Molecular cloning and expression analysis of a RGA-like gene responsive to plant hormones in Brassica napus
Molecular cloning and expression analysis of a RGA-like gene responsive to plant hormones in Brassica napus
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DOI：
10.1007/s11033-011-0943-7
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发表时间：
2012-02-01
影响因子：
2.8
通讯作者：
Wang, Maolin
中科院分区：
文献类型：
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作者：
Gao, Yong;Chen, Jianmin;Wang, Maolin
DELLA蛋白是GA诱导生长的负调节因子。DELLA蛋白家族的特征在于DELLA结构域对于DELLA阻遏物的GA依赖性蛋白酶体降解是必需的。从甘蓝型油菜（Brassica napus）中克隆了一个与拟南芥RGA（AtRGA）具有高度同源性的DELLA蛋白的全长cDNA，命名为BnRGA。BnRGA全长cDNA包含一个1,740 bp的开放阅读框（ORF），编码579个氨基酸残基的前体蛋白。比较和生物信息学分析表明，BnRGA与DELLA蛋白具有高度的同源性，包含DELLA结构域、TVHYNP结构域、VHIID结构域和RVER结构域。利用实时荧光定量PCR技术，对BnRGA和我们先前分离的两个基因BnGID 1a和BnSLY 1在B中的表达模式进行了分析。通过添加外源赤霉素-3（GA（3））、GA生物合成抑制剂多效唑（PAC）和脱落酸（阿坝）对油菜幼苗的生长发育进行研究。结果表明，GA（3）处理及PAC和阿坝诱导后，BnGID 1a和BnSLY 1的表达均下调。这些结果表明，在B中BnGID 1a和BnSLY 1的表达可能受到内源GA水平的负调控。油菜。低浓度的GA（3）、PAC和阿坝对BnRGA的调控作用不明显。这表明GA-GID 1-SCF-DELLA复合物可能具有自我调节机制，从而保持BnRGA在B中表达水平的稳定性。油菜。
DELLA proteins are negative regulators of GA-induced growth. DELLA protein family is characterized by a DELLA domain essential for GA-dependent proteasomal degradation of DELLA repressors. A full-length cDNA encoding a putative DELLA protein with high sequence homology to Arabidopsis thaliana RGA (AtRGA), designated as BnRGA, was isolated from Brassica napus. The full-length cDNA of BnRGA contained a 1,740 bp open reading frame (ORF) encoding a precursor protein of 579 amino acid residues. Comparative and bio-informatics analyses revealed that BnRGA showed a high degree of homology with DELLA proteins and contained the DELLA domain, TVHYNP domain, VHIID domain and RVER domain. Using real-time PCR, the expression patterns of BnRGA and two our previously isolated genes, BnGID1a and BnSLY1 in B. napus, were analyzed by adding exogenous gibberellins acid-3 (GA(3)), GA biosynthetic inhibitor paclobutrazol (PAC) and abscisic acid (ABA). The results showed that the expression of BnGID1a and BnSLY1 was down-regulated after treated by GA(3) and induced by PAC and ABA. These results suggest that the expression of BnGID1a and BnSLY1 may be negatively regulated by the level of endogenous GA in B. napus. Moreover, BnRGA was not significantly regulated by GA(3), PAC and ABA in the low concentrations. These suggest that GA-GID1-SCF-DELLA complex may have a mechanism of self-regulation, thereby preserving the stability of the expression level of BnRGA in B. napus.