玉米茎秆抗倒伏遗传的研究进展

茎秆倒伏严重影响玉米产量、品质和机械化收获,是当前玉米生产和育种亟待解决的主要问题之一。加强对玉米茎秆抗倒伏性的研究,对提高品种抗倒伏能力具有重要意义。本文综述了玉米茎秆倒伏的主要影响因素及其遗传特征。茎秆倒伏与茎秆自身的强度密切相关。茎秆强度越高,抗倒伏性越强。茎秆强度受茎秆所处的发育阶段、茎秆内部结构和外部形态,及其细胞壁成分等影响。处于分生组织的茎秆细胞分裂旺盛,较易折断,而进入生殖生长后,茎秆表皮、厚壁组织增厚,维管束发育成熟,对茎秆的支撑作用增强。茎秆细胞壁的主要成分——纤维素、半纤维素、木质素、可溶性糖、无机物等均可提升茎秆强度。目前,研究者借助高通量表型平台,利用玉米连锁群体和自交系群体,采用各种定位方法,鉴定到一系列影响茎秆形态、强度、细胞壁成分的相关QTL和候选基因。研究表明,基于单倍型的QTL定位方法比基于单个SNP的定位效果好。一致性QTL分析将不同遗传群体的研究整合到一起,能够提高QTL结果的通用性。茎秆强度的遗传基础复杂,受微效多基因控制,位点间具有加性效应。茎秆成分QTL中的候选基因涉及细胞壁代谢、转录因子、蛋白激酶等。MAIZEWALL是玉米细胞壁相关基因的重要数据库。目前该数据库包含1 156个玉米细胞壁生物学相关的候选基因,为该领域的深入研究提供强大的资源。已鉴定到一系列影响玉米茎秆细胞壁成分、茎秆形态和强度的基因,其功能涉及纤维素合成路径,如纤维素合成酶类、Cobra类、糖基转移酶和核糖转运蛋白类;苯丙烷路径基因,如控制bm1—bm5的相关基因;植物激素类,如赤霉素、生长素、油菜素甾醇相关基因;转录因子如NAC、MYB;miRNA（ZmmiR528）以及F-box基因（stiff1）等。今后应积极探索不同发育时期玉米茎秆倒伏的力学机制;广泛发展自然群体或育种群体进行遗传分析;采取多种定位策略,提高抗倒伏相关基因鉴定的功效;针对优良等位基因,开发各类分子标记,加强抗倒伏分子标记辅助选择。本文将为玉米茎秆抗倒伏遗传机制解析及抗倒伏玉米品种的分子育种提供参考。     

Effects of DHRS3 in C2C12 Myoblast Differentiation and Mouse Skeletal Muscle Injury

Myoblast differentiation is an essential process during skeletal muscle development. C2 C12 myoblast is a commonly used experimental model to study muscle cell differentiation in vitro. Dehydrogenase/reductase(SDR family) member 3(DHRS3) is a highly conserved member in short-chain alcohol dehydrogenase/reductase superfamily and has been shown to be involved in the metabolism of retinol. Previous experimental results showed that the expression of DHRS3 increased significantly during the differentiation of myoblasts differentiation. However, the effect of DHRS3 on mouse muscle cell differentiation was unclear. The objective of current study was to determine if DHRS3 affected muscle cell differentiation, and if DHRS3 was involved in muscle regeneration. Protein expression was determined by western blot and immunofluorescence analysis. The activation and inhibition of DHRS3 increased and decreased C2 C12 myoblast differentiation respectively, which indicated that DHRS3 could affect C2 C12 myoblast differentiation. DHRS3 expression was significantly changed during muscle regeneration, with the regeneration of muscle injury, the expression of DHRS3 tended to increase first and then decrease. It suggested that DHRS3 might be involved in muscle regeneration. In summary, this study confirmed the involvement of DHRS3 in C2 C12 myoblast differentiation and mouse skeletal muscle regeneration and provided a theoretical basis for further elucidating the molecular mechanism of muscle development.     

谷氨酰胺对斜带石斑鱼GF-1细胞中C-Myc蛋白的表达与神经坏死病毒复制的影响

为探讨C-Myc表达、谷氨酰胺代谢和神经坏死病毒复制三者之间的关系,本研究首先克隆了斜带石斑鱼鳍条细胞(GF-1)中的C-Myc基因(GF-1-C-Myc),结果显示GF-1-CMyc基因cDNA全长814 bp,开放阅读框(ORF)为285 bp,编码95个氨基酸(aa),有亮氨酸拉链结构域与螺旋-环-螺旋(HLH)结构域。实验表达和纯化了GF-1-C-Myc蛋白,并制备其多克隆抗体。采用实时定量PCR技术(qRT-PCR)与免疫印迹法(WB)检测了GF-1-C-Myc基因的表达和神经坏死病毒的复制。结果显示,缺乏谷氨酰胺会同时抑制GF-1-C-Myc基因的表达和神经坏死病毒(NNV)的复制,添加谷氨酰胺可同时促进GF-1-C-Myc的表达和NNV的复制;此外,NNV感染可上调GF-1-C-Myc基因的表达,并显著消耗GF-1细胞培养液中的谷氨酰胺。研究表明,GF-1-C-Myc基因可调控宿主谷氨酰胺代谢,从而有利于神经坏死病毒的复制。本结果为防控NNV的感染提供了参考。     

Identification of a cell-wall peptidase (NlpC/P60) from Nocardia seriolae which induces apoptosis in fathead minnow cells

Nocardia seriolae, a Gram-positive bacterium, is the main pathogen of fish nocardiosis. Protein NlpC/P60 is a cell-wall peptidase and a potential virulence factor of N. seriolae. Subcellular localization research revealed that both NlpC/P60-GFP and NlpC/P60Δsig-GFP fusion proteins were evenly distributed in the whole cell of fathead minnow (FHM) cells. Furthermore, typical apoptotic features, such as nuclear pyrosis and apoptotic bodies, were observed in the transfected FHM cells and grouper spleen cells by the overexpression of protein NlpC/P60. Then, quantitative assays of mitochondrial membrane potential (ΔΨm) value, caspase-3 activity and apoptosis-related gene (Bax, BNIP3, TNF1 and TNF6) mRNA expression were conducted. The results showed that ΔΨm was decreased, caspase-3 was significantly activated, and the mRNA expression of pro-apoptotic genes (Bax and BNIP3) and tumour necrosis factors (TNF1 and TNF6) was up-regulated in NlpC/P60-overexpressed cells. Taken together, the results indicated that the protein NlpC/P60 of N. seriolae might involve in apoptosis regulation. This study may lay the foundation for further study on the function of N. seriolae NlpC/P60 and promote the understanding of the virulence factors and pathogenic mechanism of N. seriolae.     

FAM3C activates Akt to promote viability of oral squamous-cell carcinoma cells

AIM: To investigate the expression and roles of family with sequence similarity 3, member C (FAM3C) in oral squamous-cell carcinoma cells. METHODS: The mRNA and protein expression levels of FAM3C in dysplastic oral keratinocyte (DOK) and oral squamous-cell carcinoma WSU-HN6 cells were detected by RT-qPCR and Western blot. The WSU-HN6 cells were treated with siFAM3C or FAM3C antibody. After 24, 48 and 72 h, the viability of WSU-HN6 cells was measured by CCK-8 assay, and the activation of protein kinase B (Akt) was detected by Western blot. Adenovirus was used to mediate over-expression of FAM3C in the DOK cells. The DOK cell viability was measured by CCK-8 assay after adenovirus infection for 24, 48 and 72 h, and the activation of Akt was detected by Western blot. RESULTS: Compared with the DOK cells, the mRNA and protein levels of FAM3C were significantly increased in the WSU-HN6 cells (P<0.05). The viability of WSU-HN6 cells transfected with siFAM3C was significantly inhibited at 48 h and 72 h (P<0.05). siFAM3C treatment inhibited the activation of Akt (P<0.05). FAM3C antibody treatment also suppressed the viability of the WSU-HN6 cells at 48 h and 72 h and the activation of Akt (P<0.05). Over-expression of FAM3C in the DOK cells promoted the cell viability at 48 h and 72 h and activated Akt (P<0.05). CONCLUSION: FAM3C might promote oral squamous-cell carcinoma cell growth by activating Akt.     

Effect of beclin-1 gene silencing on TuAKe-injured gastric cancer SGC-7901 cells

AIM: To observe the effect of beclin-1 silencing by the technique of RNA interference on the injury of human gastric cancer SGC-7901 cell by Sheliugu extract (the extract from tuber of Amorphophallus konjac, TuAKe). METHODS: To knock down the expression of beclin-1 gene, SGC-7901 cells were transfected with lentiviral vector carrying beclin-1-shRNA. The beclin-1 gene knock-down and non-knock-down SGC-7901 cells were treated with TuAKe. The cell viability was analyzed by CKK-8 assay. The percentages of apoptotic cells were detected by flow cytometry. The expression of beclin-1 and LC3 was detected by Western blot. RESULTS: The beclin-1 gene silencing decreased the protein expression of beclin-1 and increased the protein expression of LC3 in the SGC-7901 cells, leading to the decrease in cell viability and the increase in apoptotic rate (P<0.05). TuAKe increased the protein expression of beclin-1 and LC3 in the SGC-7901 cells, and decreased the protein expression of LC3 in the SGC-7901 cells with beclin-1 gene silencing, thus inhibiting the cell viability and increasing the apoptotic rate (P<0.05). CONCLUSION: Beclin-1 gene silencing inhibits the activation of beclin-1-related signaling pathway in gastric cancer SGC-7901 cells, and aggravates the injury of cell viability induced by TuAKe.     

高抗氧化活性lager型啤酒酵母选育

采用过氧化氢刺激lager型啤酒酵母,Tadpoling法筛选细胞活力恢复较快突变菌,根据各菌株线粒体膜电位、胞内氧自由基(ROS)水平筛选获得4株活性较高菌株。比较突变菌与原始菌传代发酵性能、胞内ROS、活力指标及每一代细胞死亡率水平,最终获得一株抗氧化能力提高且活力稳定性较高啤酒酵母菌株。分析验证突变菌线粒体DNA修复相关基因MHR1发现,该基因发生部分碱基突变。     

Effect of CHOP expression knock-down on apoptosis of renal tubular epithelial cells

AIM:To study the effect of C/EBP homologous protein (CHOP) on the apoptosis of renal tubular epithelial HK2 cells. METHODS:The serum mRNA levels of CHOP in the patients with acute kidney injury and healthy controls were detected by qPCR. In vitro, renal tubular epithelial HK2 cells were divided into control group, negative group (transfected with negative control siRNA), si-CHOP group (transfected with CHOP siRNA), and induced by transforming growth factor-β1 (TGF-β1). The viability of the cells was measured by MTT assay, and the apoptotic rate was analyzed by flow cytometry. The protein levels of nuclear antigen Ki-67, proliferating cell nuclear antigen (PCNA), caspase-3 and cleaved caspase-3 were determined by Western blot. RESULTS:Compared with the healthy controls, the serum mRNA levels of CHOP in the patients with acute kidney injury were increased significantly (P<0.05). Transfection with CHOP siRNA significantly decreased the expression of CHOP in the renal tubular epithelial HK2 cells (P<0.05). Knock-down of CHOP expression by siRNA significantly increased the viability of renal tubular epithelial HK2 cells (P<0.05), decreased the apoptotic rate (P<0.05), increased the expression of Ki-67 and PCNA (P<0.05), and down-regulated the protein level of cleaved caspase-3 (P<0.05). CONCLUSION:The serum mRNA levels of CHOP were increased in the patients with acute kidney injury. Knock-down of CHOP expression inhibits the apoptosis of renal tubular epithelial cells by regulating the expression of proliferation-and apoptosis-related proteins.     

The M43 domain-containing metalloprotease RcMEP1 in Rhizoctonia cerealis is a pathogenicity factor during the fungus infection to wheat

Wheat(Triticum aestivum L.) is an important staple crop for global human. The necrotrophic fungus Rhizoctonia cerealis is the causal pathogen of sharp eyespot, a devastating disease of wheat. Herein, we identified RcMEP1, a zinc metalloproteaseencoding gene from R. cerealis genomic sequences, and characterized its pathogenesis function. RcMEP1 expressed at markedly-high levels during R. cerealis infection process to wheat. The predicted protein RcMEP1 comprises of 287 amino acid residues and contains a signal peptide and a M43 metalloprotease domain harboring the active site motif(HEVGHWLGLYH). The assays of Agrobacterium tumefaciens-mediated transient expression in Nicotiana benthamiana leaves indicated that RcMEP1 is an apoplastic elicitor of cell death, and that the predicted signal peptide functions and is required for secretion and cell death-induction. The purified RcMEP1 protein and its M43 domain peptide were individually able to induce plant cell death and H2 O2 accumulation, and to inhibit expression of host chitinases when infiltrated into wheat and N. benthamiana leaves, while the M43 domain-deleting peptide and negative control lacked the capacity. Moreover, compared with the control pretreatment, the purified RcMEP1 protein or its M43-domain peptide resulted in enhanced pathogenesis in the inoculated wheat, whereas the M43 domain-deleting peptide failed. These results suggest that RcMEP1 acted as an important pathogenicity factor during R. cerealis infection to wheat and that its signal peptide and M43 domain are required for the secretion and pathogenesis of RcMEP1. This study provides insights into pathogenesis role of M43 domain-containing metalloproteases during R. cerealis infection to wheat.     

EPS364, a Novel Deep-Sea Bacterial Exopolysaccharide,Inhibits Liver Cancer Cell Growth and Adhesion

The prognosis of liver cancer was inferior among tumors. New medicine treatments are urgently needed. In this study, a novel exopolysaccharide EPS364 was purified from Vibrio alginolyticus 364, which was isolated from a deep-sea cold seep of the South China Sea. Further research showed that EPS364 consisted of mannose, glucosamine, gluconic acid, galactosamine and arabinose with a molar ratio of 5:9:3.4:0.5:0.8. The relative molecular weight of EPS364 was 14.8 kDa. Our results further revealed that EPS364 was a β-linked and phosphorylated polysaccharide. Notably, EPS364 exhibited a significant antitumor activity, with inducing apoptosis, dissipation of the mitochondrial membrane potential (MMP) and generation of reactive oxygen species (ROS) in Huh7.5 liver cancer cells. Proteomic and quantitative real-time PCR analyses indicated that EPS364 inhibited cancer cell growth and adhesion via targeting the FGF19-FGFR4 signaling pathway. These findings suggest that EPS364 is a promising antitumor agent for pharmacotherapy.