Analysis of miRNAs Targeted Storage Regulatory Genes during Soybean Seed Development Based on Transcriptome Sequencing.

Soybeans are an important cash crop and are widely used as a source of vegetable protein and edible oil. MicroRNAs (miRNA) are endogenous small RNA that play an important regulatory role in the evolutionarily conserved system of gene expression. In this study, we selected four lines with extreme phenotypes, as well as high or low protein and oil content, from the chromosome segment substitution line (CSSL) constructed from suinong (SN14) and ZYD00006, and planted and sampled at three stages of grain development for small RNA sequencing and expression analysis. The sequencing results revealed the expression pattern of miRNA in the materials, and predicted miRNA-targeted regulatory genes, including 1967 pairs of corresponding relationships between known-miRNA and their target genes, as well as 597 pairs of corresponding relationships between novel-miRNA and their target genes. After screening and annotating genes that were targeted for regulation, five specific genes were identified to be differentially expressed during seed development and subsequently analyzed for their regulatory relationship with miRNAs. The expression pattern of the targeted gene was verified by Real-time Quantitative PCR (RT-qPCR). Our research provides more information about the miRNA regulatory network in soybeans and further identifies useful genes that regulate storage during soy grain development, providing a theoretical basis for the regulation of soybean quality traits.

Identification of Soybean Genes Related to Soybean Seed Protein Content Based on Quantitative Trait Loci Collinearity Analysis.

Increasing the protein content of soybean seeds through a higher ratio of glycinin is important for soybean breeding and food processing; therefore, the integration of different quantitative trait loci (QTLs) is of great significance. In this study, we investigated the collinearity of seed protein QTLs. We identified 192 collinear protein QTLs that formed six hotspot regions. The two most important regions had seed protein 36-10 and seed protein 36-20 as hub nodes. We used a chromosome segment substitution line (CSSL) population for QTL validation and identified six CSSL materials with collinear QTLs. Five materials with higher protein and glycinin contents in comparison to the recurrent parent were analyzed. A total of 13 candidate genes related to seed protein from the QTL hotspot intervals were detected, 8 of which had high expression in mature soybean seeds. These results offer a new analysis method for molecular-assisted selection (MAS) and improvement of soybean product quality.

Identification of Major QTLs Associated With First Pod Height and Candidate Gene Mining in Soybean.

First pod height (FPH) is a quantitative trait in soybean [Glycine max (L.) Merr.] that affects mechanized harvesting. A compatible combination of the FPH and the mechanized harvester is required to ensure that the soybean is efficiently harvested. In this study, 147 recombinant inbred lines, which were derived from a cross between 'Dongnong594' and 'Charleston' over 8 years, were used to identify the major quantitative trait loci (QTLs) associated with FPH. Using a composite interval mapping method with WinQTLCart (version 2.5), 11 major QTLs were identified. They were distributed on five soybean chromosomes, and 90 pairs of QTLs showed significant epistatic associates with FPH. Of these, 3 were main QTL × main QTL interactions, and 12 were main QTL × non-main QTL interactions. A KEGG gene annotation of the 11 major QTL intervals revealed 8 candidate genes related to plant growth, appearing in the pathways K14486 (auxin response factor 9), K14498 (serine/threonine-protein kinase), and K13946 (transmembrane amino acid transporter family protein), and 7 candidate genes had high expression levels in the soybean stems. These results will aid in building a foundation for the fine mapping of the QTLs related to FPH and marker-assisted selection for breeding in soybean.

High Spy1 expression predicts poor prognosis in colorectal cancer.

BACKGROUND: Spy1 (SPDYA) is a new discovered cell cycle protein capable of promoting cell proliferation dependent on cyclin-dependent kinase-2 activation. However, to the best of our knowledge, the expression of Spy1 in colorectal cancer (CRC) tissues remains virtually unknown. MATERIALS AND METHODS: In this retrospective study, we investigated the mRNA and protein expression levels of Spy1 in CRC tissues and corresponding non-cancerous tissues with the analyses of quantitative real-time polymerase chain reaction, western blotting, and immunohistochemistry. In our research, the prognostic significances of Spy1 expression were further explored by univariate and multivariate survival analyses of 203 patients who were followed up. RESULTS: The results demonstrated that the levels of Spy1 mRNA were significantly higher in CRC tissues compared with corresponding non-cancerous tissues (p=0.0002). The results of immunohistochemistry demonstrated that the expressions of Spy1 were significantly associated with clinicopathological parameters, including T stage (χ2=7.126, p=0.028) and TNM stage (χ2=9.461, p=0.009). Kaplan-Meier analysis results indicated that high Spy1 expression (HR=2.573, p<0.001) and TNM stage (HR=1.494, p=0.011) were independent factors to predict poor prognosis for patients with CRC. CONCLUSION: We concluded that high Spy1 expression is significantly associated with unfavorable prognosis in CRC and could serve as a potential prognostic marker in clinical diagnosis of CRC.