Jain Mukesh, Li Qin-Bao, Chourey Prem S
Department of Plant Pathology, University of Florida, Gainesville, FL 32611-0680, USA.
Physiol Plant. 2008 Sep;134(1):161-73. doi: 10.1111/j.1399-3054.2008.01106.x. Epub 2008 Apr 19.
A full-length cDNA clone, SbSnRK1b (1530 bp, GenBank accession no. EF544393), encoding a putative serine/threonine protein kinase homologue of yeast (Saccharomyces cerevisiae) SNF1, was isolated from developing endosperm of sorghum [Sorghum bicolor (L.) Moench]. Multiple sequence alignment data showed a phylogenetic affiliation of the sorghum clone with the SnRK1b group of protein kinases that are highly expressed in cereal seed endosperm. The DNA gel blot analyses indicated that SbSnRK1b gene is present as a single- or low copy number gene in sorghum. The RNA and protein gel blot analyses confirmed the expression of SbSnRK1b in developing sorghum caryopses, overlapping with the starch biosynthesis phase, 12-24 days after fertilization. In situ hybridization and immunolocalization data resolved the spatial specificity of SbSnRK1b expression in the basal endosperm transfer cell layer, the unique port of assimilate unloading in the growing sorghum seed. Expression of SbSnRK1b was also evident in the developing sorghum microspores, coincident with the onset of starch deposition phase. As in sorghum, similar spatiotemporal specificity of SnRK1b expression was observed during maize (Zea mays L.) seed development. However, discordant in situ hybridization and immunolocalization data indicated that the expression of SbSnRK1b homologue in maize is under posttranscriptional control during endosperm development.
从高粱[双色高粱(L.)Moench]发育中的胚乳中分离出一个全长cDNA克隆SbSnRK1b(1530 bp,GenBank登录号EF544393),其编码一种假定的与酵母(酿酒酵母)SNF1同源的丝氨酸/苏氨酸蛋白激酶。多序列比对数据显示,高粱克隆与在谷物种子胚乳中高表达的SnRK1b蛋白激酶组具有系统发育关系。DNA凝胶印迹分析表明,SbSnRK1b基因在高粱中以单拷贝或低拷贝数基因形式存在。RNA和蛋白质凝胶印迹分析证实了SbSnRK1b在发育中的高粱颖果中的表达,与受精后12 - 24天的淀粉生物合成阶段重叠。原位杂交和免疫定位数据解析了SbSnRK1b在胚乳基部转移细胞层中表达的空间特异性,胚乳基部转移细胞层是生长中的高粱种子中同化物卸载的独特部位。SbSnRK1b在发育中的高粱小孢子中的表达也很明显,与淀粉沉积阶段的开始一致。与高粱一样,在玉米(Zea mays L.)种子发育过程中也观察到了SnRK1b表达的类似时空特异性。然而,不一致的原位杂交和免疫定位数据表明,玉米中SbSnRK1b同源物的表达在胚乳发育过程中受到转录后调控。