MSU-DOE Plant Research Laboratory, Michigan State University, 48824-1312, East Lansing, MI, USA.
Planta. 1990 Mar;180(4):548-54. doi: 10.1007/BF02411453.
We used RNA gel-blot analysis, in-situ hybridization, and nuclear run-on transcription to examine the effects of exogenous abscisic acid (ABA) on the spatial distribution of mRNA for the lectin wheat-germ agglutinin (WGA) in developing wheat (Triticum aestivum L. cv. Marshall) embryos and seedlings. When analyzed by RNA gel blots, both developing embryos and seedlings exhibited higher steady-state levels of WGA mRNA after ABA treatment. As determined by in-situ hybridization, incubation of developing embryos in 0.1 mM ABA resulted in accumulation of WGA mRNA in the epidermal and subepidermal cell layers of the radicle and seminal roots and throughout the rootcap and coleorhiza. This spatial distribution was identical to that in control embryos. Nuclear run-on transcription assays indicated that at least part of this increase is attributable to transcriptional induction. Thus, exogenous ABA is capable of inducing increased WGA mRNA accumulation only in cells where it is expressed during normal embryogenesis. When seeds were germinated in the absence of ABA, WGA mRNA was detected only in the rootcap. In contrast, seeds imbibed and germinated in the presence of ABA for 3 d exhibited a spatial distribution of WGA mRNA similar to that observed in developing embryos treated with ABA. In contrast, when ABA was added to 3-d-old seedlings, WGA mRNA was not detected in regions of the root beyond the rootcap. We conclude that exogenous ABA, when applied continuously from imbibition, causes retention of the embryo-specific pattern of WGA mRNA distribution and that the spatial pattern of WGA mRNA expression in roots does not change when ABA is added after germination.
我们使用 RNA 凝胶印迹分析、原位杂交和核转录跑带实验,研究外源脱落酸(ABA)对发育中的小麦(Triticum aestivum L. cv. Marshall)胚胎和幼苗中凝集素小麦胚凝集素(WGA)mRNA 空间分布的影响。用 RNA 凝胶印迹分析时,经 ABA 处理后,发育中的胚胎和幼苗的 WGA mRNA 稳态水平均升高。用原位杂交法分析时,将发育中的胚胎在 0.1 mM ABA 中孵育,导致 WGA mRNA 在胚根和初生根的表皮和表皮下细胞层以及根冠和胚根鞘中积累。这种空间分布与对照胚胎中的分布相同。核转录跑带实验表明,至少部分增加归因于转录诱导。因此,外源 ABA 仅能在正常胚胎发生过程中表达的细胞中诱导 WGA mRNA 积累增加。当种子在没有 ABA 的情况下萌发时,仅在根冠中检测到 WGA mRNA。相比之下,在 ABA 存在下吸水和萌发 3 天的种子表现出与 ABA 处理的发育胚胎中观察到的相似的 WGA mRNA 空间分布。相比之下,当 ABA 被添加到 3 天大的幼苗中时,在根冠以外的根区未检测到 WGA mRNA。我们得出结论,外源 ABA 从吸水开始连续施加时,会保留胚胎特异性的 WGA mRNA 分布模式,并且 ABA 在萌发后添加时,根中 WGA mRNA 的空间表达模式不会改变。