Key Laboratory of Biology and Genetic Improvement of Maize in Southwest Region, Maize Research Institute, Sichuan Agricultural University, Chengdu, 611130, China.
Sichuan Tourism College, Chengdu, 610100, China.
BMC Genomics. 2019 Feb 27;20(1):159. doi: 10.1186/s12864-019-5506-7.
Maize is one of the primary crops of genetic manipulation, which provides an excellent means of promoting stress resistance and increasing yield. However, the differences in induction and regeneration capacity of embryonic callus (EC) among various genotypes result in genotypic dependence in genetic transformation.
In this study, embryonic calli of two maize inbred lines with strong redifferentiation capacity and two lines with weak redifferentiation capability were separately subjected to transcriptome sequencing analysis during the early redifferentiation stages (stage I, 1-3 d; stage II, 4-6 d; stage III, 7-9 d) along with their corresponding controls. A total of ~ 654.72 million cDNA clean reads were yielded, and 62.64%~ 69.21% clean reads were mapped to the reference genome for each library. In comparison with the control, the numbers of differentially expressed genes (DEGs) for the four inbred lines identified in the three stages ranged from 1694 to 7193. By analyzing the common and specific DEGs of the four materials, we found that there were 321 upregulated genes and 386 downregulated genes identified in the high-regeneration lines (141 and DH40), whereas 611 upregulated genes and 500 downregulated genes were specifically expressed in the low-regeneration lines (ZYDH381-1 and DH3732). Analysis of the DEG expression patterns indicated a sharp change at stage I in both the high- and low-regeneration lines, which suggested that stage I constitutes a crucial period for EC regeneration. Notably, the specific common DEGs of 141 and DH40 were mainly associated with photosynthesis, porphyrin and chlorophyll metabolism, ribosomes, and plant hormone signal transduction. In contrast, the DEGs in ZYDH381-1 and DH3732 were mainly related to taurine and hypotaurine metabolism, nitrogen metabolism, fatty acid elongation, starch and sucrose metabolism, phenylpropanoid biosynthesis, and plant circadian rhythm. More importantly, WOX genes, which have an ancestral role in embryo development in seed plants and promote the regeneration of transformed calli, were specifically upregulated in the two high-regeneration lines.
Our research contributes to the elucidation of molecular regulation during early redifferentiation in the maize embryonic callus.
玉米是遗传操作的主要作物之一,为提高抗逆性和增加产量提供了极好的手段。然而,不同基因型的胚胎愈伤组织(EC)的诱导和再生能力存在差异,导致遗传转化存在基因型依赖性。
本研究分别对两种胚胎愈伤组织具有较强再分化能力的玉米自交系和两种再分化能力较弱的自交系进行了早期再分化阶段(阶段 I:1-3d;阶段 II:4-6d;阶段 III:7-9d)的转录组测序分析,并与相应对照进行了比较。共产生约 6547.2 万条 cDNA 清洁读数,每个文库的清洁读数中有 62.64%~69.21%映射到参考基因组。与对照相比,四个自交系在三个阶段鉴定的差异表达基因(DEGs)数量在 1694 到 7193 个之间。通过分析四个材料的共同和特异的 DEGs,我们发现高再生系(141 和 DH40)中鉴定出 321 个上调基因和 386 个下调基因,而低再生系(ZYDH381-1 和 DH3732)中特异表达 611 个上调基因和 500 个下调基因。DEG 表达模式分析表明,高、低再生系在 I 期均有明显变化,表明 I 期是 EC 再生的关键时期。值得注意的是,141 和 DH40 的特异共同 DEGs 主要与光合作用、卟啉和叶绿素代谢、核糖体和植物激素信号转导有关。相比之下,ZYDH381-1 和 DH3732 的 DEGs 主要与牛磺酸和羟牛磺酸代谢、氮代谢、脂肪酸延伸、淀粉和蔗糖代谢、苯丙素生物合成和植物昼夜节律有关。更重要的是,WOX 基因在种子植物胚胎发育中具有祖先作用,并促进转化愈伤组织的再生,在两个高再生系中特异性上调。
本研究有助于阐明玉米胚胎愈伤组织早期再分化过程中的分子调控机制。