The type 2C protein which belongs to the major group of protein phosphatases (PP2C) plays a vital role in abscisic acid (ABA) signaling and signal transductions processes. In the present study, 131PP2Cgenes were identified in total inBrassica rapaand categorized into thirteen subgroups based on their phylogenetic relationships. TheseB. rapaPP2C are structurally conserved based on amino acid sequence alignment, phylogenetic analysis, and conserved domains. Moreover, we utilized previously reported RNA-sequence data on various tissues (root, stem, leaf, flower, and silique), which suggests overlapping expression pattern in 29 paralogous gene pairs. The qRT-PCR validation of 15 paralogous gene pairs depicts distinct expression patterns in response to various abiotic stresses, such as heat, cold, ABA, and drought. Interestingly, stress-responsiveBraPP2Ccandidate genes were also identified, suggesting their significance in stress-tolerance mechanism inB. rapa. The evolutionary analysis for 15 paralogous gene pairs suggested that only three pairs have the positive selection and remaining were purifying in nature. The presented results of this study hasten our understanding of the molecular evolution of thePP2Cgene family inB. rapa. Thus, it will be ultimately helping in future research for facilitating the functional characterization ofBraPP2Cgenes in developing the abiotic stress tolerant plants.
To understand ubiquitination mechanism, E2s (ubiquitin conjugating enzymes) have crucial part as they play a major role in regulating many biological processes in plants. Meanwhile, Brassica rapa is an important leafy vegetable crop and therefore its characterization along with the expression pattern of E2s under various stresses is imperative. In this study, a total of 83 genes were identified in B. rapa and were classified into four different classes based on domain information. Here, we analyzed phylogenetic relationships, collinear correlation, gene duplication, interacting network, and expression patterns of E2 genes in B. rapa. Furthermore, RT-PCR analysis for 8 multiple abiotic and hormone treatments (namely, ABA, GA, JA, BR, PEG, NaCl, and heat and cold stress) illustrated striking expression pattern under one or more treatments, speculating that these might be stress-responsive genes. The cis-elements and interaction network analyses implicate valuable clues of important function of E2 genes in development and multiple stress responses in B. rapa. This study will further facilitate functional analysis of E2s for improving stress resistance mechanism in B. rapa.
The purpose of this study was to elucidate the effects of selenium-enriched probiotics on the liver of heat-27 stressed Wistar rats. Ten-week-old male rats were assigned to four groups: control (Con); high 28 temperature (HT); high temperature plus probiotics (HT+P: 10 11 CFU/mL Lactobacillus acidophilus and 29 10 9 CFU/mL Saccharomyces cerevisiae); or high temperature plus selenium-enriched probiotics 30 (HT+SeP: 0.3 mg/kg Se, 10 11 CFU/mL L. acidophilus and 10 9 CFU/mL S. cerevisiae). The HT, HT+P, 31 and HT+SeP groups were maintained at higher ambient temperature (40-42 °C), while the control group 32 were kept at room temperature (25 °C). After 42 days of thermal exposure, blood and liver tissues were 33 collected and analyzed for morphological and molecular markers of liver physiology. The body weight of 34 rats in the HT group decreased but liver weight and live index were increased. Histological examination 35 showed dilation of liver sinusoids and congestion of interstitial veins in HT group. Moreover, the 36 histomorphology of the liver in HT+P and HT+SeP groups was restored, and the serum AST, ALT, ALP, 37LDH and hepatic MDA level decreased significantly, but the serum total protein level and the liver SOD, 38 T-AOC, and GSH-PX activities were increased significantly relative to the HT group. In addition, the 39 mRNA level of Gpx1, SOD1, Nrf2, and Bcl-2 was significantly increased, while the expression level of 40 Bax, IL-6, TNF-α , COX-2, NF-κ B, α -SMA, TGF β 1, Collagen I, HSP70, and HSP90 was 41 significantly decreased in liver tissues after SeP supplementation. We concluded that SeP can protect 42 Wistar rats from oxidative stress, inflammation, apoptosis, and liver fibrosis induced by heat stress.
In recent years, more and more reports have shown that the miR156-SPL module can participate in the regulation of anthocyanin synthesis in plants. However, little is known about how this module responds to hormonal signals manipulating this process in grapes. In this study, exogenous GA, ABA, MeJA, and NAA were used to treat the ‘Wink’ grape berries before color conversion, anthocyanin and other related quality physiological indexes (such as sugar, aroma) were determined, and spatio-temporal expression patterns of related genes were analyzed. The results showed that the expression levels of VvmiR156b/c/d showed a gradually rising trend with the ripening and color formation of grape berries, and the highest expression levels were detected at day 28 after treatment, while the expression level of VvSPL9 exhibited an opposite trend as a whole, which further verifies that VvmiR156b/c/d can negatively regulate VvSPL9. Besides, VvmiR156b/c/d was positively correlated with anthocyanin content and related genes levels, while the expression pattern of VvSPL9 showed a negative correlation. Analysis of promoter cis-elements and GUS staining showed that VvmiR156b/c/d contained a large number of hormone response cis-elements (ABA, GA, SA, MeJA, and NAA) and were involved in hormone regulation. Exogenous ABA and MeJA treatments significantly upregulated the expression levels of VvmiR156b/c/d and anthocyanin structural genes in the early stage of color conversion and made grape berries quickly colored. Interestingly, GA treatment downregulated the expression levels of VvmiR156b/c/d and anthocyanin structural genes in the early color-change period, but significantly upregulated in the middle color-change and ripening stages, therefore GA mainly modulated grape berry coloring in the middle- and late-ripening stages. Furthermore, NAA treatment downregulated the expression levels of VvmiR156b/c/d and anthocyanin structural genes and delayed the peak expression of genes. Meanwhile, to further recognize the potential functions of VvmiR156b/c/d, the mature tomato transient trangenetic system was utilized in this work. Results showed that transient overexpression of VvmiR156b/c/d in tomato promoted fruit coloring and overexpression of VvSPL9 inhibited fruit coloration. Finally, a regulatory network of the VvmiR156b/c/d-VvSPL9 module responsive to hormones modulating anthocyanin synthesis was developed. In conclusion, VvmiR156b/c/d-mediated VvSPL9 participated in the formation of grape color in response to multi-hormone signals.
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