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| Status |
Public on May 26, 2012 |
| Title |
Small RNA sequencing in maize hybrids and inbreds |
| Organism |
Zea mays |
| Experiment type |
Non-coding RNA profiling by high throughput sequencing
|
| Summary |
Small RNAs (sRNAs) are hypothesized to contribute to hybrid vigor because they maintain genome integrity, contribute to genetic diversity, and control gene expression. We used Illumina sequencing to assess how sRNA populations vary between two maize inbred lines (B73, Mo17) and their hybrid. We sampled sRNAs from the seedling shoot apex and the developing ear, two rapidly growing tissues that program the greater growth of maize hybrids. We found that parental differences in siRNAs primarily originate from repeat regions. Although the maize genome contains greater number and complexity of repeats compared to Arabidopsis or rice, we confirmed that like these simpler plant genomes, 24-nt siRNAs whose abundance differs between maize parents also show a trend of downregulation following hybridization. Surprisingly, hybrid vigor is fully maintained when 24-nt siRNAs are globally reduced by mutation of the RNA-dependent RNA polymerase2 (RDR2) encoded by modifier of paramutation1 (mop1). We also discovered that 21-22nt siRNAs derived from a number of distinct retrotransposon families differentially accumulate between B73 and Mo17 as well as their hybrid. Thus, maize possesses a novel source of genetic variation for regulating both transposons and genes at a genomic scale, which may contribute to its high degree of observed heterosis.
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| Overall design |
sRNA libraries were derived from RNA isolated from the seedling shoot apex and developing ear tissues from B73, Mo17, B73xMo17 and Mo17xB73. The shoot apex was chosen because it is enriched for meristematic tissue where cell proliferation occurs, rates of organ initiation are determined, and organ size is specified. The developing ear was examined because it is enriched in meristematic tissue and is undergoing rapid growth, and also because the mature ear shows the highest degree of heterosis. Total RNA was isolated and separated on a 15% TBE-Urea polyacrylamide gel. Using a 10-bp ladder, the sRNA fraction representing 10-40-bp was excised. sRNA libraries were prepared according to Lu et al. (2007) or manufacturer's instructitions (Illumina). A combination of Perl scripts and FASTX toolkit scripts were used to remove adapters, collapse identical sequences and count reads per sequence.
Supplementary processed data text files contain the distinct sRNA sequences for all of the genotypes analyzed in that experiment. Abundance (reads per million) was calculated for each distinct sequence by dividing the number of reads of distinct sRNA in a library by the total number of sRNA reads for that library and multiplying this by 1 million.
Genome builds: B73 genome, maizesequence.org release 4a.53 (October, 2009); Mo17 whole genome shotgun clones.
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| Contributor(s) |
Moose SP, Hudson ME, Barber WT |
| Citation(s) |
22689990 |
| Submission date |
Apr 18, 2012 |
| Last update date |
May 15, 2019 |
| Contact name |
Wesley T Barber |
| E-mail(s) |
barber4@illinois.edu
|
| Organization name |
University of Illinois at Urbana-Champaign
|
| Department |
Department of Crop Sciences
|
| Lab |
Moose Maize Functional Genomics Lab
|
| Street address |
1201 W. Gregory Drive
|
| City |
Urbana |
| State/province |
IL |
| ZIP/Postal code |
61801 |
| Country |
USA |
| |
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| Platforms (2) |
| GPL9141 |
Illumina Genome Analyzer (Zea mays) |
| GPL15463 |
Illumina HiSeq 2000 (Zea mays) |
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| Samples (12)
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| GSM918104 |
Small RNAs from B73 shoot apex - 2007 expt |
| GSM918105 |
Small RNAs from Mo17 shoot apex - 2007 expt |
| GSM918106 |
Small RNAs from B73xMo17 shoot apex - 2007 expt |
| GSM918107 |
Small RNAs from Mo17xB73 shoot apex - 2007 expt |
| GSM918108 |
Small RNAs from B73 shoot apex - 2011 expt |
| GSM918109 |
Small RNAs from Mo17 shoot apex - 2011 expt |
| GSM918110 |
Small RNAs from B73 developing ear - high N |
| GSM918111 |
Small RNAs from Mo17 developing ear - high N |
| GSM918112 |
Small RNAs from B73xMo17 developing ear - high N |
| GSM918113 |
Small RNAs from B73 developing ear - low N |
| GSM918114 |
Small RNAs from Mo17 developing ear - low N |
| GSM918115 |
Small RNAs from B73xMo17 developing ear - low N |
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| Relations |
| BioProject |
PRJNA159733 |
| SRA |
SRP012317 |
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