Okubara P A, Pawlowski K, Murphy T M, Berry A M
Department of Environmental Horticulture, University of California, Davis 95616, USA.
Plant Physiol. 1999 Jun;120(2):411-20. doi: 10.1104/pp.120.2.411.
N2-fixing symbiotic root nodules of the actinorhizal host Datisca glomerata express Dgrca (D. glomerata Rubisco activase) mRNA, a transcript usually associated with photosynthetic organs or tissues. In northern blots a mature, 1700-nucleotide Dgrca mRNA was detected in green plant organs (leaves, flowers, and developing fruits) and in nodules but was not detected in roots. A second message of 3000 nucleotides was observed only in nodules. Both size classes of transcripts were polyadenylated. The larger transcript was 2- to 5-fold more abundant than the mature mRNA; it was hybridized to an intronic probe, indicating that a stable, incompletely spliced transcript was accumulating. Treatment with light on excised nodules did not alter the relative abundance of the two species. In in situ hybridizations the Dgrca message was expressed intensely in the nuclei of infected cells. The Dgrca transcripts also accumulated at lower levels in uninfected cortical cells adjacent to the periderm and the vascular cylinder. mRNA encoding the large subunit of Rubisco (DgrbcL) was abundant in mature infected cells and in the amyloplast-rich sheath of uninfected cortical cells lying between the infected cells and nodule periderm. The proteins Rubisco activase, Rubisco, and the 33-kD O2-evolving complex subunit did not accumulate to detectable levels, indicating that a functional photosynthetic apparatus was not prevalent in nodule tissue. Signals or factors required for the transcription of Dgrca appeared to be present in nodules, but efficient splicing and translation of the message were not observed in Frankia-infected tissue where transcript accumulation was highest.
放线菌根瘤植物聚花 Datisca 的固氮共生根瘤表达 Dgrca(聚花 Datisca 核酮糖-1,5-二磷酸羧化酶/加氧酶激活酶)mRNA,这种转录本通常与光合器官或组织相关。在 Northern 杂交中,在绿色植物器官(叶片、花朵和发育中的果实)以及根瘤中检测到一条成熟的、1700 个核苷酸的 Dgrca mRNA,但在根中未检测到。仅在根瘤中观察到一条 3000 个核苷酸的第二条信息。两种大小类别的转录本都进行了多聚腺苷酸化。较大的转录本比成熟 mRNA 丰富 2 至 5 倍;它与内含子探针杂交,表明一种稳定的、未完全剪接的转录本正在积累。对切除的根瘤进行光照处理不会改变这两种转录本的相对丰度。在原位杂交中,Dgrca 信息在受感染细胞的细胞核中强烈表达。Dgrca 转录本在与周皮和维管束相邻的未受感染皮层细胞中也有较低水平的积累。编码核酮糖-1,5-二磷酸羧化酶大亚基(DgrbcL)的 mRNA 在成熟的受感染细胞以及位于受感染细胞和根瘤周皮之间的富含造粉体的未受感染皮层细胞鞘中大量存在。核酮糖-1,5-二磷酸羧化酶激活酶、核酮糖-1,5-二磷酸羧化酶和 33-kD 放氧复合体亚基蛋白未积累到可检测水平,表明功能性光合装置在根瘤组织中并不普遍。Dgrca 转录所需的信号或因子似乎存在于根瘤中,但在转录积累最高的弗兰克氏菌感染组织中未观察到该信息的有效剪接和翻译。