Jalali Saakshi, Kancharla Nagesh, Yepuri Vijay, Arockiasamy Savarimuthu
Agronomy Division, Reliance Technology Group, Reliance Industries Limited, Navi Mumbai, Maharashtra India.
3 Biotech. 2020 Mar;10(3):91. doi: 10.1007/s13205-020-2082-0. Epub 2020 Feb 4.
is one of the major sources of renewable energy due to potential use of its oil as a biofuel. The genome of this crop is constituted by the high content of repetitive elements. We employed the Hi-C proximity ligation technique to re-scaffold our existing hybrid genome assembly of an elite genotype (RJC1) developed using Illumina and Pacbio technologies. We assembled 99.81% of non-truncated reads to achieve 266.80 Mbp of the genome with an N50 value of 1.58 Mb. Furthermore, we compared the efficiency of Hi-C-augmented genome assembly with the hybrid genome assembly and observed a ~ 50% reduction in scaffolds and a tenfold increase in the N50 value. The gene ontology analysis revealed the identification of terms for molecular function (45.52%), cellular component (33.47%), and biological function (20.99%). Comparative genomic analysis of 13-plant species showed the conservation of 414 lipid metabolizing genes identified in the KEGG pathway analysis. Differential gene expression (DGE) studies were conducted in the healthy and Jatropha mosaic virus-infected leaves via RNA-seq analysis and observed gene expression changes for 2185 genes. Out of these, we observed 546 genes having more than two-fold change of transcript level and among these 259 genes were down-regulated and 287 genes were up-regulated. To validate RNA-seq data, two DEGs were selected for gene expression analysis using qRT-PCR and the data was in correlation with in silico results. RNA-seq analysis further shows the identification of some of the candidate genes and may be useful to develop JMV resistant plants after functional validation. This Hi-C genome assembly provides a detailed accurate reference genome which could be utilized to improve Jatropha and other economically important Euphorbiaceae family members.
由于其油有作为生物燃料的潜在用途,它是可再生能源的主要来源之一。这种作物的基因组由高含量的重复元件构成。我们采用Hi-C邻近连接技术对我们现有的使用Illumina和Pacbio技术开发的优良基因型(RJC1)的杂交基因组组装进行重新构建。我们组装了99.81%的非截断读段,获得了266.80 Mbp的基因组,N50值为1.58 Mb。此外,我们将Hi-C增强的基因组组装效率与杂交基因组组装进行了比较,发现支架数量减少了约50%,N50值增加了十倍。基因本体分析揭示了分子功能(45.52%)、细胞成分(33.47%)和生物学功能(20.99%)相关术语的识别。对13种植物的比较基因组分析表明,在KEGG通路分析中鉴定出的414个脂质代谢基因具有保守性。通过RNA-seq分析在健康和麻风树花叶病毒感染的叶片中进行了差异基因表达(DGE)研究,观察到2185个基因的表达变化。其中,我们观察到546个基因的转录水平变化超过两倍,其中259个基因下调,287个基因上调。为了验证RNA-seq数据,选择了两个差异表达基因进行qRT-PCR基因表达分析,数据与计算机模拟结果相关。RNA-seq分析进一步显示了一些候选基因的鉴定,在功能验证后可能有助于培育抗JMV的植物。这种Hi-C基因组组装提供了一个详细准确的参考基因组,可用于改良麻风树和其他经济上重要的大戟科植物。