24 research outputs found
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
Expression of ethylene-independent genes in overexpressed lines, <i>rin</i> mutants and wild type fruits.
<p>(a). Expression of a histidine metabolism gene <i>HDC</i> in overexpressed lines, <i>rin</i> mutants and wild type fruits. (b). Expression of <i>Nor</i> in overexpressed lines, <i>rin</i> mutants and wild type fruits. RNAs were extracted for qPCR assay from B, B+4 and B+7 fruits of overexpressed lines, <i>rin</i> mutant and wild type. Three replications for each sample were performed.</p
Carotenoid accumulation and carotenoid biosynthesis genes expression in <i>CaMADS-RIN</i> overexpressed <i>rin</i> and control fruits.
<p>(a). Analysis of carotenoid accumulation at B, B+4 and B+7 fruits of transgenic lines (ov-01 and ov-03), <i>rin</i> and wild type. Standard error is indicated for a minimum of three fruits per sample. (b). Expression of <i>PSY1</i> in B, B+4 and B+7 fruits of transgenic lines (ov-01 and ov-03), <i>rin</i> and wild type. (c). Expression of <i>ZDS</i> in B, B+4 and B+7 fruits of transgenic lines (ov-01 and ov-03), <i>rin</i> and wild type. (d). Expression of <i>PDS</i> in B, B+4 and B+7 fruits of transgenic lines (ov-01 and ov-03), <i>rin</i> and wild type.</p
Anthocyanin Accumulation and Molecular Analysis of Correlated Genes in Purple Kohlrabi (Brassica oleracea var. <i>gongylodes</i> L.)
Kohlrabi (Brassica
oleracea var. <i>gongylodes</i> L.) is an
important dietary vegetable cultivated and consumed widely for the
round swollen stem. Purple kohlrabi shows abundant anthocyanin accumulation
in the leaf and swollen stem. Here, different kinds of anthocyanins
were separated and identified from the purple kohlrabi cultivar (Kolibri)
by high-performance liquid chromatography–electrospray ionization
tandem mass spectrometry. In order to study the molecular mechanism
of anthocyanin biosynthesis in purple kohlrabi, the expression of
anthocyanin biosynthetic genes and regulatory genes in purple kohlrabi
and a green cultivar (Winner) was examined by quantitative PCR. In
comparison with the colorless parts in the two cultivars, most of
the anthocyanin biosynthetic genes and two transcription factors were
drastically upregulated in the purple tissues. To study the effects
of light shed on the anthocyanin accumulation of kohlrabi, total anthocyanin
contents and transcripts of associated genes were analyzed in sprouts
of both cultivars grown under light and dark conditions
Phylogenetic tree and reported or predicted functions of SlDEAD30, SlDEAD31, and other known plant DEAD-box proteins.
<p>The phylogenetic tree analysis was constructed with MEGA 5.2 software using the neighbor-joining method and the following parameters: bootstrap analysis of 1000 replicates, poisson model and pairwise deletion. The numbers at the nodes indicate the bootstrap values. SlDEAD30 and SlDEAD31 are marked with a black triangle and a red triangle, respectively. Accession numbers and corresponding references for the proteins used are as follows: <i>Solanum lycopersicum</i>: SlDEAD30, KJ739798; SlDEAD31, KJ713393. <i>Arabidopsis thaliana</i>: AtRH3, NM_001036866 [<a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0133849#pone.0133849.ref028" target="_blank">28</a>]; AtRH9, NM_125492 [<a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0133849#pone.0133849.ref021" target="_blank">21</a>]; AtRH22, NM_104691 [<a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0133849#pone.0133849.ref026" target="_blank">26</a>]; AtRH36, NM_101494 [<a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0133849#pone.0133849.ref025" target="_blank">25</a>]; AtRH52, NM_115719 [<a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0133849#pone.0133849.ref026" target="_blank">26</a>]; STRS1, AY080680 [<a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0133849#pone.0133849.ref016" target="_blank">16</a>]; STRS2, AY035114 [<a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0133849#pone.0133849.ref016" target="_blank">16</a>]; LOS4, BT002444 [<a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0133849#pone.0133849.ref019" target="_blank">19</a>]; RCF1, BT002030 [<a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0133849#pone.0133849.ref054" target="_blank">54</a>]; AtCAF, AF187317 [<a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0133849#pone.0133849.ref027" target="_blank">27</a>]; AtHEN2, AY050658 [<a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0133849#pone.0133849.ref055" target="_blank">55</a>]. UPF1, AF484122 [<a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0133849#pone.0133849.ref056" target="_blank">56</a>]. <i>Oryza sativa</i>: OsBIRH1, Q0DVX2 [<a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0133849#pone.0133849.ref017" target="_blank">17</a>]; OsABP, LOC_Os06g33520 [<a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0133849#pone.0133849.ref057" target="_blank">57</a>]; <i>Hordeum vulgare</i>: HVD1, AB164680 [<a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0133849#pone.0133849.ref058" target="_blank">58</a>]; <i>Glycine max</i>: GmRH, FJ462142 [<a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0133849#pone.0133849.ref059" target="_blank">59</a>]. <i>Apocynum venetum</i>: AvDH1, EU145588 [<a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0133849#pone.0133849.ref015" target="_blank">15</a>].</p
Cell wall metabolism genes in <i>CaMADS-RIN</i> overexpressed <i>rin</i> and control fruits.
<p>(a). Expression of <i>PG</i> in B, B+4 and B+7 fruits of transgenic lines (ov-01 and ov-03), <i>rin</i> and wild type. (b). Expression of <i>TBG4</i> in B, B+4 and B+7 fruits of transgenic lines (ov-01 and ov-03), <i>rin</i> and wild type. (c). Expression of <i>EXP1</i> in B, B+4 and B+7 fruits of transgenic lines (ov-01 and ov-03), <i>rin</i> and wild type.</p
Lipoxygenase genes in <i>CaMADS-RIN</i> overexpressed <i>rin</i> and control fruits.
<p>(a). Expression of <i>TomLoxB</i> in B, B+4 and B+7 fruits of transgenic lines (ov-01 and ov-03), <i>rin</i> and wild type. (b). Expression of <i>TomLoxC</i> in B, B+4 and B+7 fruits of transgenic lines (ov-01 and ov-03), <i>rin</i> and wild type.</p
Drought tolerance testing of <i>SlDEAD31-</i>overexpressing plants.
<p>A water withholding assay was performed with 8-week-old plants for up to 21 d followed by a recovery period of 5 d. (A) Representative phenotypes of wild-type and <i>SlDEAD31</i> overexpression plants at 0, 14 and 21 d after initiation of drought assay. (B) Survival rates of WT and transgenic plants after rewatering for 5 d following 21 d of drought assay. Each bar represents an average of eight plants ± SE. Asterisks indicate a significant difference from WT (p < 0.05). (C) and (D) Comparisons of chlorophyll content (C) and relative water content (RWC, D) of WT and transgenic plants at 0, 14 and 21 d after drought treatment. Values represent the means ± SE (<i>n</i> = 3). (E) Water loss rates of WT and transgenic plants. Similar leaves were excised and weighed at the indicated times. Water loss was calculated as the percentage of initial fresh weight. At least 15 leaves of similar developmental stages were excised and weighed at different times after the detachment. The data represent the means ± SE (<i>n</i> = 3).</p
Expression profiles of <i>SlDEAD30</i> and <i>SlDEAD31</i> genes in different tissues of wild-type tomato.
<p>Ro, roots; St, stems; Yl, young leaves; Ml, mature leaves; Sl, senescent leaves; Fl, flowers; Se, sepals; IMG, immature green fruits; MG, mature green fruits; B, breaker fruits; B+4, 4 day after breaker stage; B+7, 7 day after breaker stage. The relative expression levels were normalized to 1 in the roots. Bars represent mean relative expression values ± SE (<i>n</i> = 3).</p
<i>SlDEAD31</i>, a Putative DEAD-Box RNA Helicase Gene, Regulates Salt and Drought Tolerance and Stress-Related Genes in Tomato
<div><p>The DEAD-box RNA helicases are involved in almost every aspect of RNA metabolism, associated with diverse cellular functions including plant growth and development, and their importance in response to biotic and abiotic stresses is only beginning to emerge. However, none of DEAD-box genes was well characterized in tomato so far. In this study, we reported on the identification and characterization of two putative DEAD-box RNA helicase genes, <i>SlDEAD30</i> and <i>SlDEAD31</i> from tomato, which were classified into stress-related DEAD-box proteins by phylogenetic analysis. Expression analysis indicated that <i>SlDEAD30</i> was highly expressed in roots and mature leaves, while <i>SlDEAD31</i> was constantly expressed in various tissues. Furthermore, the expression of both genes was induced mainly in roots under NaCl stress, and <i>SlDEAD31</i> mRNA was also increased by heat, cold, and dehydration. In stress assays, transgenic tomato plants overexpressing <i>SlDEAD31</i> exhibited dramatically enhanced salt tolerance and slightly improved drought resistance, which were simultaneously demonstrated by significantly enhanced expression of multiple biotic and abiotic stress-related genes, higher survival rate, relative water content (RWC) and chlorophyll content, and lower water loss rate and malondialdehyde (MDA) production compared to wild-type plants. Collectively, these results provide a preliminary characterization of <i>SlDEAD30</i> and <i>SlDEAD31</i> genes in tomato, and suggest that stress-responsive <i>SlDEAD31</i> is essential for salt and drought tolerance and stress-related gene regulation in plants.</p></div