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Identification and Transcriptome Analysis of C1 Nullisomic in Allotetraploid Brassica napus L

ABSTRACT: The purposes of this study are to compare euploid B. napus cv. “Oro” and the C1 nullisomics transcriptome profiling (RNA-seq) and quantitative reverse transcription polymerase chain reaction (qRT–PCR) methods and to evaluate protocols for optimal high-throughput data analysis. Methods: Leaves mRNA profiles of 30-day-old euploid B. napus cv. “Oro” and the C1 nullisomics were generated by deep sequencing, in triplicate, using Illumina. The sequence reads that passed quality filters were analyzed at the transcript isoform level with two methods. qRT–PCR validation was performed. Results: After the high-throughput sequencing, each sample generated 7.2G Clean data on average and the 44.6-53.8 million clean reads were generated. Our study represents detailed analysis of leaves transcriptomes in euploid B. napus and nullisomic, with biologic replicates, generated by RNA-seq technology for comparative investigations of expression profiles. Our results show that Dose complementary effect exists between highly homologous genes, and partial loss of C subgenomic chromosome will lead to increased expression of A genome.

ORGANISM(S): Brassica napus

PROVIDER: GSE180585 | GEO | 2021/07/24

REPOSITORIES: GEO

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Project description:Purpose: the technology of Solexa/Illumina (RNA-seq) is an attractive alternative to the transcriptome sequencing. The goals of this study are to investigate transcriptional changes in replanted R. glutinosa leaves and identify genes responding to the disease. Methods:Totally 6.01and 6.15 million raw tags were measured from leaves of the first year planted (L1) and the second year replanted R. glutinosa (L2) at the tuberous root expansion stage, respectively. mRNA profiles of leaves of the first year planted (L1) and the second year replanted R. glutinosa (L2) at the tuberous root expansion stage, were generated by deep sequencing (Illumina HiSeqâ¢ 2000 system). qRTâPCR validation was performed using SYBR Green assays. Results: we obtained a total of 6.01 and 6.15 million total tags including 377,200 and 361,659 distinct tags from the L1 and L2 samples, respectively. Removal of low quality tags, 161,298 and 149,290 distinct clean tags were remained.With reference to the 94,544 non-redundant consensus sequences defined by the RNA-seq transcriptomic procedure, 60,574 genes had the diagnostic CATG site, which were taken as reference distinct sequences for DGE analysis. 47,458 of the L1 tag library and 42,247 of the L2 tag library were perfectly matched the reference sequences. we screened differentially expressed genes in the L1 and L2 libraries by digital gene expression (DGE) technique. Finally, a set of 1,954 genes may be in differential expression of L2. By bioinformatics and qRT-PCR, the 117 most strongly differentially expressed ones were considered to be prime candidates for responsible for replanting disease. Conclusions: The study provides an important resource for further investigating the cause of replanting disease and developing the methods to control or subtract its injury. Leaf mRNA profiles of L1 (The first year planted R. glutinosa leaves ) and L2 (the second year replanted R. glutinosa leaves) root expansion stage were generated by deep sequencing (using Illumina HiSeqâ¢ 2000 system)

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Identification and Transcriptome Analysis of C1 Nullisomic in Allotetraploid Brassica napus L

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