Hsa-miR-218 inhibits growth of cervical cancer HeLa cells by targeting to LASP1

AIM: To study the effect of hsa-miR-218 on cervical cancer HeLa cell growth and the underlying molecular mechanism.METHODS: The lentivirus expression vector pmiR-218 targeting to hsa-miR-218 was constructed. pmiR-218 was transfected into HeLa cells. The number of viable HeLa cells was counted by the method of Trypan blue exclusion. The inhibitory rate of cell activity was detected by WST-8 assay. The expression of LIM and SH3 protein 1(LASP1) at mRNA and protein levels was determined by real-time PCR and Western blot. The interaction between miR-218 and LASP1 was examined using a luciferase reporter assay.RESULTS: The lentivirus expression vector pmiR-218 targeting to hsa-miR-218 was constructed successfully and confirmed by DNA sequencing. Over-expression of miR-218 inhibited the activity of HeLa cells with the inhibitory rates of 15%, 26% and 65% at 24 h, 48 h and 72 h, respectively. The difference between transfection group and blank control/negative control group was statistically significant. The luciferase activity was reduced when co-transfection with miR-218 mimics and LASP1-3,UTR plasmid. The relative expression of miR-218 was increased after transfection with pmiR-218. Over-expression of miR-218 down-regulated the LASP1 expression at mRNA and protein levels by 25% and 75% respectively. Compared with blank control group and negative control group, the difference was statistically significant(P<0.05).CONCLUSION: pmiR-218 effectively inhibits the growth of HeLa cells in a time-dependent manner. miR-218 targets to the 3,UTR of LASP1, thus down-regulating the expression of LASP1 in HeLa cells.     

基于Glb 1基因序列的西南地区玉米地方品种的系统进化

为探讨我国西南地区玉米地方品种Glb 1基因序列多态性,对40份西南地区玉米地方品种的Glb 1基因序列测序,并与Gen Bank中来自普通玉米、小颖玉米、繁茂玉米和摩擦禾的19条同源序列比对。结果表明,来自西南地区四川、重庆、云南和贵州的玉米地方品种分别有35、25、33和25个多态性位点,其遗传多态性分别是小颖玉米的87%、86%、85%和71%,8个种群遗传多态性指数的大小排序为小颖玉米、四川地方品种、重庆地方品种、云南地方品种、贵州地方品种、普通玉米、磨擦禾和繁茂玉米,Tajima’D、FuLi’D和FuLi,F检验表明Glb 1基因是中性进化基因。比较种群间DNA序列的共有多态性和固定差异,玉米地方品种与小颖玉米、繁茂玉米、磨擦禾间存在较高的固定差异和较低的共有多态性。Glb 1基因的系统进化关系及网状关系分析为玉蜀黍属的分类系统提供了佐证,四川玉米地方品种有较高的遗传多态性支持我国玉米由印度经西藏传入四川的论点。     

豌豆品系X9002抗白粉病基因鉴定

白粉病是豌豆的主要病害之一,在全球范围内引起严重经济损失。防治豌豆白粉病最有效、经济和环境友好的方法是利用抗病品种。迄今,2个隐性抗白粉病基因er1、er2和一个显性抗白粉病基因Er3已在豌豆中被鉴定,其中er1基因在世界上被广泛应用于抗病品种培育。er1基因隶属MLO基因家族,其抗性由豌豆Ps MLO1基因座位功能丧失产生。X9002是甘肃省农业科学院培育的一个半无叶(afila)抗白粉病豌豆品系。本研究对X9002抗白粉病基因进行鉴定,开发用于抗白粉病基因选择的分子标记。遗传分析表明X9002对白粉病抗性由1个隐性单基因控制,SSR标记将该基因定位到豌豆第VI连锁群er1座位区域,标记AD60和c5DNAmet与其连锁。Ps MLO1基因序列分析发现,X9002存在一个未知大小和身份的片段插入,该类型突变也发生在含有er1-2等位基因的豌豆品种Stratagem和Franklin,表明X9002抗白粉病基因为er1-2。一个鉴定er1-2等位基因的功能标记Ps MLO1-650被开发,该标记为互引相标记,仅在感病植株中扩增,可以有效用于分子辅助选择。     

Glu-1位点缺失对小麦麦谷蛋白聚合体粒度分布及面团特性的影响

以Glu-1位点正常和部分缺失的小麦品系为材料,探讨HMW-GS和LMW-GS组成与谷蛋白聚合体粒度分布和面团特性的关系,为利用HMW-GS缺失系改良小麦品质提供理论依据。在20个供试硬白冬麦品系中,1个品系为Glu-A1位点缺失,5个品系为Glu-D1缺失,3个品系为Glu-A1和Glu-D1双缺失。所有品系的蛋白质含量皆较高(13.39%~14.12%),品系间无显著差异,缺失系与非缺失系间也无显著差异。Glu-1位点缺失显著降低了高分子量谷蛋白/低分子量谷蛋白比(HMW/LMW)、不溶性谷蛋白大聚体的含量和百分比。谷蛋白/醇溶蛋白比(GLU/GLI)在基因型间变幅较小,且在缺失系和非缺失系间无显著差异。Glu-1位点缺失显著降低了面团弹性,但显著提高了面团的延展性。部分Glu-1位点缺失系仍具有较高的面团强度和突出的延展性,谷蛋白聚合体粒度分布和面团特性受谷蛋白亚基组成和表达量的共同影响。研究结果表明,利用Glu-1位点亚基缺失可能是改善面筋延展性,提高食品加工品质的方法之一。     

水稻籼粳杂种一代稻米加工和外观品质性状研究

采用不同稻米品质3个粳稻和4个籼稻按完全双列模型设计进行杂交，配组了24个籼粳杂种一代F1，对其主要稻米加工和外观品质性状进行研究。加工品质中糙米率和精米率均接近粳稻品种，与籼稻无关，无胞质效应；整精米率差异很大，但大多介于籼粳双亲之间，并且随粳稻亲本不同，其胞质效应表现不一致。外现品质中垩白度和透明度，大多数介于双亲之间．而垩白率多倾向于亲本的高值。     

棉花烂铃病流行趋势超长期预报模型及其应用研究

本文对ＧＭ（１，１）模型参数估计方法进行了改进和简化，据此组建了苏北沿海棉区棉花烂铃病流行趋势超长期预报模型，经１９８０～１９８９年回报和１９９０～１９９５年预报，棉花烂铃病流行轻、重发生的符合率达１００％；１９９６～２０００年预报结果为１９９８年和２０００年为重发生，其余３ａ为轻发生。这将为棉花烂铃病的研究和防治规划提供科学决策依据，也为ＧＭ（１，１）模型能在基层病虫测报站推广应用创造了条件。     

A simple method to control the seed purity of japonica hybrid rice varieties using PCR-based markers

Cytoplasmic male sterility (CMS) by the cms‐bo cytoplasm and its restoration by the nuclear restorer gene, Rf‐1, are used for seed production of japonica hybrid rice varieties. To produce pure hybrid seeds, a prerequisite is to properly manage the seed purity of parental lines, especially CMS lines. In this study, three dominant polymerase chain reaction (PCR)‐based markers (M1, M2 and M3) were developed to detect mutual contamination in seed batches of CMS lines, maintainer lines, restorer lines and hybrids. M1 detected the mitochondrial sequence that was present in the cytoplasm of common japonica varieties and absent in the cms‐bo cytoplasm. M2 and M3 detected the chromosomal sequence related to the Rf‐1 allele in restorer lines and the rf‐1 allele in common japonica varieties, respectively. By the strategic use of these markers, japonica hybrids and their parental lines could be efficiently distinguished from each other. Furthermore, sensitivity tests for the three markers with a series of crude DNA samples prepared from polished grains demonstrated that these markers could detect one contaminating grain among 500 or 1000 grains. Therefore, the bulk PCR analyses with the markers developed here probably make it possible to control the seed purity of japonica hybrids properly by selecting appropriate seed batches of their parental lines quickly and efficiently.     

RFLP/AFLP mapping of a brown planthopper (Nilaparvata lugens Stål) resistance gene Bph1 in rice

We have constructed a linkage map of the rice brown planthopper (BPH)resistance gene, Bph1. RFLP and AFLP markers were selected by thebulked segregant analysis and used in the mapping study of 262 F₂sthat were derived from a cross of `Tsukushibare', a susceptible japonica cultivar, and `Norin-PL3', an authentic japonicaBph1-introgression line. Twenty markers were mapped within a 28.9-cMregion containing the Bph1 locus on the long arm of rice chromosome12. Combining the result of segregation analysis of BPH resistance by themass seedling test and that of the markers, the Bph1 locus wasmapped within a 5.8-cM region between two flanking markers. The closestAFLP markers, em5814N and em2802N, was at 2.7 cM proximal to theBph1 locus. Together with the previously constructed high-resolutionmap of bph2 locating the locus at ca. 10 cM proximal to the Bph1 locus, this improved version of the linkage map would facilitatepyramiding these two important BPH resistance genes.     

臭氧防霉、杀虫和去毒效果的探讨

用臭氧机产生臭氧，进行高水分粮的防霉试验、几种主要储粮害虫的防治试验以及油脂的去毒试验。结果表明：较低浓度的臭氧对水分为16．6％和17．7％的玉米，15．5％和16．6％的稻谷，15．6%的糙米有显著的防霉效果；对玉米象、谷蠹的成虫有良好的杀灭作用；对油脂中的黄曲霉毒素B1有较好的降解功能。     

Pre-maturity α-amylase and incipient sprouting in UK winter wheat,with special reference to the variety Rialto

In 1998 and 1999 the UK winter wheat variety Rialto produced unexpected low Hagberg falling numbers that could not be directly linked to sprouting. It was proposed that these reductions in quality could be due to pre-maturity α-amylase activity (PMAA). The problem was not identified during the selection and commercial development stages. Our study tested the hypothesis that the variety Rialto is PMAA-susceptible. Analysis was done on 13 year-location combinations of field grown Rialto. Together, visual and chemical assessments of sprouting and iso-electric focusing of α-amylase isozymes identified several samples with significant α-amylase activity in the apparent absence of sprouting. In addition, tests with α-amylase sensitive Phadebas gel revealed distinctive PMAA discoloration patterns in 10–44% of the grain from the 13 samples, leading to the conclusion that Rialto is PMAA-susceptible. Diurnal temperature range accumulated for an 11 day period during a warm spell in early simulated grain development displayed a significant but negative correlation with the number of grains showing clear PMAA discoloration patterns on Phadebas gel. The number of clear PMAA grains correlated positively with rainfall accumulated over 11 days. These results suggest that PMAA can increase under conditions similar to those conducive to pre-harvest sprouting. It is however also possible that in some instances both PMAA and incipient sprouting could have produced similar patterns of α-amylase activity. In addition to tests with Rialto, Phadebas gel tests were therefore also done with the known high Hagberg varieties Option and Malacca, sprouted in a controlled environment. Results from the additional gel tests suggest that visual and chemical assessments of sprouting in the grain combined with Phadebas gel analysis could identify PMAA more reliably in grain sub-samples than Phadebas gel analysis alone.