红豆杉科及相关类群rbc L基因PCR-RFLP分析

运用RFLP方法对红豆杉科及相关类群14种植物叶绿体rbcL基因PCR产物进行限制酶酶切分析,共获29个酶切变异位点。采用PHYLIP 软件包对限制位点变异数据进行极大简约法分析得到18个步长为6的最简约树并求得一致树,结果显示：（1）红豆杉科和三尖杉科属单系群;(2) 穗花杉属Amentotaxus以置于红豆杉科内为宜,不支持将穗花杉属独立成科的处理方式;(3)白豆杉应为红豆杉科内一个属Pseudotaxus;(4) 三尖杉属内篦子三尖杉地位特殊,可设篦子三尖杉组;(5) 不赞同将竹柏类从罗汉松属中分离出去成立新科;(6) 红豆杉科、三尖杉科和罗汉松科三者间,前两者的关系更为接近。     

红豆杉科、三尖杉科和罗汉松科植物叶片结构的比较观察

扫描电镜和光镜下的叶表皮特征以及叶片解剖结构的研究结果,支持在红豆杉科内建立白豆杉属的处理方式;赞同在三尖杉属内建立篦子三尖杉组,叶片解剖结构方面,三尖杉科和红豆杉科的穗花杉属的相似之处颇多。而叶表皮形态特征方面又表现为三尖杉科和罗汉松科更加接近。反映了出了红豆杉科、三尖杉科和罗汉松科之间的密切而又复杂的系统关系。     

四种裸子植物叶精油化学成分的研究

采用气相色谱—质谱—计算机联用仪对穗花杉、南方红豆杉、三尖杉和罗汉松叶精油化学成分的研究发现,三尖杉和穗花杉叶精油的组成特点极为相似,相同成分１３个,占各自精油组成的４８．０７％和３３．３２％．三尖杉和南方红豆杉叶精油的相同成分４个,占各自精油组成的１６．１４％和４０．５９％．在一定程度上支持三尖杉科和红豆杉科的亲缘关系接近,红豆杉科可能是通过穗花杉属和三尖杉科相联系的观点．罗汉松和穗花杉叶精油的相同成分４个,占各自叶精油组成的比例为２４．０９％和２０．８２％,比罗汉松和南方红豆杉、三尖杉之间组分的相似性要高．反映出罗汉松科和红豆杉科之间有一定联系,穗花杉属是认识红豆杉科、三尖杉科和罗汉松科之间系统关系的关键属     

部分裸子植物假种皮微形态特征及其分类学意义

应用扫描电镜对红豆杉科、三尖杉科和罗汉松科植物假种皮的微形态特征进行了研究,结果表明：可以把红豆杉科植物分为两类,即具规则型网状纹饰的红豆杉属和具无规则型网状纹饰的白豆杉属、穗花杉属和榧树属。在后三属中,穗花杉属的穗花杉和白豆杉属的纹饰特征更为相近,同属厚网脊亚型。而云南穗花杉和榧树属的特征相近,属薄网脊亚型。假种皮表面纹饰特征显示不宜将红豆杉（T．chinensis）再分为两个独立的种,认为设立红豆杉和变种南方红豆杉（T．chinensisvar.mairei）的处理方式更加合理。三尖杉科（属）蓖子三尖杉（C．oliveri）假种皮纹饰的网脊、网底分化程度、细胞大小、单位面积细胞数和本届的其它植物差异很大,为建立蓖子三尖杉组提供了进一步的佐证。假种皮的微形态特征还支持把宽叶粗榧（C.sinensisvar.latifolia）上升为种C．latifolia的观点。罗汉松科假种皮纹饰类型的观察结果表明罗汉松属和陆均松属是自然分类群。尽管罗汉松属竹相组假种皮纹饰的网脊变异范围相对较大,但是变异范围仍在属内,不支持将其独立为新科的观点。假种皮的表面纹饰特征还表明三尖杉科和红豆杉科之间的联系密切。     

三尖杉属(科)植物的双黄酮成分及其化学分类的研究

本文采用薄层层析和提取、分离、光谱分析等方法分析研究了三尖杉属植物叶的双黄酮类化合物,根据获得的化学分析结果,我们同意：(1)将三尖杉属分为二个组：篦子三尖杉组Sect．1 Pectinaea,只包括蓖子三尖杉一种;三尖杉组Sect．2 Cephalotaxus,包括本属所有其它种。(2)支持三尖杉科只包括三尖杉属1属,不支持将红豆杉科中的穗花杉属(Amentotaxus)归入三尖杉科的主张。(3) 认为三尖杉科与南洋杉科、松科、杉科和柏科的亲缘关系较远,而与红豆杉科及罗汉松科的亲缘关系密切。     

红豆杉科及三尖杉科的分子系统发育——兼论竹柏属的系统位置

测定了红豆杉科、三尖杉科、罗汉松科及松科共9属13种植物的matK基因序列,发现3′端的单碱基插入造成mat k基因在这些类群间有较大的长度变异(1488∽1548bp),其它插入或缺失的长度为3、6或9bp(白豆杉中有一个长为27bp的缺失),且绝大多数插入或缺失事件具有系统发育信息。matK基因中,密码子第1、2及3位的变异率较为相近,平均同义替代率约为平均非同义替代率的2倍。以松科2属3种植物为外类群,运用PAUP软件进行的系统发育分析仅得到一棵最简约树(步长为895,CI=0．850,RI=0．876),该简约树的各分支均得到bootstrap分析的极强支持。结果表明：红豆杉科与三尖杉科均为单系群,二者互为姐妹群;白豆杉属与穗花杉属为红豆杉科中两个很自然的属,白豆杉属为红豆杉属的姐妹群,穗花杉属为榧树属的姐妹群。竹柏属与罗汉松属聚为一支,二者间的亲缘关系得到boot-strap分析的100％支持,从分支分类学及遗传距离角度,作者不赞同将竹柏属提升为科。     

裸子植物的生化系统学(三)——从种子蛋白多肽和针叶过氧化物酶探讨红豆杉科的系统位置

SDS聚丙烯酰胺凝胶电泳揭示红豆杉属和白豆杉的种子蛋白主要多肽的分子量为31、 22和20千道尔顿(K)。穗花杉的多肽谱与上述两属相差较大,没有22K多肽,代之以一个33K 主要多肽。榧树属和穗花杉有很多共同的多肽,但没有44K这个红豆杉科多数种类共有的中 等含量的多肽,并出现一个36K主要多肽。三尖杉属一些种的多肽谱十分接近红豆杉属,竹 柏的种子蛋白与上述分类群也有一定程度的近似。 红豆杉科各属之间针叶过氧化物酶谱差别很大,但三尖杉属一些种与红豆杉属却有些类 似。 两种蛋白质资料一致说明红豆杉科内的进化趋势是从红豆杉属、经白豆杉属和穗花杉属 至榧树属。红豆杉属和三尖杉属之间蛋白质的近似,说明红豆杉科和三尖杉科之间的关系,可 能通过红豆杉属相联系的,述资料也说明红豆杉科应置于松柏目之下。     

部分裸子植物假种皮微形态特征及共分类学意义

应用扫描电镜对红豆杉科、三尖杉科和罗汉松科植物假种皮的微形态特征进行了研究,结果表明：可以把红豆杉科植物分为两类,既具规则型网状纹饰的经豆杉属和具夫规则型网状纹饰的白豆杉属、穗花杉属和榧树属。在后三属中,穗花杉属的穗花帮和白豆杉属的纹饰特征更为相近,同属厚网脊亚型。而云南穗花杉和榧树属的特征相近,属薄网脊亚型。假种皮表面纹饰特征显示不宜将红豆杉（Ｔ．ｃｈｉｎｅｎｓｉｓ）再分为两个独立的种,认为设立     

部分裸子植物茎次生韧皮部和木材的比较解剖

茎次生韧皮部和木材的比较解剖支持红豆杉科、三尖杉科和罗汉松科自然类群、不主张建立穗花杉科和竹柏科     

穗花杉传粉和受精作用的研究

穗花杉于5月25日至6月15日(1980—1982)传粉,成熟花粉包括2个细胞。7月20日至29日受精。两者相隔约2个月。在花粉管和卵细胞中,分别具有许多特殊的核仁状颗粒,它是穗花杉属的明显特点之一。从传粉、受精和胚胎发育的特点来看,穗花杉和澳洲红豆杉属在红豆杉科中可能占居比较原始的位置,它们的演化顺序是:穗花杉属、澳洲红豆杉属、红豆杉属、白豆杉属和榧树属。而且,三尖杉科可能通过穗花杉属过渡到红豆杉科。     

部分裸子植物叶片总蛋白分析

采用ＳＤＳ－ ＰＡＧＥ 技术, 分析了红豆杉科（Ｔａｘａｃｅａｅ） 植物南方红豆杉（ Ｔａｘｕｓ ｃｈｉｎｅｎｓｉｓｖａｒ－ ｍａｉｒｅｉ （Ｌｅｍｅｅ ｅｔ Ｌｅｖｌ－） Ｃｈｅｎｇ ｅｔ Ｌ－Ｋ－Ｆｕ） 、穗花杉（ Ａｍｅｎｔｏｔａｘｕｓ ａｒｇｏｔａｅｎｉａ （Ｈａｎｃｅ） Ｐｉｌ ｇｅｒ） 、云南穗花杉（ Ａ－ ｙｕｎｎａｎｅｎｓｉｓ Ｌｉ） 、白豆杉（ Ｐｓｅｕｄｏｔａｘｕｓｃｈｉｅｎｉｉ（Ｃｈｅｎｇ） Ｃｈｅｎｇ） 以及三尖杉科（Ｃｅｐｈａｌｏｔａｘａｃｅａｅ） 、植物三尖杉（ Ｃｅｐｈａｌｏｔａｘｕｓ ｆｏｒｔｕｎｅｉ Ｈｏｏｋ－ｆ－） 、粗榧（ Ｃ－ｓｉｎｅｎｓｉｓ （Ｒｅｈｄ－ｅｔＷｉｌｓ－） Ｌｉ） 、海南粗榧（ Ｃ－ｈａｉｎａｎｅｎｓｉｓ Ｌｉ） 、篦子三尖杉（ Ｃ－ｏｌｉｖｅｒｉ Ｍａｓｔ－） 和罗汉松科（Ｐｏｄｏｃａｒｐａｃｅａｅ） 、植 物罗汉松 （ Ｐｏｄｏｃａｒｐｕｓ ｍａｃｒｏｐｈｙｌｌｕｓ （ Ｔｈｕｎｂ－ ） Ｄ－Ｄｏｎ） 、鸡毛 松（ Ｐ－ｉｍｂｒｉｃａｔｕｓ Ｂｌ－） 、竹柏（ Ｐ－ ｎａｇｉ（Ｔｈｕｎｂ－） Ｚｏｌｌ） 、陆均松（ Ｄａｃｒｙｄｉｕｍ ｐｉｅｒｒｅｉ Ｈｉｃｋｅｌ） 共１２ 种植物的叶片蛋白, 在蛋白质水平上采用     

Studies on Pollen Morphology and Exine Ultrastructure in Cephalotaxaceae

The pollen morphology of Cephalotaxaceae was examined with LM, SEM and TEM. Pollen grains in this family are spheroidal or subspheroidal, rounded in polar view, but usually wrinkled with irregular shape. Pollen size is 22.6- 34.8 μm in diameter. There is a distinct or indistinct tenuity on distal face. The tenuity occasionally slightly rises above the outline of pollen grains, but often sukened. Exine rather thin, 1—1.5μm thick, layers obscure, surface of pollen grains is nearly psilate or weakly granulate. Under SEM exine is covered with fine and dense granules, and sparse Ubisch bodies are found on the granular layer. The Ubisch bodies are provided with minute gemmate processes on the surface. Acorrding to our observation under TEM, exine consists of ectexine and lamellate endexine, with the former divided into outer ectexine of granules densely arranged and inner ectexine of loosely arranged microgranules. Granules of the outer ectexine are relatively thick, and connected with each other, forming a structure just like tectum or separate from each other. Microgranules of the inner ectexine are distinct or indistinct. Endexine is provided with 5- 7 lamellae. As far as information of pollen morphology is concerned, Cephalotaxus oliveri is rather special in the Cephalotaxaceae. First, the tenuity in pollen grains occupies one half of the distal part, much larger than that of the other species in the family. Second, the ectexine in Cephalotaxus oliveri may be divided into two distinct layers, outer ectexine and inner ectexine. The former is made of a layer of sporopollenin masses, which are connected with each other to form tectumlike structure, while the latter consists of a layer of loosely arranged granules or small segments of sporopollenin. The inner ectexine is different from that of other species by having a thicker layer of sporopollenin granules. Based on these two features, we support the division of Cephalotaxus into two Sections, Sect. Pectinatae and Sect. Cephalotaxus. Pollen grains of Cephalotaxaceae are similar to those of the Taxaceae in having spheroidal shape and the tenuity on its distal face. These characteristics strengthen the evidence for a close relationship between the Cephalotaxaceae and Taxaceae. Although pollen grains of the Cephalotaxaceae and Taxaceae are similar in some characteristics, they have obvious differences in , for example, size of tenuity, the fine structure of Ulbisch bodies and of the outer and inner ectexine. On the basis of pollen morphology, the present author considers theCephalotaxaceae slightly more primitive than the Taxaceae.     

PCR-RFLP Analysis on Phylogeny of Allium

The molecular phylogeny of the genus Allium which includes eighteen species selected from nine sections was investigated through PCR-RFLP analysis of two chloroplast DNA fragments, including trak gene (approximately 2 520 bp) and rpL16 gene (approximately 1 230 bp). Digestion of these two fragments by 26 restriction endonucleases yielded 303 polymorphic recognition sites, of which 163 were informative sites. The restriction site data matrix were analyzed following the parsimonious Wagner and parsimonious Dollo principle of PAUP ( version 3.1.1 ). Topologically, the most parsimonious Wagner tree constructed by branch-and-bound and heuristic search was similar to the most parsimonious Dollo tree. All the taxa of Allium form a monophyletic group, and five sections based on morphological characters were supported strongly by this result. Sect. Augninum is closely related to Sect. Bromatorrhiza, Sect. Molium is closely related to Sect. Caloscordum. Their reliability was farther confirmed by the bootstrap test very well. In morphology, A. pallasii is closely related to A. caeruleum and belongs to Sect. Haplostemen, A. cepa is closely related to A. galanthum and belongs to Sect. Cepa. But evidence from cladistics of parsimonious tree based on 163 informative sites of PCR-RFLPs showed that they are neither confined to a monophyletic group nor to a natural taxon.     

Chloroplast matK gene phylogeny of Taxaceae and Cephalotaxaceae,with additional reference to the systematic position of Nageia

Reported in the present paper is a robust chloroplast matK gene phylogeny of Taxaceae, Cephalotaxaceae and Podocarpaceae represented by 10 species of seven genera, with three species of the Pinaceae as outgroups. The matk length of the 13 species ranges from 1488 bp to 1548 bp, which results from indels, in particular, 1-bp(base pair) insertion near the 3’ end of the gene in some groups. A 27 bp deletion was found at the nucleotide position 213 from the 5’ end of the matk gene of Pseudotaxus chienii. The aligned sequences used in PAUP and MEGA analyses were 1568 bp and 1494 bp respectively. In the matK gene, the rates of variation at the first, second and third codon positions are similar although the mean frequency of synonymous substitution is approximately twice as high as that of nonsynonymous substitution. Branch-and-Bound search found only one most parsimonious tree (tree length = 895, CI = 0.850, RI = 0. 876), in which all clades were strongly supported by bootstrap test. According to the tree, Taxaceae and Cephalotaxaceae are monophyletic groups, and the sister group relationship between the two families was confirmed. Taxus is closely related to Pseudotaxus while Torreya is the sister group of Amentotaxus. In addition, the close relationship between Nageia and Podocarpus was resolved. The present study supports the generic status of Pseudotaxus and Amentotaxus in point of cladistic analysis and genetic distance, but contra-dicts the establishment of the family Nageiaceae.     

葱属系统发育的PCR—RFLP分析初报

葱属（Ａｌｉｕｍ）为广义百合科（Ｌｉｌｉａｃｅａｅ）葱族（Ａｌｉｅａｅ）的一个重要类群。该属种类丰富、分布极广，关于它的系统发育和进化问题，目前尚存在不少分歧［１］。我国有葱属１１０种（含变种和引进外来种），主要分布于东北、华北、西北和西南地区。我们...     

Phylogeny of taxaceae and cephalotaxaceae genera inferred from chloroplast matK gene and nuclear rDNA ITS region

Phylogeny of the Taxaceae genera and the monotypic family Cephalotaxaceae has been extraordinarily controversial. In this paper chloroplast matK genes and nuclear ITS sequences were determined for all six genera of the two families and representatives of other conifer families. Analysis using either the nonsynonymous sites or the deduced amino acid sequences of matK genes strongly indicates that taxad genera and Cephalotaxaceae are monophyletic, with the Taxodiaceae/Cupressaceae clade as their sister group. Cephalotaxus is basal to the taxad genera, among which two clades, Torreya/Amentotaxus and Taxus/Pseudotaxus/Austrotaxus, are resolved. They correspond to Janchen's two tribes, Torreyeae and Taxeae. In Taxeae, Austrotaxus is the first to branch off. Analyses of the nuclear ITS sequence data corroborated the topology of the matK gene tree. These results refute the views that Cephalotaxaceae has no alliance with Taxaceae and that Austrotaxus and Amentotaxus should be excluded from the Taxaceae. We estimated the divergence time between the Taxodiaceae/Cupressaceae and the Cephalotaxaceae/Taxaceae clades to be 192-230 Myr ago and the divergence time between taxads and Cephalotaxus to be 149-179 Myr ago. Soon after the latter divergence event, within 6-8 Myr, the two taxad tribes originated. In conclusion, our data do not support Florin's claim that taxads could be traced to Devonian psilophytes (359-395 Myr ago).     

Phylogeny of Taxaceae and Cephalotaxaceae Genera Inferred from Chloroplast matK Gene and Nuclear rDNA ITS Region

Phylogeny of the Taxaceae genera and the monotypic family Cephalotaxaceae has been extraordinarily controversial. In this paper chloroplast matK genes and nuclear ITS sequences were determined for all six genera of the two families and representatives of other conifer families. Analysis using either the nonsynonymous sites or the deduced amino acid sequences of matK genes strongly indicates that taxad genera and Cephalotaxaceae are monophyletic, with the Taxodiaceae/Cupressaceae clade as their sister group. Cephalotaxus is basal to the taxad genera, among which two clades, Torreya/Amentotaxus and Taxus/Pseudotaxus/Austrotaxus, are resolved. They correspond to Janchen's two tribes, Torreyeae and Taxeae. In Taxeae, Austrotaxus is the first to branch off. Analyses of the nuclear ITS sequence data corroborated the topology of the matK gene tree. These results refute the views that Cephalotaxaceae has no alliance with Taxaceae and that Austrotaxus and Amentotaxus should be excluded from the Taxaceae. We estimated the divergence time between the Taxodiaceae/Cupressaceae and the Cephalotaxaceae/Taxaceae clades to be 192–230 Myr ago and the divergence time between taxads and Cephalotaxus to be 149–179 Myr ago. Soon after the latter divergence event, within 6–8 Myr, the two taxad tribes originated. In conclusion, our data do not support Florin's claim that taxads could be traced to Devonian psilophytes (359–395 Myr ago).