中国特有种四合木种群遗传多样性的RAPD分析

采用随机扩增多态性 DNA(RAPD)技术检测了西鄂尔多斯高原特有种四合木种群 5个斑块的遗传多样性。2 2个扩增引物产生 1 1 6条带 ,Shannon信息指数和 Nei指数对 RAPD数据的分析表明 :四合木种群存在较高的遗传多样性 ,其中千里山斑块的遗传多样性和多态位点比例较高 ,石嘴山斑块的最低。遗传多样性的 86.5 %存在于斑块内 ,斑块间的遗传变异为 1 3 .5 %。遗传距离与地理距离无直接相关关系。这些结果说明 ,遗传多样性反映了四合木种群基因组 DNA存在较高的变异性 ;同时各斑块间存在一定的基因流 ,四合木各斑块可看成是处于同一种群的半隔离状态 (meta-种群 ) ,对四合木应注意保护遗传多样性丰富的 meta-种群。     

High genetic differentiation and variation as revealed by ISSR marker in Pseudotaxus chienii (Taxaceae), an old rare conifer endemic to China

Pseudotaxus chienii (Taxaceae) is an endangered conifer species endemic to China, with a historically disjunct distribution pattern. Eleven populations sampled throughout its range were examined using inter-simple sequence repeats (ISSR) markers. Twenty primers generated 242 bands, and each detected polymorphic loci, with 73.14% of polymorphic loci overall. The estimate for θ^B was 0.5306, whereas G_ST and Φ_ST values were 0.6146 and 0.6401, respectively. 50.62% of the total diversity based on Shannon's index of phenetic diversity was attributed to among populations, which was consistent with the θ^B estimation. Compared with other conifers, remarkable genetic differentiation occurred among P. chienii populations. A Mantel test indicated that pairwise values of θ^B were significantly related with geographical distances between populations (r = 0.676, P = 0.001). In terms of the above results, conservation strategies and maintenance plans of P. chienii were recommended.     

Genetic Differentiation and Conservation of 14 Surviving Individuals of Euryodendron excelsum Endemic to China

Random amplification polymorphic DNA (RAPD) markers were used to assess the genetic variations and the evolutionary relationships among all 14 individuals of a critically endangered Euryodendron excelsum (Theaceae) population distributed in Ba Jia Zhen, Yangchun, Guangdong, China. Twenty-three random primers detected 156 sites, out of which 95 (60.26%) were polymorphic loci. The number of the observed alleles was 1.6090, and the number of the effective alleles was 1.3471. Nei’s gene diversity was 0.1993, and Shannon index was 0.1534. A relatively high level of genetic variation was identified in E. excelsum. An unweighted pair group method with arithmetic mean (UPGMA) tree established from Jaccard similarity coefficients suggested that 14 individuals were clustered into two subgroups and that the No. 2 plant was genetically distant from the rest of the individuals. The UPGMA clustering was also supported by a principle components analysis of RAPD phenotypic data. The management and conservation strategy of E. excelsum was proposed based on our results. Translated from Acta Scientiarum Naturalium Universitatis Sunyatseni, 2005, 44(1) (in Chinese)     

烟草毛状根诱导及其茄尼醇含量初探

茄尼醇是合成泛醌类药物的重要中间体。以发根农杆菌(Agrobacterium rhizogenes)W.T15834感染烟草叶片诱导产生毛状根, 探讨其茄尼醇含量变化。结果显示, 获得的毛状根能在无外源生长调节剂的MS固体和液体培养基上自主生长, 但在液体培养基中培养的毛状根生长更迅速, 也不会形成愈伤组织。甘露碱检测及PCR结果证实, 发根农杆菌Ri质粒的rolB基因已在烟草(Nicotiana tabacum)毛状根基因组中整合并得到表达。用改进的HPLC法测定烟草毛状根中的茄尼醇含量, 其结果为对照根(种子萌发产生的幼苗根)的1.12倍, 但仍比废弃烟叶中茄尼醇含量低43.2％。     

Microsatellite loci from Taxus chinensis var. mairei (Taxaceae), an endangered and economically important tree species in China

Taxus chinensis var. mairei is an endangered and economically important tree species in China. Four polymorphic microsatellite markers were developed and characterized using the fast isolation by amplified fragment length polymorphism (AFLP) of sequences containing repeats (FIASCO) protocol. Polymorphism of each locus was quantified for five natural populations from Hubei Province, China. The number of alleles per locus varied from 6 to 24. The ranges of observed and expected heterozygosity were 0.493–1.000 and 0.440–0.845, respectively. These are the first microsatellites reported for the family of Taxaceae and will be helpful for the ongoing population and conservation genetics research of Taxus chinensis var. mairei.     

蕨类植物rpoC1内含子缺失及其分子进化速率

rpoC1基因编码RNA聚合酶β°亚基蛋白, 在转录过程中与DNA模板结合, 与β亚基形成的β-β°亚基复合体构成RNA合成的催化中心。以rpoC1基因为研究对象, 在贝叶斯因子大于20的条件下, 用HyPhy软件位点模型检测到3个正选择位点和541个负选择位点; 用PAML软件位点模型检测到10个正选择位点, 其中3个位点的后验概率超过99%。此外, 基于最大似然法构建64种蕨类植物的系统发育树, 结合HyPhy软件分析rpoC1基因的转换率、颠换率、转换率/颠换率、同义替换率、非同义替换率以及同义替换率/非同义替换率, 探讨rpoC1基因内含子丢失与分子进化速率的关系。结果表明, rpoC1基因内含子缺失对转换率、颠换率以及非同义替换率有一定影响。     

Alteration of De Novo Glucose Production Contributes to Fasting Hypoglycaemia in Fyn Deficient Mice

Previous studies have demonstrated that glucose disposal is increased in the Fyn knockout (FynKO) mice due to increased insulin sensitivity. FynKO mice also display fasting hypoglycaemia despite decreased insulin levels, which suggested that hepatic glucose production was unable to compensate for the increased basal glucose utilization. The present study investigates the basis for the reduction in plasma glucose levels and the reduced ability for the liver to produce glucose in response to gluconeogenic substrates. FynKO mice had a 5-fold reduction in phosphoenolpyruvate carboxykinase (PEPCK) gene and protein expression and a marked reduction in pyruvate, pyruvate/lactate-stimulated glucose output. Remarkably, de novo glucose production was also blunted using gluconeogenic substrates that bypass the PEPCK step. Impaired conversion of glycerol to glucose was observed in both glycerol tolerance test and determination of the conversion of ¹³C-glycerol to glucose in the fasted state. α-glycerol phosphate levels were reduced but glycerol kinase protein expression levels were not changed. Fructose-driven glucose production was also diminished without alteration of fructokinase expression levels. The normal levels of dihydroxyacetone phosphate and glyceraldehyde-3-phosphate observed in the FynKO liver extracts suggested normal triose kinase function. Fructose-bisphosphate aldolase (aldolase) mRNA or protein levels were normal in the Fyn-deficient livers, however, there was a large reduction in liver fructose-6-phosphate (30-fold) and fructose-1,6-bisphosphate (7-fold) levels as well as a reduction in glucose-6-phosphate (2-fold) levels. These data suggest a mechanistic defect in the allosteric regulation of aldolase activity.     

CO2浓度升高对龙血树和春羽生长及光合生理的影响

采用开顶式气室（OTC）控制模拟环境,研究了二氧化碳（CO₂）浓度升高对龙血树和春羽叶片光合生理及叶绿素荧光参数的影响。结果表明:（1）随着CO₂浓度增加,龙血树和春羽幼苗叶面积和株高均呈显著增加趋势。（2）龙血树和春羽幼苗叶片净光合速率随着CO₂浓度的升高均呈先增加后降低的趋势,而同期蒸腾速率和气孔导度显著降低。（3）随着CO₂浓度的增加,龙血树和春羽幼苗叶片最大光量子产量、实际光能转化效率均呈先升后降的趋势,而同期非光化学猝灭系数呈先降低后升高的趋势;光化学猝灭系数总体上也呈先升高后降低的趋势,但差异不显著。研究表明,一定时间的高浓度CO₂处理提高了龙血树和春羽的净光合速率,促进了植物的生长,但随着处理时间的延长,这种促进作用逐渐降低进而消失,并以春羽幼苗表现得更突出;即高浓度CO₂可能破坏了龙血树及春羽生长后期光系统Ⅱ反应中心结构,导致叶片光合能力降低。     

EST‐SSR‐based landscape genetics of Pseudotaxus chienii,a tertiary relict conifer endemic to China

Pseudotaxus chienii, belonging to the monotypic genus Pseudotaxus (Taxaceae), is a relict conifer endemic to China. Its populations are usually small and patchily distributed, having a low capacity of natural regeneration. To gain a clearer understanding of how landscape variables affect the local adaptation of P. chienii, we applied EST‐SSR markers in conjunction with landscape genetics methods: (a) to examine the population genetic pattern and spatial genetic structure; (b) to perform genome scan and selection scan to identify outlier loci and the associated landscape variables; and (c) to model the ecological niche under climate change. As a result, P. chienii was found to have a moderate level of genetic variation and a high level of genetic differentiation. Its populations displayed a significant positive relationship between the genetic and geographical distance (i.e., “isolation by distance” pattern) and a strong fine‐scale spatial genetic structure within 2 km. A putatively adaptive locus EMS6 (functionally annotated to cellulose synthase A catalytic subunit 7) was identified, which was found significantly associated with soil Cu, K, and Pb content and the combined effects of temperature and precipitation. Moreover, P. chienii was predicted to experience significant range contractions in future climate change scenarios. Our results highlight the potential of specific soil metal content and climate variables as the driving force of adaptive genetic differentiation in P. chienii. The data would also be useful to develop a conservation action plan for P. chienii.