The Effect of Seed Cryopreservation on the Development of Protocorms by the Hybrid Orchid Bratonia

The aim of this work was to study the influence of storage in liquid nitrogen on the viability of seeds of the hybrid orchid Bratonia and further development of its protocorms in vitro. Seeds were frozen in ampoules by direct immersion in liquid nitrogen and stored in the cryobank for a month. The germination rates of cryopreserved and control (nonfrozen) seeds did not differ and remained as high as 100%. The protocorms derived were cultured on the agar-solidified Murashige and Skoog nutrient medium (MS), half-strength MS and Knop media and also in Morel liquid medium. During the first 45 days of culturing, protocorms derived from cryopreserved seeds grew faster than control protocorms on the MS and half-strength MS media but, at longer culturing (496 days), the size of control protocorms was significantly larger. After 639 days of culturing, there was no difference in the amount of perished, budding, and newly formed protocorms obtained from cryopreserved and control seeds, except half-strength MS medium where the number of budding protocorms in the case of cryopreserved seeds was a little greater than in the control treatment. After seed cryopreservation, the frequency of budding and newly formed protocorms was greater on the agarized MS and in liquid Morel media. Cryopreservation had little effect on the subsequent growth of protocorms in vitro. The preferable nutrient media for culturing the protocorms have been suggested.     

Mecardonia tenella (Plantaginaceae) Attracts Oil-, Perfume-, and Pollen-Gathering Bees in Southern Brazil

Floral characteristics often indicate the pollinators' functional group visiting the plant and the pollination syndromes associated with them. This idea has been challenged in the past decades due to increasing evidence that most plants, including those exhibiting floral syndromes, are visited by large arrays of species that differ in their effectiveness as pollinators. Our study focuses on Mecardonia tenella (Plantaginaceae) from the Araucaria forest of southern Brazil, which exhibits characteristics of the oil flower pollination syndrome. However, it is visited by three types of functional groups of bees: male orchid bees, oil-collecting bees, and pollen-seeking bees. The relative contribution of each functional group to the plant's reproductive success was estimated based on their pollen load, visitation frequency, and morphology. We assessed resources, phenology, and breeding system of M. tenella . Our results indicate that flowers lack nectar, but volatiles, lipids, and pollen are resources that can be gathered by visitors. This combination of floral traits and visitors' assemblage makes M. tenella a challenge to the concept of pollination syndromes. Our findings indicate that the current interactions may not reflect the circumstances under which some floral traits of this plant were selected.     

Asymbiotic seed germination and in vitro seedling development of <Emphasis Type="Italic">Cyrtopodium punctatum</Emphasis>: a propagation protocol for an endangered Florida native orchid

Cyrtopodium punctatum Lindley is an endangered epiphytic orchid restricted in the United States to southern Florida. Due to its ornamental value, the species was extensively collected from the wild during the past 100 years. Today, only a few plants remain in protected areas. As part of a conservation plan for the species, procedures for asymbiotic seed germination were developed. Five asymbiotic orchid seed germination media (PhytoTechnology Orchid Seed Sowing Medium, Knudson C, Malmgren Modified Terrestrial Orchid Medium, Vacin &; Went Modified Orchid Medium, and ½-strengh Murashige &; Skoog) were examined for their effectiveness in promoting seed germination and protocorm development under a 16/8 h L/D photoperiod and dark (0/24 h L/D). The influence of photoperiod on growth and development was also examined. Seeds were germinated under a 16/8 h, 12/12 h, 8/16 h L/D photoperiod, at 25 ± 3°C and allowed to develop in vitro for 10 weeks. After 10 weeks, developing seedlings were transferred to Sigma Phytatrays and returned to their assigned photoperiod treatments for continued seedling development for an additional 15 weeks. Highest germination occurred in 0/24 h L/D on PhytoTechnology Orchid Seed Sowing Medium and seedlings displayed more advanced development when cultured under 16/8 h L/D photoperiod after 15 weeks in Phytatrays. Thirty-five week old seedlings potted in coconut husk growing medium exhibited 90% survival following 5 weeks acclimatization to greenhouse conditions. This asymbiotic seed germination protocol for C. punctatum will facilitate future reintroduction projects involving this endangered species.     

Growth and viability of mycorrhizal extraradical mycelia associated with three temperate orchid species

Growth and enzymatic activities of extraradical mycelia (ERM) of native mycorrhizal symbionts associated with three orchid species, Dactylorhiza fuchsii, D. majalis and Platanthera bifolia, were studied. ERM extracted from the mycorrhizosphere of these species showed features typical for fungi that form orchid mycorrhiza. In the first pot experiment, three different treatments were applied on tubers of D. fuchsii transplanted from a natural site: control (no specific treatment), reinoculated (surface-sterilized tubers reinoculated with mycorrhizal fungi-colonised roots), and benomyl (nonsterilized tubers treated with fungicide). However, no significant differences in ERM growth and intensity of root mycorrhizal colonisation at harvest were observed among these treatments. ERM associated with reinoculated D. fuchsii plants showed significantly higher alkaline phosphatase (ALP) enzymatic activity at week 36 than at week 24, but no differences were observed for NADH diaphorase activity. Benomyl application significantly reduced ALP activity in comparison with reinoculated plants at week 36. In the second experiment, plants of all three species were either untreated (control), or repeatedly treated with benomyl. Similarly to the results of the first experiment, benomyl application did not reduce the ERM growth of mycorrhizal symbionts associated with D. majalis and D. fuchsii. The low ERM growth associated with benomyl-treated P. bifolia was probably caused by poor root system development in this treatment. Significantly higher mycorrhizal colonisation was found for D. fuchsii compared to P. bifolia in control treatments at the end of cultivation. The ERM of native symbionts of the three orchid species studied seemed to have a different growth pattern over time and responded differently to fungicide application.     

Interaction between a dark septate endophytic isolate from Dendrobium sp. and roots of D. nobile seedlings

Interactions between an isolate of dark septate endophytas (DSE) and roots of Dendroblum nobile Lindl.seedlings are reported in this paper.The isolate was obtained from orchid mycorrhizas on Dendrobium sp.in subtropical forest.The fungus formed typical orchid mycorrhiza in aseptic co-culture with D.nobile seedlings on modified Murashige-Skoog (MMS)medium.Anatomic observations of the infected roots showed that the DSE hyphae invaded the velamen layer,passed through passage cells in exodermis,entered the cortex cells,and then formed fungal pelotons of orchid mycorrhiza.D.nobile seedlings' plant height,stem diameter,new roots number and biomass were greatly enhanced by inoculating the fungus to seedlings.The fungus was identified as Leptodontidium by sequencing the polymerase chain reaction-amplified rDNA ITSt-5.8S-ITS2 (internal transcribed spacer (ITS)) regions and comparison with similar taxa.     

基于标本和分布信息评估中国虾脊兰属 植物的濒危状况

兰科植物是生物多样性保护的旗舰类群之一, 但如何客观准确地评估兰科植物的濒危状况, 一直是生物多样性保护中急需解决的问题。本文以中国虾脊兰属(Calanthe)植物为例, 探讨了基于馆藏标本和野外分布信息进行兰科植物红色名录濒危等级评估的利弊。我们的研究表明, 馆藏标本能够比较客观地反映物种地理分布信息和相应的野外居群状况, 但对于物种分布的历史变化和受威胁因素反映不足; 而野外工作对于物种的实际分布状况和受威胁因素等的反映比较客观, 但能发现的物种和居群数量有限, 部分极小种群物种在短时间野外工作中很难发现。我们的研究认为, 有2种虾脊兰属植物应被评估为野外灭绝(EW), 16种应被评估为极度濒危(CR)。可见, 中国虾脊兰属植物濒危的状况远比前人的评估结果要严重得多。     

Effects of distance from models on the fitness of floral mimics

• Rewardless plants can attract pollinators by mimicking floral traits of rewarding heterospecific plants. This should result in the pollination success of floral mimics being dependent on the relative abundance of their models, as pollinator abundance and conditioning on model signals should be higher in the vicinity of the models. However, the attraction of pollinators to signals of the models may be partially innate, such that spatial isolation of mimics from model species may not strongly affect pollination success of mimics.
• We tested whether pollination rates and fruit set of the rewardless orchid Disa pulchra were influenced by proximity and abundance of its rewarding model species, Watsonia lepida.
• Pollination success of the orchid increased with proximity to the model species, while fruit set of the orchid increased with local abundance of the model species. Orchids that were experimentally translocated outside the model population experienced reduced pollinaria removal and increased pollinator‐mediated self‐pollination.
• These results confirm predictions that the pollination success of floral mimics should be dependent on the proximity and abundance of model taxa, and thus highlight the importance of ecological facilitation among species involved in mimicry systems.

     

In vitro propagation of temperate Australian terrestrial orchids: revisiting asymbiotic compared with symbiotic germination

Using a common temperate herbaceous terrestrial orchid from Australia (Caladenia latifolia) this study investigated 19 asymbiotic media variations comprising four commonly used basal media [half‐strength Murashige and Skoog (½MS), Knudson C (KC), Pa5, and Vacin and Went (VW), with combinations of the plant growth regulators 6‐benzylaminopurine (BA) and α‐naphthalene acetic acid (NAA) or coconut water (CW) and compared their performance with germination on a standard symbiotic germination medium, oatmeal agar (OMA). Percentage germination of seeds every 2 weeks for a total of 8 weeks (five replicates per treatment), time to germination, and growth and development phases in seedlings were recorded. ½MS with 5% (v/v) fresh CW delivered 93% germination, with seedling vigour and development indistinguishable from OMA (95% germination). The same protocol was applied to a further ten species (including the endangered Caladenia huegelii), demonstrating high asymbiotic germination performance (60–93%) across a wide phylogenetic range of terrestrial orchid species. © 2014 The Linnean Society of London, Botanical Journal of the Linnean Society, 2014, 176 , 556–566.     

达摩兰组织培养适宜培养基的研究

以达摩兰的原种苗茎段为外植体进行组织培养研究,筛选出最适培养基(1)原球茎诱导:MS+6-BA 0.5 mg/L+NAA 1.0 mg/L+香蕉肉250 g/L;(2)丛生芽诱导:1/2 MS+6-BA 3 mg/L+NAA 2.0 mg/L+AC 2.5 g/L+蔗糖3%+香蕉肉200 g/L;(3)芽增殖:1/2 MS+6-BA 3 mg/L+NAA 2.0 mg/L+AC 2.5 g/L+蔗糖3%+香蕉肉150 g/L;(4)根诱导:1/2 MS+6-BA 0.3 mg/L+IBA 0.5 mg/L+NAA 0.5 mg/L+AC 2.5 g/L+蔗糖3%。采用珍珠岩、细石与树皮的混合物为培养基质,移栽成活率达95%以上。     

四川黄龙沟森林植被中兰科植物群落优势种种间联结和相关分析

四川黄龙沟内分布着十分丰富的兰科植物(19属30余种),并且部分种类在沟内形成优势群落。目前尚不清楚在群落中这些兰科植物种之间作用关系。运用种间关联分析和相关分析对黄龙沟森林植被兰科植物群落中的24个优势种的种间关系进行了研究。两种分析方法得到的结果相近,表明黄龙沟优势兰科植物可以分为两组。一组包括无苞杓兰(Cypripedium bardolphianum)、黄花杓兰(C. flavum)、二叶红门兰(Orchis diantha)、广布红门兰(Orchis chusua)、少花虾脊兰(Calanthe delavayi)和西藏杓兰(C. tibeticum),主要分布在光线充足但又具有一定遮荫条件的环境中;另一组包括筒距兰(Tipularia szechuanica)、沼兰(Malaxis monophyllos)、珊瑚兰(Corallorhiza trifida)、小斑叶兰(Goodyera repens)、布袋兰(Calypso bulbosa)、小花舌唇兰(Platanthera minutiflora)和小叶对叶兰(Listera smithii),它们主要分布于荫蔽的环境中。这些兰科植物在组内大多呈现显著的正相关关系,组间大多呈现显著的负相关关系,说明黄龙沟兰科植物在资源利用方式上可能产生了分化。