利用RAPD技术分析云南兰属部分种的种间关系

采用RAPD技术对云南兰属的11个种和3个变种中的39个样本进行了相似性分析。在相似系数0．58水平上,大花亚属的虎头兰、西藏虎头兰、长叶兰和碧玉兰聚为一支;建兰亚属的寒兰、墨兰、蜜蜂兰、蕙兰、春兰、豆瓣兰、春剑、莲瓣兰和送春聚为另一支,兔耳兰与大花亚属关系更近。春剑和莲瓣兰相似性更高,它们与春兰的关系较远,不支持送春作为蕙兰下的变种。这些结果可为开展兰属育种提供重要参考价值。     

国产七种和一变种兰属植物的核型研究

对国产7种和1变种兰属植物,即邱北冬蕙兰Cymbidium qiubeiense、春兰C. goeringii、春剑 C．longibracteatum、线叶春兰C．serratum、蕙兰C.faberi、送春C.fabri var．szechuanicum、寒兰C．kanran、莎 叶兰C．cyperifolium 的核型进行了研究。具体结果如下：邱北冬蕙兰为2n=40=24m+12sm+4st;蕙兰为2n=40=30m+8sm+2st;送春为2n=40=26m+l0sm+4st;寒兰为2n=40=26m+12sm+2st;莎叶兰为2n=40=24m+12sm+4st;春兰为2n=40=24m+l0sm+4st+2t;春剑为2n=40=24m+l0sm+6st。线叶春兰为2n=40=28m+l0sm+2st。线叶春兰中偶尔发现染色体数有2n=41,43,60,80。     

兰属中若干分类群的订正

基于对许多活植物的观察,对兰科中兰属的若干种类进行了分类学考订,结果承认菅草兰Cymbidium　tortisepalum与峨眉春蕙C. omeiense为独立的种,将春剑C. longibracteatum移至菅草兰之下作为变种,送春C. szechuanicum移至莎叶兰C. cyperifolium之下作为变种。由于春剑、峨眉春蕙和送春的模式标本及引证标本已在30多年前全部丢失,本文指定了新模式。     

兰属植物种间杂交亲和性研究

以10个中国兰品种和8个大花蕙兰品种为材料开展了兰属植物间的杂交试验,比较不同组合的杂交亲和性。结果表明:(1)54个杂交组合,共授粉582朵,获得杂交果实131个,平均结实率为22.51%;中国兰与大花蕙兰正交结实率(23.19%)高于反交结实率(9.27%)。(2)墨兰与大花蕙兰无论正交或反交均可获得较高的结实率,其亲和性高于春兰、建兰与大花蕙兰杂交的亲和性。(3)中国兰种间杂交亲和性较高,3类组合结实率均大于30%,但不同品种间杂交亲和性不一,春兰‘绿萼’,墨兰‘企黑’、‘白墨’,建兰‘金荷’,大花蕙兰‘黄金岁月’比较适宜做母本;春兰‘大富贵’,墨兰‘富贵’、‘白墨’,大花蕙兰‘月影’、‘日本绿’和‘黑珍珠’比较适宜做父本。     

大花蕙兰与兰属植物种间杂交研究

进行了大花蕙兰、墨兰、建兰、纹瓣兰、兔耳兰等兰属植物的种间远缘杂交实验。在52个杂交组合中, 86.5%的杂交组合具明显的子房膨大和果荚发育, 但授粉4个月后未变黄落果的杂交果仅占组合数32.7%, 经胚胎培养获得植株的杂交组合只占组合数的19.2%。大花蕙兰×墨兰与墨兰×大花蕙兰的杂交成功率有较大的差异。     

大花蕙兰与兰属植物种间杂交研究

进行了大花蕙兰、墨兰、建兰、纹瓣兰、兔耳兰等兰属植物的种间远缘杂交实验.在52个杂交组合中,86.5%的杂交组合具明显的子房膨大和果荚发育,但授粉4个月后未变黄落果的杂交果仅占组合数32.7%,经胚胎培养获得植株的杂交组合只占组合数的19.2%.大花蕙兰×墨兰与墨兰×大花蕙兰的杂交成功率有较大的差异.     

基于ISSR标记的96份兰属种质资源遗传多样性分析及指纹图谱构建

为加强我国兰属种质资源的保护利用,本研究通过ISSR分子标记对96份兰属种质进行多样性分析和指纹图谱构建。结果显示,共筛选出11条可扩增清晰条带的多样性引物,在96份材料共检测67条多态性条带,平均多态性条带比例为73.63%;等位基因数（Na）为1.925,有效等位基因数（Ne）为1.450,Nei′s遗传多样性指数（H）为0.277,Shannon多样性指数（I）为0.427,多态性位点百分比（PPL, percentage of polymorphic loci）为92.54%;种群内基因多样性（Hs）为0.1934,基因分化度（Gst）为0.3009,总遗传多样性指数（Ht）为0.2767,种群间的平均基因流（Nm）为1.1619,种群间的两两遗传分化固定指数值范围为0.002～0.527,平均值为0.325。系统聚类结果表明,兰属种群间遗传分化程度高,8个种群可分为3类,春兰和墨兰为一大类,寒兰、春剑、蕙兰、莲瓣兰、建兰为第二类,杂交种独为一类,与其他两类种群之间的遗传距离较大。主坐标分析表明,莲瓣兰和春兰表现出较远的亲缘关系。本研究筛选出6对引物构建了96个品种的指纹图谱二...     

基于nrDNA ITS序列数据的兰属系统发育关系的初步分析

现存的兰属分类系统是基于宏观形态学性状、尤其是花粉块的数目以及唇瓣与蕊柱的愈合程度而建立的。兰属因此而划分为3个亚属：兰亚属（subgenus Cymbidium),大花亚属（subgenus Cyperorchis)和建兰亚属（subgenus Jensoa）。本文运用PCR扩增和直接测序的方法分析兰属（Cymbidium）27种、3个栽培品种以及3个外类群的核DNA ITS区段序列,通过最简约性分析产生的ITS系统发育树表明,兰属的3个亚属均可能为不自然的类群,大花亚属表现为一复系群,兰亚属的冬凤兰（C.dayanum）隐藏于基点;建兰亚属为一并系群,它的成员之一兔耳兰（C.lancifolium）偏离出去而成为兰属一最基部的分支;兰亚属为一复系群,它分为几支而分别与另两个亚属组合在一起,由于兰属ITS序列位点变异率较低,最简约性分析产生的几支主要分支均得不到Bootstrap分析的高度支持,各亚属内组之间的关系也不明确,研究兰属的系统发育关系还需要新的数据。     

基于nrDNAITS序列数据的兰属系统发育关系的初步分析(英)

现存的兰属分类系统是基于宏观形态学性状、尤其是花粉块的数目以及唇瓣与蕊柱的愈合程度而建立的。兰属因此而划分为 3个亚属 :兰亚属 (subgenusCymbidium) ,大花亚属 (subgenusCyperorchis)和建兰亚属 (subgenusJensoa)。本文运用PCR扩增和直接测序的方法分析兰属 (Cymbidium) 2 7种、3个栽培品种以及 3个外类群的核DNAITS区段序列。通过最简约性分析产生的ITS系统发育树表明 ,兰属的 3个亚属均可能为不自然的类群。大花亚属表现为一复系群 ,兰亚属的冬凤兰 (C .dayanum )隐藏于其中 ;建兰亚属为一并系群 ,它的成员之一兔耳兰(C .lancifolium)偏离出去而成为兰属一最基部的分支 ;兰亚属为一复系群 ,它分为几支而分别与另两个亚属组合在一起。由于兰属ITS序列位点变异率较低 ,最简约性分析产生的几支主要分支均得不到Bootstrap分析的高度支持 ,各亚属内组之间的关系也不明确。研究兰属的系统发育关系还需要新的数据。     

基于nrDNA ITS 序列数据的兰属系统发育关系的初步分析

现存的兰属分类系统是基于宏观形态学性状、尤其是花粉块的数目以及唇瓣与蕊柱的愈合程度而建立的.兰属因此而划分为3个亚属:兰亚属 (subgenus Cymbidium),大花亚属(subgenus Cyperorchis) 和建兰亚属 (subgenus Jensoa).本文运用PCR扩增和直接测序的方法分析兰属 (Cymbidium) 27种、3个栽培品种以及3个外类群的核DNA ITS 区段序列.通过最简约性分析产生的ITS系统发育树表明,兰属的3个亚属均可能为不自然的类群.大花亚属表现为一复系群,兰亚属的冬凤兰 (C.dayanum) 隐藏于其中;建兰亚属为一并系群,它的成员之一兔耳兰 (C.lancifolium) 偏离出去而成为兰属一最基部的分支;兰亚属为一复系群,它分为几支而分别与另两个亚属组合在一起.由于兰属ITS序列位点变异率较低,最简约性分析产生的几支主要分支均得不到Bootstrap分析的高度支持,各亚属内组之间的关系也不明确.研究兰属的系统发育关系还需要新的数据.     

兰属Cymbidium植物ISSR遗传多样性分析

将简单重复序列区间扩增多态标记(inter-simple sequence repeats, ISSR)技术应用于16种兰属的遗传多样性分析, 15个引物共扩增出836条带, 其中有227条多态带, 多态百分比为27.2%。UPGMA聚类结果显示: 春兰与春剑的亲缘关系最近, 而兔耳兰与其他15种兰属植物的距离最远。ISSR聚类结果与传统分类结果基本相似, 表明该技术能在分子水平上对传统兰属分类进行必要补充。     

兰属(Cymbidium)5种植物叶结构特征

采用光学显微镜和体视镜,对兰属5种植物（春兰、蕙兰、建兰、寒兰、墨兰）的叶结构特征（叶脉结构、叶表皮结构、解剖结构）进行了观察和测量。结果显示,兰属5种植物的叶脉结构相似,叶表皮结构和解剖结构存在较大差异,其中最主要的差异是皮下纤维束的多少和边缘角质层锯齿形态。依据叶结构特征聚类分析结果,5种兰可分为春兰组、建兰组和墨兰组3组。本研究表明叶结构特征对兰属5种植物的分类具有重要意义。     

Pollination biology of Cymbidium goeringii (Orchidaceae) in China

The genus Cymbidium comprises three subgenera with ca. 50 species. Interactions between pollinators and plants have been studied in the two subgenera Cymbidium and Cyperorchis, but only a few studies in the subgenus Jensoa have been reported. Here we report on the reproductive characteristics of C. goeringii (in sub-genus Jensoa) in three populations in the southwest of Hunan Province, China, during the winter from December 2005 to March 2006. Floral phenological and morphological features, behavior of visitors, the breeding system, and fruit sets under natural conditions were studied. The flowers of C. goeringii were strongly fragrant, but did not present any rewards to the visitors. The flowering period of C. goeringii in the studied populations lasted about 40 days, and most flowers (about 60%) opened within 30 days. Flowers opened immediately when temperature increased obviously and reached to about 16 ℃. The flowering time of the pollinated flowers, unpollinated flow-ers and the flowers with pollinarium removed were similar. Two bees of Apidae, Apis cerana cerana (honeybee) and Anthophora melanognatha, were observed visiting flowers of C. goeringii, but only the honeybees performed as pollinators. The honeybees mostly visited the orchid flowers at 10:00–17:00 on sunny days with temperature above 10 ℃. A total of only thirteen visits were observed during 20 days of observation, indicating pollinators were rare. Honeybees directly landed on the upper surface of the labellum and inserted their heads into the flowers between column and labellum, while the hind legs trod on the surface of the curved downward mid-lobe of label-lum. When a honeybee landed on the labellum of a flower, the labellum moved up and down slightly. After the honeybee entered the flower further, its head might touch the foot of the column. At this time, the body of the honeybee was parallel with the upper surface of the labellum. Then the honeybee used its front legs to scratch on the callus ridges on the upper surface of the labellum, and its hind legs hooked the edges of the side lobe of label-lum, trying to exit forcibly from the flower. During exiting process, sometimes the honeybee’s body was arched owing to tension. In this case, the surface of the scutellum came into close contact with the viscidium and then pollinaria, together with the anther cap, were removed. When this honeybee visited next flower, the pollinaria would be adhered to the stigma when it arched its body during the exiting process. Three plant species flowered synchronously with C. goeringii in the studied areas, but their flowers were different with C. goeringii in color or shape. Because C. goeringii is rewardless to pollinators, the flowers probably attract visitors by olfactory stimu-lus. Breeding system experiments showed no spontaneous autogamy and pollination success relying on pollinators in C. goeringii. Artificial pollination resulted in 90% fruit set by induced autogamy, 100% by pollinating within clone, and 100% by xenogamous pollination, respectively. These results indicate that C. goeringii is highly self-compatible and the fruit production is pollen limited. Because pollinators are essential for fruit set of C. goeringii in natural habitats, protection of wild honeybee populations or apiculture is likely a simple but effective strategy to maintain the orchid populations.     

莲瓣兰形态变异研究

莲瓣兰是分布于云南西北部、四川西南部和台湾的濒危兰花.本研究采用引种不同种群的莲瓣兰栽培在相同环境中的植株为实验材料,测定其形态性状并进行统计计算,分析莲瓣兰种群之间的形态变异以及亲缘关系.巢氏方差分析结果显示,莲瓣兰19个表型性状居群间方差百分比为22.162%,居群内方差百分比为77.838%,莲瓣兰形态变异主要来源于居群内;聚类分析显示保山施甸(SD),大理云龙形态(YL)较为接近,春剑贵州兴隆(XL)居群形态与莲瓣兰各居群差别较大,距离较远.研究结果可为莲瓣兰的类群划分、良种选育及野生资源的保护提供基础数据.     

兰属春剑叶艺突变体叶片结构的研究

以兰属春剑(Cymbidium tortisepalum Fukuy.var.longibracteatum)叶色突变体‘隆昌素’及其亲本为材料,采用石蜡切片、扫描电镜及透射电镜对叶片的组织结构、扫描结构和超微结构进行了观察。结果显示,春剑叶色突变体叶片的表皮厚度和叶肉细胞面积均大于亲本,但导管数量少于亲本;叶片上、下表皮纹饰与亲本具有明显差异;气孔密度和面积均小于亲本,且气孔闭合度较大,占总气孔数的56%;突变体的叶绿体呈高密度的囊泡状结构,内部严重解体,体积较小的嗜锇滴聚集分布,无基粒片层,无淀粉粒。本研究表明经组培诱变的春剑叶色突变体的叶片结构与亲本存在明显差异。