Pollination biology of four sympatric species of Habenaria (Orchidaceae: Orchidinae) from southern Brazil

The pollination process and breeding system of the sympatric Habenaria johannensis, H. macronectar, H. megapotamensis and H. montevidensis was documented for native populations from Rio Grande do Sul, Brazil. All species investigated offer a nectar reward (mean values of total sugars ranging from 18 to 26%) concealed in a spur. Habenaria montevidensis is pollinated by butterflies (Hesperiidae, especially of the genus Urbanus) that carry pollinaria on their eyes; the other three species are pollinated by Sphingidae. Habenaria johannensis is pollinated by the moths Manduca rustica and M. sexta that carry the pollinaria at the base of the proboscis. Habenaria macronectar is pollinated by the moths Eumorpha labrusca and M. cf. lucetius, and these bear pollinaria between the palpi. Habenaria megapotamensis is pollinated by moths of M. cf. lucetius that bear the pollinaria on the proboscis. All species studied are self‐compatible, but pollinator dependent. They also displayed high reproductive success (ranging from 69.48 to 97.40%) and male efficiency factors slightly higher than 1, suggesting that at least one flower was pollinated for each flower acting as pollen donor. At the study sites, the investigated Habenaria spp. are isolated (in terms of pollination) by a set of factors that includes differing floral morphologies, different pollinators and/or different pollinarium placement on the pollinator. © 2012 The Linnean Society of London, Botanical Journal of the Linnean Society, 2012, ??, ??–??.     

Three new species,four new records and an updated checklist of Habenaria (Orchidaceae) from Rio Grande do Sul,Brazil

Three new species of Habenaria (Orchidaceae) section Nudae from Rio Grande do Sul, southern Brazil, are described and illustrated: Habenaria australis, H. kleinii and H. sobraliana. These are the first records of H. sect. Nudae for the state. Based on the examination of living and dried specimens, a total of 33 species and 400 collections of Habenaria were recorded for Rio Grande do Sul. Based on this survey, an updated checklist of the genus Habenaria for Rio Grande do Sul has been compiled. Four species are known only from this state, while seven other species are restricted to southern Brazil, Argentina and Uruguay. Habenaria hieronymi, previously known only from Argentina, is recorded for the first time from Brazil, and H. brachyphyton, H. ekmaniana and H. melanopoda are new records for Rio Grande do Sul.     

Enhanced production of tropane alkaloids in transgenic <Emphasis Type="Italic">Scopolia parviflora</Emphasis> hairy root cultures over-expressing putrescine <Emphasis Type="Italic">N</Emphasis>-methyl transferase (PMT) and hyoscyamine-6β-hydroxylase (H6H)

Scopolia parviflora adventitious roots were metabolically engineered by co-expression of the two gene putrescine N-methyl transferase (PMT) and hyoscyamine-6β-hydroxylase (H6H) cDNAs with the aid of Agrobacterium rhizogenes. The transformed roots developed into morphologically distinct S. parviflora PMT1 (SpPMT1), S. parviflora PMT1 (SpPMT2), and S. parviflora H6H (SpH6H) transgenic hairy root lines. Consequent to the introduction of these key enzyme genes, the production of the alkaloids hyoscyamine and scopolamine was enhanced. Among the transgenic hairy root lines, SpPMT2 line possessed the highest growth index. The treatment of transgenic hairy roots with growth regulators further enhanced the production of scopolamine. Thus, the results suggest that PMT1, PMT2, and H6H genes may not only be involved in the metabolic regulation of alkaloid production but also that these genes may play a role in the root development.     

Nectar, nectaries, flower visitors, and breeding system in five terrestrial Orchidaceae from central Argentina

Floral nectar sugar composition, nectary anatomy, and visitors are studied in five Argentine Orchidaceae, from 18 populations. Hand-pollinations were performed to evaluate their breeding system. We found two different types of perigonal nectaries located either in the spur (Habenaria gouriieana, H. hieronymi, Habenariinae), or in the basal lateral parts of the labellum (Beadlea dutraei, Pelexia bonariensis, Stenorrhynchos orchioides, Spiranthinae). The spur ofHabenaria is a nonvascularised and nonstructural nectary. The inner epidermis bears one-celled long papillae. In bud stage, the papillae are filled with starch grains, but when the flower opens and nectar secretion starts, they show no starch grains. This fact may indicate that starch is a source for some of the secreted nectar. In the remainder genera, the lateral basal parts of the labellum are secretory. The two glands are located in the adaxial basal lateral faces of the labellum. These nectaries are structural and nonvascularised.Stenorrhynchos produces abundant, concentrated nectar (40–50%).Habenaria gourlieana accumulates copious nectar in a lower concentration (<20%), whereas the other species produce small quantities of concentrated nectar (ca. 50%). Three of the studied species have sucrose predominant nectar (Beadlea dutrael, Habenaria gourlieana, andPelexia bonariensis) whileH. hieronymi, Stenorrhynchos orchioides have hexose predominant ones. Nectar removal and/or pollination induce flower senescence.H. gouriieana is visited by sphingids,S. orchioides by hummingbirds, andB. dutrael by bees. For the two other species we did not record flower visitors.Pelexia bonariensis, B. dutrael, andS. orchiodes are self-compatible species but a pollinator is needed.     

中国兰科玉凤花属一新记录种——岩生玉凤花

报道了中国云南兰科植物一新记录种,岩生玉凤花(Habenaria vidua E．C．Parish &Rchb．f).该种主要特点是生长在腐殖质比较丰富的石灰岩缝隙中,不同于属内其它物种.在植株和花部特征相似于丛叶玉凤花H．tonkinensis,但侧萼片与唇瓣绿色、唇瓣侧裂片明显上举而区别于后者.     

DNA CONTENT OF SEVEN SPECIES OF ASTEREAE AND ITS SIGNIFICANCE TO THEORIES OF CHROMOSOME EVOLUTION IN THE TRIBE

Relative amounts of nuclear DNA were determined in root tip cells of seven species of Astereae: Aster hydrophilus Greene, A. oblongifolius Nutt., A. riparius H.B.K., Machaeranthera boltoniae (Greene) Turner and Home, M. brevilingulata (Sch-Bip.) Turner and Home, M. parviflora Gray, and M. tenuis (S. Wats.) Turner and Home. The results show that A. hydrophilus and M. brevilingulata, with a chromosome number of n = 9, have less nuclear DNA than other closely related species which are either n = 4 or n = 5. Cytological analyses of meiosis in the intergeneric hybrid M. parviflora X A. hydrophilus showed cells with two or more small chromosomes of the latter species pairing with single large chromosomes of the former. Pachytene cells of the hybrids M. parviflora X A. hydrophilus, M. parviflora X A. riparius, and M. boltoniae X M. tenuis showed some unpaired chromosome segments. The significance of these results to chromosome evolution in the tribe Astereae is discussed.     

Climate change might increase the invasion potential of the alien C4 grass Setaria parviflora (Poaceae) in the Mediterranean Basin

Aim Current climate change is supposed to be beneficial to many biological invaders, especially to C4 alien plants. While several experiments have been dedicated to measuring alien plants’ response to increased atmospheric CO₂ concentration, very few studies have been undertaken to measure the response of alien plants to warming. This study was aimed to test experimentally whether the predicted climate change in the Mediterranean Basin could be beneficial to the alien C4 grass Setaria parviflora (Poir.) Kerguélen. Location Three populations of S. parviflora from Corsica, southern France, were grown in Montpellier, southern France. Methods  The C4 alien grass S. parviflora was exposed to artificial climate change conditions for 3 years in open field and in competition with the local native community. We measured the response to artificial warming of +1.5 and +3 °C and artificial drought (?30% precipitation) versus ambient conditions for phenology, biomass and fecundity of S. parviflora. We compared the response of S. parviflora individuals to the response of the local community. Results Artificial warming strongly enhanced the biomass and the fecundity of S. parviflora, while it decreased or did not affect the biomass and fecundity of the local community. The phenology (onset of growth, first spike pollinating and fruit ripeness) of S. parviflora was advanced significantly and explained the changes observed in biomass and fecundity. Main conclusions Here, we report a positive effect of climate change on the growth and fertility of S. parviflora, a C4 alien plant. Our results suggest that climate change predicted for the next decades in the Mediterranean Basin might substantially enhance the performance of S. parviflora, potentially increasing its invasion success.     

The orchids of Timor: checklist and conservation status

A checklist of the Orchidaceae of Timor is presented, with emphasis on the eastern half of the island (East Timor), based on historical herbarium collections and recent botanical explorations. This list comprises 38 genera with 66 species, including 15 new genera and 32 new species records for this island. Moreover, four new species are described: Bulbophyllum sundaicum , Habenaria ankylocentron , Habenaria cauda‐porcelli , and Pterostylis timorensis . Of these, we consider the finding of a new species of Pterostylis to be especially noteworthy, because this species seems to be more closely related to certain Australian members of the genus than to the Malesian ones, suggesting earlier contacts of Timor with Australia. Four new synonyms are proposed: Calanthe veratrifolia var. timorensis J.J.Sm. (C. triplicata), Habenaria cornuta Span. (H. giriensis), H. grandis Benth. ex Ridl. (Peristylus goodyeroides), and H. mutica Span. (H. elongata). The best represented genus is Habenaria, with 13 species, followed by Dendrobium with four, and Bulbophyllum with three. Because of insufficient or sterile material, it was not possible to identify, or describe as new, 20 different taxa. The conservation status of the ten endemic species, plus six possible new undescribed species and two non‐endemic, but threatened, species, was assessed using the World Conservation Union (IUCN) criteria, and categories of threat were proposed. Seven endemic species are considered to be Critically Endangered and two Endangered. One of the nonendemic species is considered to be Critically Endangered, and the other Endangered. The survival of some of these species might be less insecure if an effective application of Regulation project N.2000/19 on protected areas (UNTAET/REG/2000/19) was implemented and maintained, because most of these species were collected in areas considered for protection under this Regulation. Further studies are required, however, in order to complete our knowledge of the diversity and population dynamics of this interesting part of Timor's biodiversity. © 2008 The Linnean Society of London, Botanical Journal of the Linnean Society, 2008, 157 , 197–215.     

Two new species of <Emphasis Type="Italic">Habenaria</Emphasis> (<Emphasis Type="Italic">Orchidaceae</Emphasis>) from the Brazilian cerrado and campo rupestre

Summary  Two new species, Habenaria ciliatisepala and H. egleriana (Orchidaceae, Orchideae), from the cerrado of central Brazil and campo rupestre of the Espinha?o mountain range in Minas Gerais and Bahia, are described and illustrated.     

A revision of Habenaria section Macroceratitae (Orchidaceae) in Brazil

Habenaria sect.Macroceratitae from Brazil is revised, and seven species are recognized:H. bractescens, H. gourlieana, H. johannensis, H. longicauda, H. macronectar, H. nabucoi, and the newly describedH. paulistana. A main feature of these species is the presence of long, separated, involute stigmatic processes.Habenaria bractescens, H. gourlieana, H. johannensis, andH. macronectar are distributed mainly from central and southeastern Brazil to southern Brazil and southern South America, whereasH. longicauda andH. nabucoi are distributed mainly from west central, southeastern and northeastern Brazil to northern South America.Habenaria paulistana is restricted to the state of São Paulo.Habenaria bradei, H. juergensii, andH. sartoroides are lectotypified, andH. kleyi is neotypified. The identity ofH. fastor is discussed andH. nabucoi is recognized as the valid name for this species. Seven other species previously placed in sect.Macroceratitae are sufficiently distinct and are removed from the section.     

MOSQUITO POLLINATION OF HABENARIA OBTUSATA (ORCHIDACEAE)

The floral morphology and pollination of Habenaria obtusata (Orchidaceae) are described. Habenaria obtusata occurs in Alaska, Canada, Norway, Siberia, etc., in great numbers and is pollinated by mosquitoes of the genus Aedes. The pollination process is described as observed in glass cages.     

红薯叶片浸提液对5种主要农田杂草种子萌发及幼苗生长的化感作用

以发芽率、发芽势、根长、茎长和鲜重变化为种子萌发和幼苗生长参数,研究了作物红薯叶片水浸液对云南省农田5种恶性杂草牛膝菊、藿香蓟、鬼针草、马唐和稗草的化感作用。结果表明,红薯叶片水浸液对5种受体杂草种子发芽率的影响不明显,但对发芽势有显著抑制作用。牛膝菊、藿香蓟、鬼针草和马唐的根长和生物量随红薯叶片水浸液浓度增加而显著降低,其中对马唐的抑制最强,高浓度0.1 g/m L时对根长和生物量抑制率分别为92.04%和73.33%,而低浓度0.0125 g/m L时分别为40.99%和46.67%;其次为鬼针草、藿香蓟、牛膝菊;最差的是稗草,随浓度的变化趋势均不明显。随红薯叶片水浸液浓度增加牛膝菊、鬼针草和马唐的茎长显著地降低,其中对马唐的抑制最强,高浓度0.1 g/m L和低浓度0.0125 g/m L时分别为86.85%和70.64%;其次为鬼针草和牛膝菊;相反藿香蓟和稗草的茎长随浓度增加而显著增加,高浓度0.1 g/m L和低浓度0.0125 g/m L时对藿香蓟的促进率分别为86.97%和16.03%。红薯叶片水浸液低浓度0.0125 g/m L时对牛膝菊的茎长和生物量有促进作用(低促高抑)。从化感作用响应指数和综合效应指数的综合对比来看,红薯对牛膝菊、藿香蓟、鬼针草、马唐具有显著的化感抑制作用,随浓度增加其抑制能力显著增加;其中对马唐的为最强,其次为鬼针草、牛膝菊和藿香蓟,相反对稗草具有促进作用(除了浓度0.1 g/m L)。所有这些表明,红薯叶片水浸液对5种杂草化感作用的敏感性趋势总体为:马唐鬼针草牛膝菊藿香蓟,最不敏感或者具有促进作用的是稗草。     

黑姜的化学成分、药理作用及毒理学研究进展

黑姜（Kaempferia parviflora）是东南亚地区广泛栽培的民间药用植物,被用于制造食品、化妆品和医药品。黑姜在我国西双版纳傣族自治州亦有较长的民间药用栽培历史。根茎是黑姜重要的药用部位,主要含有黄酮类物质。许多药理作用研究表明黑姜具有抗肿瘤、增强男性性能力、抗炎、抗氧化、抗真菌、抗病毒、神经保护、血管舒张和心脏保护活性、经皮促渗等作用。对黑姜的化学成分、药理作用及毒理学研究进行综述,以期为进一步深入研究和开发利用黑姜资源提供帮助。     

Micropropagation ofHesperaloe parviflora

Summary We successfully micropropagatedHesperaloe parviflora from mature plants. Shoot cultures were directly initiated from mature plants using pedicel bud explants on a modified Murashige and Skoog medium containing Nitsch and Nitsch vitamins and 1 μM zeatin riboside. Axillary shoot multiplication from established cultures was most responsive to changing concentrations of N⁶-benzyladenine (BA) with the greatest production of 6 μM BA. Growing shoots on a medium supplemented with 6 μM BA for 6 wk and then transferring cultures to a 1 μM BA medium for 6 more wk increased the number of transferable shoots, but not significantly. However, our data predicts that the maximum number of transferable shoots produced from a single microshoot would occur on media with 5.4 μM zeatin riboside. Shoots rooted easilyin vitro orex vitro and rooted shoots were easily acclimatized. The methods described in this paper are being used to commercially micropropagateH. parviflora.     

Spatial distribution of cpDNA and mtDNA haplotypes in a hybrid zone betweenPinus pumila andP. parviflora var.pentaphylla (Pinaceae)

Pinus species exhibit paternal chloroplast inheritance and maternal mitochondrial inheritance. This independent inheritance of two cytoplasmic genomes provides an exceptional environment for discriminating female (seeds) and male (pollen) components of gene flow across hybridizing species. We obtained mitochondrial genetic markers diagnostic toP. parviflora var.pentaphylla andP. pumila by PCR amplification of the intron ofnad1 on mtDNA, and examined the spatial-distribution pattern of the mtDNA haplotypes in a hybrid zone betweenP. parviflora var.pentaphylla andP. pumila in the Tanigawa Mountains of Japan. These data, in conjunction with previous information on cpDNA haplotypes and needle morphology, revealed contrastive patterns of introgression of two cytoplasmic genomes. CpDNA introgression has occurred uni-directionally fromP. parviflora var.pentaphylla toP. pumila. Conversely, mtDNA introgression has occurred in the opposite direction, fromP. pumila toP. parviflora var.pentaphylla. Levels of introgression are roughly equivalent for cpDNA and mtDNA. The contrastive spatial distribution pattern of cpDNA and mtDNA haplotypes could be caused by differential movement of seeds and pollen for interspecific genetic exchange.     

Different levels of inbreeding depression between outcrossing and selfing Serapias species

We quantified inbreeding depression for fruit production, embryo vitality and seed germination in three deceptive orchids, Serapias vomeracea, S. cordigera and S. parviflora, which do not provide any reward to their pollinators, and are predicted to experience high outcrossing. Of the three species examined only S. parviflora was autonomously selfing. Both S. vomeracea and S. cordigera showed highly significant differences in fitness between selfed and outcrossed progenies, resulting in high levels of inbreeding depression, which increased in magnitude from seed set to seed germination. Inbreeding depression may promote outcrossing in Serapias by acting as a post-pollination barrier to selfing. Cumulative inbreeding depression across three stages in S. parviflora was lower that in both outcrossing species. The large difference in germination between selfed and outcrossed seeds is an important issue in conservation biology.     

Melatonin affects the growth and cadmium accumulation of Malachium aquaticum and Galinsoga parviflora

Phytoremediation technology has become one of the main techniques for remediating soils polluted by heavy metals because it does not damage the environment, but heavy metal-tolerant plants have the disadvantages of low biomass and slow growth. A pot experiment was conducted to study the effects of melatonin (Mel) on growth and cadmium (Cd) accumulation in the Cd accumulator Malachium aquaticum and hyperaccumulator Galinsoga parviflora by spraying different concentrations of Mel on them. The results showed that shoot biomass, photosynthetic pigment content and antioxidant enzyme activity were increased in both species after Mel was sprayed on their leaves. Mel reduced the Cd content in shoots of M. aquaticum and increased it in those of G. parviflora. In general, Cd accumulation was greatest in M. aquaticum when Mel was 200 μmol L^?1 (120.71 μg plant^?1, increased by 15.97% than control) and in G. parviflora when Mel was 100 μmol L^?1 (132.40 μg plant^?1, increased by 68.30% than control). Our results suggest it is feasible to improve the remediation efficiency of lightly Cd-contaminated soil by spraying G. parviflora with100 μmol L^?1 Mel.     

Extensive Mitochondrial Introgression from Pinus pumila to P. parviflora var. pentaphylla (Pinaceae)

Pinus species exhibit a paternal chloroplast inheritance and a maternal mitochondrial inheritance. The levels and patterns of cpDNA and mtDNA introgression between the two pine species, P. pumila and P. parviflora var. pentaphylla, were examined at three mountain sites in Japan. The pine species were examined by using PCR-based diagnostic genetic markers of cpDNA and mtDNA. The survey which was carried out in multiple hybrid zones demonstrated a generality in the uni-directional pattern of cytoplasmic gene flow between the two pine species, i.e. paternal cpDNA flowed from P. parviflora var. pentaphylla to P. pumila, and in contrast, maternal mtDNA flowed from P. pumila to P. parviflora var. pentaphylla. Whenever plants which had a non-native combination of cpDNA and mtDNA were observed, they always had the cpDNA haplotype of P. parviflora var. pentaphylla and the mtDNA haplotype of P. pumila. The existence of only this type of cytoplasmic chimera may suggest that F₁ hybrids are successfully produced only in the crossing of P. pumila as the maternal parent and P. parviflora var. pentaphylla as the paternal parent. The present study also detected extensive mtDNA capture in populations of P. parviflora var. pentaphylla located in the southern and middle parts of the Ohu Mountains, Tohoku, Japan. In that area, nearly all of the plants examined had the mtDNA haplotype of P. pumila. The extensive mtDNA introgression suggests that seed flow could be an effective medium for interspecific gene exchange. Received 17 August 1998/ Accepted in revised form 7 January 1999