舟山群岛野生山茶种群遗传多样性的ISSR分析

采用ISSR分子标记,利用筛选的20条引物对舟山群岛4个野生山茶（Camellia japonica）种群的遗传多样性进行分析。结果表明：山茶种群的多态位点百分比（PPB）为64.06％,Nei’s基因多样性指数（HE）为0.2390,Shannon信息多态性指数（H）为0.3548,种群水平遗传多样性较高。基因分化系数Gst = 0.2241,表明种群间具有较高的遗传分化。地理距离与遗传距离具有显著相关性（r = 0.9653,P<0.05）,表明岛屿隔离对山茶种群的遗传分化具有重要影响。UPGMA聚类表明同岛种群间的亲缘关系更近。基于舟山群岛山茶种群遗传结构分析,建议在其自然生长地加强就地保护。     

观光木种群遗传多样性的ISSR研究

使用单因素筛选和均匀设计实验建立并优化了观光木ISSR—PCtL的最佳反应体系．从100条ISSR引物中筛选出8条多态性好、扩增稳定的ISSR引物,106份材料共扩增出12l条条带,其中109条呈现多态性,多态位点百分率（PPB）达90．08％。扩增片段为250～2000bp,平均每个引物扩增出15条片段．说明观光木种群间存在丰富的遗传多样性。观光木种群间的Nei’s基因多样性指数（He）=0．3557,Shannon信息指数（I）=0．5276,种群间的分化系数（Gst）=0．6480,而种群内的He=0．1261,I=0．1939,Gst=0．3520,说明观光木种群间的遗传多样性是观光木遗传变异的主要来源。     

利用ISSR标记分析红脂大小蠹微生境中球孢白僵菌遗传多样性

针对红脂大小蠹危害程度不同的3个地区的球孢白僵菌种群,利用ISSR（inter-simple sequence repeat）分子标记分析了各个种群的遗传多样性。从19条引物中筛选出10条多态性高、稳定性好的ISSR引物用于扩增分析。68株球孢白僵菌的Nei’s基因多样性（h）为0.2973,Shannon指数（Is）为0.4488。旬邑、宜君、古交3地白僵菌种群间的基因分化系数（GST）为0.0525,基因流（Nm）为9.0255;而来源于土壤、红脂大小蠹、蛀屑和树皮的白僵菌种群间的基因分化系数为0.1449,基因流为2.9508。各球孢白僵菌种群表现出不同的多样性水平,旬邑种群和红脂大小蠹虫种群的遗传多样性相对较高;地理分布种群间的差异不如分离基质种群间的差异明显,地理分布种群间存在明显的基因交流,而分离基质种群间的基因流较低,遗传分化明显。     

四川六个厚朴种群遗传结构

利用ISSR技术对四川6个野生厚朴种群的遗传结构进行了分析,以评估野生厚朴资源的遗传现状。10条ISSR引物共测到114个位点,其中多态位点93个,多态位点百分率(PPB)为81.58%,Nei遗传多样性指数(H)和Shannon信息指数(I)分别为0.320和0.469,表明厚朴物种水平具有较高的遗传多样性;相比之下,种群水平遗传多样性则相对较低(PPB、H、I分别平均为48.25%、0.189和0.277)。分子变异分析(AMOVA)表明,种群内部和种群间均存在极显著遗传分化(P0.001),其中,36.82%的变异存在于种群间,63.18%存在于种群内,种群间的基因分化系数(GST)为40.42%。厚朴种群间的基因流(Nm)为0.737,平均遗传距离为0.211,算术加权平均数法(UPGMA)显示厚朴6个种群被分为3大类群。Structure分析表明,厚朴种群间遗传结构具有独立性特点。     

武夷山地伯乐树种群遗传多样性的ISSR分析

伯乐树（Bretschneidera sinensis Hemsl.）为我国特有单型科珍稀濒危植物,具重要的科研价值。本研究采用ISSR分子标记对武夷山脉分布的5个伯乐树天然种群和1个移栽种群进行遗传多样性分析。结果表明伯乐树物种水平遗传多样性较高（PPB：75.70%;H_ES：0.304 5;H：0.450 1）,种群水平则较低（PPB：60.13%;H_EP：0.238 1;H：0.347 5）,MJY种群和BSZ种群分别是所有种群中遗传多样性最高和最低的。5个地理种群间遗传分化程度较高（G_st=0.218 1）,原因可能源于伯乐树的繁殖方式及生境片段化,Mantel检验也证实了地理距离与遗传距离具有显著相关性（r=0.626 7,P＜0.05）。针对伯乐树种群遗传多样性现状,建议加强现有自然种群的就地保护,促进种群自然更新。     

Study on population genetic diversity of Camellia japonica in 5 islands between China and Japan

运用ISSR分子标记法对中、日两国5个岛屿天然山茶种群共150个个体进行遗传多样性分析.结果表明:筛选出的20条引物扩增得到205条清晰条带,其中183条为多态性条带,多态位点百分比(PPB)为89.27％.经POPGENE软件分析,山茶种群平均多态位点百分比(PPB)为72.00％,Nei's基因多样性指数(HE)为0.2743,Shannon信息多态性指数(H)为0.4023,种群水平遗传多样性较高.基因分化系数Gst=0.2033,表明遗传变异主要存在于种群内个体间.Mantel检验(r=0.7989,P＜0.05)和UPGMA聚类表明岛屿地理隔离对山茶种群遗传分化具有重要影响.基于岛屿山茶种群遗传结构的分析,建议加强我国岛屿自然种群的就地保护力度.     

杜鹃红山茶遗传多样性的ISSR分析

运用ISSR分子标记技术,利用筛选的10条ISSR引物,对珍稀濒危植物杜鹃红山茶(Camellia changii Ye)2个亚种群60个单株的遗传多样性进行了研究。结果表明:物种水平上的多态位点百分率(PPB)为55.29%,Nei’s基因多样性指数(H)及Shannon多态性信息指数(I)分别为0.2191和0.3215。种群间的遗传分化系数(GST)仅为0.0922。研究结果揭示了杜鹃红山茶的遗传多样性较低,亚种群间遗传分化较小。小种群和人为活动干扰是杜鹃红山茶现存种群的主要限制因素。影响杜鹃红山茶种群发展的其它因素亟需进一步研究。     

小菜蛾不同地理种群遗传多样性的ISSR标记研究

为阐明小菜蛾Plutella xylostella不同地理种群的遗传多样性, 应用ISSR技术对我国小菜蛾8个地理种群的遗传多样性进行了研究分析。15条引物扩增出395条ISSR条带, 其中多态性条带占89.11%, 全部个体显示了各自独特的ISSR图谱。ISSR标记的遗传多样性分析结果表明： 小菜蛾无论在物种水平上（P=89.11%, H=0.2706, I=0.4286）, 还是在种群水平上（P=88.80%, H=0.2759, I=0.4349）都表现出较高的遗传多样性。其中, 北京南口种群内遗传变异最大, 海南海口和甘肃兰州种群内遗传变异最小, 南方地区（云南、 湖北）小菜蛾种群遗传多样性明显高于北方地区（北京、 天津、 山东、 黑龙江、 甘肃）种群。据种群变异来源分析, 有5.66%的遗传变异来自种群间, 94.34%的变异来源于种群内（N_m=8.3399）, 不同地理种群间没有明显的遗传分化。本文有关小菜蛾不同地理种群基因流动和遗传变异的研究为小菜蛾抗药性的控制及田间种群的综合防治提供了有价值的分子生物学依据。     

中日5个岛屿山茶种群遗传多样性研究

运用ISSR分子标记法对中、日两国5个岛屿天然山茶种群共150个个体进行遗传多样性分析。结果表明:筛选出的20条引物扩增得到205条清晰条带,其中183条为多态性条带,多态位点百分比(PPB)为89.27%。经POPGENE软件分析,山茶种群平均多态位点百分比(PPB)为72.00%,Nei’s基因多样性指数(HE)为0.2743,Shannon信息多态性指数(H)为0.4023,种群水平遗传多样性较高。基因分化系数Gst=0.2033,表明遗传变异主要存在于种群内个体间。Mantel检验(r=0.7989,P<0.05)和UPGMA聚类表明岛屿地理隔离对山茶种群遗传分化具有重要影响。基于岛屿山茶种群遗传结构的分析,建议加强我国岛屿自然种群的就地保护力度。     

基于ISSR分子标记的西藏杓兰种群遗传多样性分析

西藏杓兰(Cypripedium tibeticum)是具有很高观赏和药用价值的兰科植物,已被列为中国优先保护的濒危植物。该研究采用ISSR分子标记技术,对采自中国西部地区的7个西藏杓兰种群进行遗传结构及遗传多样性分析,为中国野生西藏杓兰种质资源的保护提供理论依据。结果显示:(1)从100条ISSR引物中共筛选出12条多态性高、重复性好的引物,共检测出136个位点,多态位点百分率(PPB)达100%,总体Nei's基因多样性指数(H_e)和Shannon信息指数(I)分别为0.318 6和0.484 3,种群间的Nei's遗传距离在0.033 3～0.170 1之间,Nei's遗传相似度在0.843 5～0.967 3之间,总体遗传分化系数(G_(st))和基因流(N_m)分别为0.222 9和1.743 0。(2)基于UPGMA法和邻接法的种群系统聚类结果均显示出四川种群与陕西种群之间存在遗传分化。总体上西藏杓兰种群间遗传分化与地理隔离之间相关性不显著。研究认为,西藏杓兰种群在分子水平上具有丰富的遗传多样性,且四川种群与陕西种群已产生了遗传分化;ISSR分子标记可用于西藏杓兰种群遗传多样性、遗传结构及种群间遗传分化等方面的研究。     

The effects of heavy metal pollution on genetic diversity in zinc/cadmium hyperaccumulator Sedum alfredii populations

The genetic diversity and population structure of seven populations of Sedum alfredii growing in lead/zinc (Pb/Zn) mine spoils or in uncontaminated soils from eastern and southern China were investigated using random amplified polymorphic DNA (RAPD) technology. Four of the sampled sites were heavily contaminated with heavy metals (Zn, Cd, Pb), and extremely high concentrations of Zn, Cd, and Pb were found among these corresponding populations. A significant reduction of genetic diversity was detected in the mining populations. The reduction of genetic diversity could be derived from a bottleneck effect and might also be attributed to the prevalence of vegetative reproduction of the mining populations. Analysis of molecular variance (AMOVA) and the unweighted pair group method with arithmetic mean (UPGMA) tree derived from genetic distances further corroborated that the genetic differentiation between mine populations and uncontaminated populations was significant. Polymorphism with the heavy metal accumulation capability of S. alfredii probably due to the genetic variation among populations and heavy metal contamination could have more impact on the genetic diversity and population structure of S. alfredii populations than geographic distance.     

Genetic diversity of SSR markers in wild populations of Tapiscia sinensis,an endangered tree species

Tapiscia sinensis is a Tertiary relict and endangered tree species with unique scientific research value and great economic value. In this study, we assessed the genetic diversity of five wild T. sinensis populations from different geographical regions using 10 polymorphic simple sequence repeat (SSR) markers. Our results reveal that the natural populations of T. sinensis have rich genetic diversity (PPL = 100%, He = 0.6904, I = 1.4368), with Shannon's index indicating that the T. sinensis populations are at a relatively stable stage. Of the genetic relationships among populations, the distance between the Hunan Yanling (YL) and Guizhou Xifeng (XF) populations is the smallest (0.4829); the genetic distance between the Shaanxi Ningshan (NS) and the Guizhou Xifeng (XF) populations is the largest (0.9821). A Mantel test shows that there is no correlation among the populations between geographic distance and genetic distance. AMOVA suggest that 33.3% of the genetic variation arose among the populations, while 66.7% of the variation arose within them. The moderate gene flow among populations (Nm = 0.7274) is not sufficient to counteract genetic drift within the populations and result in significant differentiation (Fst = 0.2987). Our results will benefit the conservation and exploitation of T. sinensis and provide a theoretical basis for further study of the evolution and phylogeography of the species.     

Phytoremediation of lead by a wild,non-edible Pb accumulator Coronopus didymus (L.) Brassicaceae

Coronopus didymus was examined in terms of its ability to remediate Pb-contaminated soils. Pot experiments were conducted for 4 and 6 weeks to compare the growth, biomass, photosynthetic efficiency, lead (Pb) uptake, and accumulation by C. didymus plants. The plants grew well having no visible toxic symptoms and 100% survivability, exposed to different Pb-spiked soils 100, 350, 1500, and 2500 mg kg^?1, supplied as lead nitrate. After 4 weeks, root and shoot concentrations reached 1652 and 502 mg Pb kg^?1 DW, while after 6 weeks they increased up to 3091 and 527 mg Pb kg^?1 DW, respectively, at highest Pb concentration. As compared to the 4 week experiments, the plant growth and biomass yield were higher after 6 weeks of Pb exposure. However, the chlorophyll content of leaves decreased but only a slight decline in photosynthetic efficiency was observed on exposure to Pb at both 4 and 6 weeks. The Pb accumulation was higher in roots than in the shoots. The bioconcentration factor of Pb was > 1 in all the plant samples, but the translocation factor was < 1. This suggested C. didymus as a good candidate for phytoremediation of Pb-contaminated soils and can be used for future remediation purposes.     

Genetic diversity and structure of Cordyceps sinensis populations from extensive geographical regions in China as revealed by inter-simple sequence repeat markers

Cordyceps sinensis is one of the most valuable medicinal caterpillar fungi native to China. However, its productivity is extremely limited and the species is becoming endangered. The genetic diversity of eighteen C. sinensis populations across its major distributing regions in China was evaluated by inter-simple sequence repeat (ISSR) markers. A total of 141 markers were produced in 180 individuals from the 18 populations, of which 99.3% were polymorphic. The low average of Shannon (0.104) and Nei index (0.07) of the 18 populations indicates that there are little genetic variations within populations. For all 18 populations, estimates of total gene diversity (H_T), gene diversity within populations (H_S), coefficient of genetic differentiation (G_ST), and gene flow (Nm) were 0.170, 0.071, 0.583, and 0.357, respectively. This pattern suggests that the genetic diversity of C. sinensis is low and most of the ISSR variations are found among populations with little gene exchange. The 18 populations are divided into five groups based on the genetic distance and the grouping pattern matches with the geographic distribution along the latitudinal gradient. The five groups show obvious difference in the G_ST and Nm values. Therefore, the genetic diversification of C. sinensis populations may be determined by geographic isolation and the combined effects of life history characters and the interaction with host insect species. The information illustrated by this study is useful for selecting in situ conservation sites of C. sinensis.     

Simultaneous extraction of soil phytoavailable cadmium,copper, and lead by chelating resin membrane

Cadmium, copper, and lead were extracted from suspensions of contaminated soils using metal chelating exchange resin membranes. Nine soils with widely varying properties and Cd, Cu and Pb levels were tested. Soil suspensions made up with 4 g in 40 mL deionized water were equilibrated with 5 cm² Ca-saturated Chelex exchange resin membrane which was retained inside a polypropylene bag and shaken at 150 rpm for 24 hrs. Resin membrane extractable Cd, Cu and Pb of the soils were correlated with Cd, Cu, and Pb uptake by young wheat seedlings grown in these soils and compared with soil Cd, Cu, and Pb extracted by 0.1 M HCl, 0.01 M CaCl₂, and 0.005 M Diethylenetriamine pentaacetic acid (DTPA). The amounts of Cd, Cu and Pb extracted by the Ca-saturated Chelex membrane from all tested soils correlated well with those absorbed by young wheat seedlings. The Ca-saturated Chelex membrane extractable Cd, Cu and Pb of the soil had the strongest correlation with plant uptake Cd, Cu and Pb among the extraction methods we tested. It was demonstrated that the Ca-saturated Chelex membrane extraction is an appropriate method in simultaneously estimating Cd, Cu and Pb phytoavailability of soil and is applicable to a wide range of soils.     

Improvement of element uptake and antioxidative defense in Brassica napus under lead stress by application of hydrogen sulfide

Heavy metal pollution is one of the major constraints in oilseed rape (Brassica napus L.) production. In this study, protective role of hydrogen sulfide (H₂S) on plant growth under lead (Pb) stress was studied in B. napus. Plants were grown hydroponically in greenhouse conditions under three levels (0, 100, and 400 μM) of Pb and three levels (0, 100 and 200 μM) of H₂S donor sodium hydrosulfide. Outcomes demonstrated that Pb stress significantly reduced the plant biomass, leaf chlorophyll contents, nutrients uptake in the leaves and roots of B. napus plants. Exogenous application of H₂S significantly improved the plant biomass, chlorophyll contents and concentration of macro- and micronutrients in the leaves and roots of B. napus plants under Pb-toxicity conditions. The data indicated that application of Pb alone significantly increased the reactive oxygen species (ROS) as well as malondialdehyde (MDA) in the leaves and roots of plants. Meanwhile, application of H₂S decreased the production of MDA and ROS in the leaves and roots by increasing antioxidant activities under Pb stress. Moreover, this study also revealed that plants treated with H₂S at different concentrations enhanced the contents of total glutathione and glutathione reduced/glutathione oxidized ratio in leaves and roots under different levels of Pb. The results depicted that H₂S improved the plant biomass, uptake of nutrients in the leaves and roots of B. napus plants and enhanced the performance of antioxidant defense system due to its ameliorative potential under Pb stress conditions.     

Analysis of the genetic diversity and relationships among and within species of Hippophae (Elaeagnaceae) based on RAPD markers

RAPD markers were used to assess the genetic diversity and inter- and intra-specific relationships of the genus Hippophae L. and to study the correlation between genetic distances and geographic distances among populations of H. rhamnoides ssp. sinensis. The results analyzed by the percentage of polymorphic loci and Shannon information index indicated that a high level of genetic diversity existed both among and within species of the genus Hippophae. In the UPGMA dendrogram, the species or subspecies were clustered into two main groups but not strictly grouped according to sect. Hippophae and sect. Gyantsensis Lian. The multiple regression analysis and Mantel test both indicated a significant correlation between genetic distance and altitude distance among populations of H. rhamnoides ssp. sinensis, and the cluster analysis suggested that the genetic variation among populations of H. rhamnoides ssp. sinensis was linked to their monophyletic origin. Moreover, some degree of genetic differentiation was found among samples collected at different times.     

Microsatellite analysis reveals the genetic structure and gene flow of the aquatic quillwort Isoetes sinensis, a critically endangered species in China

Isoetes sinensis is a critically endangered aquatic quillwort, occurring in two fragmented sites of China (Xiuning county of Anhui; Jiande county of Zhejiang). During a five-year period (2004-2009), the areas and sizes of the two populations diminished dramatically due to intensive human activities. Genetic structure of the species was investigated using simple sequence repeat makers (SSRs). For seven nuclear microsatellites, high levels of genetic diversity were found within populations (H_E = 0.324-0.447). Strong genetic differentiation was detected between populations (G_ST = 0.376), while weak genetic differentiation was found within populations (G_ST = 0.026-0.080). Notably, in contrast to the source-sink model suggested by previous study (Chen et al., 2009), the migration pattern of I. sinensis along the Xin’an River is best explained by the linear symmetrical non-adjacent flow model (LSNF), which indicates that intensive human activities of recent years have greatly affected the gene exchange pattern among I. sinensis subpopulations.