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首页> 外文期刊>Planta: An International Journal of Plant Biology >Two similar but distinct second intron fragments from tobacco AGAMOUS homologs confer identical floral organ-specific expression sufficient for generating complete sterility in plants
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Two similar but distinct second intron fragments from tobacco AGAMOUS homologs confer identical floral organ-specific expression sufficient for generating complete sterility in plants

机译：来自烟草AGAMOUS同源物的两个相似但截然不同的第二内含子片段赋予相同的花器官特异性表达，足以在植物中产生完全的不育性

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摘要

The carpel- and stamen-specific AtAGIP promoter derived from the Arabidopsis AGAMOUS (AG) second intron/enhancer is ideal for engineering complete sterility but it is highly host-specific. To ascertain whether a chimeric promoter with similar tissue specificity can be created for species other than Arabidopsis, we isolated two similar but distinct AG second intron/enhancers from tobacco (NtAGI-1 and NtAGI-2) and analyzed their ability to drive floral organ-specific expression in plants through the creation of forward- and reverse-oriented chimeric promoters, fNtAGIP1, rNtAGIP1, fNtAGIP2 and rNtAGIP2. Analyses of transgenic plants bearing each respective promoter fused to the beta-glucuronidase (GUS) reporter gene showed that all four promoters are able, like the AtAGIP, to drive very similar carpel- and stamen-specific expression without any leaky activity in vegetative tissues. These results indicate that unlike their counterparts in rice and maize, the tobacco NtAGI-1 and NtAGI-2 enhancers share a highly conserved regulatory function. Interestingly, all four promoters display additional tissue specificity in petals, and their activity is influenced by the orientation of the incorporated enhancer, with reverse-oriented enhancers exhibiting approximately double the effectiveness of forward-oriented enhancers. These properties are novel and have not been observed with the AtAGIP promoter in Arabidopsis. As expected, these highly specific promoters can also direct the expression of the DT-A cytotoxic gene exclusively in carpels, stamens and petals, resulting in complete sterility through the precise ablation of targeted floral organs. Further analyses demonstrated that the resulting trait is mitotically stable, which is critical for the long-term containment of seed-, pollen- and fruit-mediated gene flow in field conditions.
机译：拟南芥AGAMOUS（AG）第二内含子/增强子衍生的心皮和雄蕊特异的AtAGIP启动子是工程完全无菌的理想选择，但具有高度宿主特异性。为了确定是否可以为拟南芥以外的物种创建具有相似组织特异性的嵌合启动子，我们从烟草中分离了两个相似但截然不同的AG第二内含子/增强子（NtAGI-1和NtAGI-2），并分析了它们驱动花器官的能力。通过创建正向和反向定向嵌合启动子fNtAGIP1，rNtAGIP1，fNtAGIP2和rNtAGIP2在植物中的特异性表达。对携带每个分别与β-葡萄糖醛酸苷酶（GUS）报告基因融合的启动子的转基因植物进行的分析表明，与AtAGIP一样，所有四个启动子都能够驱动非常相似的心皮和雄蕊特异性表达，而在营养组织中没有任何渗漏活性。这些结果表明，烟草NtAGI-1和NtAGI-2增强剂与水稻和玉米中的对应物不同，它们具有高度保守的调节功能。有趣的是，所有四个启动子在花瓣中都显示出额外的组织特异性，并且它们的活性受掺入的增强子的方向影响，而反向增强子的功效大约是向前增强子的两倍。这些特性是新颖的，尚未在拟南芥中用AtAGIP启动子观察到。正如预期的那样，这些高度特异性的启动子还可以指导DT-A细胞毒性基因仅在心皮，雄蕊和花瓣中表达，从而通过精确消融目标花卉器官来实现完全无菌。进一步的分析表明，产生的性状是有丝分裂稳定的，这对于在田间条件下长期抑制种子，花粉和果实介导的基因流至关重要。

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