附生植物石斛的株丛生长及营养繁殖特征

石斛(Dendrobium nobile Lindl.)为兰科多年生附生性草本植物,特化的假鳞茎是其营养贮藏器官,通过假鳞茎可实现克隆生长。该研究以野外调查发现的石斛株丛为研究材料,比较不同等级株丛假鳞茎合轴生长和高位腋芽的差异,分析高位株丛的定植方式,探讨石斛株丛生长及营养繁殖对附生环境的适应策略。结果显示:(1)石斛株丛的生长和扩大通过合轴生长的营养繁殖方式进行,假鳞茎基部具有2~3个储备芽,每年萌发1~2个新芽,随着生长年限的增加,形成大小不一的株丛。(2)株丛具有典型的高位腋芽营养繁殖特性,且主要形成于假鳞茎密集和老根密布的大株丛。(3)高位株丛母茎一端附着于附主树种上,在母茎软化和高位株丛的重力作用下,缩短了高位株丛与附主的距离,使其根系能够触及附主,完成高位株丛的定植。研究表明,附生植物石斛通过假鳞茎合轴生长的营养繁殖方式来增强并延续株丛寿命,高位腋芽的频发是株丛假鳞茎对拥挤等逆境的响应,高位株丛的定植依赖于母茎,这也是石斛对附生环境的一种生态适应策略。     

铁皮石斛种植技术体系

铁皮石斛是我国濒临灭绝的珍贵中草药,关于种植铁皮石斛的系统技术研究还鲜见报道。本文研究了铁皮石斛苗期栽培基质的筛选、不同附主树种和种植高度、不同林分郁闭度对林下铁皮石斛附生生长的影响以及林下种植石斛对附主树种生长的影响等。结果表明:(1)综合瓶苗移栽成活率、株高、茎长、茎粗以及生物量等生长指标,铁皮石斛的苗期栽培的理想基质为等体积混合基质(松树皮+花生壳+马占相思树皮);(2)不同树种和附生部位对铁皮石斛的株高生长、茎粗、生物量等影响差异显著,但两者没有交互作用。附生60 d时,郁闭度大的杨桃最优,120 d开始,檀香迅速表现出绝对优势,各树种均为中部(1.2~1.5 m)最适附生,马占相思附生的石斛生物量最大。檀香、马占相思和降香黄檀的综合评价显著优于龙眼和杨桃;(3)铁皮石斛附树种植在林分郁闭度为30%~50%,光照入射强度为空地的62.5%左右对其生长最为有利;(4)附树种植铁皮石斛可以显著增加附主胸径。综合认为,在华南地区种植铁皮石斛,选择混合基质(松树皮+花生壳+马占相思树皮)为筛苗基质,附生林分郁闭度为30%~50%的檀香或降香黄檀树干中部是值得推广"以林养林"的复合经营模式。     

珍稀齿瓣石斛的生物学特性及其野生资源保护

阐述了齿瓣石斛的生物学特性及资源情况。为附生草本植物,主要分布于热带、亚热带温暖湿润地区,高温环境下茎条细长,低温（1℃以下）容易受冻,为冬季落叶型石斛。自然条件下,齿瓣石斛繁殖方式包括有性繁殖和营养繁殖,有性繁殖对环境要求严格,果实种子量大,为明显的“以量取胜”繁殖,是扩大种群分布范围的主要途径;营养繁殖分合轴生长和高芽繁殖,合轴生长形成并壮大株丛,有春秋两个萌芽峰期,而高芽是扦插、压条繁殖的保证。野生资源过度采摘、生境破坏与植树造林、繁殖特性缺陷、人工种植产业发展缓慢、供需矛盾突出等是齿瓣石斛野生资源濒危的主要原因。应制定相关政策,加强野生资源保护,推进齿瓣石斛工厂化育苗,并积极开展种子自然繁殖技术研究,降低种苗培育成本,加快规模化种植产业发展,实现齿瓣石斛资源的可持续利用。     

兰科石斛属植物菌根真菌研究进展

石斛属(Dendrobium)隶属于兰科(Orchidaceae)树兰亚科(Epidendroideae)石斛兰族(Dendrobiinae),是兰科最大的属之一,终生附生于树上或岩石上。石斛属很多种类具有很高的药用价值与观赏价值。由于人为过度采挖和野生生境的破坏,使得野生石斛资源濒临灭绝。石斛属植物为典型的兰科菌根植物,在自然条件下需要与真菌共生,才能完成生活史。菌根真菌对于石斛属植物的种子萌发和植株生长具有重要的作用。对石斛属植物菌根的形成、菌根真菌的作用、菌根真菌多样性及菌根技术在石斛属植物中的应用做了评述,并对今后的研究内容和重点提出了一些思路。     

兰科植物种子共生萌发真菌多样性及共生萌发机制研究进展

自然环境下,兰科植物种子细小无胚乳,需要和适宜的真菌共生才能萌发,因而与真菌有天然的共生关系。自身繁殖率低加之近年来栖息地环境破坏导致兰科植物资源更加濒危,而通过筛选适合的真菌进行种子的共生萌发可以有效地实现兰科植物的种质保育及濒危种类野生居群的生态恢复。本文对地生型、附生型以及腐生型等兰科植物已发现的萌发真菌的多样性进行了系统地梳理,发现担子菌门的胶膜菌科、角担菌科以及蜡壳耳目真菌为已报道共生萌发真菌的主要类群;同时对兰科植物种子的共生萌发机制,包括形态学机制、营养机制和分子机制等方面的相关研究进行了归纳论述,但是当前关于兰科植物和真菌互作机制方面的研究还相对较少,许多问题需要进一步明确。本文对共生萌发真菌在兰科植物保育和繁育中的应用以及共生萌发机制的研究等方面具有一定的参考价值。     

共生真菌对兰科植物种间杂交后代种子萌发的效应

石斛属(Dendrobium)植物在种子共生萌发过程中与真菌有着较为专一的共生关系,为探讨这种共生关系在种间杂交后代上的进化和适应,深入理解兰科植物和真菌共生关系的形成机制,该研究利用能有效促进铁皮石斛(Dendrobium officinale)和齿瓣石斛(D.devonianum)种子萌发形成幼苗,并具有较强专一性的胶膜菌属(Tulasnella)真菌SSCDO-5和瘤菌根菌属(Epulorhiza)真菌FDd1,开展真菌对铁皮石斛和D. tortile种间杂交种子萌发效应的研究。结果表明,在真菌与种子共生培养68天时,SSCDO-5菌株和FDd1菌株都能有效地促进杂交种子形成原球茎和幼苗,两个接菌处理之间无显著差异,来源于铁皮石斛的SSCDO-5菌株不但没有表现出优势,反而在杂交石斛幼苗形成率上低于来源于齿瓣石斛的FDd1菌株(SSCDO-5:(22.13±6.62)%; FDd1:(29.53±5.51)%); SSCDO-5菌株和铁皮石斛在幼苗形成和发育阶段的共生专一性并没有在杂交后代上得到遗传或表现,或者说是杂交打破了这种专一性的共生关系,使得杂交后代能够和不同的真菌建立新的共生关系。该结果不支持关于共生真菌专一性是石斛属植物杂交后代形成的重要限制因素的假设,推测石斛属植物在幼苗分化和发育阶段与真菌这种专一性的共生关系是在适应特定生态环境的过程中形成和建立的。     

共生真菌对兰科植物种间杂交后代种子萌发的效应

石斛属(Dendrobium)植物在种子共生萌发过程中与真菌有着较为专一的共生关系, 为探讨这种共生关系在种间杂交后代上的进化和适应, 深入理解兰科植物和真菌共生关系的形成机制, 该研究利用能有效促进铁皮石斛(Dendrobium officinale)和齿瓣石斛(D. devonianum)种子萌发形成幼苗, 并具有较强专一性的胶膜菌属(Tulasnella)真菌SSCDO-5和瘤菌根菌属(Epulorhiza)真菌FDd1, 开展真菌对铁皮石斛和D. tortile种间杂交种子萌发效应的研究。结果表明, 在真菌与种子共生培养68天时, SSCDO-5菌株和FDd1菌株都能有效地促进杂交种子形成原球茎和幼苗, 两个接菌处理之间无显著差异, 来源于铁皮石斛的SSCDO-5菌株不但没有表现出优势, 反而在杂交石斛幼苗形成率上低于来源于齿瓣石斛的FDd1菌株(SSCDO-5: (22.13 ± 6.62)%; FDd1: (29.53 ± 5.51)%); SSCDO-5菌株和铁皮石斛在幼苗形成和发育阶段的共生专一性并没有在杂交后代上得到遗传或表现, 或者说是杂交打破了这种专一性的共生关系, 使得杂交后代能够和不同的真菌建立新的共生关系。该结果不支持关于共生真菌专一性是石斛属植物杂交后代形成的重要限制因素的假设, 推测石斛属植物在幼苗分化和发育阶段与真菌这种专一性的共生关系是在适应特定生态环境的过程中形成和建立的。     

铁皮石斛种子的室内共生萌发

由于人为的采挖和原生境的破坏,使得铁皮石斛野生资源已濒临灭绝。因此,保护铁皮石斛野生资源及其生境,加快其野生资源的繁殖显得非常重要。以铁皮石斛种子(TTC染色显示种子生活力为77.65%)为材料,与分离自2种野生兰科植物根部的4株共生真菌 (C20来自铁皮石斛,L12,L24b 和 L28来自美花石斛)在燕麦培养基上进行共生萌发。经过18周的共生培养,4株真菌均不同程度地促进了铁皮石斛种子的萌发,其中菌株L24b (Epulorhiza)和L28 (Epulorhiza)显著提高了种子的萌发率,分别比对照高出26.51%和12.20%,但未形成幼苗,只是处于原生分生组织阶段(阶段3);菌株C20(Epulorhiza)和L12 (Alternaria) 虽没有显著提高种子的萌发率,但对原球茎的发育和幼苗的生长有明显的促进作用;而对照的种子仍然处于膨大转绿期,即萌发阶段(阶段2)。同时发现,TTC染色显示的铁皮石斛种子生活力要高于种子共生萌发的萌发率(除了菌株L24b)。研究结果表明:种子生活力染色检测的活力值只代表种子所具有的潜在的萌发能力,而不能代表实际的萌发率。在异地条件下,铁皮石斛与共生真菌间没有严格的专一性,可以与瘤菌根菌属、链格孢属真菌形成共生关系。菌株C20和L12能促进萌发后的种子进一步分化成幼苗。这两个菌株为铁皮石斛的人工优质栽培和野外种群的建立提供了可能。     

贵州北盘江喀斯特地区6种兰科植物结构及其生态适应性

【目的】分析兰科植物营养器官结构对喀斯特生境的适应特性和不同兰科植物响应生境异质的生存策略,为兰科植物研究、保护与发展提供理论支持和参考依据。【方法】该研究以贵州北盘江喀斯特地区6种不同生活型兰科植物的叶、假鳞茎、根为研究对象。应用石蜡切片法,进行结构观察并测量相关指标,通过描述和方差分析、相关性分析和主成分分析。【结果】结果表明：(1)不同兰科植物结构特征间存在显著差异,丘北冬蕙兰、莎叶兰、兔耳兰和单叶石仙桃均为等面叶,叶片较薄;栗鳞贝母兰与梳帽卷瓣兰叶片为异面叶,有栅栏组织和海绵组织的分化,叶片较厚。(2)假鳞茎主要由表皮、基本组织和维管束组成,地生兰基本组织细胞较小,维管束密度大;附生兰基本组织细胞较大,储水结构较发达。(3)地生兰根被和皮层较厚,根被细胞排列紧密,皮层细胞小层数多;附生兰根被与皮层较薄,皮层细胞较大。(4)叶片下表皮厚度与叶片厚度、叶维管束直径、叶肉厚度、叶脉厚度呈极显著(?<0.01)正相关,叶肉厚度与下角质层厚度、茎维管束直径呈显著(?<0.05)负相关;茎直径和表皮厚度、茎维管束数量、茎横截面积呈极显著正相关,茎维管束直径占比与茎直径、表皮厚度、茎维管束数量、茎横截面积均呈极显著负相关;根的各指标间呈极显著正相关,根直径、根被厚度和皮层厚度与根维管束直径占比呈极显著负相关。【结论】兰科植物根部结构具有相对稳定的趋同适应性,叶片和假鳞茎结构间相互促进协调,是兰科植物响应干旱喀斯特环境的重要结构。 关键词 生态适应性;兰科植物;结构特征;喀斯特地区     

细叶楠种子休眠与萌发对低温层积的生理响应

细叶楠(Phoebe hui Cheng ex Yang)是我国特有的珍贵树种,种子具有生理休眠特性。本文以细叶楠种子为实验材料,研究不同低温层积处理时间(0、20、40、60 d)对细叶楠种子发芽率、营养物质(淀粉、可溶性糖、可溶性蛋白)、过氧化物酶(POD)以及内源激素脱落酸(ABA)、赤霉素(GA₃)、生长素(IAA)、玉米素核苷(ZR)的影响,分析细叶楠种子萌发与其生理指标的相关性。结果显示：(1)低温层积可以有效打破细叶楠种子的休眠,且处理60 d效果最佳。(2)在低温层积过程,细叶楠种子萌发时间缩短,发芽率提高;种子内淀粉含量降低,可溶性糖和可溶性蛋白含量升高;POD活性上升;ABA含量下降,GA₃、IAA含量增加,而ZR含量先上升后下降。(3)细叶楠种子休眠的解除与萌发与其体内营养物质含量、POD活性以及内源激素含量的变化密切相关。     

Mast fruiting in a hawkmoth-pollinated orchid Habenaria glaucifolia: an 8-year survey

Aims The hypothesis of predator satiation has been proposed to explain mast fruiting in various flowering plants. It considers that the simultaneous production of large numbers of seeds by a plant population reduces the risk of seed predation for each individual. Orchids produce huge numbers of seeds per fruit and rarely experience seed predation. It remains unclear which factors may affect fluctuating fruit production in orchids, which generally suffer a widespread pollen limitation. To explore the temporal pattern of fruiting and potential factors related to fluctuation in fruit production, we investigated reproductive success of a long-spurred orchid (Habenaria glaucifolia) in an alpine meadow with thousands of individuals over 8 years.Methods To estimate reproductive success, pollinator observation was conducted by day and at night, and pollinia removal and receipt were recorded in the field population for 8 years. To examine whether fruit set and seed set are pollen limited, we conducted supplementary pollination experiments and compared fruit set, seed set and pollinia movement of open-pollinated flowers from 2011 to 2013. We measured lengths of spurs and pollinator proboscises, and nectar volume and concentration, to identify potential pollinators.Important findings Hawkmoths were seen to be effective pollinators for H. glaucifolia in 3 years, whereas in the remaining 5 years no pollinators were observed, and consequently pollinia were rarely transferred. Numerous pollinia movements were observed in 2012, 2013 and 2014 (pollinia removal: 48, 59 and 85%; pollinia receipt 51, 70 and 80%), and correspondingly fruit set was significantly higher in 2012 and 2013 (59 and 46%) than in 2011 (25%). It was fruit set, rather than seed set, that was pollen limited in this orchid in the 3 years, in that supplementary pollination increased fruit set but did not increase seed set per fruit compared to natural. Three species of hawkmoths had proboscis lengths that matched the spur length of H. glaucifolia. Fruit set in this long spurred orchid depends on the activity of long-tongued hawkmoths, resulting in significant temporal variation in fruit production. Mast fruiting in this alpine orchid could be attributed to a 'sit and wait' strategy, awaiting an abundance of effective pollinators.     

Impacts of Host Trees and Sowing Conditions on Germination Success and a Simple Ex Situ Approach to Generate Symbiotic Seedlings of a Rare Epiphytic Orchid Endemic to Hainan Island,China

For species relying on seeds for population regeneration, knowledge on seed germination behaviors in relation to environmental factors is critical in designing species recovery strategy. Dendrobium sinense is an orchid endemic to Hainan Island of China and listed as Endangered by the IUCN Redlist. It reproduces primarily via seeds in its natural habitat. However, how germination is impacted by major environmental factors is poorly known. This study aimed to examine germination success of D. sinense seeds using two approaches, i.e. in situ and ex situ, using host tree barks as germination media. The latter was intended to generate symbiotic seedlings in a simple and economic approach which could be used for reintroduction efforts. In addition, three factors of in situ symbiotic seed germination success, including different sowing time, location (distance from an adult plant), and host tree were investigated. Our results showed that seeds sown ex situ and in situ in July had the highest rates of germination. Seed germination was significantly higher ex situ using bark as medium than in situ. Seeds sown directly on Rhododendron moulmainense, the most common host tree, with naturally occurring conspecific orchids had the highest rate of germination. In contrast, ex situ seeds sown on the bark of Cyclobalanopsis blakeii, a non-host species, had the highest rate of seed germination. In situ a positive correlation was found between the seed germination rate and the distance of the seeds from the adult D. sinense. Based on these results, it is likely seedling recruitments are determined by host tree species, the presence of and the distance from an adult conspecific orchid, which imply the importance of the mycorrhizal fungi, which were not reported here. This study provided important information on the optimal environmental conditions for population augmentation and reintroduction, which can be used as part of the species recovery strategy.     

Seed-deposition and recruitment patterns of Clusia species in a disturbed tropical montane forest in Bolivia

Spatial patterns of seed dispersal and recruitment of fleshy-fruited plants in tropical forests are supposed to be driven by the activity of animal seed dispersers, but the spatial patterns of seed dispersal, seedlings and saplings have rarely been analyzed simultaneously. We studied seed deposition and recruitment patterns of three Clusia species in a tropical montane forest of the Bolivian Andes and tested whether these patterns changed between habitat types (forest edge vs. forest interior), distance to the fruiting tree and consecutive recruitment stages of the seedlings. We recorded the number of seeds deposited in seed traps to assess the local seed-deposition pattern and the abundance and distribution of seedlings and saplings to evaluate the spatial pattern of recruitment. More seeds were removed and deposited at the forest edge than in the interior. The number of deposited seeds decreased with distance from the fruiting tree and was spatially clustered in both habitat types. The density of 1-yr-old seedlings and saplings was higher at forest edges, whereas the density of 2-yr-old seedlings was similar in both habitat types. While seedlings were almost randomly distributed, seeds and saplings were spatially clustered in both habitat types. Our findings demonstrate systematic changes in spatial patterns of recruits across the plant regeneration cycle and suggest that the differential effects of biotic and abiotic factors determine plant recruitment at the edges and in the interior of tropical montane forests. These differences in the spatial distribution of individuals across recruitment stages may have strong effects on plant community dynamics and influence plant species coexistence in disturbed tropical forests.     

Endophytic fungi from Pecteilis susannae (L.) Rafin (Orchidaceae), a threatened terrestrial orchid in Thailand

Eight endophytic fungi were isolated from roots of the threatened terrestrial orchid, Pecteilis susannae (L.) Rafin. Phylogenetic analysis based on an alignment of internal transcribed spacer regions of nuclear rDNA indicated that seven isolates belonged to the genus Epulorhiza and one to Fusarium. All fungal isolates were cultured with orchid seeds collected from three field sites near Doi Suthep-Pui National Park, Chiang Mai, Thailand. Seed germination and protocorm development were evaluated up to 70 days after sowing. Percent symbiotic seed germination was highest (86.2%) when seeds were cultured with Epulorhiza (CMU-Aug 013). The protocorm development was the most advanced up to stage 2, continued embryo enlargement, or rupture of the testa, and the highest percentage was 17.8% when seeds were cultured with Epulorhiza (CMU-Aug 007). Without fungi, seed germination and protocorm development were 62.1% and 11.1%, respectively. The dependency of P. susannae on fungal symbionts for early seedling development is yet to be determined. Optimizing seed germination and seedling fitness will assist the conservation of this threatened orchid in Thailand.     

Effect of seed depth on slug damage to winter wheat

In a field experiment drilled at two depths on three dates in autumn 1988, with or without methiocarb pellets broadcast on the soil surface immediately after drilling, 26% of seeds of winter wheat sown at c. 20 mm depth were killed by slugs compared with only 9% of seeds sown at c. 40 mm. The protection from slug damage provided by this additional 20 mm of depth was comparable with that provided by methiocarb pellets. The effects of seed depth and pellet application did not interact and were consistent on all drilling dates. Thus, fewest seeds and seedlings were killed where methiocarb pellets were broadcast on a seed-bed with seeds sown at 40 mm depth. Intermediate damage was recorded where seeds were sown at 40 mm depth without pellets, or where pellets were broadcast on seeds sown at 20 mm depth. Most seeds and seedlings were killed where seeds were sown at 20 mm depth without pellets. Sublethal damage to seedlings was not affected by sowing depth but was reduced where pellets were broadcast immediately after sowing.     

Evaluation of the effect of three growth regulators in the germination of <Emphasis Type="Italic">comparettia falcata</Emphasis> seeds under <Emphasis Type="Italic">in vitro</Emphasis> conditions

Summary The effect of 3-indoleacetic acid (IAA), 6-furfurylaminopurine (kinetin), and gibberellic acid (GA₃) on germination of the orchid Comparettia falcata was evaluated in a factorial experiment (4×4×4) with Murashige and Skoog (1962) basal medium. It was established that seeds of this orchid could be maintained under aseptic conditions as long as the necessary nutrients and appropriate concentrations of growth regulators were provided. Of the three growth regulators used, IAA significantly decreased seed germination of Comparettia falcata. There was a synergistic effect in the kinetin:GA₃ combination that produced a positive response in both percentage seed germination and development of seedlings. This study describes a single medium-based protocol able to achieve more than 160000 seedlings within 21 wk, starting from a single capsule, sufficient for both large-scale propagation and in vitro conservation of this threatened orchid.     

Are plant populations seed‐limited? A review of seed sowing experiments

We define seed limitation to be an increase in population size following seed addition. Here, we briefly consider how theoretical models deal with seed limitation and how seed sowing experiments can be used to unravel the extent of seed limitation in natural systems. We review two types of seed addition experiments: seed augmentation studies where seeds are added to existing populations; and seed introductions where seeds are sown in unoccupied sites. Overall, approximately 50% of seed augmentation experiments show evidence of seed limitation. These studies show that seed limitation tends to occur more commonly in early successional habitats and in early successional species. Most of the studies have concentrated on simply categorising populations as seed- or microsite-limited, but we believe that seed sowing experiments could be used to reveal much more about community structure, and we discuss possible future directions. In 53% of introduction studies (where seeds were sown at sites from which the species was known to be absent) the introduced species was recorded in at least one of the experimental sites following sowing. However, of the subset of studies where both seedlings and adult plants were recorded, 64% of sites contained seedlings while only 23% contained adults. This implies that, for many species, conditions for establishment are more stringent than conditions for germination. The successful establishment of plants in unoccupied patches indicates the potential for immigration to enhance local diversity (the spatial mass effect). Few studies continued monitoring for long enough to determine whether or not self-sustaining populations were successfully established, and no study attempted to link introduction sites to a putative natural source of propagules, or considered the dynamics of the metapopulation as a whole.     

种子特征和播种深度对辽东栎种子萌发和幼苗生长的影响

该文在玻璃温室内的遮阴环境下,采用盆播方法研究了辽东栎种子特征(大小和种皮)和播种深度(0、3、6、10 cm)对种子萌发和幼苗生长的影响。结果表明:(1)辽东栎大种子萌发率、萌发速率、萌发值和萌发指数在所有播种深度均高于小种子。其中非去皮种子萌发率、萌发速率和萌发指数在0 cm播种深度的不同大小种子间差异显著,去皮种子所有萌发参数在6和10 cm播种深度的不同大小种子间均差异显著;去皮可促进大种子萌发,但抑制小种子萌发;不同大小种子所有萌发参数均在0 cm播种深度最大,在10cm播种深度最小。(2)不论有无种皮,大种子萌发幼苗的叶片数、单株叶面积、总干物质质量和根冠比在所有播种深度均大于小种子萌发幼苗;去皮种子萌发幼苗的株高、基径、叶片数、单株叶面积和总干物质质量在所有播种深度均不同程度地小于非去皮种子萌发幼苗,但前者根冠比在0、3、6 cm播种深度大于后者;随着播种深度增大,幼苗株高、叶片数、单株叶面积、总干物质质量和根冠比等生长参数均呈减小趋势,但基径随播种深度增大而增大。     

Symbiotic seed germination of Habenaria macroceratitis (Orchidaceae), a rare Florida terrestrial orchid

The rapid loss of native orchid habitat throughout ecologically important areas (e.g., Florida) has prompted researchers to develop appropriate plans for the propagation and reintroduction of many native orchid species. Ideally, symbiotic orchid seed germination methods are utilized in the production of orchid seedlings to be used in plant reintroduction programs. In the current study we (1) describe an efficient symbiotic seed germination protocol to germinate seeds of the rare sub-tropical terrestrial orchid Habenaria macroceratitis; (2) discuss the in vitro fungal specificity demonstrated by this species; and (3) describe the effects of three photoperiods (0/24 h, 16/8 h, 24/0 h L/D) on in vitro symbiotic seed germination of H. macroceratitis. Six fungal mycobionts were isolated from both vegetative and flowering plants of H. macroceratitis from two geographically distinct sites. Symbiotic seed germination percent was highest (65.7%) and protocorm development was most advanced (Stage 2) when seeds were cultured with fungal mycobiont Hmac-310. Seeds of H. macroceratitis demonstrated a degree of specificity toward fungal mycobionts isolated from plants originating from the same site where seed was collected. Continual darkness (0/24 h L/D) inhibited initial seed germination (Stage 1; 17.1%), but stimulated subsequent protocorm development (Stage 2; 53.5%). These findings will aid in developing an efficient symbiotic seed germination protocol for the conservation of this rare Florida terrestrial orchid, and may prove useful in the conservation of other sub-tropical terrestrial orchid species.     

Habitat fragmentation changes the adaptive value of seed mass for the establishment of a tropical canopy tree

To cope with the limiting light conditions in the rain forest understory, many tropical tree species have evolved large seeds that provide the emerging seedlings with nutritional reserves. Habitat fragmentation might change the adaptive value of seed size by modifying the biotic and physical conditions of the forest understory. We experimentally assessed the potential of fragmentation to alter how seed mass affects seedling survival, vigor, and attack by natural enemies of the tropical tree Nectandra ambigens. Seeds from different mother trees (families) were individually weighed and sown in experimental sites established in continuous forest and in forest fragments. Seedling survival, vigor, and damage by herbivores and pathogens were recorded periodically. While seedlings derived from larger seeds had higher survival rates in both habitats, seedling survival and vigor were significantly greater in forest fragments, and the seedlings also suffered fewer attacks by natural enemies. We found genetic variance for seed mass among families with a heritability value (h²) of 0.66, and we found evidence for selection on seed size. Average seed size differed between dead and living seedlings in three sites. In one fragment, seed size was selectively neutral in relation to survival. Overall, selection for seed size promoted survival and thus appears to affect the distribution of this trait. The maintenance of genetic variance could be related to the stochastic nature of the formation of light gaps. Our results highlight the importance of evaluating the adaptive value of traits susceptible to environmental changes for conservation purposes.