Zheng Qiaolin, Perry Sharyn E
Department of Plant and Soil Sciences, University of Kentucky, Lexington, Kentucky 40546-0312.
Plant Physiol. 2014 Mar;164(3):1365-77. doi: 10.1104/pp.113.234062. Epub 2014 Jan 30.
Somatic embryogenesis (SE) is a poorly understood process during which competent cells respond to inducing conditions, allowing the development of somatic embryos. It is important for the regeneration of transgenic plants, including for soybean (Glycine max). We report here that constitutive expression of soybean orthologs of the Arabidopsis (Arabidopsis thaliana) MADS box genes Agamous-like15 (GmAGL15) and GmAGL18 increased embryogenic competence of explants from these transgenic soybean plants. To understand how GmAGL15 promotes SE, expression studies were performed. Particular genes of interest involved in embryogenesis (abscisic acid-insensitive3 and FUSCA3) were found to be directly up-regulated by GmAGL15 by using a combination of quantitative reverse transcription-polymerase chain reaction and chromatin immunoprecipitation. To look more broadly at changes in gene expression in response to GmAGL15, we assessed the transcriptome using the Affymetrix Soybean Genome Array. Interestingly, the gene expression profile of 35Spro:GmAGL15 explants (0 d in culture) was found to resemble nontransgenic tissue that had been induced for SE by being placed on induction medium for 3 d, possibly explaining the more rapid SE development observed on 35Spro:GmAGL15 tissue. In particular, transcripts from genes related to the stress response showed increased transcript accumulation in explants from 35Spro:GmAGL15 tissue. These same genes also showed increased transcript accumulation in response to culturing nontransgenic soybean explants on the medium used to induce SE. Overexpression of GmAGL15 may enhance SE by making the tissue more competent to respond to 2,4-dichlorophenoxyacetic acid induction by differential regulation of genes such as those involved in the stress response, resulting in more rapid and prolific SE.
体细胞胚胎发生(SE)是一个尚未被充分理解的过程,在此过程中,感受态细胞对诱导条件作出反应,从而实现体细胞胚胎的发育。这对于转基因植物的再生非常重要,包括大豆(Glycine max)。我们在此报告,拟南芥(Arabidopsis thaliana)MADS盒基因类AGAMOUS15(GmAGL15)和GmAGL18的大豆直系同源基因的组成型表达提高了这些转基因大豆植株外植体的胚胎发生能力。为了了解GmAGL15如何促进体细胞胚胎发生,我们进行了表达研究。通过定量逆转录-聚合酶链反应和染色质免疫沉淀相结合的方法,发现参与胚胎发生的特定感兴趣基因(脱落酸不敏感3和FUSCA3)被GmAGL15直接上调。为了更广泛地研究响应GmAGL15的基因表达变化,我们使用Affymetrix大豆基因组阵列评估了转录组。有趣的是,发现35Spro:GmAGL15外植体(培养0天)的基因表达谱类似于在诱导培养基上放置3天诱导体细胞胚胎发生的非转基因组织,这可能解释了在35Spro:GmAGL15组织上观察到的更快的体细胞胚胎发生发育。特别是,与应激反应相关的基因转录本在35Spro:GmAGL15组织的外植体中显示出转录积累增加。这些相同的基因在将非转基因大豆外植体培养在用于诱导体细胞胚胎发生的培养基上时也显示出转录积累增加。GmAGL15的过表达可能通过对诸如参与应激反应的基因进行差异调节,使组织更能响应2,4-二氯苯氧乙酸诱导,从而增强体细胞胚胎发生,导致更快且更多产的体细胞胚胎发生。