PLoS ONEplosplosonePLOS ONE1932-6203Public Library of ScienceSan Francisco, CA USA10.1371/journal.pone.0274699PONE-D-21-38112Research ArticleBiology and life sciencesPlant sciencePlant anatomyFlowersBiology and life sciencesPlant sciencePlant anatomyFlower anatomySepalsBiology and life sciencesEcologyEcosystemsForestsTemperate forestsEcology and environmental sciencesEcologyEcosystemsForestsTemperate forestsEcology and environmental sciencesTerrestrial environmentsForestsTemperate forestsBiology and life sciencesPlant sciencePlant anatomyFlower anatomyPetalsBiology and life sciencesPlant sciencePlant anatomyLeavesBiology and life sciencesOrganismsEukaryotaPlantsFlowering plantsBiology and life sciencesEcologyEcosystemsForestsEcology and environmental sciencesEcologyEcosystemsForestsEcology and environmental sciencesTerrestrial environmentsForestsBiology and life sciencesEvolutionary biologyEvolutionary systematicsPhylogeneticsPhylogenetic analysisBiology and life sciencesTaxonomyEvolutionary systematicsPhylogeneticsPhylogenetic analysisComputer and information sciencesData managementTaxonomyEvolutionary systematicsPhylogeneticsPhylogenetic analysisA new species of Impatiens and updated checklist of Balsaminaceae in NepalNew species of Impatiens from Nepalhttps://orcid.org/0000-0002-9064-8657RaskotiBhakta BahadurConceptualizationData curationFormal analysisMethodologyResourcesWriting – original draftWriting – review & editing*AleRitaWriting – review & editingNature Research and Conservation Initiative, Banke, NepalVousdenGeorgeEditorPublic Library of Science, UNITED KINGDOM
NO authors have competing interests.
* E-mail: bbraskoti@gmail.com1910202220221710e0274699212202125820222022Raskoti, AleThis is an open access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited.
Impatiens nimspurjae (Impatiens, Balsaminaceae), a new species from Nepal is described based on molecular data and morphological characters. Notes on the diagnostic characters used to distinguish it from allied taxa are provided. I. nimspurjae is closely related with taxon belonging to sect. Racemosae (I. harae, I. radiata, I. wallichii, I. urticifolia) in having many-flowered racemose inflorescences, lateral sepals 2 (very rarely 4 with inner 2 reduced), capsule linear, seed ovoid, but differs by its sessile leaves, upper lobe of lower united petal not truncated, spur flattened at base. With the discovery of this new species and five species new records to Nepal, a checklist of Balsaminaceae having 57 species (8 endemic) in Nepal is updated. An identification key to the species of Imaptiens in Nepal is also provided.
The author(s) received no specific funding for this work.Data AvailabilityAll relevant data are available in Supporting information files.Introduction
The genus Impatiens L. is one of the largest genera of flowering plants, containing over 1000 species, which exhibit complex pattern of morphological variation [1, 2]. It is widely distributed in tropical, subtropical, temperate and alpine regions of the Old World, with the greatest diversity seen in tropical Africa, Madagascar, southern India, Sri Lanka, Himalayas and Southeast Asia [3–5]. Biogeographically, Impatiens species are of Southeast Asian origin from where it dispersed to boreal Eurasia and North America to central Asia and eastern Europe via the Himalayas to India and Africa [4]. Based on the combined molecular and morphological phylogeny, Impatiens is divided into two subgenera, Clavicarpa and Impatiens; the latter is further subdivided into different sections [6].
Most of the Impatiens species in Nepal are restricted to eastern and central Nepal [7, 8]. Various authors studied Balsaminaceae of Nepal targeting on some problematic species group [9–11]. However, Impatiens is still not well studied and documented within Nepal. During different botanical surveys in the various parts of Nepal from 2007 to 2012, some unidentified plants of Impatiens were collected. We carefully studied taxonomic characters of these plants and also performed a molecular phylogeny of Impatiens to understand systematic position of a newly collected plant.
Materials and methodsEthics statement
The collecting location reported in this work is not protected area by any law. In addition, the plant species collected here are currently neither endangered nor protected. No specific permits were required for this study.
Nomenclature
The electronic version of this article in Portable Document Format (PDF) in a work with an ISSN and ISBN will represent a published work according to the International Code of Nomenclature for algae, fungi, and plants, and hence the new names contained in the electronic publication of PLoS article are effectively published under that Code from the electronic edition alone, so there is no longer any need to provide printed copies.
In addition, new name contained in this work has been submitted to The International Plant Names Index (IPNI), from where they will be made available to the Global Names Index (http://gni.globalnames.org/). The IPNI LSIDs can be resolved and the associated information viewed through any standard web browser by appending the LSID contained in this publication to the prefix http://ipni.org/. The online version of this work is archived and available from the following digital repositories: PubMed Central and LOCKSS.
Morphological observations
Vegetative and reproductive characters were studied based on fresh and preserved specimens. Characters such as phyllotaxy, petiole, leaf shape, bracts, sepals, petals, androecium, gynoecium and fruits were carefully examined and measured. The characters were compared with related species deposited in different herbaria, and their description published in different literatures. The plants were dried and herbarium specimens were prepared following the standard procedure and deposited them in the National Herbarium and Plant Laboratories (KATH), Godawari, Lalitpur, Nepal and Tribhuvan University Central Herbarium (TUCH).
Taxon sampling
In total, 239 nucleotide sequences of 238 species of Impatiens were sampled in the phylogenetic study. We downloaded 236 sequences of 236 species from GenBank. Additionally, we sequenced three samples of the newly collected taxon for the present analyses. The ingroup taxa included a total of 236 sequences of 235 species, representing all of the major clades in the Impatiens [6, 12]. Three species namely Marcgravia umbellata, Norantea guianensis and Hydrocera triflora were selected as outgroups following previous phylogenetic studies [6, 12]. A list of sampled species, GenBank accession numbers for all sequences used in this study are available in S1 Table.
DNA extraction and marker selection
DNA extraction was carried out from fresh leaf tissue using the modified CTAB protocol [13]. Amplification of DNA regions was performed using a standard polymerase chain reaction (PCR). We selected nuclear (ITS) and plastid (atpB-rbcL) nucleotide sequences which have exhibited their efficacy for understanding phylogenetic relationships at various taxonomic levels within the Balsaminaceae [4, 6, 12, 14]. Primers and PCR protocols for atpB-rbcL and ITS are followed based on the published literatures [14, 15].
Sequence alignment and phylogenetic reconstruction
DNA sequences were initially aligned with the help of CLUSTAL X version 1.83 [16] with minor subsequent manual adjustment using the software BioEdit verion 5.0.9 [17]. To evaluate congruence between different DNA regions, we analyzed nuclear (ITS) and chloroplast (atpB-rbcL) dataset separately to see if they produce an identical topology. We observed no significant difference in topologies of ITS and atpB-rbcL (S1 and S2 Figs). We therefore analyzed ITS and atpB-rbcL dataset in combination. Phylogenetic analyses were carried out using maximum parsimony (MP) as implemented in PAUP [18]. MP analyses were conducted using heuristic searches with 1,000 replicates of random taxon addition and tree rearrangements using tree bisection-reconnection (TBR) branch swapping to obtain the parsimonious trees. Gaps were considered as missing data and all characters were treated as unordered and equal weights. The node supports for clades were assessed by bootstrap (BSMP) analyses, which were performed using 1,000 replicates with ten random taxon additions and heuristic options [19].
For the Bayesian inference (BI) analysis, posterior probabilities (PP) were conducted in MrBayes version 3.2.3 [20], implemented in CIPRES Science Gateway [21]. The best-fitting model of sequence evolution in BI analyses was chosen for each marker using ModelTest 3.7 [22] under the Akaike Information Criterion (AIC) in conjugation with PAUP [18]. Evolutionary models that best fitted ITS, atpB-rbcL and combined dataset were GTR + I + G (nst = 6, rates = invagamma). Two separate runs, each with four-Markov-Chain Monte Carlo (MCMC) analyses were performed, starting with a random tree, proceeding for 20000000 generations (for combined dataset) and sampling one tree every 1000 generations. Majority rule (>50%) consensus trees were reconstructed after removing the ‘burn-in phase’ samples (the first 25% of the sample trees).
Checklist preparation
To prepare an updated checklist of Balsaminaceae, we reviewed all scientific names of Balsaminaceae, which had been recorded in Nepal. All the scientific names were checked according to Shenzen code of International Association for Plant Taxonomy [23] together with online consulted from World Flora Online, The Plant List and International Plant Names Index.
Results
The length of aligned matrix of ITS was 857 bp, whereas atpB-rbcL matrix consisted of 1112 bp. The combined (nuclear and chloroplast) data matrix consisted of 1969 characters of which 296 were variable and 934 were parsimony informative. The topologies from ITS, atpB-rbcl and combined analysis were congruent (S1 and S2 Figs and Fig 1). The BI tree based on combined dataset is shown for the discussion of phylogenetic relationships (Fig 1).
10.1371/journal.pone.0274699.g001
Bayesian inference tree of <italic>Impatiens</italic> based on the combined nuclear (ITS), and chloroplast (<italic>atpB-rbcl</italic>) markers.
Numbers above branches indicate bootstrap percentages (BSMP) for MP analyses and posterior probabilities (PP) for BI analysis, respectively. Asterisk (*) indicates BSMP = 100 and PP = 1.0, a dash (−) indicates support at a node < 50%. Scientific name marked in blue color indicates species occurring in Nepal.
The phylogenetic tree inferred from single and combined dataset are congruent with the previous phylogenetic analysis [6, 12, 24]. Impatiens is divided into two major clades (i.e. Clade 1 and Clade II). Clade I is further divided into strong to weakly supported different subclades (Fig 1).
Our molecular results based on nuclear (ITS) and chloroplast (atpB-rbcL) DNA sequence data indicated that newly sampled taxon nested within Clade I (Fig 1). This species form a separate lineage within Clade I/Sublcade III (BSMP = 80%, PP = 1.00), which is strongly supported as sister to I. harae and its allies (BSMP = 90%, PP = 1.00).
The major morphological similarities within the clade belonging to newly sampled species and allied taxa consist of many-flowered, racemose inflorescences, lateral sepals 2, rarely 4 with inner 2 reduced, capsule linear, seed ovoid.
Line drawings of Impatiens nimspurjae: A. Flowering plant; B. Leaf; C. flower, side view; D. Flower (front view); E. Dorsal sepal; F. lateral sepals; G. Lateral united sepals (front view); H. Lateral united sepal (back view); I. Lower sepal; J. Ovary, pistil, pedicel and bract; K. Capsule; L. Seeds. Line drawings assisted by Neera Joshi Pradhan.
10.1371/journal.pone.0274699.g003
Impatiens nimspurjae: A. Flowering plant; B. Leaf; C. flower, side view; D. Dorsal petal (dorsal view); E. lateral united sepals (ventral view); F. lateral united sepals (dorsal view); G. Lateral sepal; H. Lower sepal; I. Ovary, pistil, pedicel and bract; J. Capsule; K. seeds. Photographs by Bhakta Bahadur Raskoti.
Type:—NEPAL. Gandaki Province: Myagdi District, above Moreni, at an elevation range of 2800–2900 m, 25 Aug. 2011, Bhakta B. Raskoti 201190 (holotype, designated here: KATH); isotype Bhakta B. Raskoti 201190 (TUCH).
Diagnosis
Impatiens nimspurjae is closely related to I. harae, I. radiata, I. wallichii but differ from these species by having sessile oblong-lanceolate leaves, flowers less than 3 in each peduncle, base of the spur flattened, apex of dorsal petal rounded, basal lobe of lateral united petal widely ovate, not truncated.
Description
Annual herbs, 12–99 cm tall. Stems glabrous, succulent, erect or creeper, slender, often branched above. Leaves alternate, sessile; leaf blades oblong to lanceolate, 1.5–11 × 0.5–3 cm, glabrous, setose between teeth, base cuneate and tapering, margin crenate, apex acuminate, lateral veins 5–7 pairs, whitish green. Inflorescences axillary, 2–3-flowered, peduncle 1.5–6 cm long, 1 mm diam, glabrous, pedicel 0.5–1 cm long, 1–1.5 mm diam, bracteate above base. Bracts persistent, lanceolate, 1–2 mm long. Flowers 1.5–2 cm long, white, flushed with yellowish and tinge purple on dorsal petal and lower sepal. Lateral sepals 2, falcately ovate, 2–3 × 1.5–2 mm, entire, apex mucronulate. Lower sepal bucciniform, 1–1.5 cm (excluding spur), apex with a small crest, basal portion narrowed into a hooked spur; spur stout with flat base, longitudinal canal on dorsal and ventral surface, obtuse. Dorsal petal orbicular, ca. 1 × 1 cm, apex rounded; lateral united petals ca. 1.5–2 cm long, basal lobe widely ovate, entire; apical lobe bilobulate. Stamens 5, filaments linear, anther small, apex obtuse. Ovary 5–7 mm long, erect, apex acute, glabrous. Capsule erect, linear, 1.5–2.5 cm long, 5-angled, sparsely verruculose along ribs; seeds ovoid, brownish.
Phenology
Flowering and fruiting occur from July to October.
Etymology
The species epithet ‘nimspurjae’ refers to the name of Mr. Nirmal Purja (nickname Nimsdai) for his initiation on conservation through climate change champion. Mr. Purja is a world-renowned mountaineer from Nepal and has dozens of Guinness book world records in the field of mountaineering.
Distribution and habitat
Impatiens nimspurjae is known from the type locality in western Nepal where it occurs in the temperate forest and forest margins at an elevation range of 2800–2900 m. It grows on moist humus rich slopes.
Taxonomic notes
Impatiens nimspurjae is closely related to I. radiata, I. harae, I. wallichii and I. urticifolia having many-flowered, racemose inflorescences, lateral sepals 2, rarely 4 with inner 2 reduced, capsule linear, seed ovoid but I. nimspurjae can be distinguished by its sessile, oblong to lanceolate leaves, less than three flowers in each inflorescences, flowers flushed with yellow and tinged purple, base of spur much flat (Fig 4A). The apex of dorsal petal of I. nimspurjae is rounded (vs. retuse in I. wallichii and mucronate in I. urticifolia). Moreover, basal lobe of lateral united petal is not truncated and base of spur is much flattened. Impatiens nimspurjae is also similar to I. harae and I. radiata by having white flower with purple tinge but latter two have clustered leaves and radiated inflorescences.
10.1371/journal.pone.0274699.g004
Flowers of Impatiens nimspurjae and its allied species: A. I. nimspurjae; B. I. urticifolia; C. I. radiata; D. I. harae. Photographs by Bhakta Bahadur Raskoti.
Conservation status
Impatiens nimspurjae is endemic to western Nepal and restricted in only three populations. During the fieldworks in 2008 and 2011 a total of about 100 individuals were found in an area of < 9 km2. It is expected that size of its population may be larger in the type locality and surrounding areas. However, the extent of occurrence and exact population size are unknown. According to these preliminary information I. nimspurjae is assessed as Data Deficient (DD) following the IUCN Red List Categories and Criteria [25]. The major threat for this species is habitat disturbance from anthropogenic activities such as deforestation, trampling, over grazing etc.
New addition to flora of Nepal
During the different botanical surveys in different parts of Nepal, we discovered following five species of Impatiens as new records for flora of Nepal.
Impatiens brachycentra Kar. & Kir.
NEPAL. Central Nepal: Myagdi district, at an elevation of 2400 m, Bhakta B. Raskoti 20103 (KATH).
Impatiens cathcarthii Hook.f.
NEPAL. Eastern Nepal: Ilam district, at an elevation of 1300 m, Bhakta B. Raskoti 20117 (KATH).
Impatiens gammieiHook.f.
NEPAL. Eastern Nepal: Taplejung district, at an elevation of 3400 m, Bhakta B. Raskoti 20109 (KATH).
Impatiens infundibularis Hook. f.
NEPAL. Eastern Nepal: Ilam district, Kolbung, at an elevation of 1400 m, Bhakta B. Raskoti 20101 (KATH).
Impatiens sikkimensis Govaerts & Chakrabarty
NEPAL. Eastern Nepal: Ilam district, Kolbung, at an elevation of 1500 m, Bhakta B. Raskoti 201145 (KATH).
Identification key to the species of <italic>Impatiens</italic> in Nepal
2a. Inflorescence fascicled in leaf axils, peduncle very short (less than 5 mm)...................... 1) I. arguta [sect. Fasciculatae]
2b. Inflorescence not fascicled, peduncle longer than 5 mm (except in I. tripetala)………...….3
3a. Inflorescence subscorpioid cymes, flowers white tinge with pink............................4 [sect. Scorpioidae]
3b. Inflorescence not scorpoid, flowers pinkish, yellow, white...........................6 [sect. Uniflorae]
4a. Spur distinctly involute, peduncle short (upto 5 cm), crest of dorsal petal 3–6 mm long................................................................................................................................2) I. discolor
4b. Spur not distinctly involute, peduncle more than 5 cm long …………..…..............................5
5a. Leaves glabrous, bracts broadly ovate-orbicular, lateral petals often orbicular, crest of dorsal petal 3–6 mm long,.…..............................................................................................3) I. cathcarthii
5b. Leaves hairy, bracts ovate, lateral petals usually ovate (not orbicular), crest of dorsal petal 6–16 mm long …………………..........................................................................................4) I. jurpia
6a Capsule ovoid, seeds rounded, upper lobe of lateral united petals distinctly smaller than lower lobe (plant cultivated or naturally grows in abandoned land)……………………..5) I. balsamina
6b. Capsule fusiform, seeds ovoid or ellipsoid……………………..………….…………...……..7
7a. Peduncle short (not exceeding 1 cm long), spur curved, lower sepal saccate, spur curved...………………………………………………………………...…………….6) I. tripetala
7b. Peduncle long (more than 1 cm)...............................................................................................8
8a. Lower sepal infundibuliform with long spur.............................................................................9
8b. Lower sepals saccate, bucciniform, navicular, spur usually not very long..............................10
9a. Dorsal petal with crest, upper lobe of lateral united petal smaller than the lower lobe, spur curved, flower deeply pink………………………………...…………………..7) I. infundibularis
9b. Dorsal petal without a crest; upper lobe and lower lobe of lateral united petals nearly equal in size, flower light pink, spur filliform.................................................................................8) I. exilis
10a. Flowers reddish purple, lower sepal navicular or bucciniform………………………….….11
10b. Flower yellow or orange, lower sepal bucciniform.............................................................. 12
11a. Spur coiled, lower sepal bucciniform...................................................................9) I. spirifera
11b. Spur not coiled, lower sepal deeply navicular...................................................10) I. puberula
12a. Spur not coiled, lower sepal obliquely navicular................................................11) I. porrecta
12b. Spur coiled, peduncles more than 10 mm long; flowers yellow or orange, lower sepal bucciniform….............................................................................................................12) I. pulchra
13a. Fruits 4-lobed, ovary 4-carpellate……………….……13) I. laevigata [sect. Semeiocardium]
22b. Flowers pink, yellow, purple or whitish purple.....................................................................24
23a. Apex of distal lobe of lateral united petals long tailed, flower tinge with purple.……………….…………………………………………………………………21) I. harae
23b. Flower not tinge with purple (except in I. arunensis)...........................................................27
24a. Leaves alternate and aggregated at the apical part of the stem, spur curved downwards, 17–36 mm long…………………………………………………………………...….22) I. graciliflora
24b. Spur (if present) not curved downwards but straight.............................................................25
25a. Lower sepal spurless...................................................................................23) I. brachycentra
25b. Lower sepal navicular (7–15 mm long)……………………………………........................26
26a. Lower sepals with spur, spur straight and tapering gradually…………………..24) I. radiata
26b. Lower sepals spurless, other characters similar as in I. radiata………………25) I. gammiei
33b. Spur not incurved forward……………………………………………….….………….....35
34a. Stem with several branches, flowers yellow with reddish veins, spur incurved forward.....................................................................................................................33) I. urticifolia
34b. Stem unbranched, spur S-shaped; flowers pale yellow with reddish dots……34) I. wallichii
44a. Lower sepal navicular or infundibuliform, tapering into upwardly or downwardly curved spur............................................................................................................................42) I. scabrida
44b. Lower sepal saccate........................................................................................43) I. bajurensis
45a. Apex of upper lobe of lateral united petals obtuse..............................................44) I. bicolor
45b. Apex of upper lobe of lateral united petals falcate.………………..…..….45) I. edgeworthii
57a. Basal lobe of lateral united petal narrowly triangular, lower sepal with abruptly constricted erect spur and curved distally…….………………………………………………....56) I. laxiflora
57b. Basal lobe of lateral united petal orbicular, spur straight, abruptly constricted……….…57) I. sunkoshiensis
Updated checklist of Balsaminaceae in Nepal
Impatiens arguta Hook. f. & Thomson, J. Linn. S. B. 4: 137 (1860).
Type: Hooker J.D. s.n. (lecto, designated in [26] K K000694618!), Sikkim Himalaya, Darjeeling, 7500 ft.; Hooker J.D. s.n. (syn, K K000694619!), Darjeeling.
Impatiens gagei Hook. f. Hooker’s Icon. Pl. 30: t. 2951 (1911).
Type: Burkill 27744 (lecto, designated in [26], K K000694617!), Eastern Himalaya, Darjeeling, Tonglo slopes, below Simana, at an elevation range of 6000–8000 ft.
Habitat: Temperate forest at an elevation range of 2100–2900 m.
Distribution: Eastern Nepal, China, India and Myanmar.
Specimen examined: Solukhumbu district, Naamkhali, 2240 m, 21 Jul. 1995, Miyamoto F. et al. 9592063 (KATH); Sankhuwasabha district, Arun Valley, Maghang Khola, 2550 m, 01 Jul. 1956, Stainton J.D.A. 817 (TI); Nigale, 5500 ft., 11 Sept. 1964, Banerjee, Shrestha & Upadhya 2633 (US); Phaplos, 8000–9000 ft., 1930, Lall Dhwoj 79 (E); Tinjure Danda, 2400 m, 06 Sept. 1967, Williams & Stainton 8402 (TI); Dhankuta district, Bilbatay Bhanjang, 2200 m, 25 Oct. 1963, Hara H. et al. (TI); Dhankuta district, Chitre, 2400 m, 07 Jul. 1972, Kanai H. et al. 721162 (TI); Dhankuta district, Shidua-Tute, 13 Jul. 1991, Ohba H. et al. 9120029 (TI); Panchthar district, Prangbung-Bhanduke, 2300 m, 25 Jun. 1992, Noshiro S. et al. 9241070 (TI); Ramechhap district, Bhandar-Sete, 1880 m, 29 Jul. 1997, Wakabayashi M. et al. 9710032 (TI); Rasuwa district, Syabru-Dhunche, 2300 m, 25 Jul. 1992, Takayama H. et al. 9241088 (TI); Solukhumbu district, Bandar-Namkhli, 20 Jul. 1995, Miyamoto F. et al. 9584017 (TI); Solukhumbu district, Fera-Nunthala, 2780 m, 26 Jul. 1995, Miyamoto F. et al. 9596393 (TI).
A, B. Impatiens arunensis; C. Impatiens wallichii, D. Impatiens gammiei. Photographs by Bhakta Bahadur Raskoti.
Type: Grey-Wilson et al. 4376 (holo, K K000694616!), Nepal, Arun Khola, Upper Kashwa Khola, above Hedangha, elevation 2800 m, 08 Sept. 1981.
Habitat: Temperate forest at an elevation about 2800 m.
Distribution: Eastern Nepal (endemic).
Specimen examined: Arun khola, upper kashwa khola, above Hedangha, 2800 m, 08 Sept. 1981, Bhandary & Taylor 4376 (K); Sankhuwasabha district, Arun khola, banking, Sekidim, 2600 m, 15 Aug. 1981, Grey-Wilson, Christopher and Henderson 4376 (E).
Note: Impatiens arunensis is morphologically similar to Impatiens harae; a comprehensive study is necessary for the taxonomic identity of Impatiens arunensis.
Impatiens bajurensis S. Akiyama & H. Ohba, Journ. Jap. Bot. 68: 157 (1993).
Type: Suzuki M. et al. 9171054 (holo, TI!; iso KATH!), Far West Nepal, Bajura district, Kaudegaon-Babali, 1520 m, 29 Aug. 1991.
Habitat: Subtropical forest at an elevation of 1520 m.
Distribution: Western Nepal (endemic).
Specimen examined: Doti district, Siligarhi, 4500 ft., 01 Apr. 1967, Ecker-Racz N. 38 (US); Between Munigaon and Chutta, S.E. of Jumla, 9500 ft., 24 Jul. 1952, Polunin, Sykes & Williams 4901 (E); Mugu district, Kamali Valley, near Lumsa, 7000 ft., 14 Aug. 1952, Polunin, Sykes & Williams 5186 (E); Mugu district, Ratuligaon, Bis Ram 350 (NY); Mugu district, Samla, 7000 ft., 04 Oct. 1952, Polunin, Sykes & Williams 5560 (E).
Impatiens balsamina L., Sp. Pl. 938 (1753).
Type: Anonymous s.n. (lecto, designated in [27], LINN-HL1053.5).
Balsamina cornuta (L.) DC., Prodr. 1: 686 (1824).
Type: Paul Hermann 3:9, n. 316 (lecto, designated in [28], BM BM000621820), Sri Lanka.
Impatiens coccinea Sims, Bot. Mag. 31: t. 1256 (1810).
Type: Sims (1810) t.1256 (lecto, designated in [26], illustration of specimen cultivated in Sloane street by Mr. Salisbury, seeds from Mr. Roxburgh, east India.
Type: Duthie’s collector (Inayat) s.n. (lecto, designated in [29], upper specimen of K K000694732!; isolecto DD, lower specimen of K K000694732!), India, Western Himalaya, Uttarakhand, Kumaon, 1900.
Impatiens edgeworthii var. toppinii Hook. f. ex Toppin, Bull. Misc. Inform. Kew 1920: 348 (1920).
Type: Toppin S.M. 605 (lecto, designated in [29], larger left hand side specimen of K K000694738!; isolecto, smaller right hand side specimen of K K000694738!), Pakistan, Chitral, Ziarat, 7000–8000 ft., Aug. 1908.
Impatiens pallens Edgew., Trans. Linn. Soc. London 20(1): 39 (1846).
Type: Edgeworth M.P. 334 (lecto, designated in [29], left hand side specimen of K K000694729!; isolecto right hand side specimen of K K000694729!), India, Uttarakhand, Garhwal, 4000–8000 ft., 1844.
Type: Burtt B.L. & Kazmi M.A. 1253 (holo, E E00841614!), Pakistan, Hazara, Hill territory, 7500 ft., 18 Sept. 1958.
Impatiens umbrosa Edgew., Trans. Linn. Soc. London 20(1): 39 (1846).
Type: Edgeworth M.P. 334 (lecto, designated in [29], larger left hand side specimen of K K000694727), India, Himachal Pradesh, Kangra, 3000–8000 ft., 1844.
Impatiens nepalensis Hook. f., Rec. Bot. Surv. India 4: 14, 20. 1905 & Hooker’s Icon. Pl. 30: t. 2961 (1911).
Type: Wallich cat. no. 4729 (designated in [29], K K001039776), Nepal, 1821.
Habitat: Lower temperate forest at an elevation range of 2100–2400 m.
Distribution: Western Nepal, India and Pakistan.
Specimen examined: Dhaulakot, 7500 ft., 18 Aug. 1952, Polunin, Sykes and Williams 498 (E).
Impatiens bicornuta A. Flowering plant; B. Front view of flower; C. Flower (side view). Photographs by Bhakta Bahadur Raskoti.
Type: Wallich 4765 (lecto, designated in [30], K, two specimens K001039849! and K001039850!), Nepal; Wallich cat. no. 4765 (lecto, designated in [29], K K001039849!), Nepal, Chandaghery, Jun. 1821.
Impatiens bicornuta var. micrantha H. Hara, Fl. E. Himalaya 3rd Rep. 78 (1975).
Type: Kannai H., Hara H. & Ohba H. 723214 (lecto, designated in [29], TI TI00013971!), Nepal, Rasuwa district, Dunche-Trisuli Khola-Singum Gompa, 22 Aug. 1972.
Habitat: Subtropical to temperate forest at an elevation range of 1600–3000 m.
Distribution: Eastern, Central, Western Nepal, China and India.
Specimen examined: Ramechhap district, Sivalaya, 1800 m, 17 Aug. 1995, Ohaba H. et al. 61345 (KATH); Manang district, Chame, 12 Aug. 1983, Manandhar N.P. 977 (KATH); Lalitpur district, Phulchoki, 2200–2700 m, 09 Aug. 1969, Kani H. 673398 (KATH); Thade, 2140 m, 18 Jul. 1971, Shakya P.R. & Adhikari 597 (TI).
Type: Hooker & Thomson 56 (lecto, designated in [32], K K000694682!; isolecto K K000694683!, K K000694684!, K K000694685!, K K000694686!, K K000694687!, L L0388881), India, Khasia Hills, 06 Jul. 1850, 6000 ft.
Habitat: Temperate forest at an elevation of 2000–2200 m.
Distribution: Nepal, China, India and Myanmar.
Impatiens edgeworthii Hook. f., Fl. Brit. Ind. 1: 476 (1875); in Rec. B. Surv. Ind. 4: 5 & 9 (1904).
Type: Edgeworth 1067 (holo, K!), India, Kundau and Beas Valley, shady and open places, 1800–3000 m.
Habitat: Temperate forest at an elevation range of 2000–3000 m.
Distribution: Central Nepal, India and Pakistan.
Specimen examined: Kaski district, Panchase forest, 2340 m, 09 Sept. 2014, Bhandari P. & Budhamagar S. P832 (KATH).
Type: Burkill I. H. 27689 (neo, designated in [29], K K000694668!), Darjiling, Sabarkum, 11000 ft., 01 Oct. 1906.
Impatiens minimiflora Hook. f. Rec. Bot. Surv. India 4: 16, 21 (1905).
Type: Clarke C.B. 9874 B (lecto, designated in [29] K K000694788!), Sikkim, Yakla, 11000 ft., 15 Oct. 1869.
Habitat: Temperate forest at an elevation of 3000–3600 m.
Distribution: Eastern Nepal and India
Specimen examined: Taplejung district, 3400 m, 08 Sept., 2011, Bhakta B. Raskoti 201079 (KATH).
Note: Impatiens gammiei is very close to Impatiens radiata in overall morphology; only a notable difference is spurless lower sepal in former. More robust study is necessary for the confirmation of taxonomic identity of Impatiens gammiei.
Impatiens glandulifera Royle, Ill. B. Him. t. 28, f. 2 (1834); 151 (1835). Fig 13.
Type: Mts. t. 28, f. 2. (designated in [32], Illustration Royle, Ill. Bot. Himal), 1835.
Habitat: Temperate forest at an elevation of 3000 m.
Distribution: Western Nepal, India and Pakistan.
Specimen examined: Harigaun, 3000 m, 12 Jul. 1952, Polunin O., Sykes W.R. & Williams L.H.J., 143 (TI).
Impatiens glauca Hook.f. & Thomson, J. Linn. Soc. Bot. 4: 155. (1860).
Type: Strachey & Winterbottom 14 (holo, K K000694742!), India, Kumaon ad Dwali, 9500 ft.
Habitat: Temperate forest at an elevation range of 2200–3000 m.
Distribution: Western Nepal and India.
Specimen examined: Bajura district, Damkane-Jilli, 1700 m, 13 Aug. 1991, Rajbhandari K.R. 14957 (KATH); Bajura district, Damkane, 2300 m, 13 Aug. 1991, Suzuki M., Hatta H., Kurosaki N., Mikage M., Miyamoto F., Rajbhandari K.R. et al. 70555 (TI); Bajura district, Porakya-Serigaon, 2200 m, 13 Aug. 1991, Suzuki M. et al. 9170555 (KATH); Below Garjigoth, Dori Lekh, N.W. of Jumla, 10000 ft., 09 Aug. 1952, Polunin O., Sykes W.R. & Williams L.H. 5052 (E); Ratamata Chakure Lekh, 10000 ft., 12 Aug. 1952, Polunin O., Sykes W.R. & Williams L.H. 405 (E).
Impatiens gorepaniensis A. Flowering plant; B. Flower (side view); C. Lateral sepal; D. Dorsal sepal; E. Lateral united petal; F. Lower sepal; G. Capsule. Photographs by Bhakta Bahadur Raskoti.
Type: Grey-Wilson, Phillips C. 868 (K K000694667!), Nepal, west of Ghorepani, 10 Sept. 1973.
Habitat: Temperate forest at an elevation of 2000 m.
Distribution: Western Nepal (endemic).
Specimen examined: West of Gorepani, 2000 m, 10 Sept. 1973, Grey-Wilson and Philips 868 (K).
Note: Impatiens gorepaniensis is morphologically similar to Impatiens cymbifera; further study including morphology and molecular data is required for the taxonomic identity of Impatiens gorepaniensis.
Impatiens graciliflora Hook.f., Rec. Bot. Surv. India 4(2): 15 (1905).
Type: Pantling 64I (lecto, designated in [34] K K000694665!), India, Sikkim, Lenchul, 6000 ft., Aug. 1903.
Habitat: Temperate forest at an elevation range of 2000–2500 m.
Distribution: Eastern Nepal, Bhutan, India and Myanmar.
Specimen examined: Ilam district, Maipokhari, 4500 ft., Collector name not mention in sheet 15425 (KATH); Panchther district, Parangbung-Namle Phedi, 1700–2000 m, 25 Jun. 1992, Noshiro S., Akiyama S., Acharya N. 9241084 (TI).
Note: Based on morphological characters; Impatiens graciliflora is close to Imapties radiata; further study is necessary for the confirmation of taxonomic identity of Impatiens graciliflora.
Impatiens harae H. Ohba & S. Akiyama, Journ. Jap. Bot. 62 (12): 368 (1987). Fig 15A.
10.1371/journal.pone.0274699.g015
A, Impatiens harae; B. Impatiens urticifolia. Photographs by Bhakta Bahadur Raskoti.
Type: Ohba H. et al. 8530680 (holo, TI!), Nepal, Ramechhap district, Neju-Choarma, 02 Aug. 1985.
Habitat: Temperate forest at an elevation range of 2000–2500 m.
Distribution: Eastern, Central Nepal and China.
Specimen examined: Ramechhap district, Neju-Choarma, 3651–2760 m, 02 Aug. 1985, Ohba H. et al. 61078 (KATH); Ramechhap district, Choarma, 2760 m, 02 Aug. 1985, Ohba H., Wakayabashi M., Suzuki M., Kurosaki N., Rajbhandari K.R. and Wu S.K. 8530680 (TI).
Impatiens hobsonii Hook. f., Rec. B. Surv. Ind. 4: 15 & 20 (1905), ’hobsoni’. H. Hara in H. Hara & Williams, Enum. Flow. Pl. Nep. 2: 79 (1979). Fig 8D.
Type: Hobson H. E. s.n. (lecto, designated in [29], K K000694824!), India, Sikkim, Lachen valley, Yatung, 1897.
Habitat: Temperate forest at an elevation range of 3300–3500 m.
Impatiens infundibularis A. Flowering plant; B. Flower (front view); C. Flower (side view); D. Capsule and seeds. Photographs by Bhakta Bahadur Raskoti.
Type: Hooker J.D. s.n. (lecto, designated in [29], left hand side specimen of K K000694661!, India, Sikkim, 2000–5000 ft.; isolecto CAL, right hand side specimen of K K000694661).
Habitat: Sub-tropical forest at an elevation of 1400 m.
Distribution: Eastern Nepal, Bhutan and India.
Specimen examined: Ilam district, Kolbung, 1400 m, Bhakta B. Raskoti 20121 (KATH).
Habitat: Tropical to temperate forest at an elevation range of 900–2000 m.
Distribution: Eastern, Central Nepal and India.
Specimen examined: Taplejung district, Heydewa-Garhi danra, 03 Nov. 1963, Hara H. et al. 6300502 (KATH); Kule khani, 1450 m, 02 Sept. 1970, Kani H. & Shrestha T.B. 672705 (KATH); Baju danda to Tamor river, 1550 m, 31 Aug. 1989, Kew-Edinburgh-Kathmandu Expedition to N.E. Nepal 143 (E); Terhathum district, 1650 m, 11 Sept. 1967, Williams & Stainton 8453 (TI); Kathmandu district, Sundarijal Waterfall, 1600 m, 20 Sept. 1963, Hara H. et al. 6306724 (TI); Lalitpur district, Godavari, 1500 m, 18 Sept. 1963, Hara H. et al. 6306723 (TI); Phulchoki, 1800 m, 28 Aug. 1969, Miss Manandhar & party 12915 (TI); Makwanpur district, Okhre Danra–Deorali, 1600 m, 02 Sept. 1970, Kanai H. & T.B. Shrestha 672705 (TI); Sankhuwasabha district, Dumhan, 1200 m, 30 Oct. 1963, Hara H. et al. s.n. (TI); Sindhupalchok district, Charikot–Mokaibari, 30 Sept. 1970, Kanai H. et al. 674699 (TI); Taplejung district, Dumhan–Taplejung, 1400 m, 01 Nov. 1963, Kanai H. et al. 6306725 (TI); Papung-Sangrati Pati, 1500 m, 26 Aug. 1977, Ohashi H. et al. 772736 (TI); Taplejung district, Heydewa-Garhi Danra, 03 Nov. 1963, H. Hara et al. 6300502 (TI); Taplejung district, Tuwa-Taplethok, 05 Nov. 1963, Hara H. et al. s.n. (TI).
Impatiens jurpia Buch.-Ham., J. Proc. Linn. Soc., Bot. 4: 140 (1859). Fig 8A.
Type: Wallich Herb. no. 4761 (lecto, designated in [35], A, K-W K001039839!), Nepal, Morang Hills, 14 Jul. 1810.
Habitat: Subtropical forest at an elevation range of 1500–1800 m.
Distribution: Eastern, Central Nepal, India and Myanmar.
Specimen examined: Lalitpur district, Phulchoki, 2200–2700 m, 09 Aug. 1969, Kani H. 673373 (KATH); Gorkha district, Dovan-Jagat, 1220 m, 27 Jul. 2007, Ikeda H., Kawahara T., Yano O., Watson M.F., Li Z.H., Subedi M.N., et al. 20816035 (KATH); Sanguri Bhanjang-Dhara Pani, Hara H. et al. 6300490 (TI); South of Hatora, 700 m, Grey-Wilson & Philllips 67A (K).
Impatiens kharensis S. Akiyama, H. Ohba & Wakab., Himal. Pl. 2: 75 (1991).
Type: Ohba H. et al. 8320702 (holo TI!, iso KATH!), Nepal, Khare Khola, Bitta Kharka (4100 m) -a valley (3300 m)-Patale Pokhari, 4000 m, 12 Sept. 1983.
Habitat: Temperate forest to alpine slopes at an elevation range of 2500–4000 m.
Distribution: Central Nepal (endemic).
Specimen examined: Dolakha district, Vitte kharka-Valley patale pokhari, 3300–4000 m, 12 Sept. 1983, Ohba H., Wakabayashi M., Suzuki M., Akiyama S. 8320702 (TI); Dolakha district, Vitte kharka-Valley patale pokhari, 3300–4000 m, 12 Sept. 1983, Ohba H., Wakabayashi M., Suzuki M., Akiyama S. 8341278 (TI).
Impatiens laevigata Wall. ex Hook. f. & Thomson, J. Linn. Soc. Bot. 4: 146 (1860).
Habitat: Subtropical forest at an elevation range of 1000–1500 m.
Distribution: Nepal, India and Myanmar.
Note: Though, this speicies is included in the literature [8], authors are unable to track authentic specimen of Impatiens laevigata from Nepal. Further study is necessary for the existence of this species within Nepal.
Impatiens laxiflora Edgew., Tr. Linn. S. 20: 40 (1846). Fig 18.
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Impatiens laxiflora A. Flowering plant; B. Flower (front view). Photographs by Bhakta Bahadur Raskoti.
Type: Edgeworth M.P. 399 (holo, K K000694834!), India, Serain, 8000–10000 ft., 1844.
Habitat: Temperate to subalpine forest at an elevation range of 2700–3900 m.
Distribution: Eastern, Central, Western Nepal, China and India.
Specimen examined: Lumsa north west of Jumla, 9500 ft., 10 Aug. 1952, Stainton, Sykes & Williams 5117 (E); Barabar, 5000 ft., 20 Aug. 1935, Bailey’s F.M. (BM); Bajhang district, Khaptad National Park, 2900 m, 27 Aug. 1991, Suzuki M. et al. 9170978 (TI); Porakya-Serigaon, 2200 m, 13 Aug. 1991, Suzuki M. et al. 9170552 (TI); Jumla district, Below Gajigoth, Dori Sekh, 10000 ft., 09 Aug. 1952, Polumin, Sykes & Williams 5054 (BM); Rukum district, East of Chalike Pahar, 05 Aug. 1954, Stainton, Sykes & Williams 3755 (E); Sankhuwasabha district, Barun khola, Sana Pokhari, 3300 m, 22 Aug. 1981, Grey-Wilson, Henderson, Rajbhandary & Taylor 4149 (K); Sankhuwasabha district, Barun khola, W. of Yangle, 3500 m, 22 Aug. 1981, Grey-Wilson, Henderson, Rajbhandary & Taylor 423 (K).
Habitat: Temperate to lower belt of subalpine zone at an elevation ranges of 2600–3600 m.
Distribution: East Nepal and India.
Specimen examined: Sankhuwasabha district, north of sando pokhari near barun khola, 3400 m, 22 Aug. 1981, Grey-Wilson Henderson 4141 (E).
Note: This species is not well documented; it was collected from Phalut (Darjeeling, India), which is close to the boarder of eastern Nepal. Literature [36] included a photograph of Impatiens leptocarpa taken in Ramitay Odaar, Sankhuwasabha, Nepal. We recommend its further study and collections from Nepal.
Type: Gammie 414 (lecto, designated in [33] K K000694783!), India, Sikkim, 12000–13000 ft.
Habitat: Temperate forest and alpine grasslands at an elevation range of 3350–4000 m.
Distribution: Eastern, Central Nepal, China and India.
Specimen examined: Solukhumbu district, Randu Kharka, 3350 m, 09 Aug. 1997, Rajbhandari K.R. 9740196 (KATH); Ramechhap district, Neju, 3651 m, 31 Jul. 1985, Ohba H. & Wu S.K. 8580614 (KATH); Solukhumbu district, Pike dongshar, 3600 m, 09 Sept. 1985, Botanical Expedition to Himalaya (1985) 8531381 (E), Rasuwa district, Singum gumpa-Gosaikunda, 3900 m, 23 Aug. 1972, Hara H. et al. 721860 (TI); Ramechhap district, Neju, 3651 m, 31 Jul. 1985, Botanical Expidition to Himalaya (1985) (BM).
Impatiens porrecta Wall. ex Hook. f. & Thomson, J. Linn. S. B. 4: 138 (1860).
Type: Wall. Cat. no. 7275 (lecto, designated in [26], K K001127061!), Sylhet.
Impatiens bella Hook. f. & Thomson, J. Proc. Lin. Soc. Bot. 4(15):138 (1859).
Type: Griffith s.n. (lecto, designated in [26], K K000694775!), Khasia.
Habitat: Temperate forest at an elevation of 3000 m.
Distribution: Nepal, India and Manipur.
Impatiens pradhanii H. Hara, J. Jap. Bot. 40: 99 (1965).
Type: Pradhan K.C. & Hara H. 6306729 (holo TI!), India, Sikkim, Lagyap, 13 Sept. 1964.
Habitat: Temperate forest at an elevation ranges of 2000–3200 m.
Distribution: Central Nepal and India
Specimen examined: Gorkha district, Ghap to Namrung, 2280 m, 31 Jul. 2008, Ikeda H. et al. 20811094 (KATH); Dolakha district, Jagat-Samagaun, 2030 m, 15 Oct. 2007, Noshiro et al. 20710078 (KATH); Manang district, Marsyandi Khola Chame, 2630 m to Dhanagyang, 2200 m, 06 Aug. 1983, Ohba H. et al. (TI); Dolakha district, Rolwaling Khola: Simigaon, 1950 m to Kyalche, 2700 m, 31 Aug 1983, Ohba H. et al. (TI); Kyalche, 2700 m to Thandingma, 3200 m, 01 Sept. 1983, Ohba H. et al. (TI).
Type: Prain 377 (lecto, designated in [37], K K000694764!; isolecto K! sheet 1 with illustration, K sheet 2), India, eastern Himalaya, Sikkim, Lachung Valley, near Choongtang.
Impatiens mallae S. Akiyama & H. Ohba, Journ. Jap. Bot. 67: 187 (1992).
Type: Suzuki et al. no 8821106 (holo, TI!; iso, KATH!), Nepal, Koshi, Sankhuwasabha, Tashi Gaun, 2160–3500 m.
Habitat: Temperate forest at an elevation range of 2100–3500 m.
Distribution: Eastern Nepal, China and Myanmar.
Specimen examined: Solukhumbu district, Tasigaun-Kauma, 2590 m, 25 Sept., 1991, Long D.G. et al. 241 (KATH).
Habitat: Tropical to temperate forest at an elevation range of 1500–2700 m.
Distribution: Eastern, Central Nepal, Bhutan, China and India.
Specimen examined: Ramechhap district, Choarma, 2760 m, 03 Aug. 1995, Ohba H. et al. 61154 (KATH); Sankhuwasabha district, ArunValley, Chhoyang Khola, 2550 m, Stainton J.D.A. 699 (TI); Lalitpur, Phulchoki, 1800 m, 27 Aug. 1965, Shilling A. D. 618 (TI); Taplejung district, Yamphodin, 2100 m, 25 Jun. 1969, Williams L. H. J. 941 (TI); Dhankuta district, Sedua, 1740 m, 15 Sept. 1971, Ohsawa M. & Shakya P.R. 1799 (TI); Terhathum district, Tute-Tinjure Phedi, 14 Jul. 1991, Ohba H. et al. 9120057 (TI); Terhathum district, Tinjure Danda, 7500 ft., 06 Sept. 1967, Williams & Stainton 8378 (TI); Dolakha district, Jiri-Sibalaya, 2250 m, 08 Jul. 1995, Miyamoto F. et al. 9596013 (TI); Kaski district, Banthanti-Ghodepani Deurali, 2720 m, 24 Aug. 1988, Suzuki M. et al. 8842103 (TI); Kaski district, Ghandruk–Bhainsi Kharka 2160 m, 22 Aug. 1988, Suzuki M. et al. 8860585 (TI); Kathmandu district, Sheopuri, 6500 ft., 19 Aug. 1954, Stainton, Sykes & Williams 6936 (E); Lalitpur district, Phulchoki, 09 Aug. 1969, Kanai H. 673348 (TI); Myagdi district, Chitre, 2150 m, 27 Aug. 1994, Mikage M. et al. 9485557 (TI); Ghar Khola, 3000 m, Stainton, Sykes & Williams 5777 (E); Panchthar district, Prang–Goruwale Bhanjang, 25 Jun. 1992, Noshiro S. et al. 9241085 (TI); Sindhupalchok district, Thale–Thale Bisauna, 2500 m, 10 Sept. 1970, Kanai H., Ch. Chuma & T. Nagano s.n. (TI); Solukhumbu, Nunthala–Junbesi, 2620 m, 31 Aug. 1997, Wakabayashi M. et al. 9710412 (TI); Solukhumbu, Nunthala-Kharikhola, 1670 m, 27 Jul. 1995, Miyamoto F. et al. 9596158 (TI); Taplejung district, Shewaden-Mewa Khola, 2300 m, 29 Jun. 1972, Kanai H. et al. 720947 (TI).
Impatiens pulchra Hook. f. & Thomson, J. Linn. S. B. 4: 139 (1860).
Type: Hooker J. D. & Thomson T. 2281 (lecto, desiginated in [26], K K000694754!; isolecto, K K0006947530!), India, Khasia, elevation 4000–5000 ft.
Habitat: Temperate forest at an elevation range of 2400–2700 m.
Distribution: Eastern Nepal, China and India.
Specimen examined: East Nepal, Moyong valley, 4000–5000 ft., Hooker J.D. s.n. (K).
Type: Pantling s.n. (lecto, designated in [26] with illustration by Hooker, K), Sikkim, 6000 ft.
Habitat: Tropical to sub-alpine forest at an elevation range of 1300–3900 m.
Distribution: Eastern, Central, Western Nepal, Bhutan, China, India, Myanmar and Thailand.
Specimen examined: Dolakha district, Lamabagar-Hum, 2000 m, 16 Jul. 1977, Rajbhandari K.R. & Roy B. 1451 (KATH); Bajura district, Pategaon–Badigaon, 2700 m, 16 Aug. 1991, Suzuki M. et al. 9170710 (TI); Dhankuta district, Chauke-Tute, 2600 m, 01 Sept. 1977, Ohashi H. et al. 771266 (TI); Dhankuta district, Chitre, 2300 m, 05 Jun. 1972, Kanai H. et al. 720200 (TI); Gorkha district, Ekle Ghar-Sardu Khola, 2000 m, 25 Jul. 1994, Suzuki M. et al. 9470180 (TI); Gorkha district, Sardu Khola-Ripche, 2060 m, 27 Jul. 1994, Suzuki M. et al. 9480052 (TI); Kaski, Banthanti-Ghodepani Deurali, 2700 m, 24 Aug. 1988, Suzuki M. et al. 8881302 (TI); Kaski district, Banthati, 2360 m, 28 Aug. 1994, Mikage M. et al. 9485568 (TI); Pothana-Tolka, 1800 m, 20 Aug. 1988, Suzuki M. et al. 8881086 (TI); Kathmandu district, Shiwapuri Summit–Borlang Bhanjyng, 2732 m, 25 Aug. 1990, Minaki M. et al. 9011018 (TI); Lalitpur district, Phulchoki, 19 Sept. 1970, Kanai H. & Ch. Chuma 675184 (TI); Makwanpur district, Kuli Khani, 1450 m, 03 Sept. 1970, Kanai H. & T.B. Shrestha 672733 (TI); Manang district, Koto, 2440 m, 14 Aug. 1994, Mikage M. et al. 9485418 (TI); Mustang district, Kalopani–Tatopani, 1870 m, 23 Aug. 1999, Mikage M. et al. 9961317 (TI); Myagdi district, Shika–Narchang, 2200 m, 26 Aug. 1988, Suzuki M. et al. 8881498 (TI); Panchthar district, Gairi Kharka–Beteni, 21 Jun. 1992, Noshiro S. et al. 9240982 (TI); Rasuwa district, Dhunche–Grang, 2000 m, 01 Aug. 1995, Hoshino T. et al. 9539277 (TI).
Impatiens scullyi A. Flowering plant; B. Flower (Front view); C. Flower (side view). Photographs by Bhakta Bahadur Raskoti.
Type: Scully J. 272 (holo, K000694939!), Central Nepal.
Habitat: Temperate forest at an elevation of 2300 m.
Distribution: Eastern, Central, Western Nepal (endemic).
Specimen examined: Manang district, Thanchok-Dhangyung, 2400 m, 06 Aug. 1983, Rajbhandari K.R. 8993 (KATH); Bajhang district, Jimkot-Meltadi-Khaptad National Park, 1650 m, 26 Aug. 1991, Suzuki M. et al. 9170957 (TI); Bajura district, Kaudegaon-Babali, 1500 m, 29 Aug. 1991 Suzuki M. et al. 9171053 (TI); Kaski district, Ghandruk-Bhainsi Kharka, 2250 m, 22 Aug. 1988, Suzuki M. et al. 8881141 (TI); Manang district, Danake, 14 Aug. 1994, Mikage M. et al. 9485411 (TI); Mustang district, Kabre-Kalopani, 2300 m, 28 Aug. 1988, Suzuki M. et al. 8881529 (TI); Myagdi district, Shika-Narchang, 1460 m, 26 Aug. 1988, Suzuki M. et al. 8881450 (TI); Rasuwa district, Dhunche-Syabru, 29 Aug. 1991, Noshiro S. 9154506 (TI); Lingju-Tibling, 2100 m, 13 Aug. 1994, Miyamoto F. et al. 9420261 (TI).
Impatiens serrata Benth. ex Hook. f. & Thomson, J. Linn. Soc. Bot. 4: 136 (1860). Fig 25.
10.1371/journal.pone.0274699.g025
Impatiens serrata A. Flowering plant; B. Flower (Front view). Photographs by Bhakta Bahadur Raskoti.
Type: Wallich cat. no. 4771 (lecto, designated in [29], larger middle specimen of K K001039866!; isolecto other specimens of CAL, K K001039866!), Nepal, 1821.
Habitat: Temperate forest at an elevation range of 2000–3600 m.
Distribution: Central, Western Nepal, Bhutan and China.
Specimen examined: Lamjung district, Tarma-Qubara, 3528 m, 03 Sept. 2021, Thapa T.K. & Magar R. 20210939 (KATH); Bajhang district, Jimkot-Meltadi-Khaptad National Park, 2850 m, 26 Aug. 1991, Suzuki M. et al. 9170959 (TI); Kalikot district, Panipokhari–Chaukebada, 2800 m, 04 Aug. 1991, Suzuki M. et al. 9170231 (TI); Rasuwa district, Chyauche Kharka-Lingju, 3580 m, 12 Aug. 1994, Miyamoto F. et al. 9420242 (TI); Rasuwa district, Deolari–Sing Gomba, 22 Jul. 1995, Hoshino T. et al. 9539060 (TI); Sindhupalchok district, Jangdang Kharka-Tingoang, 3100 m, 24 Sept. 1970, Kanai H. et al. 672949 (TI); Sindhupalchok district, Thale–Thale Bisauna, 2600 m, 10 Sept. 1970, Kanai H., Ch. Chuma & T. Nagano 675182 (TI).
Impatiens stenantha A. Flowering plant; B. Flower (Front view); C. Flower (side view). Photographs by Bhakta Bahadur Raskoti.
Type: Hooker J.D. s.n. (lecto, designated in [29], larger upper specimen of K K000694611!, isolecto CAL, G00237315, smaller lower specimen of K K000694611!), India, Sikkim, 6000–8000 ft.
Habitat: Subtropical to temperate forest at an elevation range of 1800–2600 m.
Distribution: Eastern Nepal, Bhutan, China and India.
Specimen examined: Solukhumbu district, Bhotebas-Chichila, 1940 m, 20 Sept. 1991, Long D.G. et al. 108 (KATH); Dhankuta district, above Sedua, 7400 ft., 15 Aug. 1975, Beer L.W. 25308 (BM); Sedua-Thokpu, 2100 m, 25 Sept. 1971, Shakya P.R. & Joshi D.P. 919 (TI); Sankhuwasabha district, Tashi Gaun, 2250 m, 13 Jul. 1988, Suzuki M. et al. 8880424 (TI); Sankhuwasabha district, Arun Valley, Kasuwa Khola, 8500 ft., 11 Jun. 1956, Stainton J.D.A. 607 (BM); Panchthar district, Beteni-Chyangthapu, 2000 m, 22 Jun. 1992, Noshiro S. et al. 9240997 (TI); Panchthar district, Chyangthapu-Dabale Deurali, 2500 m, 23 Jun. 1992, Noshiro S. et al. 9241022 (TI); Taplejung district, Khokling-Thunglung, 1620 m, 12 May 1992, Suzuki M. et al. 9240047 (TI); Taplejung district, Mewa Khola-Papung, 2500 m, 29 Jun. 1972, Kanai H. et al. 720900 (TI).
Impatiens gigantea Edgew., Trans. Linn. Soc. London 20(1): 28 (1846).
Habitat: Subtropical forest to alpine grassland slopes at an elevation range of 1700–4100 m.
Distribution: Central, Western Nepal, Bhutan, China, India and Pakistan.
Specimen examined: Lamjung district, Thokyo-Nai Sima, 3429 m, 03 Sept. 2021, Thapa T.K. & R. Magar 20210921 (KATH); Bajhang district, Khaptad National Park, 2900 m, 27 Aug. 1991, Suzuki M. et al. 9170984 (TI); Bajura district, Berseni–Porakya, 2650 m, 12 Aug. 1991, Suzuki M. et al. 9170486 (TI); Dolakha district, Charikot-Kalinchok, 9500 ft., 16 Sept. 1964, Banerjee, Shrestha & Upadhya 2783 (US); Dolpa district, Balanga Pass, 11000 ft., 19 Jul. 1950, Polunin, Sykes & Williams 2512 (E); Dolpa district, Jangla Banyang, 3800 m, 13 Jul. 1973, Einarsson S., Skärby L. & Wetterhall B. 2276 (BM); Ilam district, Samtar, 12000–13000 ft., Lall Dhwoj 303 (E); Jumla district, Bhurchula Lehk, near Jumla, 12000 ft., 14 Jul. 1952, Polumin O., Sykes W.R. & Williams L.H.J. 4685 (BM); Kalikot district, Panipokhari-Chaukebada, 2680 m, 04 Aug. 1991, Suzuki M. et al. 9170232 (TI); Kaski district, Banthanti–Ghorapani, 3100 m, 24 Aug. 1988, Suzuki M. et al. 8881319 (TI); Manang district, Khangsar, 12500 ft., 15 Jul. 1950, Stainton, Sykes & Williams 1194 (E); Manang district, Gunsang–Yak Kharka, 3720 m, 17 Aug. 1994, Mikage M. et al. 9470461 (TI); Mugu district, Ghurchi Lekh, between Lumra and Murma, 11000 ft., 11 Aug. 1952, Polunin O., Sykes W.R. & Williams L.H.J. 5121 (BM); Mustang district, Ghemi, 3520 m, 17 Aug. 2002, Miyamoto F. et al. 20230087 (TI); Mustang district, Ghenkar Gompa-Ghemi, 04 Aug. 2002, Watanabe T. et al. s.n. (TI).
Impatiens sunkoshiensis S. Akiyama, H. Ohba & Wakab., Himal. Pl. 2: 77 (1991). Fig 30.
10.1371/journal.pone.0274699.g030
Impatiens sunkoshiensis A. Flowering plant; B. Flower (Front view). Photographs by Bhakta Bahadur Raskoti.
Type: Ohba H. et al. 8520277 (holo, TI), Sagarmatha Zone, Sete-Taktor, 2550–3000 m, 19 Aug. 1985.
Habitat: Temperate forest at an elevation range of 2500–3200 m.
Distribution: Central Nepal (endemic).
Specimen examined: Gorkha distict, Ghopte to Namrung, 2240 m, 32 Jul. 2008, Ikeda H., Kawahara T., Yano O., Watson M.F., Li Z.H., Subedi M.N. et al. 20811095 (KATH).
Note: Impatiens sunkoshiensis is morphologically similar to Impatiens laxiflora but molecular analysis [6] indicated that these two taxa are two distinct species.
Impatiens tricoris A. Flowering plant; B. Side view of flower; C. Front view of flower; D. Capsule. Photographs by Bhakta Bahadur Raskoti.
Type: Illustration, t. 7 published in Bot. Reg. 26 (1840), (lecto, designated in [39]).
Impatiens praetermissa Hook.f., J. Linn. Soc. Bot. 37: 29 (1904).
Type: Wallich s.n. (not found).
Impatiens cristata auct. non Wall.: Hook. f., Rec. Bot. Surv. India 4: 7 & 10 (1904); 22 (1905). Grey-Wilson, in Grierson and Long (eds.), Fl. Bhutan 2: 92, Fig 12a–12c (1991).
Distribution: Central, Western Nepal, China, India, Pakistan
Habitat: Subtropical to temperate forest and forest margins.
Specimen examined: Dadeldhura district, 2000 m, 05 May 1971, Shakya P.R. & Joshi D.P. 479 (TI); Bajhang district, Chainpur-Jimkot, 1200 m, 25 Aug. 1991, Suzuki M. et al. 9170946 (TI); Dolakha district, Gungur Khpla-Jagat, 1120 m, Noshiro S. et al. 20710213 (TI); Gorkha district, Syaule-Jagat, 1000 m, 23 Jul. 1994, Suzuki M. et al. 9470146 (TI); Jumla district, Munigaon-Chutta, 2850 m, 24 Jul. 1952, Polunin, Sykes & Williams s.n. (TI); Kalikot district, Chaukebada-Badarigaon, 05 Aug. 1991, Suzuki M. et al. 9170299 (TI); Kaski district, Tikhedhunga-Kande, 07 Sept. 1988, Suzuki M. et al. 8881795 (TI); Lalitpur district, Godavari, 18 Sept. 1963, Hara H. et al. 6306731 (TI); Makwanpur district, Deorali-Kuli Khani, 1700 m, 03 Sept. 1970, Kanai H. & Shrestha T.B. 672729 (TI); Manang, Latamanang, 2440 m, 14 Aug. 1994, Mikage M. et al. 9485416, (TI); Myagdi district, Ghodepani Deurali-Shika, 2400 m, 25 Aug. 1988, Suzuki M. et al. 8881364, (TI); Rasuwa district, Dhunche-Syabru, 29 Aug. 1991, Noshiro S. 9154503 (TI).
Impatiens tripetala A. Flowering plant; B. Flower (Front view); C. Flower (side view); D. Capsule. Photographs by Bhakta Bahadur Raskoti.
Type: ex hort. calc (lecto, designated in [42], G00218033), Silhet, India, Jul. 1818.
Impatiens tripetala Roxb. (1814) 18, nom. nud.; Wall. (1824) 453; G. Don (1831) 750; Roxb. (1832) 651; Hook.f. (1904a) 25, 27, 31; W.J. Kress et al. (2003) 170.
Type: s. coll. (lecto, designated in [42] G G00218033), India, 1818.
Impatiens multiflora Wall. ex Hook.f. & Thomson, Numer. List (Wallich) n. 4742 (1831).
Type: De Silva, Wallich Cat. no. 334742 (lecto, designated in [26], K K001039802!), Bangladesh, Pundua.
Impatiens ternifolia Buch.-Ham. ex Hook.f. & Thomson (1860) 126, nom. nud. Impatiens ternifolia Buch.-Ham. in Hook.f. (1904a) 27, nom. nud.,
Type: Based on: Hb. Ham., Wall. Cat. 4752B in Hooker (1904a) 27, 31 (Syn, K K001039802!), India, Assam State, Goalpara, 08 May 1862.
Habitat: Subtropical forest at an elevation range of 1400–1800 m.
Distribution: Eastern Nepal, Bhutan, India and Myanmar.
Specimen examined: Taplejung district, between tawa and Chiruwa, 1200 m, 02 Aug. 1989, Kew-Edinburgh-Kathmandu Expedition to N.E. Nepal 153 (E); Taplejung district, Tapethok, 05 Nov. 1963, Hara H. et al. 6300496 (TI); Taplejung district, Linkhim-Tuwa, 04 Nov. 1963, Hara H. et al. (TI); Taplejung district, Papung-Bir Gaon-Sangrati Pati, 1300 m, 26 Aug. 1977, Ohashi H. et al. 774018 (TI); Taplejung district, Saju Khola, 1400 m, 01 Jul. 1972, Kanai H. et al. 720981 (TI); Taplejung district, Taplethok, Tuwa-Kiwa-Tapethok, 05 Nov. 1963, Hara H. et al. 6300496 (KATH).
Impatiens uncipetala C. B. Clarke ex Hook. f., Rec. B. Surv. Ind. 4: 18 & 22 (1905).
Type: Clarke 9121 (lecto, designated in [43], K K000694903!), India, Darjeeling, Rungyroong, 6500 ft. 06 Sept. 1869.
The relationships in the resulting cladograms of Impatiens (Fig 1), generally agree with those recovered in the previous phylogenetic analyses [6–9, 12, 24]. Impatiens is divided into two major clades (Clade I and Clade II). Clade II is further divided into different subclades.
Based on nuclear (ITS) and chloroplast (atpB-rbcL) markers, phylogenetic analyses revealed a separate lineage of newly sampled species (Fig 1, Clade II/Subclade III) with BSMP = 80%, PP = 1.00, which is strongly supported as sister to I. harae and its allies (BSMP = 90%, PP = 1.00). Taxa belonging to newly sampled species and their closely related clade (I. harae, I. radiata, I. racemosa, I. urticifolia, I. wallichii) share similar morphological characters such as many-flowered, racemose inflorescences, lateral sepals 2, rarely 4 with inner 2 reduced; capsule linear; seed ovoid.
Impatiens bicornuta also shares similar morphological characters with the taxa belonging to newly sampled clade. But in the present molecular analysis as well as in previous studies of Impatiens [6], I. bicornuta forms a separate lineage as sister to I. cymbifera, I. purpurea and I. blinii. There are different specimens identified as I. bicornuta deposited in different herbaria (such as K, E, TI) having variations in shape of leaves, petioles, inflorescence, color of flowers, peduncles, pod etc. Similarly, such variations can be observed in the natural populations in different habitats. In the literature [29] all these polymorphic taxa were treated as I. bicornuta and synonymized several species such as I. harae, I. arunensis based on only morphological observation. In this study, we did not accept treatment by [29] because of separate phylogenetic position of I. bicornuta and I. harae, based on molecular result (Fig 1). There may be existence of cryptic speciation in I. bicornuta complex group. More robust molecular study (specially population level) is needed to confirm taxonomic position and identity of I. bicornuta having different morphology in different population.
Conclusions
With the discovery of this new species and five additional species new records to Nepal, a total 57 species of Impatiens (8 endemic species) were confirmed in the updated checklist of Balsaminaceae in Nepal. We strongly recommend that the future authors should integrate morphological and molecular data for the recognition and description of new species of Impatiens. Furthermore, we propose more robust studies (sampling population level) for the clarification of taxonomic position of I. bicornuta as well as taxonomic identity of its different polymorphic forms.
Supporting information
Bayesian inference tree generated from ITS marker.
Numbers above branches indicate bootstrap value for posterior probabilities (PP) for BI analysis. Asterisk (*) indicates PP = 1.00, a dash (−) indicates support at a node < 50%.
(PDF)
Bayesian inference tree generated <italic>atpB-rbcl</italic>.
Numbers above branches indicate bootstrap value for posterior probabilities (PP) for BI analysis. Asterisk (*) indicates PP = 1.00, a dash (−) indicates support at a node < 50%.
(PDF)
Taxa analyzed, voucher information and GenBank accession numbers for the DNA sequences.
(DOC)
We are grateful to Neera Joshi Pradhan, a botanical artist in Studio Petals, Khumaltar, Lalitpur Nepal for line drawings.
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