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SRX5416237: GSM3619434: Lotus japonicus nodules harvested from well-watered plants rep 1 [Lj.nodule.T0.1.S11]; Mesorhizobium loti; Lotus japonicus; RNA-Seq
4 ILLUMINA (NextSeq 500) runs: 21.7M spots, 1.6G bases, 613.2Mb downloads

Submitted by: NCBI (GEO)
Study: Transcriptomic changes in Medicago truncatula and Lotus japonicus root nodules during drought stress
show Abstracthide Abstract
Drought is one of the major environmental factors limiting biomass and seed yield production in agriculture. In this research we focused on plants from Fabaceae family, which have a unique ability for establishment of symbiosis with nitrogen-fixing bacteria, and are relatively susceptible to water limitation. We present the changes in nitrogenase activity and global gene expression occurring in Medicago truncatula and Lotus japonicus root nodules during water deficit. Our results prove a decrease in the efficiency of nitrogen fixation as well as extensive changes in plant and bacterial transcriptomes shortly after watering cessation. We show for the first time that not only symbiotic plant component, but also Sinorhizobium meliloti and Mesorhizobium loti bacteria residing in the root nodules of M. truncatula and L. japonicus, respectively, adjust their gene expression in response to water shortage. Although our results demonstrate that both M. truncatula and L. japonicus root nodules are susceptible to water deprivation, they indicate significant differences in plant and bacterial response to drought between tested species, which may be related to various type of root nodules formed by these species. Overall design: RNA was obtained from Medicago truncatula and Lotus japonicus non-inoculated roots (R) and root nodules either subjected to drought for two (T2) or four (T4) days, or well-watered control (T0). Two biological replicates for each sample was obtained. Each biological replicate consisted of roots or nodules harvested from 10-12 plants. RNA obtained from root nodules contained both plant and bacterial RNA. For Medicago truncatula the compatible rhizobium species is Sinorhizobium meliloti, for Lotus japonicus - Mesorhizobium loti.
Sample: Lotus japonicus nodules harvested from well-watered plants rep 1 [Lj.nodule.T0.1.S11]
SAMN10995008 • SRS4396991 • All experiments • All runs
Library:
Instrument: NextSeq 500
Strategy: RNA-Seq
Source: TRANSCRIPTOMIC
Selection: cDNA
Layout: SINGLE
Construction protocol: RNA was obtained from L. japonicus and M. truncatula non-inoculated roots and root nodules either subjected to drought for two (T2) or four (T4) days and control ones (T0). Total RNA extraction was performed using a GeneMATRIX Universal RNA Purification Kit (EURX, Gdańsk, Poland) with the additional step of on-column DNase I digestion. Per sample, an amount of 1 ug of total RNA was used as input. Using the Illumina TruSeq® Stranded Total RNA Sample Prep Kit (protocol 15031048 version E “October 2013”) rRNA is depleted from the total RNA samples using Ribo-Zero ribosomal RNA reduction chemistry. Illumina Ribo-Zero Plant (seed/root) kit was used for the plant samples, Illumina Ribo-Zero Gram negative bacteria kit was used for the bacterial samples and a mixture of both kits was used for the mixed samples. Subsequently, RNA was purified and fragmented and converted into first strand cDNA in a reverse transcription reaction using random primers. Next, double-stranded cDNA was generated in a second strand cDNA synthesis reaction using DNA Polymerase I and RNAse H. The cDNA fragments were extended with a single 'A' base to the 3' ends of the blunt-ended cDNA fragments after which multiple indexing adapters were ligated introducing different barcodes for each sample. Finally an enrichment PCR was carried out to enrich those DNA fragments that have adapter molecules on both ends and to amplify the amount of DNA in the library. For each run, libraries were equimolar pooled and sequenced in two runs using high 75 cycles NextSeq v2 kits on an Illumina NextSeq 500 instrument.
Experiment attributes:
GEO Accession: GSM3619434
Links:
Runs: 4 runs, 21.7M spots, 1.6G bases, 613.2Mb
Run# of Spots# of BasesSizePublished
SRR86173175,437,849410.2M153.3Mb2019-02-27
SRR86173185,316,177401.1M149.9Mb2019-02-27
SRR86173195,471,760412.8M155.1Mb2019-02-27
SRR86173205,460,686412M155Mb2019-02-27

ID:
7323937

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