基于ITS和trnL-F序列碱基差异的繁缕及其近缘种的亲缘关系分析

应用PCR产物直接测序法分析了繁缕[Stellaria media(L.)Villars]及其近缘种和鹅肠菜[Myosoton aquaticum (L.)Moench]的ITS和trnL-F序列的碱基差异,并以孩儿参[Pseudostellaria heterophylla(Miq.)Pax]为外类群构建N-J系统树,分析了这些种类的种间亲缘关系.结果表明,供试的繁缕属(Stellaria L.)种类及鹅肠菜的ITS和trnL-F序列长度分别为521～784和788～951 bp,各有变异位点77和59个,其中信息位点分别为18和11个,种间碱基差异百分率分别为6.5%和3.1%.ITS的碱基组成为A 22.1%、T20.9%、G 28.5%和C 28.5%,G+C含量57.0%;trnL-F的碱基组成为A 35.9%、T 33.7%、G 15.7%和C 14.8%,G+C含量30.5%.繁缕、雀舌草(S.alsine Grimm)和箐姑草(S.vestita Kurz)的ITS和trnL-F序列一致;鹅肠菜的ITS序列中有4个位点与前述种类不同,但trnL-F序列则相同;中国繁缕(S.chinensis Regel)的ITS和trnL-F序列与孩儿参较相似.在N-J系统树中,鹅肠菜与繁缕、雀舌草和箐姑草聚为一支,表明它们的亲缘关系相对较近,支持将鹅肠菜重新归入繁缕属的分类处理.研究结果显示,ITS和trnL-F序列分析均可用于繁缕及其近缘种的鉴别,且ITS是更为适宜的分子标记.     

茅山地区南苍术的特异性分子标记研究

以中国野生南苍术〔Atractylodes lancea(Thunb.)DC.〕全部分布区9个居群(包括江苏句容的茅山居群、射乌山居群和小九华山居群,安徽金寨的燕子河居群和天堂寨居群,河南嵩县的五马寺居群和纸坊居群,湖北的保康后坪居群和红安七里坪居群)为研究对象,进行rbcL、matK、psbA-trnH、ITS和nadF片段序列比对及SRAP指纹图谱比较来筛选江苏句容茅山居群南苍术的特异性分子标记.结果表明:通过5个DNA片段的序列比对,江苏句容茅山居群南苍术在nadF片段的第581位点处碱基为T,其余8个居群为A,可以将江苏句容茅山居群与其余8个居群区分开.利用SRAP随机引物筛选出1条江苏句容茅山居群南苍术特有的长度为323 bp的条带,根据该条带序列设计1对SCAR引物,正向引物序列为5′-ATCTTTGCTATTAACTATTAAGCTATTCTTCGCTATTCA-3′,反向引物序列为5′-CGATGTTGAAATTGCTGTCTCTGCTTTCTATTC-3′,该引物可在江苏句容茅山居群所有南苍术单株中扩增得到长度为268 bp的特异性条带,而在其他居群中均不能扩增出该条带.本研究筛选出的2种特异性标记可以为鉴定南苍术的道地性提供可靠的技术支持.     

野生与栽培茅苍术挥发油成分的比较分析

采用气相色谱法分析茅苍术〔Atractylodes lancea(Thunb.)DC.〕在原产地及引种栽培1年后5个主要挥发油成分的含量变化。结果表明,江苏产茅苍术挥发油成分以苍术酮和苍术素为主;湖北产茅苍术挥发油的主要成分为茅术醇和β-桉叶醇;不同产地野生茅苍术挥发油的主要成分和含量明显不同;引种栽培至同一环境(江苏南京)后,茅苍术挥发油的主要成分及含量差异明显减小,表明产地对茅苍术挥发油有效成分的组成有一定的影响。     

黑龙江省关苍术资源的调查

苍术(Rhizoma Atractyiodis)为常用中药,以根茎入药。《中华人民共和国药典》(一部1985年版)确定的苍术为茅苍术(Atractyloides llancea(Thunb.)DC.)和北苍术(Atractyloides chinensis(Bge.)Koidz.)。在我省进行中药资源普查中,了解到我省大量收购关苍术(Atractyloides japonica Koidz. ex. Kitam.),做苍术用。为了更好地利用开发关苍术,我们进行了资源调查,现将结果报告如下: 一、原植物形态及商品药材特征关苍术原植物为多年生草本,株高30—60厘米。根状茎横走,肥大而呈结节状。茎直立     

山东莴苣亚族植物的修订

对山东现有的莴苣亚族的植物标本进行了研究,确定山东莴苣亚族植物有24种,分属于12个属,即：苦苣菜属(Sonchus L.)、乳苣属(mulgedium Cass.)、山柳菊属(Hieracium L.)、黄鹌菜属(Youngia Cass.)、福王草属(Prenanthes L.)、翅果菊属(Pterocypsela Shih)、莴苣属(Lactuca L.)、苦荬菜属(Ixeris Cass.)、小苦荬属(Ixeridium (A.Gray) Tzvel.)、沙苦荬属(Chorisis DC.)、黄瓜菜属(Paraixeris Nakai)和蒲公英属(Taraxacum L.)。发现1个新记录种：华蒲公英(Taraxacum borealisinense Kitam.)。     

国产藜科14种植物种皮微形态特征比较研究

应用扫描电子显微镜和光学显微镜观察了中国产藜科(Chenopodiaceae)5族10属14种植物的种子形态和种皮微形态结构。观察结果表明:供试的14种藜科植物的种子多为卵形和圆形,少数为双凸镜形;种皮表面都具有网状纹饰,包括浅网纹、负网纹和穴状网纹。种皮可分为膜质和壳质2类,其中,膜质型种皮表面不光滑,除网状纹饰外,还有瘤状或褶皱状等纹饰,网眼无规则,纹饰排列无序,猪毛菜族(Salsoleae C.A.Mey.)的松叶猪毛菜(Salsola laricifolia Turcz.ex Litv.)及樟味藜族(Camphorosmeae Moq.)的地肤〔Kochia scoparia(L.)Schrad.〕、全翅地肤(K.krylovii Litv.)、黑翅地肤(K.melanoptera Bunge)、雾冰藜〔Bassia dasyphylla(Fisch.et Mey.)O.Kuntze〕、钩刺雾冰藜〔B.hyssopifolia(Pall.)O.Kuntze〕、樟味藜(Camphorosma monspeliaca L.)、兜藜(Panderia turkestanica Iljin.)和棉藜(Kirilowia eriantha Bunge)均属此类;壳质型种皮表面近光滑,网眼规则,排列整齐有序,少数为不规则浅穴,网脊平而不明显或凹陷,滨藜族(Atripliceae C.A.Mey.)的滨藜〔Atriplex patens(Litv.)Iljin.〕、碱蓬族(Suaedeae Reich.)的角果碱蓬〔Suaeda corniculata(C.A.Mey.)Bunge〕、纵翅碱蓬〔S.pterantha(Kar.et Kir.)Bunge〕和异子蓬(Borszczowia aralocaspica Bunge)以及藜族(Chenopodieae C.A.Mey.)的刺藜(Chenopodium aristatum L.)均属此类。各族之间种皮微形态结构也有一定的区别。根据种皮微形态,地肤属(Kochia Roth)与雾冰藜属(Bassia All.)的关系较近,属的界限和部分种类的亲缘关系需要进一步界定。藜科植物种皮表面大部分凹凸不平,这种结构可能与荒漠干旱生境下吸收和保留水分有关。     

拟南芥属的系统位置: 种皮及分子证据

拟南芥属(Arabidopsis (L.) Heynhold)拥有现代分子生物学研究的模式植物拟南芥(A. thaliana (L.) Heynhold), 但其属的系统位置及与近缘属关系争议较大。根据对拟南芥属及其相关属种的种皮微形态观察, 结合测定分析各属种叶绿体DNA的trnL内含子和trnL-F基因间隔区序列, 结果表明, 拟南芥属的近缘属种包括荠属(Capsella Medic.)、亚麻荠属(Camelina (L.)Crantz)、须弥芥属(Crucihimalaya Al-Shehbaz et al.)、无苞芥属(Olimarabidopsis Al-Shehbaz et al.)、旗杆芥属(Turritis L.)、南芥属(Arabis L.)和糖芥属(Erysimum Kitagawa)。     

基于psbA-trnH序列变异分析川明参属亲缘关系及分类地位

采用PCR直接测序法对伞形科(Apiaceae)前胡族(Trib.Peucedaneae)种类川明参(Chuanminshen violaceum Sheh et Shan)、泰山前胡〔Peucedanum wawrae(H.Wolff)Su〕和华中前胡(P.medicum Dunn)以及美味芹族(Trib.Smyrnieae)种类明党参(Changium smyrnioides H.Wolff)、宝兴棱子芹(Pleurospermum davidii Franch.)、丽江棱子芹(P.foetens Franch.)和鸡冠棱子芹(P.cristatum de Boiss.)的叶绿体基因组psbA-trnH片段进行了扩增测序,获得的序列已提交至GenBank,登录号为KF557756—KF557762。结合引自GenBank的前胡族阿魏属(Ferula Linn.)1种、大瓣芹属(Semenovia Regel et Herder)1种、当归属(Angelica Linn.)2种和美味芹族的舟瓣芹属(Sinolimprichtia H.Wolff)1种、羌活属(Notopterygium de Boiss.)1种、瘤果芹属(Trachydium Lindl.)1种以及针果芹族(Trib.Scandicineae)刺果芹〔Turgenia latifolia(Linn.)Hoffm.〕的psbA-trnH片段序列,对各种类的psbA-trnH片段信息进行分析;并以刺果芹为外类群构建了MP、ML和BI系统发育树。结果表明:川明参和明党参的psbA-trnH片段长度均为258 bp、GC含量均为23%,而其他种类的psbA-trnH片段长度为228~405 bp、GC含量为26%~35%;排序后psbA-trnH序列总长度为553 bp(包括空位),其中变异位点237个、信息位点178个。川明参与明党参间的相对遗传距离最小(仅为0.02),而川明参与其他种类间的相对遗传距离为0.10~1.34,且总体上川明参与美味芹族种类的相对遗传距离较小,表明川明参与明党参及美味芹族种类的亲缘关系较近。在3类系统树上,川明参与明党参均聚在一起,并与美味芹族的属种聚为一大支,而远离由前胡族属种构成的另一大支。结合外部形态和果实解剖结构特征,建议将川明参属(Chuanminshen Sheh et Shan)从前胡族中分出并置于美味芹族中,与明党参属(Changium H.Wolff)为姐妹类群。     

4种薯蓣属植物的分子鉴别和亲缘关系研究

为揭示薯蓣属植物的亲缘关系,根据叶绿体matK、rbcL、trnL-F和psbA-trnH序列片段,对小花盾叶薯蓣(Dioscorea sinopatviflora)、盾叶薯蓣(D.zingibiernsis)、黄独(D.bulbifera)和山药(D.polystachya)进行种间分子鉴别研究,并探讨这4个片段在薯蓣属植物系统发育上的意义。结果表明,4种薯蓣属植物共22份材料的matK、rbcL、trnL-F和psbA-trnH序列片段的长度分别为1026~1142 bp、1156~1178 bp、744~822 bp和355~599 bp。用PAUP 4.0b10和贝叶斯推断构建的系统发育树分析表明,云南的黄独与盾叶薯蓣的亲缘关系较近;小花盾叶薯蓣与盾叶薯蓣的亲缘关系很近;而非洲的黄独与云南的黄独的亲缘关系很远。但仅用这4个cpDNA片段还不能完全区分小花盾叶薯蓣和盾叶薯蓣,这说明基于这4个序列片段的系统发育证据与4种薯蓣属植物属内的分类划分并不十分吻合。     

9种观赏草苗期耐盐性评价及NaCl胁迫对芨芨草生长的影响

采用砂培法,对质量浓度0(对照)、5、10、15、20和30 g.L-1NaCl胁迫条件下长芒草(Stipa bungeana Trin.)、丽色画眉草〔Eragrostis spectabilis(Pursh)Steud.〕、狼尾草〔Pennisetum alopecuroides(L.)Spreng〕.、‘小兔子’狼尾草(P.alopecuroides‘Little Bunny’)、拂子茅(Calamagrostis brachytricha Steud.)、野古草〔Arundinella hirta(Thunb.)Tanaka〕、远东芨芨草〔Achnatherum extremiorientale(Hara)Keng〕、芨芨草〔A.splendens(Trin.)Nevski〕和须芒草(Andropogon scoparius Michx.)幼苗的存活率和死亡率进行了分析,并根据植株死亡率、植株存活或出现死亡的NaCl质量浓度和5 g.L-1 NaCl胁迫条件下6个生长指标的聚类分析结果对9种观赏草的耐盐性进行了评价;在此基础上,对NaCl胁迫条件下耐盐性最强的芨芨草生长指标的变化进行了详细分析。结果表明:采用植株死亡率、植株存活或出现死亡的NaCl质量浓度以及生长指标的聚类分析结果都可对供试9种观赏草的耐盐性进行评价,评价结果有一定的差异但表现出相似的趋势;综合3种方法的评价结果,9种观赏草的耐盐性可被分为4个等级:芨芨草最强;狼尾草、‘小兔子’狼尾草、丽色画眉草和长芒草较强;须芒草中等;拂子茅、远东芨芨草和野古草最弱。在NaCl胁迫条件下,芨芨草栽培基质及其渗出液的电导率随NaCl质量浓度的提高逐渐增大;而芨芨草的不同生长指标对NaCl胁迫的响应有差异,其中,NaCl胁迫对芨芨草根长的影响较小;随NaCl质量浓度的提高,芨芨草的苗高、分蘖数、根数、茎叶干质量和根干质量以及叶绿素相对含量均逐渐降低,而根冠比逐渐增大。在NaCl质量浓度高于10 g.L-1条件下,芨芨草幼苗可以存活;而在NaCl质量浓度不高于10 g.L-1条件下,芨芨草幼苗可以维持生长。实验期间的极端高温及极端低温以及栽培基质电导率的变化均对芨芨草幼苗的生长和耐盐性有一定的影响。     

东北产苍术属植物的细胞学研究

报导了中国东北产三种苍术属植物的核型资料,核型公式分别为;北苍术「Ａｔｒａｃｔｙｌｏｄｅｓ ｃｈｉｎｅｎｓｉｓ（ＤＣ．）Ｋｏｉｄｚ．」Ｋ（２ｎ）＝２４＝１２ｍ＋６ｓｍ＋６ｓｔ（４ＳＡＴ）,关苍术「Ａ．ｊａｐｏｎｉｃａ Ｋｏｉｄｚ．」（２ｎ）＝２４＝１４ｍ＋１０ｍ（２ＳＡＴ）朝鲜苍术「Ａ．ｋｏｒｅａｎａ（Ｎａｋａｉ）Ｋｉｔａｍ．」Ｋ（２ｎ）＝２４＝１０ｍ＋１４ｓｍ（２ＳＡＴ）,三者核型对称性均为２Ｂ     

A study on the medicinal plants of the genus Atractylodes

Genus Atractylodes (Fam. Compositae) is the main source of two important Chinese traditional drugs, “Baizhu”and “Cangzhu”, both being long used as a stomachic. After a general survey and taxonomical study, it has been found that “Baizhu” was only derived from A. macrocephala Koidz. (A. ovata auct. Fl. Orient. Asiat. non A. P. DC.), while “Cangzhu” were mainly from A. lancea (Thunb.) DC. and A. lancea DC. var. chinensis Kitam. Comparison of the components in the rhizomes of Chinese Atractylodes has been made by TLC and GLC. The results have shown to be in accordance with their morphological features and pharmaceutical merits. “Baizhu”, A. macrocephala, with its leave pinnately incised, is characterized by the presence of rich atractylon and absence or lack of atractylodin. As for “Cangzhu”, A. lancea and A. lancea var. chinensis with their leave not incised or only lobed, are characterized by high contents of atractylodin, β-eudesmol and hinesol, but poor in atractylon. The above conclusion may be of value to both the classification and utilization of this group of Chinese medicinal plants.     

On the nomenclature of Chinese drug “Cangzhu”

This paper is the documentation of specimens and literature reffered to Atractylodes lancea (Thunb.) DC. etc. As a result, it is found that, in taxonomy of the genus Atractylodes, what represcents by four long-established names, i. e. Atractylodes lancea (Thunb.) DC., A. ovata (Thunb.) DC., A. chinensis (Bunge) Koidz. and A. lyrata Sieb. et Zucc., in fact, is the one and same species. According to the law of priority in nomenclature, the first name should be given to Chinese drug “Cangzhu”, while theother three names have to be treated as its synonyms.     

山东地区药用植物染色体数目的观察(Ⅲ)

本文对17科35种药用植物体细胞染色体作了计数,并就有关的细胞学和分类学问题略作了讨论。     

磁处理白术药液对消化功能影响的药效研究

目的 :研究磁处理白术药液对消化功能影响的药效作用。方法 :用碳末法比较生理盐水、磁处理白术药液与非磁处理白术药液对小鼠肠推进运动的影响 ;用对比法 ,比较磁处理白术药液与正常台氏液及与非磁处理白术药液对离体兔小肠平滑肌收缩运动的影响。结果 :小鼠肠碳末推进率 :与生理盐水组比较 ,磁处理白术药液有促进作用 (p <0 .0 1 ) ,磁处理白术药液与非磁处理白术药液相比差异不显著 ;离体兔小肠平滑肌收缩运动 :与正常台氏液相比 ,磁处理白术药液有促进作用 (p <0 .0 1 ) ,磁处理白术药液与非磁处理白术药液相比 ,有抑制作用。结论 :磁处理白术药液对小鼠肠推进运动功能及对离体兔小肠平滑肌收缩运动有明显作用。     

侵染白术的大豆花叶病毒的全基因序列测定及分析

大豆花叶病毒(soybean mosaic virus,SMV)是影响大豆产量与质量的重要病原体,且自然寄主范围窄。本研究对疑似感病白术叶片进行非序列依赖性扩增(sequence-independent amplification,SIA)检测,结果表明,感病白术为SMV侵染,并命名为SMV-Am。为明确其基因组结构特征及系统进化关系,本研究利用双链RNA(double-stranded RNA,dsRNA)提取、RT-PCR及RACE技术对其进行全基因组扩增,并进行生物信息学分析。结果表明,SMV-Am基因组全长9 587 nt,具有显著的马铃薯Y病毒属(Potyvirus)基因组结构特征;核苷酸与氨基酸序列相似性分析表明,SMV-Am与来自中国的SMV-Liaoning分离物相似性最高,核苷酸与氨基酸序列相似性分别为96.57%和98.86%;系统进化分析也显示与SMV-Liaoning分离物亲缘关系最近;对SMV-Am蛋白序列进一步分析发现,存在与功能位点相关的氨基酸突变,经I-TASSER和PyMOL软件进行蛋白建模分析,发现SMV-Am的P1、HC-Pro、P3、6K2、NIa-Pro、NIb蛋白结构均发生不同程度的变化,且P1蛋白最明显;重组分析结果显示,在SMV-Am基因组的6 560~8 950 nt位置存在重组事件,其主要亲本和次要亲本分别为SMV-XFQ012(GenBank登录号:KP710875.1)和SMV-pCB301-SC15(GenBank登录号:MH919386.1)。这是SMV侵染白术的首次报道,研究结果有望为防范SMV病害在白术上爆发流行以及防治提供科学依据。     

Phylogeny and systematics of the genus Lophozia s. str. (Dumort.) Dumort. (Hepaticae) and related taxa from nuclear ITS1-2 and chloroplast trnL-F sequences

Nuclear ITS1-2 and chloroplast trnL-F were sequenced for 21 taxa of Lophozia s. str., two species of Protolophozia, five species of Schistochilopsis, three species of Barbilophozia and Obtusifolium obtusum. The topologies of phylogenetic trees for 49 taxa constructed from combined sequences of these regions by maximum parsimony, maximum likelihood and Bayesian methods are similar. The species of Lophozia s. str., excluding Lophozia sudetica, combine into two main clades and these contradict subdivisions of Lophozia s. str. based on morphology. The species status of Lophozia lantratoviae is confirmed, whereas Lophozia austro-sibirica is almost identical to Lophozia ventricosa var. guttulata. The genus Schistochilopsis is paraphyletic and occupies basal position to Lophozia s. str., while O. obtusum is clearly separated from Schistochilopsis. A low level of divergence was found between L. sudetica and Protolophozia debiliformes, which are closer to Barbilophozia than to Lophozia s. str. Molecular divergence between geographically remote populations of L. sudetica, Lophozia silvicoloides and Protolophozia debiliformis are low as opposed to those of Lophozia polaris, Lophozia pellucida or Lophozia excisa. Consideration of the trnL intron P8 region indels alone can adequately assign some clades revealed by tree building. A consensus secondary structure of the trnL intron P8 region could not be inferred for taxa studied mainly due to high sequence length diversity originated from deletions.     

Molecular phylogeny of selected Old World Astragalus (Fabaceae): incongruence among chloroplast trnL-F, ndhF and nuclear ribosomal DNA ITS sequences

This study reports maximum parsimony and Bayesian phylogenetic analyses of selected Old World Astragalus using two chloroplast fragments including trnL-F and ndhF and the nuclear ribosomal internal transcribed spacer (nrDNA ITS). A total of 52 taxa including 34 euploid Old World and New World Astragalus , one aneuploid species from the Neo-Astragalus clade as a representative and 14 other Astragalean taxa, plus Cheseneya astragalina and two species of Caragana as outgroups were analyzed for both trnL-F and nrDNA ITS regions. ndhF was analyzed in 30 taxa and the same number for the combination of these three datasets were examined. In general, the trnL-F dataset and the ndhF and nrDNA ITS datasets generated more or less the same clades within Astragalus . However, in the trnL-F and ndhF phylogenies, Astragalus species are not gathered in a single clade, the so-called Astragalus s.s., as indicated by the nrDNA ITS tree. Visual inspection of these three phylogenies revealed that they were inconsistent regarding the position and relationships of Astragalus hemsleyi , A. ophiocarpus , A. annularis–A. epiglottis / Astragalus pelecinus, A. echinatus and A. arizonicus . Incongruence length difference test suggested that the trnL-F , ndhF and nrDNA ITS datasets were incongruent. In spite of this, phylogenetic analyses of the combined datasets as one unit or as three partitions generated trees that were topologically similar as a mix of the cpDNA and the nrDNA ITS trees. However, the combined dataset provided more resolved and statistically supported clades. The recently described A. memoriosus appeared closely related to A. stocksii (both from sect. Caraganella ) based on both trnL-F and nrDNA ITS sequences.     

Chemical constituents of Atractylodes chinensis (DC.) Koidz.

Phytochemical investigations of the rhizomes of Atractylodes chinensis (DC.) Koidz. resulted in the isolation of four sesquiterpenes (1–4), five polyacetylenes (5–9), oleanolic acid (10), and 5-hydromethyl furaldehyde (11). Among them, seven compounds (3, 5–10) were firstly isolated from the A. chinensis, and all have been reported in the genus Atractylodes except compound 10. The chemotaxonomic significance of these compounds was summarized.     

罗布麻及其易混品的DNA分子鉴定

为从分子水平更准确地鉴别罗布麻及其易混品,本研究利用PCR产物直接测序法对罗布麻、大叶白麻和Apoacynum cannabinum的核基因组（rDNA）ITS区与叶绿体基因组（cpDNA）trnL内含子及trnL-F间隔区序列进行测序与比较。结果显示,罗布麻和大叶白麻ITS序列完全一致,与A. cannabinum在ITS1区有13个位点、在ITS2区有10个位点不同;在trnL内含子区及trnL-F间隔区,罗布麻和大叶白麻共有3个位点不同,罗布麻和A. cannabinum间共有23个位点、大叶白麻和A. cannabinum间共有20个位点不同。研究表明,依据rDNA ITS区序列可鉴别A. cannabinum和国产“罗布麻”（罗布麻与大叶白麻）;利用cpDNA trnL内含子及trnL-F间隔区序列可鉴别罗布麻及其相似种。