Molecular Characterization and Target Prediction of Candidate miRNAs Related to Abiotic Stress Responses and/or Storage Root Development in Sweet Potato

Sweet potato is a tuberous root crop with strong environmental stress resistance. It is beneficial to study its storage root formation and stress responses to identify sweet potato stress- and storage-root-thickening-related regulators. Here, six conserved miRNAs (miR156g, miR157d, miR158a-3p, miR161.1, miR167d and miR397a) and six novel miRNAs (novel 104, novel 120, novel 140, novel 214, novel 359 and novel 522) were isolated and characterized in sweet potato. Tissue-specific expression patterns suggested that miR156g, miR157d, miR158a-3p, miR167d, novel 359 and novel 522 exhibited high expression in fibrous roots or storage roots and were all upregulated in response to storage-root-related hormones (indole acetic acid, IAA; zeaxanthin, ZT; abscisic acid, ABA; and gibberellin, GAs). The expression of miR156g, miR158a-3p, miR167d, novel 120 and novel 214 was induced or reduced dramatically by salt, dehydration and cold or heat stresses. Moreover, these miRNAs were all upregulated by ABA, a crucial hormone modulator in regulating abiotic stresses. Additionally, the potential targets of the twelve miRNAs were predicted and analyzed. Above all, these results indicated that these miRNAs might play roles in storage root development and/or stress responses in sweet potato as well as provided valuable information for the further investigation of the roles of miRNA in storage root development and stress responses.

Comparative Transcriptome Analysis of Storage Root Reveals Regulatory Mechanisms of Lignin Biosynthesis During Root Development in Two Sweetpotato Cultivars

BackgroundSweetpotato(Ipomoea batatas (L.) Lam.) is one of the most important crops with high storage roots yield. Lignin affects the storage root formation. However, the molecular mechanisms of lignin biosynthesis in storage roots development have been lacking.ResultsTo reveal the molecular mechanism of lignin biosynthesis and identify new homologous genes in lignin biosynthesis during storage root development, the storage root (SR) at three different stages (D1, D2 and D3) in the two cultivars (Jishu25 and Jishu29) was investigated with full-length and second-generation transcriptome. A total of 52,137 transcripts and 21,148 unigenes were obtained after corrected with Hiseq2500 sequencing. Through the comparative analysis, 9577 unigenes were found to be differently expressed in the different stage in two cultivars. Among of them, 91 unigenes enriched in the phenylpropanoid biosynthesis, and 201 unigenes in hormone signal transduction pathway with KEGG analysis. Weighted gene co-expression network analysis of differentially expressed unigenes showed that lignin biosynthesis genes might be co-expressed with transcription factors such as AP2/ERF and MYB at the transcription level, and regulated by phytohormones auxin and GA3.ConclusionsTaken together, our findings will throw light on molecular regulatory mechanism of lignin biosynthesis involved in storage root development.

Comparative analysis of the SPL genes among four Ipomoea species and the evidence for a role of some IbSPLs in storage root development in sweet potato (Ipomoea batatas)

Background As a major family of plant-specific transcription factors, SQUAMOSA PROMOTER BINDING PROTEIN-LIKE (SPL) genes play crucial regulatory roles in plant growth, development, and stress tolerance. SPL transcription factor family has been widely studied in various plant species, however, there are no systematic studies on SPL genes in genus Ipomoea. Results In this study, a total of 29, 27, 26, 23 SPL genes were identified in Ipomoea batatas, Ipomoea trifida, Ipomoea triloba, and Ipomoea nil, respectively. Phylogenetic analysis indicated that Ipomoea SPL genes could be clustered into eight clades. SPL members within the same clade showed similar gene structures, domain organizations, and cis-acting element compositions, suggesting similarity of biological function potentially. Evolutionary analysis revealed that segmental duplication events played a major role in the Ipomoea genus-specific expansion of SPL genes. Of these Ipomoea SPL genes, 69 were predicted as the target genes of miR156, and 7 IbSPL genes were further confirmed by degradome data. Additionally, IbSPL genes showed diverse expression patterns in various tissues, implying their functional conservation and divergence. Finally, by combining the information from expression patterns and regulatory sub-networks, we found that four IbSPL genes (IbSPL16/IbSPL17/IbSPL21/IbSPL28) may be involved in the formation and development of storage roots. Conclusions This study not only provides novel insights into the evolutionary and functional divergence of the SPL genes in all available sequenced species in genus Ipomoea, but also lays a foundation for further elucidation of the potential functional roles of IbSPL genes during storage root development.

Characteristics of the AT-Hook Motif Containing Nuclear Localized (AHL) Genes in Carrot Provides Insight into Their Role in Plant Growth and Storage Root Development

The AT-hook motif containing nuclear localized (AHL) gene family, controlling various developmental processes, is conserved in land plants. They comprise Plant and Prokaryote Conserved (PPC) domain and one or two AT-hook motifs. DcAHLc1 has been proposed as a candidate gene governing the formation of the carrot storage root. We identified and in-silico characterized carrot AHL proteins, performed phylogenetic analyses, investigated their expression profiles and constructed gene coexpression networks. We found 47 AHL genes in carrot and grouped them into two clades, A and B, comprising 29 and 18 genes, respectively. Within Clade-A, we distinguished three subclades, one of them grouping noncanonical AHLs differing in their structure (two PPC domains) and/or cellular localization (not nucleus). Coexpression network analysis attributed AHLs expressed in carrot roots into four of the 72 clusters, some of them showing a large number of interactions. Determination of expression profiles of AHL genes in various tissues and samples provided basis to hypothesize on their possible roles in the development of the carrot storage root. We identified a group of rapidly evolving noncanonical AHLs, possibly differing functionally from typical AHLs, as suggested by their expression profiles and their predicted cellular localization. We pointed at several AHLs likely involved in the development of the carrot storage root.

Sweetpotato root development influences susceptibility to black rot caused by the fungal pathogen Ceratocystis fimbriata

Black rot of sweetpotato caused by Ceratocystis fimbriata, is an important reemerging disease threatening sweetpotato production in the United States. This study assessed disease susceptibility of the storage root surface, storage root cambium, and slips (vine cuttings) of 48 sweetpotato cultivars, advanced breeding lines, and wild relative accessions. We also characterized the effect of storage root development on susceptibility to C. fimbriata. None of the cultivars examined at the storage root level were resistant, with most cultivars exhibiting similar levels of susceptibility. In storage roots, Jewel and Covington were the least susceptible and significantly different from White Bonita, the most susceptible cultivar. In the slip, significant differences in disease incidence were observed for above and below ground plant structures among cultivars, advanced breeding lines, and wild relative accessions. Burgundy and Ipomoea littoralis displayed less below ground disease incidence as compared to NASPOT 8, Sunnyside and LSU-417, the most susceptible cultivars. Correlation of black rot susceptibility between storage roots and slips was not significant, suggesting that slip assays are not useful to predict resistance in storage roots. Immature, early developing storage roots were comparatively more susceptible than older, fully developed storage roots. The high significant correlation between storage root cross-section area and cross-sectional lesion ratio suggests the presence of an unfavorable environment for C. fimbriata as the storage root develops. Incorporating applications of effective fungicides at transplanting and during early storage root development when sweetpotato tissues are most susceptible to black rot infection may improve disease management efforts.

Impact of reduced rates of 2,4-D and glyphosate on sweetpotato growth and yield

Commercialization of 2,4-D–tolerant crops is a major concern for sweetpotato producers because of potential 2,4-D drift that can cause severe crop injury and yield reduction. A field study was initiated in 2014 and repeated in 2015 to assess impacts of reduced rates of 2,4-D, glyphosate, or a combination of 2,4-D with glyphosate on sweetpotato. In one study, 2,4-D and glyphosate were applied alone and in combination at 1/10, 1/100, 1/250, 1/500, 1/750, and 1/1,000 of anticipated field use rates (1.05 kg ha−1 for 2,4-D and 1.12 kg ha−1 for glyphosate) to ‘Beauregard’ sweetpotato at storage root formation (10 days after transplanting [DAP]). In a separate study, all these treatments were applied to ‘Beauregard’ sweetpotato at storage root development (30 DAP). Injury with 2,4-D alone or in combination with glyphosate was generally equal or greater than with glyphosate applied alone at equivalent herbicide rates, indicating that injury is attributable mostly to 2,4-D in the combination. There was a quadratic increase in crop injury and quadratic decrease in crop yield (with respect to most yield grades) with increased rate of 2,4-D applied alone or in combination with glyphosate applied at storage root development. However, neither the results of this relationship nor of the significance of herbicide rate were observed on crop injury or sweetpotato yield when herbicide application occurred at storage root formation, with a few exceptions. In general, crop injury and yield reduction were greatest at the highest rate (1/10×) of 2,4-D applied alone or in combination with glyphosate, although injury observed at lower rates was also a concern after initial observation by sweetpotato producers. However, in some cases, yield reduction of U.S. no.1 and marketable grades was also observed after application of 1/250×, 1/100×, or 1/10× rates of 2,4-D alone or with glyphosate when applied at storage root development.