金钱槭属植物的遗传多样性与保护策略

金钱槭属包括2个种:金钱槭(Dipteronia sinensis Oliv.)和云南金钱槭(Dipteronia dyeriana Henry),均为珍稀濒危植物,主要分布于中国的中部和南部。该研究应用18种随机扩增多态性DNA(RAPD)引物、8对扩增片段长度多态性(AFLP)引物和10种多态性叶绿体微卫星基因(cpSSR)引物,获得金钱槭和云南金钱槭的遗传多样性与种群结构的相关信息。结果表明,在物种水平上,由RAPD(金钱槭:H=0.386 4,Hsh=0.556 3;云南金钱槭:H=0.304 7,Hsh=0.445 0)和AFLP(金钱槭:H=0.331 9,Hsh=0.488 0;云南金钱槭:H=0.304 7,Hsh=0.4450)2个核标记的遗传多样性具有可比性,而cpSSR标记(金钱槭He=0.603 2,云南金钱槭He=0.671 1)的遗传多样性高于AFLP和RAPD标记的遗传多样性,遗传分化系数(GST)也显示出相似的结果,由此揭示2个种均拥有相对较高的遗传多样性和遗传分化。研究认为,人类活动破坏植物栖息地是造成2种物种处于濒危状态的主要原因;从长远来看,保护2种金钱槭属植物的主要策略是保护植物栖息地,同时应加强扩大种群规模以维持现有遗传变异水平。     

中国特有金钱槭属植物物种分化研究

金钱槭属(Dipteronia Oliv.)隶属于槭树科(Aceraceae),是北温带植物区系中古老、残遗的木本属之一,为中国特有属,属下仅包括金钱槭(D.sinensis Oliv.)和云南金钱槭(D.dyerana Henry)2个物种。本实验从形态学、分子遗传学及基因序列分析等方面对金钱槭(D.sinensis Oliv.)和云南金钱槭(D.dyerana Henry)进行对比研究,探讨其物种间分化程度。结果表明:(1)形态学研究显示两物种形态性状存在显著差异,表明其在形态上发生了显著分化;(2)基因组DNA的AFLP标记分析表明,两物种间存在极显著遗传分化(ΦST=0.390 4),在物种水平及群体水平,两物种遗传多样性及遗传变异水平皆有明显差异(金钱槭云南金钱槭);(3)从GenBank数据库中选出2物种已知的叶绿体基因(psbM-trnD、trnD-trnT、trnL、trnL-trnF、rpl16)和核基因ITS(ITS1、5.8SRNA、ITS2)进行序列进化比对分析,t-test检验表明,2物种叶绿体基因序列间有极显著差异。研究结果表明:金钱槭和云南金钱槭两物种间产生了极显著的分化,推测可能由于长期的地理隔离所致。     

封面植物简介——金钱槭

金钱槭（Dipteronia　sinensis　Ｏliv．）隶属于槭树科金钱槭属,为中国特有的寡种属植物之一。     

封面植物简介——金钱槭

金钱槭(Dipteronia sinensis Oliv．)隶属于槭树科金钱槭属,为中国特有的寡种属植物之一。金钱槭为落叶乔木,高可达10m,具裸露的冬芽。奇数羽状复叶对生．通常具7～11枚小叶。花白色,杂性,雄花与两性花同株,形成网锥花序。翅果周围环绕着圆形的翅．嫩时粉红色．成熟时棕黄色,状如占钱。金钱槭分布于陕西、     

封面植物简介——金钱槭

金钱槭(Dipteronia sinensis Oliv．)隶属于槭树科金钱槭属,为中国特有的寡种属植物之一。金钱槭为落叶乔木,高可达10m,具裸露的冬芽。奇数羽状复叶对生,通常具7～11枚小叶。花白色,杂性,     

中国特有植物金钱槭种子化学成分的研究

采用气相色谱,液相色谱,氨基酸高速自动分析仪,以及化学分析的方法,对中国特有植物金钱槭（ＤｉｐｔｅｒｏｎｉａｓｉｎｅｎｓｉＯｌｉｖ．）种子的化学成分进行了研究。首次报道金钱槭种子含油率２９．１４％。油的理化常数：折光率１．４７３２（２０℃）,皂化值１７８;７０,酸值６．９０。油的脂肪酸组成（％）：油酸３６．７,亚油酸２４．２,芥酸１７．８,花生油酸７．２．亚麻酸２．８,棕榈酸６．７,花生酸３．０,硬脂酸１．５,以及微量的棕榈油酸、十六碳二烯酸,山酸和１种未鉴定酸。种子含蛋白质２９．１６％,含游离氨基酸０．４２５％,含水解氨基酸２２;８４４％,含１８种氨基酸成分。含可溶性糖１０．８６％,其糖分组成：蔗糖２．７３９％,葡萄糖０．９０９％,果糖０．６９８,及微量木糖与１种未鉴定糖。含纤维素２．２５％,含灰分５．３７％,含干物质９２．１０％。     

金钱槭和云南金钱槭遗传多样性比较研究

金钱槭属(Dipteronia)是我国特有少种属,属下仅金钱槭(D. sinensis)和云南金钱槭(D. dyeriana)两种。该文用RAPD标记揭示了金钱槭的遗传多样性和遗传结构,并与云南金钱槭的RAPD研究结果进行了比较。同时,对两物种遗传距离与地理距离的相关性进行了分析,结果有助于阐释该属植物遗传变异的产生机制。研究显示,18条随机引物在17个金钱槭居群(226个个体)中检测到128个扩增位点,物种水平的多态位点比率为92.97%,在4个云南金钱槭居群(45个个体)中则检测到103个扩增位点,物种水平的多态位点比率为81.55%,金钱槭的多态位点比率高于云南金钱槭。相似性系数值、Shannon多样性指数和Nei基因多样性指数分析反映了与多态位点比率相一致的结果。AMOVA(Analysis of molecular variance)分析结果显示,金钱槭居群内、居群间的遗传变异分别占总变异量的56.89%和43.11%。云南金钱槭居群内、居群间的遗传变异分别占总变异量的57.86 %和42.14%。Shannon多样性指数、Nei基因多样性指数的分析结果与AMOVA分析结果趋势相同。上述特征值揭示,金钱槭和云南金钱槭居群间的遗传分化均已达到较高水平,推测居群间低水平的基因流可能是导致上述现象产生的原因之一。遗传距离与地理距离的相关分析结果显示,金钱槭居群间的遗传距离与经度差异存在极显著水平的相关性(p<0.01),云南金钱槭居群间的遗传距离与地理隔离则无显著相关关系。说明在大尺度上遗传距离与地理距离相关而在小范围内则无上述关系,该结果可能与位于不同分布区内的物种所承受的生境选择压力不同有关。建议在对该属植物进行就地保护时,应设立多个保护点,保护自然居群及其周围生境;在迁地保护时,应通过加大居群间种子和幼苗的交换,人为创造基因交流和重组的条件,保存该属植物的遗传多样性。     

封面植物简介——金钱槭

金钱槭（Dipteronia sinensis Oliv．）隶属于槭树科金钱槭属,为中国特有的寡种属植物之一。金钱槭为落叶乔木,高可达10m,具裸露的冬芽。奇数羽状复叶对生,通常具7～11枚小叶。花白色,杂性,雄花与两性花同株,形成圆锥花序。翅果周围环绕着圆形的翅,嫩时粉红色,成熟时棕黄色,状如古钱。金钱槭分布于陕西、     

封面植物简介——金钱槭

金钱槭（Dipteronia sinensis Oliv．）隶属于槭树科金钱槭属,为中国特有的寡种属植物之一。金钱槭为落叶乔木,高可达lOITI,具裸露的冬芽。奇数羽状复叶对生,通常具7～ll枚小叶。花白色,杂性,雄花与两性花同株,形成圆锥花序。翅果周围环绕着圆形的翅,嫩时粉红色,成熟时棕黄色,状如古钱。     

封面植物简介——金钱槭

金钱槭（Dipteronia sinensis Oliv．）隶属于槭树科金钱槭属,为中国特有的寡种属植物之一。金钱槭为落叶乔木,高可达10m,具裸露的冬芽。奇数羽状复叶对生,通常具7～11枚小叶。花白色,杂性,雄花与两性花同株,形成圆锥花序。翅果周围环绕着圆形的翅,嫩时粉红色,成熟时棕黄色,状如古钱。金钱槭分布于陕西、甘肃、湖北、河南、四川、贵州等省,被列为中国三级保护植物。它对于研究槭树科植物的系统演化及中国植物区系具有重要意义。金钱槭宜栽培,可引种为行道树或庭园绿化树。     

A comparison of the genetic diversity in Dipteronia sinensis Oliv. and Dipteronia dyeriana Henry

Dipteronia is an endemic genus to China and includes only two species, Dipteronia sinensis and D.dyeriana.Based on random amplified polymorphic DNA (RAPD) markers,a comparative study of the genetic diversity and genetic structure of Dipteronia was performed.In total,128 and 103 loci were detected in 17 D.sinensis populations and 4 D.dyeriana populations,respectively,using 18 random primers.These results showed that the proportions of polymorphic loci for the two species were 92.97% and 81.55%,respectively,indicating that the genetic diversity of D.sinensis was higher than that of D.dyeriana.Analysis,based on similarity coefficients,Shannon diversity index and Nei gene diversity index,also confirmed this result.AMOVA analysis demonstrated that the genetic variation of D.sinensis within and among populations accounted for 56.89% and 43.11% of the total variation,respectively,and that of D.dyeriana was 57.86% and 42.14%,respectively.The Shannon diversity index and Nei gene diversity index showed similar results.The abovementioned characteristics indicated that the genetic diversity levels of these two species were extremely similar and that the interpopulational genetic differentiation within both species was relatively high.Analysis of the genetic distance among populations also supported this conclusion.Low levels of interpopulational gene flow within both species were believed to be among the leading causes for the above-mentioned phenomenon.The correlation analysis between genetic and geographical distances showed the existence of a remarkably significant correlation between the genetic distance and the longitudinal difference among populations of D.sinensis (p＜0.01),while no significant correlation was found between genetic and geographical distances among populations of D.dyeriana.This indicated that genetic distance was correlated with geographical distances on a large scale rather than on a small scale.This result may be related to differences in the selection pressure on species by their habitats with different distribution ranges.We suggest that in situ conservation efforts should focus on establishing more sites to protect the natural populations and their habitats.Ex situ conservation efforts should focus on enhancing the exchange of seeds and seedlings among populations to facilitate gene exchange and recombination,and to help conserve genetic diversity.     

猕猴桃AFLP分析体系的建立

从64对AFLP分析引物中随机选取了19对引物组合。经过跑小板的聚丙烯酰胺凝胶,银染显带AFLP条带,从中筛选出可做荧光的8对引物。再从中选取了扩增位点丰富,带型质量好,分辨率较高,条带信号强度一致性好,条带分布均匀,且条带较完整的4对引物：E-AAC＋M-CAC、E-AAG＋M-CTG、E-AAC＋M-CAG、EAAC＋M-CTA进行荧光跑带、读带。共在156个位点上扩增出条带,4对引物共扩增出多态性条带132条,多态性比例平均为84．62％,4对引物对10份猕猴桃材料的区分率达100％。说明该4对引物用于猕猴桃属植物的AFLP分析是可行的。     

裸燕麦AFLP反应体系的优化

影响裸燕麦AFLP反应的关键因素包括基因组DNA提取过程中氯仿-异戊醇的抽提次数,酶切时间,预扩增产物稀释倍数,选择性扩增中Mg2+、dNTP、引物浓度等.本研究对这些影响因素进行了优化,初步建立了适合裸燕麦的AFLP反应体系.并将该体系应用于引物筛选,在12份材料中,共筛选出20对条带清晰、多态性好的引物组合,为裸燕麦遗传多样性分析提供了基础.     

紫茎泽兰DNA的提取及AFLP反应体系的建立

以紫茎泽兰叶片为材料,用改进的CTAB法,在提取液中加入2%PVP40(V/V)、0.4%BME(V/V),提取到了高质量的基因组DNA,OD260/OD280在1.7~1.9之间,蛋白质、多酚类、多糖、RNA等去除较彻底,适于AFLP分析。通过对紫茎泽兰DNA提取、酶切连接、预扩增、选择性扩增等实验过程中各关键因素的比较研究,建立了一套优化的AFLP分子标记体系,得到了清晰的AFLP银染指纹图谱,实验结果重复性好。     

天麻AFLP分析技术体系的建立

目的:建立一个适于天麻研究用的AFLP分析技术体系。方法:以11份天麻种质资源为试材,构建供试天麻的AFLP指纹图谱,同时对AFLP分析过程中DNA提取、酶切、连接、预扩增、引物筛选、选择性扩增、电泳和银染等多种因素进行分析。结果:AFLP分析体系要求DNA模板质量高,酶切连接可同时进行,37℃、9h效果较好,预扩增要求高质量DNA聚合酶,产物15倍稀释作为选择性扩增模板效果好,制胶前玻璃板的处理和银染时间的掌控对获得较好结果非常关键。P-GGA/M-GT引物构建的指纹图谱拥有可清晰辨认的扩增条带45条。结论:AFLP指纹技术具有稳定性高、重复性好等优点,可用于天麻DNA分析。     

Genetic diversity and geographical differentiation of Dipteronia Oliv. (Aceraceae) endemic to China as revealed by AFLP analysis

The genus Dipteronia Oliv. endemic to central and southern China consists of two species, Dipteronia sinensis Oliv. and Dipteronia dyeriana Henry, both of them are rare and endangered. AFLP markers were used to characterize the genetic diversity and geographical differentiation of the genus. Eight out of 32 PstI + 3/MseI + 3 selective primer combinations screened were applied to the analysis on 142 individuals of 17 D. sinensis and 4 D. dyeriana populations, respectively. A total of 324 fragments with 316 polymorphic were amplified. The proportion of polymorphic loci (PPB) was 97.53%. The Nei's gene diversity in D. sinensis and D. dyeriana was 0.3319 and 0.3047, respectively. About 43.6% (G_ST = 0.4356) of the genetic variation occurred among the populations, indicating a relatively high genetic differentiation among the populations. Cluster analysis grouped the 21 populations into two groups according to their species delimitation. The populations of D. sinensis were further divided into three subgroups corresponding to their geographical distributions. Correlation analysis revealed a significant correlation (p < 0.05) between geographical distance and genetic distance of these populations, suggesting that the relatively high genetic differentiation among the populations of D. sinensis might be caused by geographical isolation.     

A comparison of the genetic diversity in Dipteronia sinensis Oliv. and Dipteronia dyeriana Henry

Dipteronia is an endemic genus to China and includes only two species, Dipteronia sinensis and D. dyeriana. Based on random amplified polymorphic DNA (RAPD) markers, a comparative study of the genetic diversity and genetic structure of Dipteronia was performed. In total, 128 and 103 loci were detected in 17 D. sinensis populations and 4 D. dyeriana populations, respectively, using 18 random primers. These results showed that the proportions of polymorphic loci for the two species were 92.97% and 81.55%, respectively, indicating that the genetic diversity of D. sinensis was higher than that of D. dyeriana. Analysis, based on similarity coefficients, Shannon diversity index and Nei gene diversity index, also confirmed this result. AMOVA analysis demonstrated that the genetic variation of D. sinensis within and among populations accounted for 56.89% and 43.11% of the total variation, respectively, and that of D. dyeriana was 57.86% and 42.14%, respectively. The Shannon diversity index and Nei gene diversity index showed similar results. The abovementioned characteristics indicated that the genetic diversity levels of these two species were extremely similar and that the interpopulational genetic differentiation within both species was relatively high. Analysis of the genetic distance among populations also supported this conclusion. Low levels of interpopulational gene flow within both species were believed to be among the leading causes for the above-mentioned phenomenon. The correlation analysis between genetic and geographical distances showed the existence of a remarkably significant correlation between the genetic distance and the longitudinal difference among populations of D. sinensis (p < 0.01), while no significant correlation was found between genetic and geographical distances among populations of D. dyeriana. This indicated that genetic distance was correlated with geographical distances on a large scale rather than on a small scale. This result may be related to differences in the selection pressure on species by their habitats with different distribution ranges. We suggest that in situ conservation efforts should focus on establishing more sites to protect the natural populations and their habitats. Ex situ conservation efforts should focus on enhancing the exchange of seeds and seedlings among populations to facilitate gene exchange and recombination, and to help conserve genetic diversity. __________ Translated from Acta Phytoecologica Sinica, 2005, 29(5): 785–792 [译自: 植物生态学报, 2005, 29(5): 785–792]     

鼠尾草属部分物种AFLP指纹图谱构建及遗传多样性分析

为了建立适用于鼠尾草属植物的AFLP技术体系,并对鼠尾草属部分物种遗传多样性进行分析。作者经筛选得到24对有效AFLP引物组合,共检测1616个有效扩增位点;其中22对AFLP引物组合针对不同材料可检测到特异性扩增位点,每对引物检测强度从1到16个位点不等;依据AFLP数据计算的22个鼠尾草属植物材料间231个配对遗传相似系数介于0.5677～0.9898,显示了丰富的遗传多样性;系统树显示AFLP分析可以将11个鼠尾草属植物准确区分,其中种内不同居群的材料可有效聚为亚组,说明本文所构建的AFLP技术体系能够有效应用于鼠尾草属植物种间及种内鉴定、遗传多样性分析及系统发育研究中。     

华中五昧子AFLP反应体系的建立

目的:建立一个适于华中五味子研究用的AFLP反应体系.方法:以华中五味子硅胶干燥嫩叶为试材,采用改良CTAB法提取到高质量DNA.通过琼脂糖凝胶电泳和聚丙烯酰胺凝胶电泳对Mse I/EcoR Ⅰ双酶切、连接、预扩增和选择性扩增过程中的关键因素进行分析.结果:双酶切6 h,片段主要集中在250～2 000bp;连接产物和预扩增产物最适稀释倍数均为10倍;预扩增产物经选择性引物E-ACF/M-CAT和E-ACA/M-CAG扩增,琼脂糖电泳检测其主带分别集中在250～375 bp和500～750 bp,6%聚丙烯酰胺凝胶电泳检测及银染,条带清晰可辨.结论:该体系具有稳定性高、重复性好等优点,可用于华中五味子AFLP分析.