Assessment of genetic diversity and variety identification based on developed retrotransposon-based insertion polymorphism (RBIP) markers in sweet potato (Ipomoea batatas (L.) Lam.).

Sweet potato, a dicotyledonous and perennial plant, is the third tuber/root crop species behind potato and cassava in terms of production. Long terminal repeat (LTR) retrotransposons are highly abundant in sweet potato, contributing to genetic diversity. Retrotransposon-based insertion polymorphism (RBIP) is a high-throughput marker system to study the genetic diversity of plant species. To date, there have been no transposon marker-based genetic diversity analyses of sweet potato. Here, we reported a structure-based analysis of the sweet potato genome, a total of 21555 LTR retrotransposons, which belonged to the main LTR-retrotransposon subfamilies Ty3-gypsy and Ty1-copia were identified. After searching and selecting using Hidden Markov Models (HMMs), 1616 LTR retrotransposon sequences containing at least two models were screened. A total of 48 RBIP primers were synthesized based on the high copy numbers of conserved LTR sequences. Fifty-six amplicons with an average polymorphism of 91.07% were generated in 105 sweet potato germplasm resources based on RBIP markers. A Unweighted Pair Group Method with Arithmatic Mean (UPGMA) dendrogram, a model-based genetic structure and principal component analysis divided the sweet potato germplasms into 3 groups containing 8, 53, and 44 germplasms. All the three analyses produced significant groupwise consensus. However, almost all the germplasms contained only one primary locus. The analysis of molecular variance (AMOVA) among the groups indicated higher intergroup genetic variation (53%) than intrapopulation genetic variation. In addition, long-term self-retention may cause some germplasm resources to exhibit variable segregation. These results suggest that these sweet potato germplasms are not well evolutionarily diversified, although geographic speciation could have occurred at a limited level. This study highlights the utility of RBIP markers for determining the intraspecies variability of sweet potato and have the potential to be used as core primer pairs for variety identification, genetic diversity assessment and linkage map construction. The results could provide a good theoretical reference and guidance for germplasm research and breeding.

Combination of High Zn Density and Low Phytic Acid for Improving Zn Bioavailability in Rice (Oryza stavia L.) Grain.

BACKGROUND: Zn deficiency is one of the leading public health problems in the world. Staple food crop, such as rice, cannot provide enough Zn to meet the daily dietary requirement because Zn in grain would chelate with phytic acid, which resulted in low Zn bioavailability. Breeding new rice varieties with high Zn bioavailability will be an effective, economic and sustainable strategy to alleviate human Zn deficiency. RESULTS: The high Zn density mutant LLZ was crossed with the low phytic acid mutant Os-lpa-XS110-1, and the contents of Zn and phytic acid in the brown rice were determined for the resulting progenies grown at different sites. Among the hybrid progenies, the double mutant always displayed significantly higher Zn content and lower phytic acid content in grain, leading to the lowest molar ratio of phytic acid to Zn under all environments. As assessed by in vitro digestion/Caco-2 cell model, the double mutant contained the relatively high content of bioavailable Zn in brown rice. CONCLUSIONS: Our findings suggested pyramiding breeding by a combination of high Zn density and low phytic acid is a practical and useful approach to improve Zn bioavailability in rice grain.

Comparative de novo transcriptomics and untargeted metabolomic analyses elucidate complicated mechanisms regulating celery (Apium graveolens L.) responses to selenium stimuli.

Presently, concern regarding the effects of selenium (Se) on the environment and organisms worldwide is increasing. Too much Se in the soil is harmful to plants. In this study, Illumina RNA sequencing and the untargeted metabolome of control and Se-treated celery seedlings were analyzed. In total, 297,911,046 clean reads were obtained and assembled into 150,218 transcripts (50,876 unigenes). A total of 36,287 unigenes were annotated using different databases. Additionally, 8,907 differentially expressed genes, including 5,319 up- and 3,588 downregulated genes, were identified between mock and Se-treated plants. "Phenylpropanoid biosynthesis" was the most enriched KEGG pathway. A total of 24 sulfur and selenocompound metabolic unigenes were differentially expressed. Furthermore, 1,774 metabolites and 237 significant differentially accumulated metabolites were identified using the untargeted metabolomic approach. We conducted correlation analyses of enriched KEGG pathways of differentially expressed genes and accumulated metabolites. Our findings suggested that candidate genes and metabolites involved in important biological pathways may regulate Se tolerance in celery. The results increase our understanding of the molecular mechanism responsible for celery's adaptation to Se stress.

Comparative proteomic analysis of pepper (Capsicum annuum L.) seedlings under selenium stress.

Selenium (Se) is an essential trace element for human and animal health. Se fertilizer has been used to increase the Se content in crops to meet the Se requirements in humans and animals. To address the challenge of Se poisoning in plants, the mechanisms underlying Se-induced stress in plants must be understood. Here, to elucidate the effects of Se stress on the protein levels in pepper, we used an integrated approach involving tandem mass tag labeling, high performance liquid chromatography fractionation, and mass spectrometry-based analysis. A total of 4,693 proteins were identified, 3,938 of which yielded quantitative information. Among them, the expression of 172 proteins was up-regulated, and the expression of 28 proteins was down-regulated in the Se/mock treatment comparison. According to the above data, we performed a systematic bioinformatics analysis of all identified proteins and differentially expressed proteins (DEPs). The DEPs were most strongly associated with the terms "metabolic process," "posttranslational modification, protein turnover, chaperones," and "protein processing in endoplasmic reticulum" according to Gene Ontology, eukaryotic orthologous groups classification, and Kyoto Encyclopedia of Genes and Genomes enrichment analysis, respectively. Furthermore, several heat shock proteins were identified as DEPs. These results provide insights that may facilitate further studies on the pepper proteome expressed downstream of the Se stress response. Our data revealed that the responses of pepper to Se stress involve various pathways.

Transcriptome profiling of Gerbera hybrida reveals that stem bending is caused by water stress and regulation of abscisic acid.

BACKGROUND: Gerbera hybrida is one of the most popular cut flowers in the world; however, stem bending, which always happens when gerbera flower harvested from the field, greatly limits its vase life. To date the molecular mechanisms underlying stem bending remain poorly understood. RESULTS: In this study, we performed high-throughput transcriptome sequencing of gerbera during stem bending using the Illumina sequencing technology. Three cDNA libraries constructed from mRNAs of gerbera stem at stem bending stage 0, 2 and 4 were sequenced. More than 300 million high-quality reads were generated and assembled into 96,492 unigenes. Among them, 34,166 unigenes were functionally annotated based on similarity search with known protein. Sequences derived from plants at different stem bending stages were mapped to the assembled transcriptome, and 9,406 differentially expressed genes (DEGs) were identified. Through Kyoto Encyclopedia of Genes and Genomes (KEGG) enrichment analysis, specific pathways were identified during the stem bending process, such as phenylpropanoid biosynthesis pathway, phenylalanine metabolism pathway, starch and sucrose metabolism pathway, and plant hormone signal transduction pathway. A total of 211 transcription factors (TFs), including TF families involved in plant senescence, such as NAC, MYB, WRKY, and AP2/ERF members, as well as TFs related to water stress tolerance, were shown to be regulated during stem bending. Gene Onotology (GO) functional enrichment analysis indicated that key genes involved in responses to osmotic and oxidative stresses were also varied in expression during this process. Furthermore, analysis of DEGs involved in the hormone signaling pathways and determination of endogenous abscisic acid (ABA) content showed that stem bending may be an ethylene-independent process, but regulated by ABA. In short, our findings suggested that the stem bending of cut gerbera may be caused by the involvement of water stress and regulation of ABA during the postharvest life. CONCLUSIONS: The transcriptome sequences provide a valuable resource in revealing the molecular mechanism underlying stem bending of cut flower and offer novel genes that can be used to guide future studies for ornamental plant breeding.

Effects of osmotic stress on antioxidant enzymes activities in leaf discs of PSAG12-IPT modified Gerbera.

Leaf senescence is often caused by water deficit and the chimeric gene P(SAG12)-IPT is an auto-regulated gene delaying leaf senescence. Using in vitro leaf discs culture system, the changes of contents of chlorophylls, carotenoids, soluble protein and thiobarbituric acid reactive substance (TBARS) and antioxidant enzymes activities were investigated during leaf senescence of P(SAGl2)-IPT modified gerbera induced by osmotic stress compared with the control plant (wild type). Leaf discs were incubated in 20%, 40% (w/v) polyethylene glycol (PEG) 6000 nutrient solution for 20 h under continuous light [130 micromol/(m(2) x s)]. The results showed that the contents of chlorophylls, carotenoids and soluble protein were decreased by osmotic stress with the decrease being more pronounced at 40% PEG, but that, at the same PEG concentration the decrease in the transgenic plants was significantly lower than that in the control plant. The activities of superoxide dismutase (SOD), catalases (CAT), ascorbate peroxidase (APX), guaiacol peroxidase (GPX) and dehydroascorbate reductase (DHAR) were stimulated by PEG treatment. However, the increases were higher in P(SAG12)-IPT transgenic plants than in the control plants, particularly at 40% PEG treatment. Lipid peroxidation (TBARS content) was increased by PEG treatment with the increase being much lower in transgenic plant than in the control plant. It could be concluded that the increases in the activities of antioxidant enzymes including SOD, CAT, APX, GPX and DHAR were responsible for the delay of leaf senescence induced by osmotic stress.