posted on 2024-10-31, 02:19authored byUSDA-ARS-MWA
The bacterial metagenome in greenbug (Schizaphis graminum) aphids was investigated in response to aphid biotype, host species, host cultivar, and time post infestation. The V5-V7 hypervariable region from 16S rRNA genes was amplified from whole aphids, sequenced, and characterized against bacterial 16S sequences. Greenbug biotypes E and K were placed on potted plants of 17 accessions/cultivars in growth chambers. The accessions were: A1, Aegilops triuncialis TA1675 (Gbz), A2, Ae. triuncialis TA1695 (Gbx1); B1 to B2, barley cultivars Wintermalt, Jao (RSg1), and Post90; R1 and R2, rye cultivars Elbon and Insave F.A. (Gb2 amd Gb6); S1 to S4, sorghum cultivars TX7000, TX2737, TX2783, and DZhugara Belaya (PI 550607); and W1 through W6, wheat cultivars Newton, Custer, Amigo, Largo, C117882, and GRS 1201. Whole aphids were sampled by freezing in liquid nitrogen immediately before placement and 2, 4, and 8 days after placement. DNA was extracted using the DNeasy Blood and Tissue Kit for DNA Isolation (Qiagen, Germantown, MD) following its instructions. PCR was performed with V5-V7 primers 799F (degenerate AACMGGATTAGATACCCKG) and 1193R (ACGTCATCCCCACCTTCC). The libraries were barcoded, pooled, and sequenced on an Illumina Novaseq to generate 2x250-base paired-end reads. Demultiplexed reads were returned as fastq files with the biotype, host species, host cultivar, time point, and replicate encoded in each file name as BSC_TR, where B is biotype E or K; S is A (Aegilops triuncialis), B (barley), R (rye), S (sorghum), or W (wheat); C is a number from 1 to 6 for the cultivar as listed above; T is a number from 0 to 3 as 0, 2, 4, or 8 days; and R is biological replicate 1 to 3. This data set adds to knowledge of bacterial microfloras in the group of related genera (Aphis, Rhopalosiphum) to which Schizaphis belongs.
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