Ceropegia thaithongiae sp. nov. (Apocynaceae,Asclepiadoideae) from northern Thailand and preliminary observations on impacts of climate change on its only known population

A new pendulous herb, Ceropegia thaithongiae Kidyoo (Apocynaceae, Asclepiadoideae), is described from northern Thailand. This plant is strictly endemic to Chiang Dao Wildlife Sanctuary and critically endangered owing to it living in an ecosystem affected by frequent natural disturbances. It is here described, illustrated and compared to the similar species, C. mairei. Both species have fusiform rootstocks, 1–2‐flowered cymes, deeply bifid outer corona lobes and linear, hairy inner corona lobes. However, the new species can be distinguished by its glabrous stem, leaf covered with hairs on both surfaces and glabrous corolla tube and lobes.     

High-frequency<Emphasis Type="Italic">in vitro</Emphasis> flowering in six species of<Emphasis Type="Italic">ceropegia</Emphasis>

High-frequencyin vitro flowering is reported here fromin vitro regenerated shoots ofin vitro-raised seedlings of rare and endemicCeropegia lawii, Ceropegia maccannii, Ceropegia oculata, andCeropegia sahyadrica, as well as the widely distributedCeropegia bulbosa var.bulbosa andCeropegia hirsuta. In our first set of experiments, the MS medium contained 87 mM sucrose and was supplemented with varying concentrations of BAP (4.4 to 26.6 μM). For the second set of trials, varying concentrations of sucrose (87 to 233 mM) were tested in MS media containing a constant 4.4 p.M BAP. Sub-cultured apical as well as axillary buds flowered with similar frequencies after 30 d of incubation. For all six species, the highest percentage of flowering shoots was obtained with either 26.6 μM BAP or 175 mM sucrose. Although smaller in size, theirin vitro flowers were morphologically comparable within wVo-derived flowers. Variations among species were noted for the number of flower buds per shoot and the percentage of flower formation. Because all six species showed similar responses in both experiments, we can suggest that this protocol is applicable across the wide range ofCeropegia species.     

Ceropegia pullaiahii sp. nov. (Apocynaceae,Asclepiadoideae) from India

Ceropegia pullaiahii Kullayiswamy, Sandhyarani et Karuppusamy sp. nov. (Apocynaceae) is described from open dry deciduous forest of Anantapur district, Andhra Pradesh, India. Its tubers are eaten by the local people. It is similar to C. mahabalei Hemadri et Ansari but differs in its twining habit and 3‐flowered cyme.     

Development of micro satellite markers for a critically endangered species, Ceropegia fantastica from the Western Ghats, India

Ceropegia fantastica L. (Asclepiadaceae) is a highly endemic and endangered species in the Western Ghats of India. Fourteen microsatellite markers were developed for C. fantastica. Eight microsatellite primers screened had 2–5 alleles per locus and the observed and expected heterozygosity ranged from 0.48 to 0.83 and 0.48 to 0.62, respectively. The primers were also evaluated for their cross amplification against two related species Ceropegia hirsuta and Ceropegia oculata. The microsatellites developed for this species could be used for addressing population genetics of this endemic and critically endangered species.     

<Emphasis Type="Italic">Ceropegia bhatii</Emphasis>, a new species of Apocynaceae: Ceropegieae from Karnataka,India

Ceropegia bhatii S. R. Yadav & Shendage, discovered from Malebennur Ghat in Davangere district of Karnataka State is described and illustrated. It is similar to Ceropegia noorjahaniae M. A. Ansari but is distinct in its twining habit, two-flowered cyme, slightly inflated corolla tube and corolla lobe margins that are not recurved.     

The cost of selfing inEncyclia cochleata (Orchidaceae)

To determine whether inbreeding depression accounts for the maintenance of outcrossing in populations of the self-compatible orchidEncyclia cochleata, the estimated selective advantage of selfing was compared to a measure of inbreeding depression. Individuals from three populations ofE. cochleata and some of their progeny were phenotyped using isozyme analysis. The electrophoretic data were used to estimate the outcrossing rate and the theoretical cost of outcrossing. Inbreeding depression was estimated by comparing the fitness of the progeny resulting from both types of pollinations. The seeds from outcrossed and selfed hand-pollinations and naturally pollinated seeds from a population of the triandrous form ofE. cochleata were grown aseptically on culture media, and their development over the next three years recorded. Inbreeding was common, particularly in one population (outcrossing rate 40%). However, the level of inbreeding depression was only 1–2%, considerably less inbreeding depression than expected.     

华南吊灯花属(夹竹桃科)植物小志

通过野外调查、标本查阅及文献考证,订正了华南吊灯花属(Ceropegia Linn.)植物,将狭瓣吊灯花(C. angustilimba Merr.)从吊灯花(C. trichantha Hemsl.)的异名中予以恢复,提供了二者的形态描述,确认华南地区记录的柳叶吊灯花(C. salicifolia H. Huber)实为剑叶吊灯花(C. dolichophylla Schltr.)的错误鉴定,并提供了后者的形态描述。     

秦岭种子植物区系新记录

报道了秦岭种子植物区系4新记录属[杜若属(Pollia Thunb.)、美冠兰属(Eulophia R.Br.ex Lindl.)、常山属(Dichroa Lour.)和吊灯花属(Ceropegia Linn.)],8新记录种[川杜若Pollia miranda(Lévl.)Hara、长距美冠兰Eulophia dabia(D.Don)Hochr.、常山Dichroa febrifugaLour.、肉色土栾儿Apios carnea(Wall.)Benth.exBaker、柳叶蓬莱葛Gardneria lanceolata Rehd.et Wils.、显脉獐牙菜Swertia nervosa(G.Don)Wall.ex C.B.Clarke、皱果赤瓟Thladiantha henryi Hemsl.var.henryi和宝兴吊灯花Ceropegia paohsingensis Tsiang et P.T.Li]。     

The linkage relations of thecochleata mutant inPisum

Thecochleata mutant inPisum is a monogenic recessive characteristic. The relations between the geneCoch and 15 well known other genes were determined.Coch is linked toR and toTl and this bringsCoch intoLamprecht's linkage-group VII.Coch was also found to be linked toGp and this is not in harmony with the above conclusion, sinceGp belongs to linkage-group V. The only possibility for an explanation at hand seems to be a translocation. This possibility could not be tested in the present material.Publication 234.     

Inclusion of Tenaris and Macropetalum in Brachystelma (Apocynaceae-Asclepiadoideae-Ceropegieae) inferred from non-coding nuclear and chloroplast DNA sequences

 The inclusion of Tenaris and Macropetalum in Brachystelma as proposed by Peckover in 1996 and contested by Victor and Nicholas in 1998 is supported by molecular studies. Parsimony analysis of sequence data from two non-coding molecular markers (ITS region of nrDNA and trnT-L and trnL-F spacers as well as the trnL intron of cpDNA) suggests a well-supported Brachystelma s.l. clade (including Tenaris and Macropetalum) with little internal resolution. The Brachystelma s.l. clade occupies a sistergroup position to the Ceropegia/stapeliad clade, and both clades together are sister to an Anisotoma/Sisyranthus/Neoschumannia clade. Received January 8, 2001 Accepted April 10, 2001     

Middle Triassic radiolarians from the Southeastern Pamirs (Republic of Tajikistan)

Triassic radiolarians were studied from a limestone/chert succession of the Dzhangisu and Boztere formations, Southeastern Pamirs, Republic of Tajikistan. The middle part of the Dzhangisu Formation is characterized by a Late Anisian assemblage with Eptingium nakasekoi Kozur and Mostler, Archaeospongoprunum bispinosum Kozur and Mostler, A. mesotriassicum mesotriassicum Kozur and Mostler, Paroertlispongus multispinosus Kozur and Mostler, P. rarispinosus Kozur and Mostler and Hozmadia rotunda (Nakaseko and Nishimura) that belongs either to the upper part of the Tetraspinocyrtis laevis Zone or to the Spongosilicarmiger transitus Zone (Kozur, 2003). The lowermost part of the Boztere Formation is characterized by a Late Ladinian assemblage with Muelleritortis cochleata (Nakaseko and Nishimura), M. expansa (Kozur and Mostler), Spongoserrula rarauana Dumitrica that corresponds to the M. cochleata Zone. Pamirian localities have a transitional position between the Western Tethys (Alpine–Mediterranean region) and the Eastern Tethys–Pacific junction (Southern China and Thailand). The assemblages are Tethyan in affinity.     

Total phenolics,flavonoids and antioxidant properties of three Ceropegia species from Western Ghats of India

The aim of present work was to assess the total phenolic content (TPC), total flavonoid content (TFC), phenolic compounds and antioxidant properties of various extracts of three Ceropegia spp.: Ceropegia spiralis, Ceropegia panchganiensis and Ceropegia evansii from Western Ghats of India. TPC of the samples varied from 0.3 ± 0.2 to 28.5 ± 0.3 mg TAE/g FW, whereas, TFC of the samples ranged between 0.1 ± 0.1 and 15.3 ± 0.3 mg RE/g FW. The major phenolic compounds identified were gallic acid, vanillin, cathechol and ferulic acid. All the extracts possess 1,1-diphenyl-2-picryl hydrazyl (DPPH) radical scavenging activity, ferric reducing antioxidant power (FRAP) as well as metal chelating ability and this was also supported by significant correlation with TPC and TFC. To the best of our knowledge, this is the first paper presenting comprehensive data on TPC, TFC, phenolic profile and antioxidant properties of the Ceropegia spp.     

Acid epimerization of 20-keto pregnane glycosides is determined by 2D-NMR spectroscopy

Carbohydrates influence many essential biological events such as apoptosis, differentiation, tumor metastasis, cancer, neurobiology, immunology, development, host-pathogen interactions, diabetes, signal transduction, protein folding, and many other contexts. We now report on the structure determination of pregnane glycosides isolated from the aerial parts of Ceropegia fusca Bolle (Asclepiadaceae). The observation of cicatrizant, vulnerary and cytostatic activities in some humans and animals of Ceropegia fusca Bolle, a species endemic to the Canary Islands, encouraged us to begin a pharmacological study to determine their exact therapeutic properties. High resolution ¹H-NMR spectra of pregnane glycosides very often display well-resolved signals that can be used as starting points in several selective NMR experiments to study scalar (J coupling), and dipolar (NOE) interactions. ROESY is especially suited for molecules such that ωτ_c ~ 1, where τ_c are the motional correlation times and ω is the angular frequency. In these cases the NOE is nearly zero, while the rotating-frame Overhauser effect spectroscopy (ROESY) is always positive and increases monotonically for increasing values of τ_c. The ROESY shows dipolar interactions cross peaks even in medium-sized molecules which are helpful in unambiguous assignment of all the interglycosidic linkages. Selective excitation was carried out using a double pulsed-field gradient spin-echo sequence (DPFGSE) in which 180° Gaussian pulses are sandwiched between sine shaped z-gradients. Scalar interactions were studied by homonuclear DPFGSE-COSY and DPFGSE-TOCSY experiments, while DPFGSE-ROESY was used to monitor the spatial environment of the selectively excited proton. Dipolar interactions between nuclei close in space can be detected by the 1D GROESY experiment, which is a one-dimensional counterpart of the 2D ROESY method. The C-12 and C-17 configurations were determined by ROESY experiments.     

夹竹桃科2种引种植物分泌结构的解剖学研究

为了解夹竹桃科(Apocynaceae)植物乳汁管的发生发育,对爱之蔓(Ceropegia woodii)和百万心(Dischidia ruscifolia)营养器官中的分泌结构进行了显微观察。结果表明,爱之蔓和百万心营养器官中均有无节分枝乳汁管的分布,茎皮层中的乳汁管大部分具有明显的分枝,叶中乳汁管具明显分枝,分布与走向多与叶脉维管组织平行。另外,爱之蔓营养器官中的分泌结构除乳汁管外,还有分泌腔。这为夹竹桃科植物的系统分类研究提供了解剖学依据。     

<Emphasis Type="Italic">In vitro</Emphasis> propagation of the rare medicinal plant <Emphasis Type="Italic">Ceropegia candelabrum</Emphasis> L. through somatic embryogenesis

Summary Efficient in vitro propagation of Ceropegia candelabrum L. (Asclepidaceae) through somatic embryogenesis was established. Somatic embryogenesis depended on the type of plant growth regulators in the callus-inducing medium. Friable callus, developed from leaf and internode explants grown on Murashige and Skoog (MS) medium supplemented with 4.52μM2,4-dichlorophenoxyacetic acid (2,4-D), underwent somatic embryogenesis. Compared to solid media, suspension culture was superior and gave rise to a higher number of somatic embryos. Transfer of the friable callus developed on MS medium containing 4.52μM 2,4-D to suspension cultures of half- or quarter-strength MS medium with lower levels of 2,4-D (0.23 or 0.45 μM) induced the highest number of somatic embryos, which developed up to the torpedo stage. Somatic embryogenesis was asynchronous with the dominance of globular embryos. About 100 mg of callus induced more than 500 embryos. Upon transfer to quarter-strength MS agar medium without growth regulators, 50% of the somatic embryos underwent maturation and developed into plantlets. Plantlets acclimatized under field conditions with 90% survival.     

Chemotaxonomic studies onArachniodes (Dryopteridaceae) II. Phloroglucinol derivatives and taxonomic evaluation

Seven species ofArachniodes from Japan and the Philippines were examined for phloroglucinol derivatives and glandularity of rhizome and petiole bases. For comparison 3 species ofDryopteris, 2 species ofNothoperanema, 3 species ofPolystichum, 1 species ofAcrophorus and 2 species ofCtenitis were investigated as well. Phloroglucinol derivatives were detected for the first time in the following taxa:Arachniodes assamica, A. cavalerii, A. tripinnata, A. yasu-inouei, Dryopteris pulvinulifera, D. cochleata, Polystichum rigens, Acrophorus nodosus, Ctenitis setosa andC. subglandulosa. The phloroglucinols ofDryopteris, Arachniodes andPolystichum are structurally related in agreement with the taxonomic relationships of these genera. On the other hand, the less related generaAcrophorus andCtenitis contain several unknown phloroglucinols not identical with those occurring in the former genera. The phloroglucinol containing species ofArachniodes, Polystichum, Acrophorus andCtenitis contain external glands, whereasDryopteris pulvinulifera andD. cochleata have both internal and external glands. The species of the genusNothoperanema seem to completely lack phloroglucinols and glands. The phloroglucinol composition of most taxa investigated is almost constant except for that inArachnoides amabilis and its varieties,A. aristata (A. exilis) andPolystichum tsus-simense. These species show varying phloroglucinol spectra in materials collected from different geographical sources.     

Regulation of stipule development by <Emphasis Type="Italic">COCHLEATA</Emphasis> and <Emphasis Type="Italic">STIPULE</Emphasis>-<Emphasis Type="Italic">REDUCED</Emphasis> genes in pea <Emphasis Type="Italic">Pisum sativum</Emphasis>

Pisum sativum L., the garden pea crop plant, is serving as the unique model for genetic analyses of morphogenetic development of stipule, the lateral organ formed on either side of the junction of leafblade petiole and stem at nodes. The stipule reduced (st) and cochleata (coch) stipule mutations and afila (af), tendril-less (tl), multifoliate-pinna (mfp) and unifoliata-tendrilled acacia (uni-tac) leafblade mutations were variously combined and the recombinant genotypes were quantitatively phenotyped for stipule morphology at both vegetative and reproductive nodes. The observations suggest a role of master regulator to COCH in stipule development. COCH is essential for initiation, growth and development of stipule, represses the UNI-TAC, AF, TL and MFP led leafblade-like morphogenetic pathway for compound stipule and together with ST mediates the developmental pathway for peltate-shaped simple wild-type stipule. It is also shown that stipule is an autonomous lateral organ, like a leafblade and secondary inflorescence.     

FOLIAR TRICHOMES OF PLEUROTHALLIDINAE (ORCHIDACEAE): FUNCTIONAL SIGNIFICANCE

Foliar surfaces of nine pleurothallid orchids (Cryptophoranthus lepidotus L. O. Wms., Octomeria sp., Pleurothallis pidax Luer, P. poeppigii Lindl., P. revoluta (Ruiz & Pav.) Garay, Restrepiella ophiocephala (Lindl.) Garay & Dunsterv., Restrepia muscifera Rchb. f. ex Lindl., Scaphosepalum rapax Luer, Stelis endresii Rchb. f.), four nonpleurothallid orchids (Cyrtopodium punctatum (L.) Lindl., Encyclia cochleata (L.) Lemee, E. tampensis (Lindl.) Small, Paphiope-dilum hybrid) and five tillandsioid bromeliads (Catopsis nutans (Sw.) Griseb., Tillandsia fas-ciculata Sw., T. streptophylla Scheidw., T. stricta Soland., T. tectorum E. Morr.) were assayed for permeability to Ca, S and P ions. Compared to leaves of the atmospheric bromeliads, those of the orchids proved less permeable to Ca and S. Moisture exchange profiles were determined for two of the bromeliads and three orchids; bromeliad leaves were able to rehydrate completely, orchid leaves only partially. Absorptive capacities of trichomes borne by five taxa were tested by autoradiographic analysis. Unlike bromeliad trichomes, those of the orchids exhibited no capacity to accumulate ³H-leucine. These results and other considerations described here suggest that, unlike tillandsioid bromeliads, neither the pleurothallid nor the nonpleurothallid orchids examined in this survey rely heavily on shoots for mineral and moisture procurement.