烟草丛顶病毒编码的沉默抑制子初步鉴定

 烟草丛顶病毒（Tobacco bushy top virus,TBTV）为幽影病毒属成员,能在烟草上造成严重危害。TBTV基因组编码4个ORF,其中ORF3和ORF4几乎完全重叠,可能在TBTV的致病性中发挥重要作用。分别将TBTV的ORF3和ORF4与载体pGR107连接,构建植物表达载体pGR107 ORF3,pGR107 ORF4。经农杆菌与含外源GFP基因的pGREEN208共浸润转GFP基因的本氏烟16c叶片,2～5d后（days post inoculation,dpi)在紫外灯下观察发现浸润区有明显绿色荧光,其中pGR107 ORF4在12dpi后浸润区绿色荧光区域扩大,推断ORF4编码产物可能具有局部抑制本氏烟16c对外源GFP基因进行沉默的功能;而pGR107 ORF3浸润区绿色荧光逐渐恢复红色,GFP基因发生沉默,说明ORF3编码产物不具有抑制基因沉默的功能。进一步利用real time PCR定量检测发现,pGR107 ORF4浸润15 dpi 的本氏烟叶片中GFP RNA的含量要比仅接种pGR107的对照高,比未接种的本氏烟16c稍低。推断是因为ORF4编码蛋白抑制了植物对外源GFP RNA产生的沉默,使GFP得以在本氏烟内表达。由此进一步证实ORF4编码产物是TBTV的基因沉默抑制子。     

水稻瘤矮病毒S11基因沉默抑制子的原核表达及抗血清制备

以采自广东省水稻瘤矮病的典型病株为材料,通过RT-PCR法扩增和克隆了水稻瘤矮病毒(Rice gall dwarf virus,RGDV)S11组分核苷酸序列全长。序列分析结果表明,RGDV广东分离物S11(登录号EF532326)核苷酸序列全长1168bp,含有一个1068bp的开放阅读框,编码一条355个氨基酸的多肽,推测分子量约40kD,与泰国分离物基因结构基本一致,其核苷酸与氨基酸序列相似性分别为94.3%和98.8%。将广东分离物S11基因克隆至pBV221温敏表达载体上,在大肠杆菌DH5α中实现了高效表达。以诱导蛋白为抗原免疫家兔获得了抗血清。利用ELISA法测定抗血清的效价为1∶4096,Western blot分析结果表明,抗血清能与S11蛋白发生特异血清学反应。这可为进一步研究S11蛋白的结构和功能,揭示其抑制基因沉默的作用机制提供参考。     

Emerging roles of p53 in ferroptosis and its potential application

p53 is a widely recognized tumor suppressor. Recent studies show that its classical functions such as inducing apoptosis and arresting cell cycle cannot be the essential role in explaining its effects on tumors properly. Instead, its role as a cell metabolism regulator is more important to suppress tumor development. Ferroptosis is a unique cell death form and is associated with many diseases, such as tumors, neurodegenerative diseases and ischemic diseases. The sensitivity of cells to ferroptosis is closely related to the intracellular metabolic state, suggesting that induction or inhibition of ferroptosis may be significant ways in which p53 plays a role in tumor pathology. Meanwhile, the potential mechanism of p53 involved in neurodegenerative diseases and ischemic diseases should also be noticed. Therefore, our study aims to review the latest development in theoretical and practical knowledge of p53's role in ferroptosis, thus widening the understanding of the tumor suppression mechanism, finding new therapeutic targets for tumors that are resistant to chemotherapy and providing new insights into the treatment of neurodegenerative diseases and ischemic diseases.     

Expression of hairpin RNA (hpRNA) targeting the three CTV-silencing suppressor genes confers sweet orange with stem-pitting CTV tolerance

Silencing the silencing suppressor genes of viruses by RNA interference (RNAi) has been demonstrated to be an efficient way to combat viral diseases in plants. Citrus tristeza virus (CTV) has three silencing suppressor genes, p20, p23, and p25. To silence them, the splicing by overlap extension by PCR (SOE-PCR) was applied in this study for construction of two RNAi vectors, pCAMBIA2301G-hp2520 (to target p25 and p20) and pCAMBIA2301G-hp252023 (to target p25, p20 and p23). Following Agrobacterium-mediated transformation, seven and one independent sweet orange transgenic lines were obtained for expressing the fusion transgenes, hp2520 (hairpin RNA of p25 and p20) and hp252023 (hairpin RNA of p25, p20 and p23), respectively. Challenging the transgenic plants by graft-inoculation with severe stem-pitting CTV strain CT14A showed that no transgenic line was immune to the strain. However, lower to much lower CTV titer was detected in two lines of hp2520 and the only line of hp252023. Vegetative progenies of the hp252023-expressing line showed no stem-pitting symptoms 2 years post CT14A inoculation, although a low level of CT14A was still detectable in their leaves. Our results show that expressing the hpRNAs of CTV silencing suppressor genes conferred sweet orange with high tolerance to stem-pitting CTV.     

UvSMEK1, a Suppressor of MEK Null,Regulates Pathogenicity,Conidiation and Conidial Germination in Rice False Smut Fungus Ustilaginoidea virens

Rice false smut, which is caused by Ustilaginoidea virens, is an emerging disease of rice spikelets in rice-growing areas worldwide. However, the infection mechanism of U. virens on rice spikelets is still unclear. Here, we characterized a suppressor of mitogen-activated protein kinase kinase or ERK kinase(MEK) null(UvSMEK1) in U. virens that is conserved among filamentous fungi. Compared with wild type U. virens strain P-1, Uv SMEK1 deletion mutants were defective in pathogenicity and conidial germination. In addition, conidiation of UvSMEK1 deletion mutants was significantly reduced on yeast extract tryptone(YT) plates, but increased in YT broth compared with the wild type. Compared with Uv SMEK1 expression level during the vegetative mycelia and conidiation stages, UvSMEK1 dramatically increased during infection of rice florets. Surprisingly, the Uv SMEK1 deletion mutants exhibited higher tolerance to H2 O2 and NaCl. In summary, presented evidence suggested that UvSMEK1 positively regulated pathogenicity, conidial germination and conidiation in YT broth, and negatively regulated conidiation on YT medium and tolerance to oxidative and osmotic stresses. The results enhance our understanding of the regulatory mechanism of pathogenicity of U. virens, and present a potential molecular target for blocking rice infection by U. virens.     

Characterization of HOX gene expression in canine mammary tumour cell lines from spontaneous tumours

Spatial/temporal controls of development are regulated by the homeotic (HOX) gene complex and require integration with oncogenes and tumour suppressors regulating cell cycle exit. Spontaneously derived neoplastic canine mammary carcinoma cell models were investigated to determine if HOX expression profiles were associated with neoplasia as HOX genes promote neoplastic potential in human cancers. Comparative assessment of human and canine breast cancer expression profiles revealed remarkable similarity for all four paralogous HOX gene clusters and several unlinked HOX genes. Five canine HOX genes were overexpressed with expression profiles consistent with oncogene‐like character (HOXA1, HOXA13, HOXD4, HOXD9 and SIX1) and three HOX genes with underexpressed profiles (HOXA11, HOXC8 and HOXC9) were also identified as was an apparent nonsense mutation in HOXC6. This data, as well as a comparative analysis of similar data from human breast cancers suggested expression of selected HOX genes in canine mammary carcinoma could be contributing to the neoplastic phenotype.     

The effect of the lateral suppressor gene on seed germination in the tomato

Summary A reduction in germination of tomato seed due to the lateral suppressor mutant (ls) is demonstrated. This is shown to be an effect of the maternal genotype rather than of the embryo. Poor germination is not determined until comparatively late in seed development and the effect can be avoided by embryo culture. Cuticular necrosis of the mutant seed appears to be associated with poor germination but the former symptom may be suppressed independently of the latter.     

Biochemical and functional characterization of lymphocytes from a horse with lymphosarcoma and IgM deficiency

Neoplastic lymphocytes from a horse with lymphosarcoma and IgM deficiency were analyzed for ability to grow in culture; surface and cytoplasmic IgM; functional activity in blastogenesis, cytoxicity, and suppressor assays; and activities of six enzymes involved in purine and pyrimidine metabolism. The cells lacked surface and cytoplasmic IgM. They had elevated activity of adenosine deaminase and reduced activity of purine nucleoside phosphorylase. Neoplastic cells were nonresponsive in blastogenesis assay and did not kill allogeneic lymphocyte target cells or YAC-1 targets in a lectin-dependent cytotoxicity assay, however, the cells were active in a suppressor assay. They were grown for 16 weeks in cultures supplemented with interleukin 2, during which time the cells retained suppressive activity. These results are consistent with a T cell lymphoma of suppressor cell origin, and may explain the deficiency of IgM observed in some horses with lymphoreticular neoplasms.     

p53基因研究进展

转录调节因子p53作为一种抑癌基因,可诱导细胞生长阻滞,细胞凋亡,细胞分化以及DNA修复。但p53突变体可能会使野生型p53基因的抑癌功能失活,甚至发挥癌基因的功能。随着分子生物学技术的发展,人们对p53基因调控网络有很多新的认识。笔者就p53的调节通路以及在肿瘤治疗方面的新进展进行综述。     

二倍体及四倍体中抗叶锈病基因在双二倍体中的表达与抑制

 通过杂交将近缘植物中的抗病基因导入普通小麦是抗病育种的常用方法。在利用二倍体和四倍体杂交合成双二倍体小麦过程中, 二倍体或四倍体携带的抗叶锈病基因在双二倍体中大多数情况下可以完全表达或部分表达其固有的抗病性, 但部分抗叶锈病基因则不能表达。四倍体波斯小麦Ps5、Ps8和野生二粒小麦D s3含有相同的抗叶锈病基因LrPs (暂定名), 在双二倍体Am1、Am2、Am3、Am5和Am7中可以表达其抗病性, 但在Am4中不能表达;二倍体粗山羊草Ae37含有Lr41和未知基因, 但Lr41在双二倍体Am2中不能表达;四倍体硬粒小麦Dr147携带Lr23和未知基因, 在双二倍体Am6中不能充分表达。抑制基因的存在是导致抗病基因不能表达或部分表达的主要原因之-。抑制基因位于AB染色体组或D染色体组上, 其抑制作用对抗病基因和病菌致病类型具有专化性, 还可能受温度等环境条件和寄主遗传背景等因素的影响。遗传分析结果表明, 在常温下, 双二倍体Am1、Am2、Am3和Am5对叶锈菌致病类型DGS/HB的抗病性均由1对相同的隐性抗病基因LrPs (暂定名)控制, 与它们具有共同的四倍体亲本Ps5有关。Am4不具有苗期抗叶锈病基因, 但含有来自粗山羊草A e39的1对隐性抑制基因SuLrPs (暂定名), 可抑制Am1、Am2、Am3和Am5中隐性抗叶锈病基因的表达。对抗病抑制基因存在原因和遗传分析验证方法等进行了讨论。