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1.
The difficulty in adult tissue genetic transformation in woody species is still an obstacle to be overcome, including in most sweet orange cultivars of the Brazilian citrus industry. This work reports that, after in vitro culture adjustments, transgenic adventitious buds of ‘Hamlin’, ‘Pêra’, and ‘Valencia’ sweet oranges (Citrus sinensis L. Osbeck) were recovered using adult material as explant source, in genetic transformation experiments via Agrobacterium tumefaciens. The transgenic buds were identified by the GUS histochemical analysis and confirmed by PCR analysis, which indicated the presence of an amplified fragment of 817 bp corresponding to the uidA gene sequence. The efficiencies of genetic transformation for ‘Hamlin’, ‘Pêra’, and ‘Valencia’ sweet orange cultivars were 2.5, 1.4, and 3.7%, respectively. Media supplemented with auxins and cytokinins during co-culture, and media with high concentrations of cytokinins (3 mg L−1) during transgenic selection led to the transformation and, consequently, the regeneration of adequate number of adventitious buds for the three cultivars. The use of sonication during the explant disinfection was not effective to reduce endophytic contamination and reduced transformation efficiency.  相似文献   

2.
Stable co-existence of similar species should be facilitated by mechanisms impairing, besides exploitative, interference competition. We investigated avoidance of intra- and interspecific conflicts in a four-species community of shrews [Sorex minutus Linnaeus, 1766,S. araneus Linnaeus, 1758,Neomys anomalus Cabrera, 1907, andN. fodiens (Pennant, 1771)], using the method of dyadic encounters in a neutral arena. We tested whether the use of passive (habituation, reduction of mobility, increase of inter-individual distance, and stillness) and active (‘to-and-fro’ and ‘keeping distance’ behaviours) forms of conflict avoidance depends on species, size or domination rank. The duration of conflicts was positively correlated with mobility and negatively with inter-individual distance, whereas it was unrelated to time of stillness and the active forms. The repertoire of conflict avoidance mechanisms was not species-specific and the display of these mechanisms depended rather on the size and domination rank of animals participating in a given interaction. In contrast to rodents, shrews did not avoid conflicts by the most passive forms: freeze and stillness reactions. All other forms were used with a higher or lower efficiency by all species. However, consistent with our predictions, large shrews (asN. fodiens) used mainly the passive mechanisms of conflicts avoidance (‘wait-and-see’ strategy), whereas small shrews (asS. minutus) invest proportionally more time in active forms (‘escape’ strategy).  相似文献   

3.
4.
Genetic data were used to examine the diversity in some allocreadiid trematodes. Nuclear ribosomal DNA (ITS2 and partial sequences of 5.8S and 28S) was sampled from sexual adult and ‘larval’ stages. From these and previous reference datasets phylogenetic trees were constructed. The results uncovered genetically distinct lineages within Bunodera luciopercae (Müller, 1776), suggesting that the two Palaearctic subspecies, B. l. luciopercae and B. l. acerinae Roitman & Sokolov, 1999, and Nearctic B. luciopercae from Perca flavescens may represent distinct species with a restricted host-specificity. Identical rDNA was revealed for the sexual adult of B. l. acerinae and ‘larval’ B. luciopercae described by Wiśniewski (1958). An unexpected match between the rDNA sequences of adult B. l. luciopercae and ‘larval’ Allocreadium isoporum (sensu Wiśniewski, 1958) was also detected. The adult A. isoporum (Looss, 1894) differs significantly from the ‘larval’ A. isoporum, the level of rDNA sequence divergence between them (8.6 % for 5.8S-ITS2-28S sequences and 6.26% for 28S) being consistent with the level expected for intergeneric variation. These results revealed the possible existence of a cryptic species complex within the nominal species B. luciopercae and a clear need for reconsideration of some of the accepted, but largely untested, tenets regarding allocreadiid life-cycles.  相似文献   

5.
Mating disruption treatments for the tufted apple bud moth (TABM),Platynota idaeusalis (Walker), were tested in small plot trials in apple orchards in Pennsylvania. Treatments were evaluated by fruit injury and by capture of male TABM in traps baited with synthetic pheromone sources or virgin females. The TABM pheromone is a two component isomeric blend ofE-11-tetradecen-1-ol (E11-14:OH) andE-11-tetradecenyl acetate (E11-14:Ac). A 50∶50 ratio of these two components was used in standard monitoring septa and in mating disruption treatments released from either hollow fibers (‘fiber’) or PVC tubes (‘PVC’). Other pheromone blends tested included a 90∶10 ratio of E11-14: Ac and E11-14:OH (‘EAc’) and its reverse (‘EOH’), mixture ‘EAc’ with 30% of the Z-isomers (‘low AEc’), and a blend similar to the preceding with 2% Z9-12:Ac (‘generic’). These other blends were released from multi tube tape (‘tape’) or Shin-Etsu type rope (‘rope’) dispensers. Seasonal dispenser release rate in mg ha−1 h−1 was ca. 30 for the ‘rope’ dispensers, 14 for ‘PVC’ and 6 for ‘fiber’. ‘EAc-tape’ and ‘EOH-tape’ were equally effective in reducing catches of males in traps baited with synthetic lures and in traps baited with virgin females. Both treatments also reduced fruit injury. ‘EAc-rope’, ‘fiber’ and ‘PVC’ also were generally effective; whereas, the ‘low EAc’ and ‘generic’ treatments reduced trap capture less than 90% and did not reduce fruit injury. Dispenser density was positively correlated with reduction in trap capture for the ‘low EAc-rope’ and ‘genericrope’ treatments. Traps loaded with ‘fiber’ dispensers captured more male TABM than the other treatments in non-pheromone permeated environments. Trap capture of other tortricids was reduced in pheromone treatments. ‘EAc-rope’ and the ‘TABM’ treatments provided mean (s.e.) percent reduction in trap catch of 99.5 (0.4) and 42.9 (10.1), respectively, for the redbanded leafroller,Argyrotaenia velutinana (Walker), and 90.4 (6.8) and 90.4 (1.3), respectively, for the obliquebanded leafroller,Choristoneura rosaceana (Harris).  相似文献   

6.
 We used graphical genotyping and linkage analyses with molecular markers to determine the chromosomal location of the rice stripe disease resistance gene, Stv-b i . The stripe resistance gene from the indica rice (Oryza sativa) cv ‘Modan’ was introgressed into several Japanese rice varieties. We found 4 RFLP markers in ‘Modan’, five susceptible parental rice varieties (‘Norin No. 8’, ‘Sachihikari’, ‘Kanto No. 98’, ‘Hokuriku No.103’ and ‘Koganebare’) and four resistant progeny varieties (‘St. No. 1’, ‘Aichi No. 6’, ‘Aoisora’ and ‘Asanohikari’). Graphical genotyping of the resistant progeny revealed a chromosomal segment ascribable to ‘Modan’ and associated with stripe resistance. The chromosomal segment from ‘Modan’ was located at 35.85 cM on chromosome 11. Linkage analysis using 120 F2 individuals from a cross between ‘Koshihikari’ (susceptible) and ‘Asanohikari’ (resistant) revealed another 8 RFLP markers in the same chromosome. We performed a bioassay for rice stripe resistance in F3 lines of the F2 individuals using infective small brown planthoppers and identified an 1.8-cM segment harboring the rice stripe disease resistance gene, Stv-b i , between XNpb220 and XNpb257/ XNpb254. Furthermore, Stv-b i was linked by 0.0 cM to a RFLP marker, ST10, which was developed on the basis of the results of RAPD analysis. These DNA markers near the Stv-b i locus may be useful in marker-assisted selection and map-based cloning of the Stv-b i gene. Received: 26 September 1997 / Accepted: 4 November 1997  相似文献   

7.
This article argues that organisms, defined by a semi-permeable membrane or skin separating organism from environment, are (must be) semiotically alert responders to environments (both Innenwelt and Umwelt). As organisms and environments complexify over time, so, necessarily, does semiotic responsiveness, or ‘semiotic freedom’. In complex environments, semiotic responsiveness necessitates increasing plasticity of discernment, or discrimination. Such judgements, in other words, involve interpretations. The latter, in effect, consist of translations of a range of sign relations which, like metaphor, are based on transfers (carryings over) of meanings or expressions from one semiotic ‘site’ to another. The article argues that what humans describe as ‘metaphor’ (and believe is something which only pertains to human speech and mind and, in essence, is ‘not real’) is, in fact, fundamental to all semiotic and biosemiotic sign processes in all living things. The article first argues that metaphor and mind are immanent in all life, and are evolutionary, and, thus, that animals certainly do have minds. Following Heidegger and then Agamben, the article continues by asking about the place of animal mind in humans, and concludes that, as a kind of ‘night science’, ‘humananimal’ mind is central to the semiotics of Peircean abduction.  相似文献   

8.
Summary An efficient and simple plant regeneration system via organogenesis from leaf segments of persimmon (Diospyros kaki Thunb.) cultivars ‘Fuyu’ and ‘Nishimurawase’ has been developed. The regeneration capacity was influenced by the culture vessels, gelling agents, plant growth regulators, and light conditions. Leaf explants taken from in vitro shoots were cultured on a modified Murashige and Skoog medium (MS1/2N), for 16 wk without transfer to fresh medium. Adventious shoots appeared after 4 and 8 wk in culture of ‘Nishimurawase’ and ‘Fuyu’ tissues, respectively. The culture of leaf explants in Erlenmeyer flasks with medium containing 4 g l−1 agar enhanced shoot formation in comparison to media with increased agar concentrations. Optimal shoot regeneration was obtained with 5 mg l−1 (22.8 μM) zeatin and 0.1 mg l−1 (0.05 μM) indole-3-butyric acid (IBA) for ‘Nishimurawase’, and 10 mg l−1 (45.6 μM) zeatin and 0.1 mg l−1 (0.05 μM) IBA for ‘Fuyn’. Shoot regeneration frequencies in both cultivars were 100%, and shoot numbers per explant reached up to 9.2 for ‘Nishimurawase’ and 2.2 for ‘Fuyu’. Dark incubation during the first 4–5 wk was the most effective condition to successfully influence shoot regeneration in both cultivars. While dark incubation was essential for adventitious shoot formation by ‘Fuyu’, it was only slightly beneficial to ‘Nishimurawase’. More than 80% of the regenerated shoots rooted within 4 wk on hormone-free MS1/2N demium after having been dipped for 30 s in 250 mg l−1 (1.1. mM) IBA solution.  相似文献   

9.
Phytophthora fragariae var. fragariae is the causal agent of red stele (red core) root rot in strawberry (Fragaria spp.). The inheritance of resistance to one isolate of this fungus was studied in 12 segregating populations of F.×ananassa derived from crosses between four resistant cultivars (‘Climax’, ‘Redgauntlet’, ‘Siletz’, and ‘Sparkle’) and three susceptible cultivars (‘Blakemore’, ‘Glasa’, and ‘Senga’ Sengana’). The analysis clearly supports the hypothesis of a single segregating dominant resistance gene. It is proposed that this gene be designated Rpf2. Received 12 November 1996 / Accepted: 22 November 1996  相似文献   

10.
Reported crystallographic data and calculated molecular models indicated that chlorophyll (Chl) a and bacteriochlorophyll (BChl) a tend to bind the fifth ligand on the side of the macrocycle where the C132-(R)-methoxycarbonyl moiety protrudes (denoting the ‘back’ side). The crystal structures of 34 photosynthetic proteins possessing (B)Chl cofactors revealed that most of Chl a and BChl a (and b) are coordinated by any peptidyl residue (e.g., histydyl-imidazolyl group), peptidyl backbone or water from the ‘back’ side. Almost all the cofactors that bind a water molecule as the fifth ligand in these proteins have a ‘back’ configuration. Theoretical model calculations for methyl chlorophyllide a (MeChlid a) and methyl bacteriochlorophyllide a (MeBChlid a) bound to an imidazole molecule indicated that the ‘back’ side is energetically favored for the ligand binding. These results are consistent with the fact that ethyl chlorophyllide a (EtChlid a) dihydrate crystal consists of the ‘back’ complex. The modeling also showed that both removal and stereochemical inverse of the C132-methoxycarbonyl group affect the relative stability between the ‘back’ and ‘face’ complexes. The effect of the C132-moiety on the choice of the macrocycle side for the ligand binding is discussed in relation to the function of P700. This revised version was published online in June 2006 with corrections to the Cover Date.  相似文献   

11.
Summary An in vitro protocol has been developed for callus indiction, somatic embryogenesis, and plant regeneration from stigma-style culture of grapevine. Four different grapevine cultivars (Vitis vinifera L.: cvs. ‘Bombino Nero’, ‘Greco di Tufo’, ‘Merlot’, and ‘Sangiovese’) were tested. Exlants were cultured on Nitsch and Nitsch medium (NN) supplemented with various combinations of 6-benzylaminopurine (BA: 4.5 and 9.0 μM) and β-naphthoxyacetic acid (NOA; 5.0 and 9.9 μM). Sucrose (88 mM) was used as the carbon source. Somatic embryogenesis was induced within 3–7 mo. after culture initiation. Even though explants of different origin (unfertilized ovules and anthers) regenerated somatic embryos, the higher embryogenic potential was observed in stigma and style explants, with the exception of ‘Merlot’, which regenerated somatic embryos only from unfertilized ovules. The percentages of stigma-style explants producing somatic embryos was 7% in ‘Bombino Nero’ (cultured on NN medium supplemented 9.0 μM BA and 9.9 μM NOA). 14% in ‘Greco di Tufo’ (4.5 μM BA and 9.9 μM NOA), and 8% in ‘Sangiovese’ (9.0 μM BA and 9.9 μM NOA). The presence of growth regulators (BA and NOA) in the medium was essential for induction of somatic embryogenesis. Plants were regenerated on hormone-free NN medium containing 88 mM sucrose.  相似文献   

12.
Experiments were conducted to quantify parasitism of Colorado potato beetle,Leptinotarsa decemlineata (Say), by the egg parasitoid,Edovum puttleri Grissell, on 3 different cultivars of eggplant,Solanum melongena L. Levels of parasitism were higher (P<0.05) on ‘Black Pride’ than on other cultivars. The percentage of egg masses that were parasitized was 1.2-fold higher (P<0.05) on ‘Black Pride’ than on ‘Harris Special’ and ‘White’. The number of eggs per mass that were parasitized was 1.3- and 1.4- fold greater (P<0.05) on ‘Black Pride’ than on ‘Harris Special’ and ‘White’, respectively. The percentage of eggs that were parasitized per mass and percentage of emerged adult parasitoids did not differ (P>0.05) among cultivars; between 2.1- to 2.6- fold more females than males emerged from eggs on all cultivars during the growing season.Edovum puttleri suppressed the 2nd generation ofL. decemlineata on ‘Black Pride’ and ‘Harris Special’, but did not suppress populations on ‘White’.   相似文献   

13.
In the Western Cape region of South Africa, dormancy release and the onset of growth does not occur normally in apple (Malus x domestica Borkh.) trees during spring due to the mild winter conditions experienced and fluctuations in temperatures experienced during and between winters. In this region, the application of chemicals to induce the release of dormancy forms part of standard orchard management. Increasing awareness of the environmental impact of chemical sprays and global warming has led to the demand for new apple cultivars better adapted to local climatic conditions. We report the construction of framework genetic maps in two F1 crosses using the low chilling cultivar ‘Anna’ as common male parent and the higher chill requiring cultivars ‘Golden Delicious’ and ‘Sharpe’s Early’ as female parents. The maps were constructed using 320 simple sequence repeats, including 116 new markers developed from expressed sequence tags. These maps were used to identify quantitative trait loci (QTL) for time of initial vegetative budbreak (IVB), a dormancy related characteristic. Time of IVB was assessed four times over a 6-year period in ‘Golden Delicious’ x ‘Anna’ seedlings kept in seedling bags under shade in the nursery. The trait was assessed for 3 years on adult full-sib trees derived from a cross between ‘Sharpe’s Early’ and ‘Anna’ as well as for 3 years on replicates of these seedlings obtained by clonal propagation onto rootstocks. A single major QTL for time of IVB was identified on linkage group (LG) 9. This QTL remained consistent in different genetic backgrounds and at different developmental stages. The QTL may co-localize with a QTL for leaf break identified on LG 3 by Conner et al. (1998), a LG that was, after the implementation of transferable microsatellite markers, shown to be homologous to the LG now known to be LG 9 (Kenis and Keulemans 2004). These results contribute towards a better understanding regarding the genetic control of IVB in apple and will also be used to elucidate the genetic basis of other dormancy related traits such as time of initial reproductive budbreak and number of vegetative and reproductive budbreak.  相似文献   

14.
An incompletely dominant gene conferring resistance to Puccinia hordei, Rph14, identified previously in an accession of Hordeum vulgare, confers resistance to all known pathotypes of P. hordei in Australia. Knowledge of the chromosomal location of Rph14 and the identification of DNA markers closely linked to it will facilitate combining it with other important leaf rust resistance genes to achieve long lasting resistance. The inheritance of Rph14 was confirmed using 146 and 106 F3 lines derived from the crosses ‘Baudin’/‘PI 584760’ (Rph14) and ‘Ricardo’/‘PI 584760’ (Rph14), respectively. Bulk segregant analysis on DNA from the parental genotypes and resistant and susceptible DNA bulks using DArT markers located Rph14 to the short arm of chromosome 2H. DArT marker bPb-1664 was identified as having the closest genetic association with Rph14. PCR based marker analysis identified a single SSR marker, Bmag692, linked closely to Rph14 at a map distance of 2.1 and 3.8 cm in the ‘Baudin’/‘PI 584760’and ‘Ricardo’/‘PI 584760’ populations, respectively.  相似文献   

15.
The compositions of leaf monoterpenoids from 11 species in the Juniperus section (Juniperus chinensis var. chinensis,J. virginiana, J. communis var. montana,J. rigida, J. chinensis var. globosa,J. chinensis var. sargentii,J. chinensis ‘Kaizuka Variegata’,J. squamata ‘Wilsonii’,J. x media ‘Shimpake’,J. x media ‘Plamosa Aurea’, andJ. squamata ‘Aloderi’) were comparatively analyzed by GC-MS. Of the 24 compounds identified, α-pinene, myrcene, limonene, terpinolene, and bornyl acetate were common to all, but particular combinations differed remarkably among taxa. The simplest composition (eight compounds) was found in.J, chinensis var. chinensis; the most complex (19 compounds), inJ. x media ‘Shimpake’. Cluster analysis generated four distinctive clades within the Juniperus section. The minimum spanning network revealed thatJ. squamata ‘Wilsonii’ andJ. x media ‘Shimpake’ were most similar in their chemical makeup.  相似文献   

16.
Marker-assisted selection (MAS) offers quick and reliable prediction of the phenotypes of seedlings in large populations and thus opens new approaches for selection to breeders of apple (Malus x domestica Borkh.). The development of framework maps enables the discovery of genetic markers linked to desired traits. Although genetic maps have been reported for apple scion cultivars, none has previously been constructed for apple rootstocks. We report the construction of framework genetic maps in a cross between ‘M.9’ (‘Malling 9’) and ‘R.5’ (‘Robusta 5’) apple rootstocks. The maps comprise 224 simple sequence repeat (SSR) markers, 18 sequence-characterised amplified regions, 14 single nucleotide polymorphisms and 42 random amplified polymorphic DNAs. A new set of 47 polymorphic SSRs was developed from apple EST sequences and used for construction of this rootstock map. All 17 linkage groups have been identified and aligned to existing apple genetic maps. The maps span 1,175.7 cM (‘M.9’) and 1,086.7 cM (‘R.5’). To improve the efficiency of mapping markers to this framework map, we developed a bin mapping set. Applications of these new genetic maps include the elucidation of the genetic basis of the dwarfing effect of the apple rootstock ‘M.9’ and the analysis of disease and insect resistance traits such as fire blight (Erwinia amylovora), apple scab (Venturia inaequalis) and woolly apple aphid (Eriosoma lanigerum). Markers for traits mapped in this population will be of direct use to apple breeders for MAS and for identification of causative genes by map-based cloning.  相似文献   

17.
Summary Callus was initiated from in vitro-grown plants of Gladiolus cultivars ‘Jenny Lee’ and ‘Florida Flame.’ The age of callus used for regeneration of plants was either 9 mo. old or 8 yr old from ‘Jenny Lee,’ and 4 mo. old from ‘Florida Flame.’ Regeneration medium consisted of Murashige and Skoog’s basal salts medium supplemented with 2.0 mg/l (9.3 μM) kinetin. This medium was supplemented with various concentrations of either bialaphos (Meiji Seika, Tokyo, Japan) or phosphinothricin (Hoechst-Roussell, Frankfurt, Germany). Bialaphos was more effective than phosphinothricin at stimulating plant regeneration. Plants regenerated from 8-yr-old callus of ‘Jenny Lee’ only when the regeneration medium was supplemented with 0.10 mg/l bialaphos. A bialaphos concentration of 0.01 mg/l stimulated regeneration from 9-mo.-old callus of cultivar ‘Jenny Lee’ and 4-mo.-old callus of ‘Florida Flame.’  相似文献   

18.
Vitis vinifera ‘Kishmish vatkana’, a cultivated grapevine from Central Asia, does not produce visible symptoms in response to natural or artificial inoculation with the fungus Erysiphe necator Schwein., the casual agent of powdery mildew. ‘Kishmish vatkana’ allowed pathogen entry into epidermal cells at a rate comparable to that in the susceptible control Vitis vinifera ‘Nimrang’, but was able to limit subsequent hyphal proliferation. Density of conidiophores was significantly lower in ‘Kishmish vatkana’ (33.6 ± 8.7 conidiophores mm−2) than in ‘Nimrang’ (310.5 ± 24.0 conidiophores mm−2) by 120 h after inoculation. A progeny of 310 plants from a ‘Nimrang’ × ‘Kishmish vatkana’ cross were scored for the presence or absence of visible conidiophores throughout two successive seasons. Phenotypic segregation revealed the presence of a single dominant allele termed Resistance to Erysiphe necator 1 (REN1), which was heterozygous in ‘Kishmish vatkana’. A bulked segregant analysis was carried out using 195 microsatellite markers uniformly distributed across the entire genome. For each marker, association with the resistance trait was inferred by measuring in the bulks the ratio of peak intensities of the two alleles inherited from ‘Kishmish vatkana’. The phenotypic locus was assigned to linkage group 13, a genomic region in which no disease resistance had been reported previously. The REN1 position was restricted to a 7.4 cM interval by analyzing the 310 offspring for the segregation of markers that surrounded the target region. The closest markers, VMC9H4-2, VMCNG4E10-1 and UDV-020, were located 0.9 cM away from the REN1 locus. Electronic supplementary material The online version of this article (doi:) contains supplementary material, which is available to authorized users.  相似文献   

19.
Eight native Iranian almond species from three sections, ‘Euamygdalus’ (Prunus communis; Prunus eleagnifolia and Prunus orientalis); ‘Lycioides’ (Prunus lycioides and Prunus reuteri) and ‘Spartioides’ (Prunus arabica, Prunus glauca and Prunus scoparia) were in vitro screened for drought tolerance using sorbitol and polyethylene glycol (PEG) as an osmoticum. Different levels of water stress were induced using five concentrations of either sorbitol or polyethylene glycol in Woody Plant Medium (WPM). Water potential of various media ranged from −0.80 to −2.05 MPa and water stress in culture medium adversely affected plantlet growth. Wild species from ‘Spartioides’ were less affected than ‘Lycioides’ and ‘Euamygdalus’. At the same level of water potential, sorbitol had lower adverse effects than PEG; the latter being severe. Prunus × sorbitol and Prunus × PEG interactions were significant. At 0.2 M sorbitol and 0.003 M PEG, ‘Spartioides’ produced significantly more roots with higher total root length and root volume, as well as root-dry weight than those of ‘Lycioides’ and ‘Euamygdalus.’ It is concluded that in vitro screening of native Iranian almond species under specific and limited water-stress conditions may provide a system for effectively differentiating the wild species of almond for their expected root mass production under field conditions.  相似文献   

20.
The phyllodocid polychaete Notophyllum foliosum occurs in two colour morphs in Swedish and Norwegian waters, one palish yellow to grey form with black patches that is restricted to deeper waters and often associated with reefs of the deep-water coral Lophelia pertusa, and one usually yellow-orange form with black patches and white spots that is usually encountered on more shallow bottoms. We have sampled the two forms from sympatric occurrences in Norway, and the shallow form from the Swedish west coast. Phylogenetic and haplotype analyses based on the mitochondrial cytochrome c oxidase subunit I (COI) gene and the nuclear internal transcribed spacer region (ITS1-5.8SrDNA-ITS2) unequivocally indicate that the two forms represent different species. We apply the name N. foliosum (Sars, 1835) to the ‘shallow form’, and propose N. crypticum n. sp. for the ‘deep form’. A lectotype is fixed for N. foliosum.  相似文献   

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