生长素和细胞分裂素对拟南芥主根细胞分裂和伸长的影响
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摘要
从细胞生物学出发,所有生物体器官、组织的发育都可以划分为细胞分裂和细胞生长这两个基本事件,因此细胞分裂和细胞生长调控机制研究一直是生物学界一个令人着迷的热门研究领域。在植物界,作为模式植物的拟南芥其主根结构相对简单,从形态学上可以分为分生区、伸长区、成熟区等几个区域。不同于其它组织器官,细胞的分裂、伸长(生长)和分化分别在相应的分区内进行,分生区主要进行细胞分裂,伸长区进行细胞生长,因此拟南芥主根是研究细胞分裂和细胞生长的理想材料。而植物激素,特别是生长素和细胞分裂素都参与调节细胞的分裂、伸长和分化,关于生长素调控细胞的分裂、伸长方面的研究,目前也有许多的的报道,但进一步的机制仍有待于进一步的研究。为了进一步探讨生长素对细胞的分裂和伸长的调控机制,我们以拟南芥的主根为实验材料,通过外源施加不同浓度的IAA、NAA、2,4-D,分析比较各种处理下的主根根长及各个分区的变化,结果表明:虽然外源施加生长素都可以抑制拟南芥主根生长,但三种生长素对主根的抑制效应可分为两类:IAA、NAA主要影响了伸长区的细胞伸长;2,4-D主要影响了分生区的细胞分裂。另外通过比较6-BA对主根的抑制效应,可以发现2,4-D的这种抑制效应类似于6-BA作用。通过Real Time-PCR,对生长素和细胞分裂素信号途径中相关基因的表达进行分析,还发现:生长素极性运输载体PIN1、PIN3、PIN7、AUX1的表达受到IAA和NAA的上调,但受到2,4-D和6-BA的下调;同时细胞分裂素信号途径中的A型ARRs家族的一些成员受到IAA和NAA下调,受到2,4-D和6-BA的上调。这些结果表明:施加四种不同的激素时,拟南芥主根的生长都受到抑制,而这种抑制效应对于不同的激素来说可能通过不同的调节机制起作用的,IAA和NAA可能作用于伸长区,调节伸长区细胞的伸长,2,4-D和6-BA作用于分生区,调节分生区细胞的分裂,并且这种调节机制可能是通过调节生长素运输载体基因或A型ARRs的表达来实现的。
     这些研究结果加深我们对激素之间的相互作用的理解,通过对细胞的分裂和伸长与根长之间的关系的理解,有利于我们解释器官大小的不同和个体大小的差异。
From the angle of cell biology, the organ and organization development of all organisms can be divided into two basic process which inclued cell division and cell growth. Therefore, the mechanism of regulating cell division and cell growth has been a fascinating research fields in biology. In the plant kingdom, as a mode plant, the morphology structure of Aarabidopisis thaliana priamary root is relatively simple and can be divided into the meristematic zone, elongation zone and mature zones. Unlike other organs, cell division, elongation (growth) and differentiation were occured in different districts, which provide an ideal material for studying cell division, elongation and differentiation. Plant hormones,in particular,auxin and cytokinin are involved in regulating cell division, elongation and differentiation. Although there are some studies about the mechanism of auxin regulation of cell division, elongation and differentiation, it also need further study.
     To further investigate the mechanism how auxin control cell division and elongation, we use the primary root of Arabidopsis thaliana as experimental material. By applying different concentrations of exogenous IAA, NAA,2,4-D, we analysis and comparison the primary root length and changes of each parti-tion in various treatments,while applying exogenous auxin can inhibit the grow-th of Arabidopsis primary root, the inhibitory effect of auxin on the primary root can be divided into two categories:IAA, NAA mainly affects the cell elongation of elongationzone, and 2,4-D major affects the ability to divide of meristem zone. Furthermore,comparing with 6-benzylaminopurine(6-BA) on the inhibitory effect of the primary root, the effect of 2,4-D is similar to 6-BA. We analysis the expression genes of auxin and cytokinin signaling pathway using Realtime-PCR, and found the expression of auxin polar transport carrier genes PIN1,PIN3, PIN7, AUX1 were upregulated by IAA and NAA, but reduced by 2,4-D and 6-BA; while A-type ARRs family members of cytokinin signaling pathway were downregulated by the IAA and NAA, and upregulated by 2,4-D and 6-BA. These results show that by applying exogenous IAA,NAA,2,4-D and 6-BA,the growth of primary root was inhibitied and the mechanisms of regulation was different. Based on their effects on cell division, elongation, Auxins and cytokinin can be divided into two classes:IAA and NAA may act on elongation zone inhibiting root growth primarily through regulating cell elongation; 2,4-D and 6-BA inhibit root growth by regulating meristematic zone cell division. This process may be through regulating the expression of polar auxin transport carrier or A-type ARRs.
     These findings deepen our understanding of the interaction between plant hormones, through the relationship between cell division,cell enlongtion and root length will help us to explain the differences of different organ size and body size.
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