De novo transcriptome analysis and identification of defensive genes in garlic (Allium sativum L.) using high-throughput sequencing.

IF 3.6 Q2 BIOTECHNOLOGY & APPLIED MICROBIOLOGY Journal, genetic engineering & biotechnology Pub Date : 2023-05-10 DOI:10.1186/s43141-023-00499-5
Malyaj R Prajapati, Jitender Singh, Pankaj Kumar, Rekha Dixit
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Abstract

Background: Garlic (Allium sativum L.) is the second most widely cultivated Allium which is mainly grown in temperate regions and used as a flavoring agent in a wide variety of foods. Garlic contains various bioactive compounds whose metabolic pathways, plant-pathogen interactions, defensive genes, identify interaction networks, and functional genomics were not previously predicted in the garlic at the genomic level. To address this issue, we constructed two garlic Illumina 2000 libraries from tissues of garlic clove and leaf.

Results: Approximately 43 million 125 bp paired-end reads were obtained in the two libraries. A total of 239,973 contigs were generated by de novo assembly of both samples and were compared with the sequences in the NCBI non-redundant protein database (Nr). In total, 42% of contigs were matched to known proteins in public databases including Nr, Gene Ontology (GO), and Cluster Orthologous Gene Database (COG), and then, contigs were mapped to 138 via functional annotation against the Kyoto Encyclopedia of Genes and Genomes pathway database (KEGG). In addition, a number of regulatory genes including the CCHC (Zn) family, followed by WD40, bromodomain, bZIP, AP2-EREBP, BED-type (Zn) proteins, and defense response proteins related to different conserved domains, such as RGA3, NBS-LRR, TIR-NBS-LRR, LRR, NBS-ARC, and CC-NBS-LRR were discovered based on the transcriptome dataset. We compared the ortholog gene family of the A. sativum transcriptome to A. thaliana, O. sativa, and Z. mays and found that 12,077 orthologous gene families are specific to A. sativum L. Furthermore, we identified genes involved in plant defense mechanisms, their protein-protein interaction network, and plant-pathogen interaction pathways.

Conclusions: Our study contains an extensive sequencing and functional gene-annotation analysis of A. sativum L. The findings provide insights into the molecular basis of TFs, defensive genes, and a reference for future studies on the genetics and breeding of A. sativum L.

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大蒜(Allium sativum L.)防御基因的高通量测序分析与鉴定。
背景:大蒜(Allium sativum L.)是第二大广泛种植的葱属植物,主要生长在温带地区,在各种食品中用作调味剂。大蒜含有多种生物活性化合物,这些化合物的代谢途径、植物与病原体的相互作用、防御基因、鉴定相互作用网络和功能基因组学在基因组水平上没有被预测到。为了解决这一问题,我们利用大蒜瓣和大蒜叶组织构建了两个大蒜Illumina 2000文库。结果:在两个文库中共获得约4300万个125 bp的成对末端reads。两个样本的从头组装共产生239,973个contigs,并与NCBI非冗余蛋白数据库(Nr)中的序列进行比较。总共有42%的contigs与Nr、Gene Ontology (GO)和Cluster Orthologous Gene Database (COG)等公共数据库中的已知蛋白质匹配,然后通过京都基因与基因组百科全书路径数据库(KEGG)的功能注释将contigs映射到138个。此外,基于转录组数据发现了CCHC (Zn)家族、WD40、bromodomain、bZIP、AP2-EREBP、BED-type (Zn)蛋白以及RGA3、NBS-LRR、TIR-NBS-LRR、LRR、NBS-ARC和CC-NBS-LRR等与不同保守结构域相关的防御反应蛋白。我们将sativum转录组的同源基因家族与拟南芥(a.thaliana)、O. sativa和Z. mays进行了比较,发现sativum l特有的同源基因家族有12077个。此外,我们还鉴定了涉及植物防御机制、蛋白质-蛋白质相互作用网络和植物-病原体相互作用途径的基因。结论:本研究对sativum L.进行了广泛的测序和功能基因注释分析,为进一步了解sativum L. TFs和防御基因的分子基础提供了依据,为今后sativum L.的遗传育种研究提供了参考。
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