Conservation ecology of endangered species Paphiopedilum armeniacum (Orchidaceae)

Paphiopedilum armeniacum is an endangered orchid species, endemic to China. During the period of April 2000 to October 2005, 66 observation sites were selected in Luoshapo of Nushan Mountains in Yunnan, China, to carry out the conservation ecological research on P. armeniacum. A total of 443 genets (1302 ramets in total) of P. armeniacum were sampled, their biological characteristics such as reproductive pattern, phenology, and life cycle were observed, and the ecological habits of the species such as the habitat and the structure of communities were studied. Experiments on ex-situ conservation were conducted, and the cloned ramets were replanted to their original habitat after ex-situ reproduction in Shenzhen, Guangdong, China. The relationships between P. armeniacum and climate, vegetation, other environmental factors in the original habitat, and the biological characteristics of asexual offsprings of P. armeniacum, which were replanted to the original habitat after ex-situ cultivation and reproduction, were investigated. The studies show that P. armeniacum in Luoshapo grows very well in secondary shrub boskets or in tussocks on limestone hills. It has both sexual and asexual reproduction. Asexual reproduction serves to complete the sexual reproduction and to extend the lifetime of genets, while it does not reduce sexual reproduction. There are two modes of asexual reproduction-by tillering or by producing rhizomes. The litter of shrub boskets or tussocks provides P. armeniacum with humus, and the rhizome reproduction of P. armeniacum is an adaptation to the litter-covered condition, i.e. to escape from the unfavorable environment. Blooming rate of ramets is 7.39% 1.02%, and fruit set rate from the blooming ramets is 32.23% 12.08%. P. armeniacum is able to invade the moderately destroyed forests and those in early restoring but is unable to grow in large dense forests. P. armeniacum also grows very well in artificial spare woods in Shenzhen and can reproduce many cloned ramets, which can normally bloom and yield fruits after being replanted to the original habitat. The results of this study show that P. armeniacum can be conserved by ex-situ conservation and by replanting the ex-situ reproduced ramets to original habitat. On the basis of the analysis of endangered mechanisms of P. armeniacum, it can be concluded that P. armeniacum has strong capability of both asexual and sexual reproductions, and an emergency mechanism consisted of massive production of rhizomes to cope with damage. Because highly effective pollinating insects that facilitate pollination in P. armeniacum are present in the habitat, flowering ramets produce fruits with large quantity of seeds, many of which in turn grow into new genets that can reproduce many cloned ramets. P. armeniacum makes very effective use of its environment and has distinct characteristics of enduring harsh environmental conditions; therefore, rather than its own inherent biological defects, the main threats facing this species are the destruction of its survival space and the wipe-out collecting of the plants as a result of trading. Based on the analysis mentioned above, certain appropriate strategies have been proposed for the conservation of P. armeniacum.     

云南野生兜兰的引种栽培

1997～ 2 0 0 1年引种云南野生兜兰 1 3种 ,存苗 2万余株 ,在昆明园林植物园 (金殿 )兰圃栽培。具有引种成活率高、容易栽培、观赏价值高的优点 ,在养护管理实践中确立了一套栽培管理技术措施     

麻栗坡兜兰及其近缘植物的分类研究

对麻栗坡兜兰Paphiopedilum malipoense及其近亲种作了对比研究。它们被作为一个种之下的4个变种：麻栗坡兜兰（原变种）Paph.malipoense var.malipoense,浅斑兜兰（变种）var.jackii,钩唇兜兰（变种）var.hiepii;窄瓣兜兰（变种）var.angustatum（变种,新等级）。其鉴别特征已列入表中,还附有一幅图版。     

三种兰科植物的保护遗传学研究初探

采用随机扩增多态 DNA(RAPD)分析研究了中国3种珍稀濒危兰科植物硬叶兜兰(Paphiopedilum micranthum Tang et Wang)、麻栗坡兜兰(P. malipoense S.C.Chen et Tsi)和独花兰(Changnienia amoena Chien)的遗传多样性与群体遗传结构.12个RAPD引物在2种兜兰中共扩增出131条带.对4个硬叶兜兰群体的检测表明其物种水平的多态条带百分率(PPB)为 71.6%,Nei 的基因多样度(h)为 0.217 1,Shannon多样性指数 (I) 为 0.330 1;4个群体的平均多样性水平为 PPB = 45.2%,h = 0.145 7,I = 0.220 4,低于远交兰花的平均水平.在总遗传变异中,群体间遗传变异占20.31%,略高于远交物种的平均水平.在物种水平上,麻栗坡兜兰的PPB为49.5%,h为0.117 4,I为0.176 4,均大大低于硬叶兜兰.对11个独花兰群体采用16个RAPD引物共扩增出119条带.物种水平PPB=76.5%,h=0.194 1,I=0.305 8;在群体水平上,上述3个指标的平均值则分别为37.2%、0.119 7和0.181 0,均低于远交兰花的平均水平.群体间的遗传变异占45.27%,遗传分化明显高于远交物种的平均水平.导致3个物种遗传多样性偏低而群体间遗传分化较高的主要原因在于人为的过度采挖和生境的片断化.研究结果为兰花保护策略和措施的制定提供了理论基础.     

珍稀濒危植物硬叶兜兰的遗传多样性及遗传结构研究

由于人为采集、走私贩卖以及生境的破坏,分布于中国西南石灰岩地区的野生硬叶兜兰居群受到严重的干扰与威胁。为有效地保护这种珍稀野生植物,本研究采用ISSR和SRAP两种分子标记对15个硬叶兜兰野生居群进行遗传多样性及遗传结构的研究。结果表明,硬叶兜兰在物种水平上具有较高的遗传多样性（ISSR：PPB=91．66％,He=0．3839;SRAP：PPB=99．29％,Hc=0．2806）。硬叶兜兰居群间存在一定程度的遗传分化（ISSR：Gs1： 0．2577;SRAP：Gst=0．2383）,可能由于较低的基因流（ISSR：Nm=0．7201;SRAP：Nm=0．7991）所致。UPGMA聚类分析以及主成分分析均把15个居群分成2个主要分支。居群间的地理距离和海拔差距是引起居群遗传分化的自然因素。     

小叶兜兰的种子发育和无菌萌发

为建立小叶兜兰的繁育技术体系,本研究通过无菌播种的方法,辅以TTc生活力测定等方法,比较了小叶兜兰种子在授粉后不同发育时期和培养条件下的萌发率,对小叶兜兰种胚的发育过程进行了显微观察,探讨种胚发育程度与萌发的关系。结果表明,小叶兜兰种胚的发育阶段对萌发的影响最大,授粉后255d的种子萌发率最高（90．71％）,该阶段种子仍呈白色但微干燥,种胚刚发育至球形胚阶段,胚柄尚存。1／4MS和1／2MS为小叶兜兰适宜的基本培养基,添加100mg·L^-1的土豆汁对小叶兜兰的无菌萌发有良好的促进作用。     

巧花兜兰——越南中部兰科一新种

在今年年初的一次访问云南南部的旅行中,笔者曾购得数苗小型的兜兰,据兰商称系采自越南中部,又叫越南黄花仔。此种兜兰的叶甚似小叶兜兰ＰａｐｈｉｏｐｅｄｉｌｕｍｂａｒｂｉｇｅｒｕｍＴａｎｇｅｔＷａｎｇ和海伦兜兰Ｐａｐｈ.ｈｅｌｅｎａｅＡｖｅｒｙａｎｏｖ,但花却有着明显的不同。小叶兜兰的花大,直径达5.5～7ｃｍ;合萼片狭窄,其宽度仅为中萼片的1/2～3/5。海伦兜兰花纯黄色;花瓣线形或宽线形。而此种兜兰的花直径3～3.5ｃｍ,呈淡黄绿色,唇瓣表面、合萼片中脉下半部和花瓣的中脉及中脉上方一侧均有枣红色晕;合萼片宽度为中萼片的4/5;…     

翡翠兜兰,中国云南兰科一新种

对兰科新种翡翠兜兰(Paphiopedilum smaragdinum)作了描述与绘图。新种产云南西部高黎贡山,与虎斑兜兰有亲缘关系,但花淡黄绿色无斑点和斑纹,退化雄蕊矩圆形,先端有短尖,短尖长1～1．5mm,易于区别。     

In vitro propagation of Paphiopedilum orchids

Paphiopedilum is one of the most popular and rare orchid genera. Members of the genus are sold and exhibited as pot plants and cut flowers. Wild populations of Paphiopedilum are under the threat of extinction due to over-collection and loss of suitable habitats. A reduction in their commercial value through large-scale propagation in vitro is an option to reduce pressure from illegal collection, to attempt to meet commercial needs and to re-establish threatened species back into the wild. Although they are commercially propagated via asymbiotic seed germination, Paphiopedilum are considered to be difficult to propagate in vitro, especially by plant regeneration from tissue culture. This review aims to cover the most important aspects and to provide an up-to-date research progress on in vitro propagation of Paphiopedilum and to emphasize the importance of further improving tissue culture protocols for ex vitro-derived explants.     

小叶兜兰的果实生长和雌配子体发育

为了解濒危兰科植物小叶兜兰(Paphiopedilum barbigerum Tang et Wang)胚珠和雌配子体的发育过程,采用常规石蜡切片技术对其果实的生长动态进行了研究。结果表明,授粉后60~75 d的蒴果内种子数量迅速增加,到授粉后120 d时种子充满整个蒴果。授粉后40 d的胎座上分化形成多数由1层表皮细胞包被1列细胞的胚珠原基;授粉后60 d时位于胎座指状结构末端处紧靠表皮细胞下方的孢原细胞分化为大孢子母细胞。之后,大孢子母细胞经过减数分裂和有丝分裂最终形成成熟胚囊;授粉后135 d胚囊发育成熟,附着在胎座上的种子个体分化明显。小叶兜兰胚囊的发育类型为双孢子葱型,胚珠为倒生胚珠,薄珠心,单珠被,成熟胚囊为8核。这为小叶兜兰的生殖生物学及繁殖体系的建立提供理论依据。