approach and cDNA cloning, the precise gene and putative protein domain structures were determined. Based on phylogenetic analysis, four AHK2 (BnCHK1-BnCHK4) homologues and one AHK3 (BnCHK5) homologue were defined. It is further suggested that BnCHK1 and BnCHK3, and BnCHK5 are orthologues of AHK2 and AHK3, originally from the B. rapa A genome, respectively. BnCHK1, BnCHK3, and BnCHK5 displayed high affinity for trans-zeatin (1-3 nM) in a live-cell competitive receptor assay, but not with other plant hormones (indole acetic acid, GA3, and abscisic acid), confirming the prediction that they are genuine CK receptors. It is shown that BnCHK1 and BnCHK3, and BnCHK5 display distinct preferences for various CK bases and metabolites, characteristic of their AHK counterparts, AHK2 and AHK3, respectively. Interestingly, the AHK2 homologues could be divided into two subfamilies (BnCHK1/BnCK2 and BnCHK3/BnCHK4) that differ in putative transmembrane domain topology and CK binding specificity, thus implying potential functional divergence.
机译：已知细胞素素（CK）信号传导在植物生长和发育，作物产量和对非生物胁迫和病原体防御的调节中起关键作用，但涉及的机制表征在二坐子作物中的特征不佳。这里提出了对亚洲油菜中的拟南芥CK受体AHK2和AHK3同源的传感器组氨酸激酶的鉴定和功能表征。在Brassica Napus Var中鉴定了含有其激酶的五个追逐。通过拟南芥与基因特异性探针的基因组文库的特点杂交所识别的细菌人工染色体（BAC）克隆是指指纹的，并测序五个不同组中的代表克隆。使用生物信息化方法和cDNA克隆，确定精确基因和推定的蛋白质结构域结构。基于系统发育分析，定义了四个AHK2（BNCHK1-BNCHK4）同源物和一个AHK3（BNCHK5）同源物。进一步表明BNCHK1和BNCHK3和BNCHK5分别是AHK2和AHK3的直噻吩，分别来自B. RAPA A基因组。 BNCHK1，BNCHK3和BNCHK5在活细胞竞争受体测定中对Trans-Zeatin（1-3nm）显示出高亲和力，但与其他植物激素（吲哚乙酸，Ga3和脱钙酸）没有，确认预测它们是真正的CK受体。结果表明，BNCHK1和BNCHK3，BNCHK5分别显示出各种CK基础和代谢物，其AHK对应物，AHK2和AHK3的特征的不同偏好。有趣的是，AHK2同源物可以分为两个亚属（BNCHK1 / BNCK2和BNCHK3 / BNCHK4），其具有推定的跨膜域拓扑结构和CK结合特异性，因此暗示潜在的功能性分歧。

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