Seed storage protein (SSP) acts as one of the main components of seed storage reserves, of which accumulation is tightly mediated by a sophisticated regulatory network. However, whether and how gibberellin (GA) signaling is involved in this important biological event is not fully understood. Here, we show that SSP content in Arabidopsis (Arabidopsis thaliana) is significantly reduced by GA and increased in the GA biosynthesis triple mutant ga3ox1/3/4. Further investigation shows that the DELLA protein RGA-LIKE3 (RGL3), a negative regulator of GA signaling, is important for SSP accumulation. In rgl3 and 35S:RGL3-HA, the expression of SSP genes is down- and upregulated, respectively, compared with that in the wild-type. RGL3 interacts with ABSCISIC ACID INSENSITIVE3 (ABI3), a critical transcription factor for seed developmental processes governing SSP accumulation, both in vivo and in vitro, thus greatly promoting the transcriptional activating ability of ABI3 on SSP genes. In addition, genetic evidence shows that RGL3 and ABI3 regulate SSP accumulation in an interdependent manner. Therefore, we reveal a function of RGL3, a little studied DELLA member, as a coactivator of ABI3 to promote SSP biosynthesis during seed maturation stage. This finding advances the understanding of mechanisms in GA-mediated seed storage reserve accumulation.
Cytoplasmic male sterility (CMS) lays a foundation for the utilization of heterosis in soybean. The soybean CMS line SXCMS5A is an excellent CMS line exhibiting 100% male sterility. Cytological analysis revealed that in SXCMS5A compared to its maintainer SXCMS5B, its tapetum was vacuolated and abnormally developed. To identify the genes and metabolic pathways involving in pollen abortion of SXCMS5A, a comparative transcriptome analysis was conducted between SXCMS5A and SXCMS5B using flower buds. A total of 372,973,796 high quality clean reads were obtained from 6 samples (3 replicates for each material), and 840 differentially expressed genes (DEGs) were identified, including 658 downregulated and 182 upregulated ones in SXCMS5A compared to SXCMS5B. Among them, 13 DEGs, i.e., 12 open reading frames (ORFs) and 1 COX2, were mitochondrial genome genes in which ORF178 and ORF103c were upregulated in CMS lines and had transmembrane domain(s), therefore, identified as CMS candidate mitochondrial genes of SXCMS5A. Furthermore, numerous DEGs were associated with pollen wall development, carbohydrate metabolism, sugar transport, reactive oxygen species (ROS) metabolism and transcription factor. Some of them were further confirmed by quantitative real time PCR analysis between CMS lines with the same cytoplasmic source as SXCMS5A and their respective maintainer lines. The amount of soluble sugar and adenosine triphosphate and the activity of catalase and ascorbic acid oxidase showed that energy supply and ROS scavenging decreased in SXCMS5A compared to SXCMS5B. These findings provide valuable information for further understanding the molecular mechanism regulating the pollen abortion of soybean CMS.
Abstract The lncRNA biomarkers in melanoma remain to be further explored. The lncRNAs with different expression levels in melanoma tissue were identified by microarray analysis. To investigate the biological functions of target lncRNA, several in - vivo and in - vitro studies were performed. Potential mechanisms of competitive endogenous RNAs (ceRNAs) were predicted by using bioinformatics analysis and explored by western blot assay, fluorescence in situ hybridization assay, real-time quantitative PCR (RT-qPCR) array, RNA pull-down analysis, AGO2-RIP assay, and dual-luciferase reporter assay. The results demonstrated decreased LINC00459 in melanoma cell lines and tissues. According to the in-vitro and in-vivo experiments, up-regulated LINC00459 had inhibitory effect on cell proliferation and invasion. Bioinformatics analyses suggested that miR-218 could be a direct target of LINC00459. In addition, miR-218 was proved to be able to directly target the dickkopf-related protein 3 (DKK3) gene. In conclusion, our analysis suggested that the LINC00459 could sponge miR-218 and increase the expression of DKK3 gene, thus inhibiting the invasion and proliferation of melanoma cells, which indicated that the LINC00459 could be an effective biomarker for melanoma and its potential as the therapeutic target.
Background: To construct a prediction model based on the clinical characteristics of epidermoid cysts to identify pathologic infections, evaluate the diagnostic accuracy of the model, and conduct preliminary verification. Patients and Methods: We conducted a retrospective analysis of 314 patients diagnosed with epidermoid cysts that had been removed surgically. The clinical and pathologic data of all patients were collected. The patients were divided randomly into modeling group and verification group in a 75:25 ratio. In the modeling group, the multifactor logistic regression method was used to construct a prediction model for identifying epidermoid cyst pathologic infection, and the receiver operating characteristic (ROC) curve was used to evaluate the diagnostic accuracy of the model, which was then validated in the verification group. Results: All 314 patients with epidermoid cysts were divided into non-infected group (183 cases) and infected group (131 cases) according to the pathologic results. Logistic regression analysis showed that the disease course, growth trend, redness, and texture of epidermoid cysts were independent factors affecting pathologic infection. The above four indicators were selected to construct the prediction model of epidermoid cyst pathologic infection. In the modeling group, the prediction model showed an area under the curve (AUC) of 0.898, with the sensitivity of 0.830, specificity of 0.890, positive likelihood ratio of 7.523, and negative likelihood ratio of 0.191. The AUC of the prediction model in the verification group was 0.919, which was not significantly different from that of the modeling group (p = 0.886). Conclusions: The prediction model based on the clinical characteristics of epidermoid cysts had good diagnostic accuracy and high specificity; it can be used to identify pathologic infections of epidermoid cysts.
Abstract To facilitate the pseudochromosomes assembly and gene cloning in rapeseed, we developed a reference genetic population/map (named BnaZNF 2 ) from two sequenced cultivars, Zhongshuang11 and No.73290, those exhibit significant differences in many traits, particularly yield components. The BnaZNF 2 genetic map exhibited perfect collinearity with the physical map of B. napus , indicating its high quality. Comparative mapping revealed several genomic rearrangements between B. napus and B. rapa or B. oleracea . A total of eight and 16 QTLs were identified for pod number and seed number per pod, respectively and of which three and five QTLs are identical to previously identified ones, whereas the other five and 11 are novel. Two new major QTL respectively for pod number and seed number per pod, qPN.A06-1 and qSN.A06-1 ( R 2 = 22.8% and 32.1%), were colocalised with opposite effects and only qPN.A06-1 was confirmed and narrowed by regional association analysis to 180 kb including only 33 annotated genes. Conditional QTL analysis and subsequent NILs test indicated that tight linkage, rather than pleiotropy, was the genetic causation of their colocalisation. Our study demonstrates potential of this reference genetic population/map for precise QTL mapping and as a base for positional gene cloning in rapeseed.
The authors employed the polymerase chain reaction (PCR) to amplify the full coding region of the BDNF gene from lesser panda (Ailurus fulgens) genomic DNA directly with the primers based on the sequence of human BDNF gene. The amplified fragment was cloned and sequenced. Sequence analysis showed that the coding region of BDNF gene from lesser panda was highly homologous with that of human and giant panda counterpart genes (93% and 98% homology, repectively). It was found that there were only four amino acid residues showing difference in the prosequence region by comparing the deduced amino acid sequence of BDNF gene of lesser panda with that of human. However, the amino acid sequence of mature BDNF from lesser panda is identical to that of alll the reported mammalian BDNFs.
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