Third-generation sequencing found LncRNA associated with heat shock protein response to heat stress in Populus qiongdaoensis seedlings.

BACKGROUND: As air temperatures increase globally, more and more plants are exposed to heat-stress conditions. Although many studies have explored regulation networks in plants with the aim of improving their heat-stress tolerance, only few have revealed them in trees. Here, individuals of Populus qiongdaoensis seedlings, which grows naturally in tropical areas, exposed to heat at 40 °C and the non-coding regulation networks were explored using the PacBio RSII and the Illumina sequencing platform. RESULTS: In total, we obtained 88,161 full-length transcripts representing 39,343 genes using 5,498,988 long reads and 350,026,252 clean reads, and also 216 microRNAs (miRNAs) via 95,794,107 reads. We then identified 928 putative long non-coding RNAs (lncRNAs), consisting of 828 sense lncRNAs (89.22%), 34 long intergenic non-coding RNAs (3.66%), 16 antisense (1.72%), and 50 sense intronic lncRNAs (5.39%). Under the dual criteria of |log2fold-change| ≥ 1 and P-value < 0.05, 1690 genes, 25 lncRNAs, and 15 miRNAs were found differentially expressed under the heat stress treatment. Furthermore, 563 and 595 mRNAs were detected as target genes of 14 differently expressed miRNAs and 26 differentially expressed lncRNAs. Functional annotation analysis of these target genes demonstrated they were related to cell membrane stability, plant hormone signal transduction, antioxidation, and aldarate metabolism. Lastly, we uncovered a key interaction network of lncRNAs, miRNAs and mRNAs that consisted of miR1444d, miR482a.1, miR530a, lncHSP18.2, HSP18.1, and HSP18.2. Expression level analysis showed that miRNAs in the network were up-regulated, while mRNAs and lncRNA were down-regulated, and also found that lncHSP18.2 may cis-regulate HSP18.2. CONCLUSIONS: Functional enrichment analysis of target genes of miRNAs and lncRNAs indicated that miRNAs and lncRNAs play an important role in the response to heat stress P. qiongdaoensis. Lastly, by investigating the miRNA-lncRNA-mRNA network of this species, we revealed that miRNAs may negatively regulate both lncRNAs and mRNAs in tree responses to heat stress, and found that lncHSP18.2 may cis-regulate HSP18.2.

Phenotypic and molecular marker analysis uncovers the genetic diversity of the grass Stenotaphrum secundatum.

BACKGROUND: Stenotaphrum secundatum is an important grass with a rich variety of accessions and great potential for development as an economically valuable crop. However, little is known about the genetic diversity of S. secundatum, limiting its application and development as a crop. Here, to provide a theoretical basis for further conservation, utilization, and classification of S. secundatum germplasm resources, we used phenotypic and molecular markers (single-nucleotide polymorphisms, SNPs; sequence-related amplified polymorphism, SRAP; inter-simple sequence repeat, ISSR) to analyze the genetic diversity of 49 S. secundatum accessions. RESULTS: Based on seven types of phenotypic data, the 49 S. secundatum accessions could be divided into three classes with great variation. We identified 1,280,873 SNPs in the 49 accessions, among which 66.22% were transition SNPs and 33.78% were transversion SNPs. Among these, C/T was the most common (19.12%) and G/C the least common (3.68%). Using 28 SRAP primers, 267 polymorphic bands were detected from the 273 bands amplified. In addition, 27 ISSR markers generated 527 amplification bands, all of which were polymorphic. Both marker types revealed a high level of genetic diversity, with ISSR markers showing a higher percentage of polymorphic loci (100%) than SRAP markers (97.8%). The genetic diversity of the accessions based on SRAP markers (h = 0.47, I = 0.66) and ISSR markers (h = 0.45, I = 0.64) supports the notion that the S. secundatum accessions are highly diverse. S. secundatum could be divided into three classes based on the evaluated molecular markers. CONCLUSIONS: Phenotypic and molecular marker analysis using SNP, SRAP, and ISSR markers revealed great genetic variation among S. secundatum accessions, which were consistently divided into three classes. Our findings provide a theoretical basis for the genetic diversity and classification of S. secundatum. Our results indicate that SNP, SRAP and ISSR markers are reliable and effective for analyzing genetic diversity in S. secundatum. The SNPs identified in this study could be used to distinguish S. secundatum accessions.