Morphogenesis of the protocorm ofCypripedium acaule (Orchidaceae)

The nature of the protocorm of theOrchidaceae has fascinated morphologists for more than a century. In the present study, the development of the protocorm was followed using in vitro germination of seeds on a culture medium containing sugar, but without a symbiont. Inside the seed, the embryo consists of about a hundred cells. In the embryo, cells are arranged along a longitudinal axis according to size; these cells contain protein and lipid reserve material. In the first stages of seedling development, the embryo is transformed into a protocorm and meristematic tissue becomes organized into a meristematic dome (promeristem) at the anterior pole. This meristematic dome will give rise to a scale and the apex of the seedling. At first, the apex and the scale leaf develop synchronously. The development of the root always follows that of the apex. The study of the development of the seed ofCypripedium acaule showed that the protocorm is a distinct morphological system with respect to the rest of the cormus. The protocorm may be interpreted as an extension of the proembryonic stage.     

Rare plant translocation between mineral islands in Biebrza Valley (northeastern Poland): effectiveness and recipient site selection

We carried out translocations of three rare plants that inhabit mineral islands in the marshy Biebrza Valley in order to create alternative populations and facilitate connectivity between existing subpopulations. The species chosen were Iris aphylla and two orchids: Cypripedium calceolus and Cephalanthera rubra. Thirty soil monoliths with vegetative orchid plants or parts of I. aphylla rhizomes were dug out and transplanted to three different sites on new mineral islands (half in 2012 and half in 2013). Prior to translocation, we measured soil moisture and pH and took phytosociological characteristics in 68 potential recipient sites for orchids and 15 for I. aphylla. Then, we monitored the number of shoots for 4 years and retrospectively conducted principal component analysis (PCA) in order to compare the similarity of donor sites and chosen recipient sites. Three years after transplantation we found new C. calceolus populations in good condition, I. aphylla populations in moderate, while C. rubra transplants emerged only in the first and second season. All newly established populations of C. calceolus and I. aphylla survived. In the case of the first species, fruiting was observed in two populations, while a marked increase in shoot number was observed in one population. The most successful C. calceolus translocation site was also the most similar to the donor sites according to PCA.     

On Paphiopedilum malipoense sp. nov.—An Intermediate Form Between Paphiopedilum and Cypripedium,with a Discussion on the Origin of the genus

Paphiopedilum malipoense S. C. Chen et Tsi is a very interesting new species with its flower similar to that of Cypripedium, especially section Cypripedium. It belongs to subgenus Brachypetalum, the most primitive group of Paphiopedilum, but differs from its allied species in hgniva elliptic-lanceolate sepal with cuspidately acuminate apex, rather narrow petals and horizontal lip, which are of common occurrence in many cypripediums, but very rare in paphiopedilums. Apparently, this is an intermediate form, or a link, between Paphiopedilum and Cypripedium, but it does not seem to arise from hybridization between them, because no Cypripedium has been found wherever Paphiopedilum occurs. The new species is distributed in southeastern Yunnan of China. In this area, as well as in river valleys of western Yunnan or the Hengduan Mountains, there have been four species of the same genus reported before. As we know, the Hengduan Mountains and their adjacent areas are rich in Cypripedium. The differentiation of the genus there is remarkable. All five sections it contains occur there and three of them are quite distinctive. For example, the general appearance of the section Bracleosa is dissimilar to that of any other cypripediums, but closely resembles that of Listera. It appears that the difference between sect. Bracteosa of Cypripedium and sect. Brachypetalum of Paphiopedilum is not necessarily wider than that between sect. Bracteosa and sect. Cypripedium of the same genus. Apparently, it is reasonable to consider Paphiopedilum to be an evolutional branch of Cypripedium extending into tropical area, with its primitive group (subgenus Brachypetalum) still remaining in its northern fringe area. This primitive subgenus has eight species, distributed from western Yunnan to the Malay Peninsula. Five of them, including the intermediate and primitive form published here, are found in the hilly land of southeastern Yunnan and the river valleys of western Yunnan. All these facts suggest its area of origin: the river valleys of the Hengduan Mountains and the lower hilly land contiguous to the southof them.     

Seed biology and in vitro seed germination of Cypripedium

Abstract

Cypripedium orchids have high horticultural value. The populations of most species are very geographically restricted and they are becoming increasingly rare due to the destruction of native habitats and illegal collection. Reduction of the commercial value through large-scale propagation in vitro is a preferable option to reduce pressure from illegal collection. Cypripedium species are commercially propagated via seed germination in vitro. This review focuses on in vitro seed germination and provides an in-depth analysis of the seed biology of this genus.     

扇脉杓兰和无距虾脊兰的核型分析

为了解扇脉杓兰(Cypripedium japonicum Thunb.)和无距虾脊兰(Calanthe tsoongiana T. Tang et F. T. Wang)的核型,采用根尖压片法对扇脉杓兰和无距虾脊兰的染色体数目和核型进行了研究。结果表明,扇脉杓兰体细胞的染色体数为22,核型公式为2n=2x=22=16m+2sm+2st+2t,染色体相对长度组成为2n=22=2L+6M2+12M1+2S,核不对称系数为60.01%,核型分类为2B型;而无距虾脊兰体细胞的染色体数为40,核型公式为2n=2x=40=28m+10sm+2st,染色体相对长度组成为2n=40=8L+10M2+16M1+6S,核不对称系数为59.84%,核型分类为2B型;两者核型都较为对称。其中,无距虾脊兰的核型为首次报道。这为扇脉杓兰和无距虾脊兰的进化地位和种质保护提供了细胞学证据。     

Post‐anthesis changes in the labellum of Cypripedium macranthos var. rebunense (Orchidaceae) and a speculation on functional design of the labellum

In bumblebee‐pollinated Cypripedium macranthos var. rebunense, some plants have small labella that do not allow pollinating queens to enter, but they occasionally produce capsules for reasons that remain unclear. We found that after anthesis, the labellum gradually droops, and its width and length (but not height) distinctly increase, eventually producing an elongate and dorsoventrally compressed shape. The rate of increase in labellum size was significantly greater in flowers that initially had a smaller labellum, so that the post‐anthesis changes actually enabled large‐bodied bumblebees to enter those labella. Therefore, we conclude that C. macranthos var. rebunense relied on only bumblebee queens for pollination, irrespective of labellum size at the time of anthesis. Observations of in‐labellum bumblebee behavior suggested that the elongate and dorsoventrally compressed shape and the inclination seemed to result in more predictable and desirable bumblebee movements into the labellum.     

Effect of light on photosynthetic capacity during cold acclimation in a cold-sensitive and a cold-tolerant potato species

The effect of low temperature acclimation at various light levels on the photosynthetic capacity of Solanum species was examined. Two species, Solanum tuberosum L. cv. Red Pontiac and Solanum acaule Bitt., which differ significantly in degree of frost-tolerance and in their ability to acclimate to low temperature stress, were compared. Acclimation conditions included 5/2°C (day/night) temperatures, and either moderate (400 · mol · m⁻²· s⁻¹) or low (40 · mol · m⁻²· s⁻¹) photosynthetic photon flux densities. Several parameters of photosynthesis were measured in tissue pieces during acclimation treatments including chlorophyll content, chlorophyll a/b ratios and carbon dioxide-saturated photosynthetic oxygen evolution during light-limited and light-saturated assays.
Most measured photosynthetic parameters of low temperature-grown plants of both species showed greater declines under the moderate light than the low light conditions. Chlorophyll a/b ratios were unchanged after low temperature exposures in both light level treatments. At low temperatures, the cold-sensitive S. tuberosum demonstrated a greater inhibition of photosynthetic capacity in light- and carbon dioxide-saturated assays than S. acaule at all light levels. In addition to a pronounced inhibition at the higher light level, S. tuberosum demonstrated a very strong inhibition of photosynthetic capacity at very low light levels. Our results suggest a correlation between ability to maintain essential metabolic processes during low temperature stress in the presence of moderate light levels and the ability to increase cold tolerance.     

Studies in the embryology of the diandrous orchidCypripedium cordigerum (Cypripedieae,Orchidaceae)

The anther wall layers ofCypripedium cordigerum are six to eight. The glandular tapetum is 2- or 3-layered and its cells are uninucleate. Simultaneous cytokinesis results in decussate, isobilateral and tetrahedral pollen tetrads. Ripe pollen grains are 2-celled. The mature ovules are anatropous, bitegmic and tenuinucellate. Both the integuments are dermal in origin and 2-layered. The inner integument alone forms the micropyle. The female gametophyte is 6-nucleate and bisporic. The reduction of nuclei is due to the strike phenomenon. Double fertilization occurs. The primary endosperm nucleus divides to form two free endosperm nuclei. The mature embryo is undifferentiated. The cells ca, m and n contribute to the embryo. The suspensor is single-celled. The seed coat is formed entirely by the outer layer of the outer integument. There are three sterile and three fertile valves in the ovary. In the prefertilization stages these valves consist of parenchymatous cells with starch and raphides. After fertilization, the sterile valves develop sclerotic cells whereas the fertile valves remain parenchymatous. The pericarp structure and embryological features support the retention of tribeCypripedieae within theOrchidaceae.     

Surface features of the adaxial epidermis in the conduplicate-leaved Cypripedioideae (Orchidaceae)

Epidermal cells from adaxial leaf surfaces of 42 species of Paphiopedilum and 6 species of Phragmipedium were surveyed with the SEM. The surfaces of the cells are flat to papillose and often have various sculpturing patterns. To designate two orders of papilla size the terms 'macropapilla' and 'micropapilla' are proposed. Species exhibiting unornamented flat to macropapillose epidermal cells appear to be correlated with high light environments, whereas those species exhibiting micoropapillae and various degrees of sculpturing appear to be correlated with low light environments. Sculpturing features are often characteristic of a single species, but they may vary considerably between species. Epidermal characters are of some utility in identifying sterile plants which are otherwise indistinguishable.