中国蛇根草属(茜草科)几种植物的分类学订正

根据对中国蛇根草属Ophiorrhiza植物标本的研究和野外调查,将琼崖蛇根草O.aureolina f.qiongyaensis Lo和溪畔蛇根草O.humilis Tseng归并入短小蛇根草O.pumila Champ.ex Benth.,将海南蛇根草O.hainanensis Tseng归并入垂花蛇根草O.nutans C.B.Clarke,将广西蛇根草O.kwangsiensis Merr.exLi和变黑蛇根草O.nigricans Lo归并入日本蛇根草O.japonica Bl.,将龙州蛇根草O.longzhouensis Lo和圆锥蛇根草O.paniculiformis Lo归并入广州蛇根草O.cantoniensis Hance。     

国产尖叶蛇根草(茜草科)的订正

中国文献中记载的尖叶蛇根草（Ophiorrhiza hispida Hook.f.）以及采自中国鉴定为该种的标本均实为近簇花蛇根草（O.pseudofasciculata Schanzer）,在此予以纠正。     

中国纤细蛇根草(茜草科)的订正

通过标本考证和野外研究,证实中国文献记载的纤细蛇根草（Ophiorrhiza gracilis Kurz）实为尾瓣蛇根草（O. caudipetala Deb & Monda）错误鉴定,而后者之前仅被报道分布于印度,因此为中国新记录。尾瓣蛇根草具有明显不等大的对生叶片, 花冠裂片背部具长0.6~1 mm的角状突起,易与中国蛇根草属其它种类区分。     

日本蛇根草DFR3基因的克隆分析及其编码蛋白的分离纯化

二氢黄酮醇4-还原酶(DFR)作为花色苷代谢途径下游的关键酶,对植物花色苷的合成具有重要调控作用。该研究以日本蛇根草(Ophiorrhiza japonica)为材料,采用RT-PCR方法克隆获得一个DFR基因(OjDFR3),利用生物信息学方法对OjDFR3蛋白的性质进行了分析,通过实验完成该基因原核表达载体的构建及其重组蛋白的制备与纯化,为深入揭示日本蛇根草DFR基因的功能以及花色苷的合成与调控研究奠定基础。结果表明:(1)成功克隆获得一个DFR基因(OjDFR3);序列分析显示,OjDFR3基因cDNA全长为1 071 bp,可编码356个氨基酸,蛋白分子量为39.52 kD。(2)生物信息学分析表明,OjDFR3基因编码形成的蛋白由20种氨基酸组成,其中亮氨酸含量最多,不存在信号肽,是一种亲水性蛋白,定位在细胞质的可能性最大,三级结构由α螺旋、延伸链、无规则卷曲组成。(3)原核表达分析显示,重组质粒pET32a-OjDFR3可在大肠杆菌BL21(DE3)中表达,其最佳诱导表达条件为37℃、4 h、IPTG浓度为0.8 mmol/L,同时在100和200 mmol/L咪唑洗脱下的蛋白纯度最好。(4)按照上述最佳条件,制备并获得了大量浓度和纯度较好的蛋白。     

Phylogeography of Ophiorrhiza japonica (Rubiaceae) in continental islands,the Ryukyu Archipelago,Japan

Aim Phylogeographical patterns in the Ryukyu Archipelago have been explained primarily by landbridge formation and the opening of two straits in the Pliocene, namely the Tokara and Kerama gaps. These old straits have been considered to be the barriers most likely to determine genetic boundaries. To test this, we conducted a molecular analysis of the herb Ophiorrhiza japonica. We discuss the causes of and processes involved in its phylogeographical structure and explore aspects of island separation other than the duration of the straits to explain genetic boundaries at the gaps. Location Ryukyu Archipelago, Japan. Methods Plants were collected from 40 localities in the archipelago and vicinity. Non‐coding regions of chloroplast DNA were sequenced. The genealogical relationships among haplotypes were estimated using a statistical parsimony network. To examine the phylogeographical structure, we compared two parameters of population differentiation, namely G_ST and N_ST, and conducted correlation analysis of genetic and geographical distances. Genetic boundaries were identified using Monmonier’s maximum difference algorithm. To test vicariance–dispersal hypotheses, that is, vicariance after migration via the Pliocene landbridge or over‐sea dispersal in the Pleistocene, molecular dating analysis was conducted. Results A statistical parsimony network revealed that the haplotypes from the Ryukyu Archipelago and northwards coalesce to one ancestral haplotype in Taiwan. A clear phylogeographical structure was observed: plants within the same population and populations in geographical proximity were phylogenetically close. A genetic boundary was recognized across the Kerama Gap, but not across the Tokara Gap. Dating analysis suggested that population divergence across the Kerama Gap occurred in the early to late Pleistocene. Main conclusions The statistical parsimony network suggests migration from Taiwan and northward range expansion in the archipelago. Based on the divergence time, over‐sea dispersal in the Pleistocene is likely, although migration via a Pliocene landbridge is not totally rejected. Negligible genetic differentiation across the Tokara Gap suggests recent over‐sea dispersal, possibly facilitated by the small geographical width of the gap. Conversely, the large genetic differentiation across the Kerama Gap is probably explained by the large geographical distance across it. The past splitting of a landbridge would have had a significant influence on population differentiation after a certain geographical distance was reached.