The role of epiphytism in architecture and evolutionary constraint within mycorrhizal networks of tropical orchids

Characterizing the architecture of bipartite networks is increasingly used as a framework to study biotic interactions within their ecological context and to assess the extent to which evolutionary constraint shape them. Orchid mycorrhizal symbioses are particularly interesting as they are viewed as more beneficial for plants than for fungi, a situation expected to result in an asymmetry of biological constraint. This study addressed the architecture and phylogenetic constraint in these associations in tropical context. We identified a bipartite network including 73 orchid species and 95 taxonomic units of mycorrhizal fungi across the natural habitats of Reunion Island. Unlike some recent evidence for nestedness in mycorrhizal symbioses, we found a highly modular architecture that largely reflected an ecological barrier between epiphytic and terrestrial subnetworks. By testing for phylogenetic signal, the overall signal was stronger for both partners in the epiphytic subnetwork. Moreover, in the subnetwork of epiphytic angraecoid orchids, the signal in orchid phylogeny was stronger than the signal in fungal phylogeny. Epiphytic associations are therefore more conservative and may co‐evolve more than terrestrial ones. We suggest that such tighter phylogenetic specialization may have been driven by stressful life conditions in the epiphytic niches. In addition to paralleling recent insights into mycorrhizal networks, this study furthermore provides support for epiphytism as a major factor affecting ecological assemblage and evolutionary constraint in tropical mycorrhizal symbioses.     

Effects of local density and flower colour polymorphism on pollination and reproduction in the rewardless orchid Dactylorhiza sambucina (L.) Soò

The rewardless orchid Dactylorhiza sambucina shows a stable flower colour polymorphism, with both yellow- and red-flowered morphs growing sympatrically. Pollination biology and breeding system were investigated to examine the effects of density of plants, colour polymorphism, inflorescence dimension, and flower position within inflorescence on male and female reproductive success in three natural populations of D. sambucina. There were significant differences among sites in the number of pollinia removed and in fruit set per inflorescence. Number of removed pollinia and capsule production in D. sambucina were independent from flower and inflorescence size or flower position. As a whole, the red morphs showed the highest number of capsules produced, while the yellow morphs had the greatest male success. The relative male and female reproductive success were independent from plant density but were significantly correlated with the yellow morph frequency at the population level. Overall, our findings show that the contribution to the total reproductive success deriving from the two colour morphs does not conform with the predictions of negative frequency-dependent selection.     

鹤顶兰胚囊发育过程中微管变化的共焦显微镜观察

光镜的观察确定了鹤顶兰（Ｐｈａｉｕｓ ｔａｎｋｅｒｖｉｌｌｉａｅ （Ａｉｔｏｎ） Ｂｌ．）胚囊发育属单孢子蓼型。应用免疫荧光标记技术及共焦镜观察了胚囊发育过程中微管分布的变化。当孢原细胞初形成时,细胞内的微管呈网状分布。之后,孢原细胞体积增大发育为大孢子母细胞。大孢子母细胞延长,进入减数分裂Ⅰ。微管由分裂前的网状分布变为辐射状排列。二分体的两个细胞内的微管分布一样,呈辐射状。四分体的近珠孔端的３ 个大孢子解体,细胞内的微管消失。靠合点端的功能大孢子内有许多微管呈网状分布。当功能大孢子进入第一次有丝分裂时,细胞内的微管由网状变为辐射状,从核膜伸展至周质。再经两次有丝分裂形成八核胚囊。在核分裂之前微管一般是呈网状分布并紧包围着核。在分裂期间二核和四核胚囊都呈极性现象,微管系统也呈极性分布。微管在八核胚囊内的分布变化情形特别复杂。首先,八核分别作不同程度的移动,其中两个核移向胚囊中央,珠孔端和合点端的３ 个核分别互相靠拢,形成３ 个区,即中央区、反足区和卵器区。胚囊未形成区时,８ 个核都被网状分布的微管包围着。当胚囊明显分成区时,反足区内的微管仍作网状分布。中央区的微管分布则趋疏松,形成篮形结构,包围着液泡和两个极核。在     

Fruit set in a deceptive orchid: The effect of flowering phenology,display size,and local floral abundance

We investigated the effect of flowering time, display size, and local floral density on fruit set in Tolumnia variegata, a pollination-limited orchid that offers no reward to its pollinator(s). During 1990, natural variation in flowering time, display size, and fruit set were monitored in 508 plants at one locality in Puerto Rico. The following season, orchid floral abundance per host tree (Randia aculeata) was manipulated to investigate its effect on fruit set. Four floral abundance treatments were established (700, 500, 300, and 100), each replicated four times. Flowering time was the most important trait affecting fruit set. The proportion of plants setting at least one fruit was significantly high early and late in the season, but low during the flowering peak. Thus, strong disruptive selection differential on flowering phenology was found. Display size had little effect on fruit set. A weak, but significant disruptive selection differential on display size was found. Orchid floral abundance per host tree had a significant effect on fruit set. Early in the season, T. variegata flowers with intermediate number of conspecific flowers exhibited a greater probability of setting fruit than those in host trees with fewer or more flowers. Our results show that flowering phenology may be evolutionarily unstable, possibly a consequence of the deception pollination system. Furthermore, a deception strategy would be relatively unsuccessful in populations where plants are found in either very dense or sparse patches.     

Pollination of the orchidEpipactis thunbergii by syrphid flies (Diptera: Syrphidae)

The behavior of visitors to the flowers of the orchidEpipactis thunbergii was studied, with special attention to the role of the epichile in the pollination process. Only four species of syrphid flies legitimately pollinated the flower, among whichSphaerophoria macrogaster was regarded as the most effective pollinator. The movable epichile, possessing a furrow at its base, played a critical role in the pollination process: it threw the syrphid fly onto the stigmatic surface when both sides of the basal slanting surface of the furrow were presumably pressed in the direction of the hypochile by the fore (and middle) legs of the retreating syrphid fly. At this moment, the fly received a set of pollinia on the thorax.     

Use of VIRAZOLE® to eradicate odontoglossum ringspot virus from in vitro cultures of Cymbidium Sw.

Odontoglossum Ringspot Virus has been eradicated from Cymbidium Sw. through chemotherapy based on incorporation of ribavirin (VIRAZOLE^®) into the in vitro culture medium of protocorms. Applications of the virustatic agent for several consecutive subcultures freed protocorms of the virus. Acclimated plantlets regenerated from those protocorms are healthy as determinated by enzyme-linked immunosorbent assay (ELISA). No resurgence of virus occurred over a period of 5 years. Besides, trueness to type was total at flowering level and the batch grown was perfectly homogeneous.To secure fast and effective eradication of the virus during the consecutive subcultures of protocorms with ribavirin, three factors proved to be of prime importance: accurate isolation of new growths from initial tissues, VIRAZOLE^® concentration and frequency of transplanting in new media.     

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。     

Foliar regeneration of the endangered Red Vanda,Renanthera imschootiana Rolfe (Orchidaceae)

An in vitro method was developed to regenerate large numbers of phenotypically uniform plants from the basal parts of the leaves of flowering plants of Renanthera imschootiana Rolfe. Differentiation of up to 10 shoot buds free of callus and protocorm-like bodies occurred in 10–12 weeks from the base of a single leaf implanted in Mitra et al. (1976) medium supplemented with 2% sucrose, 2 g l^-1 peptone, 44.4 M benzyladenine (BA) and 10.7 M naphthaleneacetic acid (NAA). Subculture of the tissues in medium enriched with 10% coconut water and 35 g l^-1 ripe banana pulp resulted in the production of highest average number of 40 shoots in 12 weeks. No difference in the regeneration potential was observed among the three young leaves while mature leaves did not respond. All the leaves of the regenerated shoots were easily recultured to increase shoot multiplication. Shoots readily formed roots on transfer to a medium containing 4.4 M BA, 10.7 M NAA and 1% activated charcoal. All regenerated plants examined were normal diploids with 2n=38. Foliar meristem culture appears to have great potential for ex situ conservation and propagation of this extremely endangered orchid.Abbreviations BA benzyladenine - NAA naphthaleneacetic acid     

Fungal coil formation ofRhizoctonia repens in seedlings ofGaleola septentrionalis (orchidaceae)

Protocorms or protocorms with roots of an achlorophyllous orchidGaleola septentrionalis were inoculated with isolates ofRhizoctonia repens, R. solani, andRhizoctonia spp. The seedlings were infected with eight of twelve isolates ofR. repens. Fungal coils were formed in the cells, which was suggestive of a symbiotic association. The other isolates caused soft rot or no infection to the protocorms or the protocorms with a root. Contribution No. 97, Laboratories of Plant Pathology and Mycology, Institute of Agriculture and Forestry, University of Tsukuba, Tsukuba, Ibaraki 305, Japan.     

菌根真菌与兰科植物互作的组学研究进展

兰科植物因其具有丰富的物种多样性和重要的社会经济价值,多年来一直是植物学及生态学界的重点研究对象。菌根真菌对兰科植物的种子萌发、营养吸收和种群动态等多个方面都具有十分重要的作用,因而近年来受到越来越多的关注。探究菌根真菌与兰科植物互作的内在机制是目前兰科菌根研究的一大热点领域,同时也为兰科植物野生资源保护和种群恢复提供了许多新方法与新思路。随着新一代测序技术以及多种组学数据库的发展与建立,菌根真菌与兰科植物互作机制研究已然进入了一个全新的时代。纵观近年来的研究,通过基因组学、转录组学、蛋白质组学和代谢组学等相关技术,人们对兰科菌根共生过程中的种子萌发、营养物质转运、信号转导、宿主免疫和逆境抗性等方面的内在机制有了一定的了解。本文对近十年来国内外采用组学技术研究菌根真菌与兰科植物互作机制方面的研究成果进行了总结和梳理,并对未来该领域的研究进行了展望。