Global and local perturbation of the tomato microRNA pathway by a trans-activated DICER-LIKE 1 mutant.
Global and local perturbation of the tomato microRNA pathway by a trans-activated DICER-LIKE 1 mutant.
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DOI：
10.1093/jxb/ert428
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发表时间：
2014-02
影响因子：
6.9
通讯作者：
Arazi T
中科院分区：
文献类型：
--
作者：
Kravchik M;Sunkar R;Damodharan S;Stav R;Zohar M;Isaacson T;Arazi T
DICER-like 1 (DCL1) 是 microRNA (miRNA) 生物发生的主要参与者，因此，其少数已知的功能丧失突变体要么是致命的，要么表现出发育停滞。因此，通过反向遗传学产生 dcl1 突变体并在晚期发育器官中进行 DCL1 功能分析具有挑战性。在这里，这些挑战通过反式激活 RNA 干扰的独特使用得到了解决。通过将生成的响应线 (OP:SlDCL1IR) 与适当的驱动线交叉，诱导全局以及器官特异性番茄 DCL1 (SlDCL1) 沉默。组成型反式激活使 SlDCL1 水平降低约 95%，导致严重异常，包括发芽后生长停滞，并伴有 miRNA 和 21 核苷酸小 RNA 水平降低，但 22 核苷酸小 RNA 水平显着升高。 22 核苷酸小 RNA 的增加与 SlDCL2b 和 SlDCL2d 的特异性上调相关，这可能参与其生物发生。叶和花特异性 OP:SlDCL1IR 反式激活分别抑制叶片生长、诱导花芽过早衰老并产生苍白花瓣，这强调了 SlDCL1 依赖性小 RNA 在这些过程中的重要性。总之，这些结果确立了 OP:SlDCL1IR 作为分析番茄中 SlDCL1 介导的基因调控过程的有效工具。
DICER-like 1 (DCL1) is a major player in microRNA (miRNA) biogenesis and accordingly, its few known loss-of-function mutants are either lethal or display arrested development. Consequently, generation of dcl1 mutants by reverse genetics and functional analysis of DCL1 in late-developing organs are challenging. Here, these challenges were resolved through the unique use of trans-activated RNA interference. Global, as well as organ-specific tomato DCL1 (SlDCL1) silencing was induced by crossing the generated responder line (OP:SlDCL1IR) with the appropriate driver line. Constitutive trans-activation knocked down SlDCL1 levels by ~95%, resulting in severe abnormalities including post-germination growth arrest accompanied by decreased miRNA and 21-nucleotide small RNA levels, but prominently elevated levels of 22-nucleotide small RNAs. The increase in the 22-nucleotide small RNAs was correlated with specific up-regulation of SlDCL2b and SlDCL2d, which are probably involved in their biogenesis. Leaf- and flower-specific OP:SlDCL1IR trans-activation inhibited blade outgrowth, induced premature bud senescence and produced pale petals, respectively, emphasizing the importance of SlDCL1-dependent small RNAs in these processes. Together, these results establish OP:SlDCL1IR as an efficient tool for analysing processes regulated by SlDCL1-mediated gene regulation in tomato.
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影响因子：
11.6
作者：
ERRAMPALLI, D;PATTON, D;MEINKE, D
通讯作者：
MEINKE, D
影响因子：
7.4
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Fernandez, Ana I.;Viron, Nicolas;Hilson, Pierre
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影响因子：
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作者：
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Cline, MG
影响因子：
5.1
作者：
Hendelman, A.;Buxdorf, K.;Arazi, T.
通讯作者：
Arazi, T.