Expression patterns of <i>SlDEAD30</i> and <i>SlDEAD31</i> genes under various stress treatments including NaCl (200 mM, A, D), heat (40°C) and cold (4°C, B, E), dehydration, and wounding (C, F).

<p>Gene expression was detected by RT-PCR using total RNA from leaves or roots of 35-day-old tomato plants. Bars represent mean relative expression values ± SE (<i>n</i> = 3). Values are presented relative to untreated plants (0 h). Asterisks indicate a significant difference (P<0.05) between untreated and treated plants. The twofold threshold is indicated by a dotted line.</p

Primers used for construction of Pull-down assay.

(DOCX)</p

Table_2_Molecular and Phylogenetic Analyses of the Mediator Subunit Genes in Solanum lycopersicum.xlsx

The Mediator complex is a multi-subunit protein assembly that serves as a central scaffold to help regulate DNA-binding transcription factors (TFs) and RNA polymerase II (Pol II) activity controlled gene expression programmed in response to developmental or environmental factors. However, litter information about Mediator complex subunit (MED) genes in tomato is available, although it is an essential model plant. In this study, we retrieved 46 candidate SlMED genes from the genome of tomato, and a comprehensive analysis was conducted, including their phylogenetic relationship, chromosomal locations, gene structure, cis-regulatory elements prediction, as well as gene expression. The expression profiling of 46 SlMED genes was analyzed using publicly available RNA-seq data. Furthermore, we selected some SlMED genes to evaluate their expression patterns in various tissues and under different abiotic stress treatments by quantitative reverse transcription PCR experiments. This is the first detailed report to elucidate the molecular and phylogenetic features of the MED genes in tomato, and it provides valuable clues for further functional analysis in order to clarify the role of the SlMED genes in diverse plant growth, development and abiotic stress response.</p

A Non-Climacteric Fruit Gene <i>CaMADS-RIN</i> Regulates Fruit Ripening and Ethylene Biosynthesis in Climacteric Fruit

<div><p>MADS-box genes have been reported to play a major role in the molecular circuit of developmental regulation. Especially, <i>SEPALLATA</i> (<i>SEP</i>) group genes play a central role in the developmental regulation of ripening in both climacteric and non-climacteric fruits. However, the mechanisms underlying the regulation of <i>SEP</i> genes to non-climacteric fruits ripening are still unclear. Here a <i>SEP</i> gene of pepper, <i>CaMADS-RIN</i>, has been cloned and exhibited elevated expression at the onset of ripening of pepper. To further explore the function of <i>CaMADS-RIN</i>, an overexpressed construct was created and transformed into <i>ripening inhibitor</i> (<i>rin</i>) mutant tomato plants. Broad ripening phenotypes were observed in <i>CaMADS-RIN</i> overexpressed <i>rin</i> fruits. The accumulation of carotenoid and expression of <i>PDS</i> and <i>ZDS</i> were enhanced in overexpressed fruits compared with <i>rin</i> mutant. The transcripts of cell wall metabolism genes (<i>PG</i>, <i>EXP1</i> and <i>TBG4</i>) and lipoxygenase genes (<i>TomloxB</i> and <i>TomloxC</i>) accumulated more abundant compared to <i>rin</i> mutant. Besides, both ethylene-dependent genes including <i>ACS2</i>, <i>ACO1</i>, <i>E4</i> and <i>E8</i> and ethylene-independent genes such as <i>HDC</i> and <i>Nor</i> were also up-regulated in transgenic fruits at different levels. Moreover, transgenic fruits showed approximately 1–3 times increase in ethylene production compared with <i>rin</i> mutant fruits. Yeast two-hybrid screen results indicated that CaMADS-RIN could interact with TAGL1, FUL1 and itself respectively as SlMADS-RIN did in vitro. These results suggest that <i>CaMADS-RIN</i> affects fruit ripening of tomato both in ethylene-dependent and ethylene-independent aspects, which will provide a set of significant data to explore the role of <i>SEP</i> genes in ripening of non-climacteric fruits.</p></div

Anthocyanin Accumulation and Transcriptional Regulation of Anthocyanin Biosynthesis in Purple Bok Choy (Brassica rapa var. <i>chinensis</i>)

Bok choy (Brassica rapa var. chinensis) is an important dietary vegetable cultivated and consumed worldwide for its edible leaves. The purple cultivars rich in health-promoting anthocyanins are usually more eye-catching and valuable. Fifteen kinds of anthocyanins were separated and identified from a purple bok choy cultivar (Zi He) by high-performance liquid chromatography–electrospray ionization tandem mass spectrometry. To investigate the molecular mechanisms underlying anthocyanin accumulation in bok choy, the expression profiles of anthocyanin biosynthetic and regulatory genes were analyzed in seedlings and leaves of the purple cultivar and the green cultivar (Su Zhouqing). Compared with the other tissues, BrTT8 and most of the anthocyanin biosynthetic genes were significantly up-regulated in the leaves and light-grown seedlings of Zi He. The results that heterologous expression of BrTT8 promotes the transcription of partial anthocyanin biosynthetic genes in regeneration shoots of tomato indicate that BrTT8 plays an important role in the regulation of anthocyanin biosynthesis

Co-immunoprecipitation (Co-IP) studies.

(TIF)</p

The JJW (SlJAZ10-SlJAV1-SlWRKY51 or SlJAZ10-SlJAV1-SlWRKY51).

(a) Y2H assay shows that SlJAV1 interacts with SlJAZ10 and SlJAZ11. (b) Subcellular localization of SlJAZ10 and SlJAZ11 in the epidermal cells of N. benthamiana leaves. Bars = 40μm. (c) BiFC assay confirms the interactions among SlJAZ10, SlJAZ11, SlJAV1 and SlWRKY51 in N. benthamiana leaves. (d) Y1H assay shows the transcriptional binding activity of SlJAV1 and SlWRKY51 with SlAOC promoter. (e) Transient transcriptional activation assays show that SlJAZ10 and SlJAZ11 have transcriptional repression activity. (f) SlJAZ10-SlJAV1-SlWRKY51 and SlJAZ11-SlJAV1-SlWRKY51complex effectively suppresses the expression of SlAOCPro-LUC in N. benthamiana transient expression assay. Relative ratio of LUC/REN are means ± SEM (n≥6); *p SlAOC promoter, with the W-box intacted or mutated.</p

Primers used for qRT-PCR analysis.

(DOCX)</p

Salt sensitivity of <i>SlDEAD31</i> overexpressing plants at postgermination stage.

<p>(A) <i>SlDEAD31</i>-overexpressing vector. RB, right border; Pro, promoter; Ter, terminator; LB, left border. (B) Quantitative RT-PCR analysis of <i>SlDEAD31</i> expression in the leaves of WT and transgenic lines OE-2, OE-7, and OE-8. Values represent the means ± SE (<i>n</i> = 3). Asterisks indicate a significant difference from WT (p<0.05). (C) and (D) Representative growth performance (C) and shoot and root length (D) of WT and <i>SlDEAD31</i> overexpressing plants in MS medium containing 0 and 100 mM NaCl at postgermination stage. The shoot and root length of seedlings (n≥20 each) were counted after growth for 10 d. Values represent the means ± SE (<i>n</i> = 3). Asterisks indicate a significant difference from WT (p<0.05).</p