光皮桦miR164及其靶基因NAC1在低氮胁迫中的表达分析

发布时间：2018-05-21 00:45

本文选题：光皮桦 + 低氮胁迫　；参考：《遗传》2016年02期

【摘要】：氮是植物生长发育所必需的大量营养元素,植物缺氮后严重影响地上部分生物量的积累,因此,揭示植物如何抵抗或适应低氮胁迫的分子机制具有重要意义。杨树(Populus tremula×P.alba)NAC1(NAM,ATAF,CUC1)基因位于调控网络上游,在低氮环境下调控下游关键基因的表达,进而调控根系生长以抵抗低氮胁迫。本文以光皮桦(Betula luminifera)G49-3无性系组培苗为材料,探讨了miR164及其靶基因NAC1对低氮胁迫的响应。通过RACE技术克隆了光皮桦NAC1基因(Gen Bank登录号:KT900889),全长1497 bp,编码358个氨基酸,N端具有高度保守的NAM结构域;运用5′-RACE验证了NAC1为miR164靶基因,切割位点在第10和11位碱基之间;采用q RT-PCR分析miR164与靶基因NAC1在低氮胁迫时的表达模式,发现miR164表达在根中的低氮处理前期(4 d)受到抑制,而后升高,而茎叶中表达模式与根不同;靶基因NAC1与miR164表达水平呈负相关,且在恢复实验组(重新添加全营养液)中,根中miR164表达上升,NAC1显示出相应的表达变化,暗示miR164及其靶基因NAC1可能在低氮胁迫响应中发挥调控功能。本研究结果有助于揭示miR164对NAC1在低氮胁迫响应中转录后水平的分子调控机制,为进一步研究miR164-NAC1在低氮胁迫响应中的功能提供有价值的信息。
[Abstract]:Nitrogen is a large amount of nutrient elements necessary for plant growth and development, and plant nitrogen deficiency seriously affects the accumulation of aboveground biomass. Therefore, it is important to reveal how plants resist or adapt to the molecular mechanism of low nitrogen stress. The gene of poplar (Populus tremula x P.alba) NAC1 (NAM, ATAF, CUC1) is located in the upstream of the regulation network and in low nitrogen environment. The expression of the downstream key genes was regulated and the root growth was regulated to resist low nitrogen stress. In this paper, the response of miR164 and its target gene NAC1 to low nitrogen stress was investigated with the tissue culture of Betula luminifera G49-3 clone. The NAC1 gene of birch (Gen Bank login number: KT900889) was cloned by RACE technology, and the whole length was 1497 BP. With 358 amino acids, the N terminal has a highly conserved NAM domain, and 5 '-RACE is used to verify that NAC1 is a miR164 target gene and the cutting site is between tenth and eleventh bases. Q RT-PCR is used to analyze the expression pattern of miR164 and target gene NAC1 at low nitrogen stress, and it is found that miR164 expression is inhibited in the early stage of low nitrogen treatment (4 d) in the root and then increases. The expression pattern of the stem and leaf was different from the root, and the target gene NAC1 was negatively correlated with the expression level of miR164, and the expression of miR164 in the root of the experimental group was increased and the NAC1 showed a corresponding expression change, suggesting that miR164 and its target gene NAC1 may play a regulatory function in the response to low nitrogen stress. The results of this study are helpful. The molecular regulatory mechanism of the post transcriptional level of miR164 in the response of NAC1 to low nitrogen stress provides valuable information for further study of the function of miR164-NAC1 in the response to low nitrogen stress.
【作者单位】：浙江农林大学亚热带森林培育国家重点实验室培育基地;
【基金】：国家自然科学基金项目(编号:31300566) 浙江省竹木新品种选育重大科技专项(编号:2012C12908-8)资助~~
【分类号】：S792.15

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