Abstract
Replanting disease is a major factor limiting the artificial cultivation of the traditional Chinese medicinal herb Salvia miltiorrhiza. At present, little information is available regarding the role of miRNAs in response to replanting disease. In this study, two small RNA libraries obtained from first-year (FPR) and second-year plant (SPR) roots were subjected to a high-throughput sequencing method. Bioinformatics analysis revealed that 110 known and 7 novel miRNAs were annotated in the roots of S. miltiorrhiza. Moreover, 39 known and 2 novel miRNAs were identified and validated for differential expression in FPR compared with SPR. Thirty-one of these miRNAs were further analyzed by qRT-PCR, which revealed that 5 miRNAs negatively regulated the expression levels of 7 target genes involved in root development or stress responses. This study not only provides novel insights into the miRNA content of S. miltiorrhiza in response to replanting disease but also demonstrates that 5 miRNAs may be involved in these responses. Interactions among the differentially expressed miRNAs with their targets may form an important component of the molecular basis of replanting disease in S. miltiorrhiza.
Highlights
Salvia miltiorrhiza Bunge is a very popular traditional Chinese medicinal plant
Two sRNA libraries were constructed from the root tissues of FPR and SPR to identify miRNAs responding to replanting disease in S. miltiorrhiza roots
Their target genes including SPL13, ARF18-like, NAC100-like, athb-14-like and GRF3-like which were related to root growth were downregulated and might be presented a growth inhibition (Fig 5)
Summary
Salvia miltiorrhiza Bunge is a very popular traditional Chinese medicinal plant. The economic importance of S. miltiorrhiza results from the medicinal activity of extracts of its tuberous roots, which include tanshinones and salvianolic acid, and the herb has been used extensively to treat a variety of conditions, especially cardiovascular and cerebrovascular diseases [1,2,3]. While several studies have described how to improve the major medicinal bioactive constituents of S. miltiorrhiza [4,5,6,7,8], little attention has been paid to S. miltiorrhiza yields and quality. The cultivation area of S. miltiorrhiza in China has expanded over the years because of increased demands for the plant in the domestic and international markets. PLOS ONE | DOI:10.1371/journal.pone.0159905 August 2, 2016
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