Using antisense nucleic acid technology to study the influence of POLκ on genetic stability

AIM:To establish cell line FL-POLκ^- and to study the role ofPOLκ(polymerase kappa) on genetic stability.METHODS:A mammalian expression vector expressing antisense POLκ gene fragment pMAMneo -amp^--POLκ was constructed by cloning the 1 690-1 918 fragment of POLκ gene into the mammalian expression vector pMAMneo-amp^- in antisense orientation. FL cells were fransfected with this antisense RNA expressing vector and selected by G418. Based on the shuttle-plasmid pZ189, the mutation assay was made.RESULTS:The spontaneous mutation frequency of supF tRNA gene in the plasmid replicated in the FL- POLκ^- was 11.2×10^-4, while it was 4.9×10^-4 and 3.7×10^-4 in the control cells FL and FL-M, respectively.CONCLUSION: POLκ playes an important role in maintenance of genetic stability.     

The effect of populations of Xanthomonas campestris pv. phaseoli in bean reproductive tissues on seed infection of resistant and susceptible bean genotypes

Surface and internal populations of Xanthomonas campestris pv. phaseoli, causal agent of common bacterial blight of bean, on and in flower buds, blossoms and pods of seven bean (Phaseolus vulgaris) genotypes were studied. Bean plants were grown in the field and artificially inoculated at the seedling stage (18 days old). The pathogen was recovered in high numbers from flower buds, blossoms, pods and seed of both resistant and susceptible bean genotypes. Significant differences (P = 0.05) in population levels of X. c. pv. phaseoli between stages of reproductive tissue development were observed. Infected seed from resistant bean genotypes had no visible symptoms. Such seed may play an important role in the epidemiology of common bacterial blight because they are difficult to detect and may occur at low frequency in seed lots, as was the case in the current study.     

Variation and distribution of virulence phenotypes of Bremia lactucae in natural populations of Lactuca serriola

During 1997–2000 the virulence variation and distribution of virulence phenotypes of Bremia lactucae (lettuce downy mildew) in natural populations of Lactuca serriola (prickly lettuce) were studied. Altogether 139 isolates of B. lactucae originating from the Czech Republic, France and Germany were examined for the presence of 27 virulence factors (v-factors) and their combinations. In the Czech population, 37 different v-phenotypes (P1–P37) of B. lactucae were found to occur on L. serriola . Most v-phenotypes were characterized by v-factors that match resistance ( Dm genes/R-factors) carried by L. serriola . A wide diversity of v-phenotypes was recorded every year, but most were rare and did not reappear in other populations of B. lactucae . The three v-phenotypes P21, P28 and P29 were most frequent and widely distributed. There was variation both between spatially isolated populations and within populations of the pathogen. Geographic differences in virulence were found for the southern parts of Moravia, where the phenotypic composition of pathogen populations was completely different from the remaining part of the area investigated. However, some populations had v-phenotypes similar to those of spatially distant populations in Moravia.     

土壤pH值对极小种群毛枝五针松生理特性的影响

幼苗建成期是毛枝五针松(Pinus wangii)天然更新的关键阶段,为找出影响毛枝五针松幼苗存活的关键生态因子,采用植物生理学方法,研究在7个酸碱度土壤培育下,毛枝五针松松针超氧化物歧化酶(SOD)、过氧化物酶活性(POD)活性和脯氨酸、可溶性糖质量分数及丙二醛(MDA)摩尔质量浓度的变化。结果表明:毛枝五针松在弱碱土壤中抗逆性更强,形态观察得知其在弱碱性土壤中生长更好;生理指标测定结果表明,土壤在p H=7.69~8.42时最适宜毛枝五针松幼苗的生长发育。     

极小种群野生植物八蕊单室茱萸的种群状况研究

八蕊单室茱萸为云南省特有的极小种群野生植物和濒危物种。通过查阅相关文献并结合访问调查,确定八蕊单室茱萸可能分布的区域,开展种群资源调查。结果显示:八蕊单室茱萸目前仅在普洱市和西双版纳州有10个天然种群,其种群规模已低于最小可存活种群,属极小种群野生植物和濒危物种,亟需开展保护工作。生境破坏和生境片段化是导致八蕊单室茱单室茱萸濒危的人为因素。提出收集种质资源、营建和管护近地保护种群、建立迁地和回归种群等保育建议。     

An investigation of links between metabolic rate and feed efficiency in European sea bass Dicentrarchus labrax

Feed efficiency (FE) is the amount of body weight gain for a given feed intake. Improving FE through selective breeding is key for sustainable finfish aquaculture but its evaluation at individual level is technically challenging. We therefore investigated whether individual routine metabolic rate (RMR) was a predictor of individual FE in the European sea bass Dicentrarchus labrax, a major species in European mariculture. The European sea bass has three genetically distinct populations across its geographical range, namely Atlantic (AT), West Mediterranean (WM), and East Mediterranean (EM). We compared FE and RMR of fish from these three populations at 18 or 24 °C. We held 200 fish (62 AT, 66 WM, and 72 EM) in individual aquaria and fed them from ad libitum down to fasting. FI was assessed for an ad libitum feeding rate and for a fixed restricted ration (1% of metabolic body weight·day⁻¹, with metabolic body weight = body weight^0.8). After being refed 12 wk in a common tank, individual RMR was measured over 36 h by intermittent flow respirometry. There was a significant effect of temperature whereby fish at 18 °C had greater mean FE (P < 0.05) and lower RMR (P < 0.001). There was also a significant effect of population, where AT fish had lower FE (P < 0.05) and greater RMR (P < 0.001) than WM and EM, at both temperatures. Despite these differences in temperature and population means, individual FE and RMR were not significantly correlated (P > 0.05). Therefore, although the results provide evidence of an association between metabolic rate and FE, RMR was not a predictor of individual FE, for reasons that require further investigation.     

极小种群植物“川柿”生物学特性及种子萌发方法研究

摘 要：【目的】为了保护和发展极小种群植物川柿,开展此研究。【方法】采用观测法,连续3年对其生物学特性研究;种子萌发方法：（1）后熟0、30、45、60天,剥离清洗播种;（2）后熟60天,以GA300、600、1000 mg/L溶液浸种1h;播种基质为河沙（保持一定湿度）,放置室内,统计萌发时间和出苗率。【结果】（1）川柿花期4-5月,10月为果熟期,单果直径32.7±5.80mm,单果重23.0±5.30g;种子肾形、千粒重1070g;成年植株呈隔年开花、结果现象;（2）种子经“后熟+河沙播种”,可萌发,其中以后熟60天的出苗率较高,为65%;直接播种种子,出苗率5%;“后熟+GA溶液浸种+河沙播种”,出苗率均高于65%,其中以GA600mg/L浸泡1h处理,出苗率最高为83.3%。【结论】极小种群植物川柿为常绿叶乔木,有隔年开花、结果现象; “果后熟60天+GA600mg/L浸泡种子1h +室内河沙播种”可提高种子萌发率,为川柿种子繁殖提供参考。     

大白菜DH0与DH1代群体的遗传稳定性分析

以游离小孢子培养获得的HY大白菜DH0代及其自交后代DH1群体与HY(F1),共33份材料为试验材料,调查了其株高、球茎和外叶数3个农艺性状,发现不同的株系性状分离较大。同时采用RAPD技术分析了2代之间的遗传稳定性。从106条RAPD随机引物中筛选出33条重复性好并且具有多态性的引物,进行扩增。从33份材料中扩增了206条带,其中多态带为136条(66.1%)。通过系统聚类分析,DH0代及其子代DH1代遗传关系近,绝大部分能优先聚在一起。说明纯合的小孢子植株单株性状在遗传中很少发生变异,具有很好的遗传稳定性。     

不同地理群体三疣梭子蟹mtDNA本底甲基化水平分析

利用重亚硫酸盐测序法,对适于三疣梭子蟹(Portunus trituberculatus)线粒体基因组甲基化分析的BSP(Bisulfite sequencing PCR)引物进行了筛选,在设计的21对引物中,获得了12对BSP引物。利用这12对引物分析了三疣梭子蟹海州湾群体(江苏连云港)和莱州湾群体(山东东营)线粒体基因组本底甲基化水平,结果表明:海州湾群体不存在甲基化现象,莱州湾群体发现了2个个体在ND2基因(NADH脱氢酶亚基2)位点存在甲基化现象;在存在甲基化的个体中,甲基化类型主要为CHG和CHH类型,还有少量Cp G类型。该研究结果说明,莱州湾群体和海州湾群体线粒体基因组本底水平上的甲基化特征存在一定差异,相关研究值得进一步深入探讨。     

草鱼野生与选育群体遗传变异微卫星分析

为探究经过2个选育世代后选育群体遗传多样性和遗传结构的变化,实验采用多重PCR技术对4个野生草鱼群体(邗江、九江、石首、吴江)和2个选育群体(F1和F2)进行了微卫星序列遗传变异分析。结果显示,6个草鱼群体遗传多样性水平较高,2个选育群体除了平均等位基因数外,其他遗传多样性参数均小于4个野生群体。哈迪—温伯格平衡(Hardy-Weinberg equilibrium)检测显示,在120个群体—位点组合中有62个位点显著偏离哈迪—温伯格平衡,62个群体—位点组合中只有11个组合其近交系数值为负值,其余的51个组合的Fis均为正值。6个草鱼群体AMOVA分析结果显示,3.75%的变异来自于群体间,96.25%的变异来自于群体内,整体的遗传分化指数值为0.038。进一步分析各个群体间Fst,只有石首群体与F1、F2群体之间的Fst大于0.05,处于中等分化,其余群体间分化程度较低,且F2群体与4个野生群体之间Fst比F1群体与4个野生群体之间的Fst大。奈氏标准遗传距离分析结果显示,2个选育群体与野生群体之间的遗传距离大于野生群体之间的遗传距离。基于Dn建立的UPGMA系统发育树得出了相同的结果,即2个选育群体与野生群体之间的亲缘关系比4个野生群体之间的亲缘关系要远。研究表明,经过2个世代选育后,相比4个野生群体,2个选育群体遗传多样性虽有部分下降,但仍处于较高的水平,2个选育群体的遗传结构已发生变化,但其遗传分化程度尚不明显。本研究结果为制定出更加完善有效的选育方案提供了重要参考。