Journal of Botany

Volume 2013 (2013), Article ID 248943, 14 pages

http://dx.doi.org/10.1155/2013/248943

Research Article

Ethnobotany and Ethnomedicinal Uses, Chromosomal Status and Natural Propagation of Some Plants of Lahaul-Spiti and Adjoining Hills

Puneet Kumar and Vijay Kumar Singhal

Department of Botany, Punjabi University, Patiala, Punjab 147 002, India

Received 18 July 2013; Accepted 22 September 2013

Academic Editor: Philip J. White

Copyright © 2013 Puneet Kumar and Vijay Kumar Singhal. This is an open access article distributed under the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited.

Abstract.

The present study documented the ethnobotanical and medicinal uses of plants from an ecologically fragile cold desert area of Lahaul-Spiti (Himachal Pradesh, India). Local people use plants for curing the stomach troubles, pain reliever, cough, gastric disorders, and aphrodisiac and other household purposes. In addition, chromosome numbers, male meiosis, and natural propagation were also investigated in these ethnobotanically used plants. Present investigations also form the basis for exploitation of intraspecific chromosomal variation/new cytotypes recorded in some of the presently studied species to detect biochemical diversity in the medicinally important plants. For documentation of ethnobotanical information, personal observations and interviews were conducted with medicine men, hakims, farmers, shepherds, local healers, and old aged people. This study identified 40 plant species under 33 genera belonging to 17 families which have been used locally for curing various diseases and other purposes. All the chromosome counts are new to the study area. On worldwide basis, meiotic chromosome counts of and in Rosularia alpestris and Corydalis govaniana, respectively, are the first ever reports. The present study indicates that the people of the area possess good knowledge about the different uses of plants in the area. It has been noticed that due to the lack of interest among younger generations in the preservation of invaluable ethnic knowledge, there is every possible chance of losing such a rich heritage of knowledge. It is very urgent to conserve such invaluable ethnic knowledge before it gets lost.

1. Introduction

Present study was conducted in Lahaul-Spiti, a cold and desolate alpine region in northwest Himalayas which is known for its seclusion, Buddhist culture, hostile climate, and unexplored, formidable, and breathtaking scenic beauty of the high snow clad mountains. It is a part of the Indian cold deserts which is situated between 31°44′57′′ and 32°59′57′′N latitudes and between 76°29′46′′ and 78°41′34′′E longitudes. Climatic conditions are typical of dry temperate and alpine zones. The vegetation which is mainly composed of perennial herbs and prostrate shrubs survives extreme variation in temperature including months buried under several feet of snow. Using plants for different purposes and in curing and healing is as old as man himself [1]. It was very well known to primitive people that plants contain beneficial and medicinal properties which they used in some form or the other. Even in the ancient time, people classified plants on the basis of their medicinal uses. No comprehensive attempt has been made so far to gather ethnobotanical information and to assess the cytological diversity and natural propagation of the plants growing in the higher altitude and cold desert regions of Lahaul-Spiti. Presently an attempt has been made to collect information on the ethnobotanical uses and to examine the cytomorphological diversity and natural propagation in the species of Lahaul-Spiti and adjoining high hills.

1.1. Topography and Vegetation

The topography of the area is entirely different from the adjoining regions of northwest Indian Himalayas. The cold desert plants of Lahaul-Spiti show a wide range of habitat diversity which includes alpine pastures and slopes, moist shady and exposed places, forests and forest undergrowth, water courses, scree slopes, rock crevices, rocks/boulders, roadsides, riverbeds, and sandy areas (Figures 1(a)–1(d)). The Lahaul-Spiti which is the largest district of Himachal Pradesh with an area of ca.13835 km² consists of two regions (Lahaul Valley and Spiti Valley) which are different in many aspects. One such aspect is the vegetation which is quite different. The Lahaul Valley is on the greener side compared to the barren Spiti region where climate is much drier. Since the district possesses a great range of elevation, ranging from 2400 to 6500 m, the vegetation is diverse, unique, and plentiful. It comprises dry temperate to high alpine type except for some parts of Pattan Valley (Lahaul) adjoining the Pangi area of Chamba district, where it is of dry temperate to moist temperate type (Figure 1(c)).

(a)

(b)

(c)

(d)

Figure 1: (a) Alpine pasture. (b) A part of Lahaul Valley near Baralacha Pass with scanty vegetation. (c) Dry temperate forests near Keylong. (d) Sheep grazing on alpine moist slopes.

2. Materials and Methods

2.1. Information Collection, Fixation of Material for Cytological Studies, Identification of Plants, and Male Meiotic Chromosome Counts

As the valley remains cut off from other parts of country for more than six months due to heavy snow fall, observations on ethnobotanical information and natural propagation and collection of material for cytological studies were made between June and October in the years 2006–2009. During the intensive cytomorphological and ethnobotanical surveys, 40 plant species used by the local people of Lahaul-Spiti had been collected from different localities (Figure 2). For documentation of ethnobotanical information, personal observations and interviews were conducted with medicine men, hakims, farmers, shepherds, local healers, and old aged people. Plant identification was done by consulting the Flora of Lahaul-Spiti [2], the Flora of Kullu district [3], and Flora of Chamba district [4]. Besides, the plant accessions were also compared to the samples lying in the Herbaria of the Department of Botany, Punjabi University, Patiala (PUN), Botanical Survey of India (BSI), and Forest Research Institute (FRI) at Dehra Dun. Voucher specimens of the cytologically worked-out specimens were deposited in the Herbarium, Department of Botany, Punjabi University, Patiala (PUN). For male meiotic studies, floral buds of suitable sizes from the wild plants were fixed in Carnoy’s fixative (mixture of ethanol, chloroform, and glacial acetic acid in volume ratio of 6 : 3 : 1) for 24 h and preserved in 70% ethanol in a refrigerator until analysed. For chromosome counts and detailed meiotic behavior in pollen mother cells (PMCs) at early prophase-I, metaphase-I (MI), anaphase-I/II (AI/II), telophases-I/II (TI/II), and sporad stage developing anthers were squashed in 1% acetocarmine. Pollen fertility was estimated through stainability tests by smearing mature anthers from different flowers in glycerol-acetocarmine mixture (1 : 1) and aniline blue dye (1%). Best plates of chromosome counts, meiotic abnormalities, sporads, and pollen grains were photographed from the temporary mounts with digital imaging system of Nikon Eclipse 80i microscope and Leica QWin. Field photographs of plants and landscape were taken with Nikon Coolpix L3 camera.

Figure 2: Study area: Lahaul Spiti, Chamba, and Kullu districts of Himachal Pradesh (India).

3. Results

3.1. Ethnobotany

In the present investigation, 40 plant species under 33 genera belonging to 17 families have been reported to be used for ethnobotanical and medicinal purposes. Data given in Table 1 includes the name of the family to which a particular species belongs, their meiotic chromosome number, ploidy level, vernacular name, locality and its altitude, voucher specimen number, habit and habitat, natural regeneration, flowering and fruiting period, plant part used, and ethnobotanical uses (Figures 3(a)–3(g)). Out of the 40 species, the maximum number of species belongs to Ranunculaceae and Leguminosae with 7 species each and 4 species to Apiaceae and Rosaceae each. The most common plant parts used in different preparations are the roots and leaves. Other plant parts used as herbal medicine are whole plant, seeds, fruits, flowers, and rhizomes. Based on the information gathered from different sources, the local people used a good number of plant species for curing the stomach troubles, pain reliever, cough, gastric disorders, and aphrodisiac; as fodder and fuel; to obtain different dyes; and in religious ceremonies and as offerings to deities (Figures 4(a)–4(d)).

Table 1: Family, species and its vernacular name, meiotic chromosome number, ploidy level, locality and altitude, habit and habitat, natural regeneration, flowering and fruiting period, plant part used and ethnobotanical uses in the wild plants of Lahaul-Spiti, and adjoining high altitude hills.

Family/species (vernacular name)/meiotic chromosome number “”/ploidy level
Locality and altitude in meters (m)
Voucher specimen number (PUN)
Habit and habitat
Natural regeneration
Flowering and fruiting period
Plant part used
Ethnobotanical and ethnomedicinal uses

Apiaceae 

(1) Angelica glauca Edgew.

  (Choru, Chora) 

(2x)
Dhancho, 3,030 m
51529 PUN
Tall perennial herb; shady moist places
Seeds, rootstock
July–September
Whole plant, roots
Stomach troubles and stimulant

Spices and condiments

Bronchitis

(2) Bupleurum falcatum L.

(2x)
Bharmour, 2,130 m
51524 PUN
Perennial herb; dry slopes
Seeds, rootstock
June–September
Roots
Liver troubles

(3)  Chaerophyllum aromaticum L.

(2x)
Koksar, 3,140 m
51546 PUN
Herb; moist slopes
Seeds, rootstock
June–September
Roots
Plant eaten raw or used as pot herb

Along with mint is used as flavouring agent

(4) Heracleum lanatum Michx. (Raswal)

(2x)
Trilokinath, 3,020 m
51542 PUN
Perennial herb; along water courses and amongst stones and open slopes
Seeds, rootstock
June–September
Leaves, tender shoots, and fruits
Check the excessive menses and bleeding, abdominal pain and gastric disorders

Fodder

Aphrodisiac

Balsaminaceae  

(5) Impatiens sulcata Wall.

(Don)

(2x)
Sissu, 3,170 m
51627 PUN
Annual herb; along streams
Seeds
June–August
Whole plant
Paste made by mixing it with lichens and Rumex sp. is used as nail polish

(6) I. brachycentra Royle

(2x)
Jhalama, 3,070 m
51632 PUN
Annual herb; glacial streams and moist shady places
Seeds
July–August
Leaves
Red dye obtained is used as nail paint

Biebersteiniaceae 

(7) Biebersteinia odora Steph. ex Fish. (Figure 3(a))

(2x)
Baralacha Pass,

5,030 m
51587 PUN
Perennial herb; alpine meadows and dry rocky and scree slopes
Seeds, rootstocks
July–October
Rootstocks
Antiseptic and blood purifier

Brassicaceae 

(8) Lepidium latifolium L.

(Tharag-Thokpa)

(2x)
Bahu, 2,830 m
51618 PUN
Perennial herb; open slopes and around cultivated fields
Seeds, rootstocks
July–September
Aerial parts
Rheumatic pain

Caryophyllaceae  

(9) Cerastium cerastoides (L.)

Britt.

(2x)
Batal, 3,960 m
51479 PUN
Perennial herb; moist slopes and meadows
Seeds, stolon
June–September
Whole plant
Decoction for cough

(10) Silene vulgaris (Moench) Garcke

(Ghandoli)

(2x)
Udaipur, 2,740 m
51713 PUN
Perennial herb; grassy slopes and meadows
Seeds, rootstocks
June–September
Fresh young leaves and twigs
Vegetable

Crassulaceae 

(11) Rosularia alpestris (Kar

& Kir) Boriss.

(Pyau Chakti)

(2x)
Kukumsheri, 2800 m
51163 PUN
Perennial herb; rock crevices and dry sandy slopes
Seed, offsets
June–September
Whole plant
Juice—considered good for health

Datiscaceae

(12) Datisca cannabina L.

(2x)
Jahlama, 3,070 m
49418 PUN
Perennial robust herb; dry sandy slopes
Seeds, rootstocks
July–September
Roots
Yellow dye

Fumariaceae

(13)  Corydalis govaniana Wall.

(2x)
Gauri Kund, 3,930 m
51673 PUN
Perennial herb; rock crevices and glacial moraines
Seeds, rootstocks
July–September
Whole plant
Leprosy and rheumatism

Geraniaceae 

(14) Geranium pratense L.

(Porlo, Likatur)

(4x)
Lossar, 3,900 m
51573 PUN
Perennial herb; moist places
Seeds, rootstocks
July–September
Whole plant,

roots

flowers
Cough, jaundice, and gastric disorders

Poultice to bruises. Black dye obtained by boiling roots along with Naktsur (sulfurous raw mineral)

Offered to deities

(15) G. wallichianum D. Don

ex Sweet

  2x)
Rohtang Pass, 3,980 m
51190 PUN
Perennial herb; moist slopes
Seeds, rootstocks
July–September
Whole plant
Stomachaches, cough, cold, and throat infections

Leguminosae 

(16)  Astragalus bicuspis Fischer

(2x)
Gondhla, 3,160 m
51179 PUN
Under shrub: open, dry, and sandy slopes
Seeds, rootstocks
July–August
Whole plant
Soil binder

(17) A. himalayanus Klotzsch

(Kayabachhutup)

(2x)
Zingzingbar, 4,350 m
51288 PUN
Herb; meadows, dry slopes, and around cultivated areas
Seeds
July–September
Seeds, flowers
Strangury

(18) A. rhizanthus Royle ex

Benth. (Figure 6(a)) 

(Zomoshing)

(2x)
Chandartal Lake, 4,300 m
51276 PUN
Perennial herb; dry slopes, riverbeds, and among stones
Seeds, rootstock
June–September
Roots
Fodder

(19)  Cicer microphyllum Benth. (Figure 3(b))

(Nayarcha, Chiri)

(2x)
Chandartal Lake, 4,300 m
51219 PUN
Perennial herb; sandy and dry slopes
Seeds, rootstocks
June–September
Aerial parts, seeds
Treatment of sore mouth in cattle

“Khur” disease of sheep and goats

Dry as well as fresh leaves are used as vegetable or eaten raw

(20) Indigofera gerardiana Wall. ex Baker

(6x)
Tindi, 2,600 m
51312 PUN
Shrub: shady and sometimes also on open moist sloes
Seeds, new sprouts, and suckers begin from dormant buds on stems, rootstocks
May–August
Leaves and young shoots
Fodder and fuel

(21) Medicago falcata L.

(Kathoama)

(2x)
Keylong, 3,340 m
51328 PUN
Perennial herb; moist shady places
Seeds
July–September
Aerial parts
Used as a good fodder

Cuts and wounds

(22) Trigonella emodi Benth.

(Kuchona, Buksup)

(2x)
Sundrasi, 3,370 m
51160 PUN
Herb; moist places, riverbeds, and around cultivated fields
Seeds
June–August
Young leaves, shoots, and flowers
Vegetable

Plant considered being poisonous

if fed before flowering

Insect repellent

Dried plants as perfume and fresh flowers worn as garland

Malvaceae

  (23) Malva neglecta Wall.

(2x)
Keylong, 3,340 m
51696 PUN
Perennial herb; moist places and roadsides
Seeds, rootstocks
June–September
Whole plant
Decoction for malaria

Dried plants for kidney and bladder disorders

(24) M. verticillata L.

(Mikanchi)

(polyploid)
Kaza, 3,800 m
51492 PUN
Perennial herb; moist places
Seeds, rootstocks
June–September
Seeds
Kidney and bladder disorders

Onagraceae 

(25)  Epilobium angustifolium L.

(Dharshak)

(4x)
Miyar, 3,000 m
51665 PUN
Perennial herb; moist places, amidst stones, and boulders
Seeds, rootstocks
July–September
Roots
Crushed roots used as a detergent

(26)  E. latifolium ssp.

Speciosum (Decne.) Raven

(4x)
Batal, 3,960 m
51654 PUN
Perennial herb; dry slopes and on rocks
Seeds, rootstocks
July–September
Flowers
Ornamental

Papaveraceae 

(27)  Meconopsis aculeata Royle (Figure 3(c))

(Chharbongcha Chharmen)

(4x)
Gauri Kund, 3,930 m
49361 PUN
Herb; among boulders in alpine rocky areas
Seeds
June–September
Aerial parts roots
Swelling of legs after long distance walking

Narcotic and poisonous

Podophyllaceae 

(28)  Podophyllum hexandrum Royle (Figure 3(d))

(Omo-Shey)

(2x)
Gulaba 3,400 m
51708 PUN
Perennial herb; moist slopes
Seeds, rootstocks
July–September
Roots, fruits
Purgative

Tuberculosis and cough

Ranunculaceae 

(29)  Aconitum heterophyllum Wall. ex Royle (Figure 3(e))

(Boa)

(2x)
Manimahesh Lake, 4,300 m
51398 PUN
Herb; subalpine and alpine grassy meadows and moist slopes
Seeds
June–September
Roots
Diarrhoea, coughs, astringent, and aphrodisiac

(30)  A. violaceum Jacq. Ex

Stapf.

(Yangtso)

(2x)
Manimahesh Lake, 4,300 m
51402 PUN
Small herb; alpine Himalayan regions
Seeds, rootstocks
June–September
Roots
Cough, high fever stomach problems, snake and scorpion bites, contagious infections, and intestine inflammation

(31) Aquilegia fragrans Benth.

(Lande, Kumuk)

Chhatru, 3,360 m
51377 PUN
Perennial herb; moist slopes
Seeds, rootstocks
July–September
Flowers
Hair follicle infection

(32) Caltha palustris L.

(4x) (Figure 3(f))
Trilokinath, 3,020 m
51333 PUN
Perennial herb; marshy areas along water streams and glacial moraines
Seeds, rootstocks
June–September
Leaves

Roots
To keep maggots out of cattle wounds. To clean the hands

Gonorrhoea

(33) Delphinium denudatum Wall. ex Hook. f. & Thomson

(2x)
Dibri, 2,300 m
51384 PUN
Perennial herb; margins of Cedrus and Pine forest, dry scrubby slopes
Seeds, rootstocks
July–September
Roots
Toothache

(34)  Ranunculus hirtellus Royle

(2x), 16 (4x)
Keylong, 3,340 m
51374 PUN
Perennial herb; moist places and alpine meadows
Seeds, rootstocks
June–September
Whole plant
Counter irritant

Swelling in testes

(35) Thalictrum foetidum L.

(6x)
Trilokinath, 3,020 m
51335 PUN
Perennial herb; dry slopes, among stones and boulders
Seeds, rootstocks
June–September
Flowers
Religious ceremonies

Rosaceae  

(36) Cotoneaster microphyllus Wall. ex Lindl.

(Rogthali)

(2x)
Kothi, 2,400 m
49218 PUN
Trailing evergreen shrub on rocky slopes, rocky mountain areas, thickets, and river valleys
Seeds, new shoots from dormant bud on old stem
June–September
Whole plant, fruits
Eaten raw

Walking sticks and baskets

Used for fuel

(37)  Potentilla atrosanguinea Lodd. var. argyrophylla (Wall. ex Lehm.) Griers. & Long 

(6x), 42 (12x)
Kilingsarai, 4,500 m
51101 PUN
Perennial herb; alpine meadows and glacial moraines
Seeds, rootstock
June–October
Roots
Toothache and gingivitis

(38)  Rosa webbiana Wall. ex

Royle (Figures 3(g), 4(a), and 4(b))

(Shaybala)

(2x)
Kaza, 3,800 m
51450 PUN
Deciduous prickly shrub; open sandy and rocky slopes
Seeds, rootstocks
June–September
Fruits, stems, and flowers
Eaten

Used as biofence

Brooms made—used to remove stones and debris from roads

Used as fuel

Offered in gompas, stomach ache

(39) Spiraea canescens D. Don

(2x)
Kothi, 2,400 m
51421 PUN
Shrub: dry slopes
Seeds, rootstocks
June–September
Slender branches
Rim of baskets

Saxifragaceae  

(40) Bergenia stracheyi Engl.

(Figures 6(b), 6(c), and 6(d))

(Shilpada)

(2x)
Gramphoo, 3,160 m
51682 PUN
Perennial herb; moist rocky slopes, among boulders
Seeds, rootstocks
May–September
Leaves, rootstocks, and roots
Used as fuel wood

Diuretic

(a)  

(b)

(c)

(d)

(e)

(f)

(g)

Figure 3: (a) Biebersteinia odora plants in a group, coming out of withered remains of stems. (b) Cicer microphyllum growing on rock. (c) An individual of Meconopsis aculeata growing among the boulders. (d) Podophyllum hexandrum. (e) Aconitum heterophyllum with greenish blue flowers having purple veins growing on moist slopes. (f) Caltha palustris with bright yellow flowers and large reniform leaves growing near water. (g) A shrub of Rosa webbiana with pink flowers.

(a)

(b)

(c)

(d)

Figure 4: (a) Spiny branches of Rosa webbiana wrapped around the stem of Salix tree as a biofence. (b) A broom made of twigs of Rosa webbiana to remove stones and debris from road. (c) An old person carrying a bundle of young twigs of Indigofera gerardiana on his back to be used as a fodder and fuel. (d) Timber of some species used in house construction (arrowed).

3.2. Chromosomal Status

In addition to the gathering of ethnobotanical information, chromosome number and details of male meiosis have also been investigated in all the species. Of the 40 cytologically studied species, 30 (75%) exist at diploid level with basic chromosome number ranging between (Biebersteinia odora Steph. ex Fish., ) and 18 (Cerastium cerastoides (L.) Britt., ), while nine (22.5%) of them were found to be at polyploid level (4x, 6x, 12x). One species, Ranunculus hirtellus Royle (), existed at both diploid and tetraploid levels based on . All the chromosome counts are new to the study area. On worldwide basis, meiotic chromosome counts of (Figure 5(a)) and (Figure 5(b)) in Rosularia alpestris (Kar and Kir) Boriss. and Corydalis govaniana Wall., respectively, are the first ever chromosomes counts. While in some other species new cytotypes have been added to the already existing diploid (Thalictrum foetidum L.,  new, (Figure 5(c)), previous, ; Geranium pratense L., new (Figure 5(d)), previous ) or polyploid cytotypes (Malva verticillata L., new,  , previous, ). New/additional intraspecific cytotypes for India have also been established in Indigofera gerardiana Wall. ex Baker (, 6x), Epilobium angustifolium L. (, 4x), Cerastium cerastoides (L.) Britt. (, 2x), Geranium wallichianum D. Don ex Sweet (, 2x, Figure 5(e)), and Impatiens sulcata Wall. (Don) (, 2x, Figure 5(f)). Present study also recorded additional intraspecific diploid/polyploid cytotypes from India in Ranunculus hirtellus Royle (, 2x; , 4x) and Potentilla atrosanguinea Lodd.var. argyrophylla (Wall. ex Lehm.) Griers. and Long (, 6x; , 12x). As many as eight species, namely, Angelica glauca Edgew. (), Astragalus bicuspis Fischer (), Biebersteinia odora Steph. ex Fish.(, Figure 5(g)), Bergenia stracheyi Engl.(), Chaerophyllum aromaticum L. (), Datisca cannabina L. (), Rosa webbiana Wall. ex Royle (), and Trigonella emodi Benth. () have been counted chromosomally for the first time from India.

Figure 5: Meiotic chromosome number. (a) Rosularia alpestris, at metaphase-I. (b) Corydalis govaniana, at metaphase-I. (c) Thalictrum foetidum, at metaphase-I. (d) Geranium pratense, 28 : 28 chromosomes distributions at metaphase-II. (e) Geranium wallichianum, at diakinesis. (f) Impatiens sulcata, at metaphase-I. (g) Biebersteinia odora, at metaphase-I. Meiotic abnormalities in pollen mother cells (PMCs) and sterile pollen grains. (h) A PMC showing univalents (arrowed) and multivalents (arrowhead). (i) PMCs in a group showing inter-PMC chromatin transfer (arrowed) and chromatin stickiness (arrowhead). (j) A PMC showing laggards at anaphase-II. (k) A PMC showing a chromatin bridge at anaphase-II. (l) Pycnotic chromatin material scattered in the cytoplasm of PMCs. (m) A PMC showing unequal distribution of chromatin material at poles. (n) Micronuclei at telophase-I. (o) Micronuclei at sporad stage. (p) Apparently sterile (arrowed) and fertile pollen grains (arrowhead).

3.3. Abnormal Cytological Events during Male Meiosis and Pollen Fertility

Besides new chromosome reports and additional cytotypes, in some of the species meiosis was abnormal and was characterized by the presence of multivalents and/or univalents, impaired synapsis, cytomixis, and its associated meiotic aberrations such as laggards and bridges, chromatin stickiness, pycnotic chromatin, irregular distribution of chromosomes, and micronuclei at telophases/sporads consequently leading to abnormal microsporogenesis and variable amount of pollen sterility (Figures 5(h)–5(p)).

Among the abnormal taxa, 12x cytotype of P. atrosanguinea Lodd.var. argyrophylla (Wall. ex Lehm.) Griers. and Long is an imbalanced polyploid. Owing to the presence of high chromosome number (), it depicted a typically irregular meiotic behaviour (presence of multivalents and/or univalents, laggards, and irregular distribution of chromosomes), abnormal microsporogenesis, and pollen sterility (14–46%) which is the characteristic feature in such kind of imbalanced polyploids. Cytomixis which involves the interpollen mother cell transfer of chromatin material through narrow and broad cytomictic channels is existent in Astragalus bicuspis Fischer (), A. himalayanus Klotzsch (), A. rhizanthus Royle ex Benth. (), Caltha palustris L. (), Geranium pratense L. (), Meconopsis aculeata Royle (), P. atrisanguinea Lodd. var. argyrophylla (Wall. ex Lehm.) Griers. and Long (, 42), Trigonella emodi Benth. (), Rosularia alpestris (Kar and Kir) Boriss. (), Silene vulgaris (Moench) Garcke (), Thalictrum foetidum L. (), and Ranunculus hirtellus Royle (, 16). Impaired synapsis owing to the lack of pairing of homologous chromosomes in Rosularia alpestris (Kar and Kir) Boriss. resulted in the presence of all chromosomes as univalents during early prophase stages of meiosis-I and consequently abnormal microsporogenesis and pollen sterility (28%). Besides, in Bergenia stracheyi Engl. high pollen sterility (22%) was recorded without the occurrence of any meiotic irregularity which might be attributed to irregularities in postmeiotic events during pollen development.

3.4. Natural Propagation

Natural propagation observations made on all the 40 species revealed that majority of the species are perennial and propagate through asexual (vegetative propagation) means such as rootstocks, offsets, root suckers, and stolons (Figures 6(a)–6(d)).

(a)

(b)

(c)

(d)

Figure 6: Natural propagation. (a) A plant grown from old rootstock (arrowed) in Astragalus rhizanthus. (b) New plants of Bergenia stracheyi coming out of the rootstocks (encircled) underlying withered leaves. (c) B. stracheyi plants in full bloom with white pinkish flowers. (d) A new sprout can be seen growing on the old rootstock (arrowed) of B. stracheyi.

4. Discussion

A steady stream of papers over the years have recognized the fact that the reproductive stage in the life cycle of plants is highly sensitive to different abiotic stresses [5–8] especially the low temperature conditions [9–16]. It has been observed that when plants of this area are about to enter the flowering stage, the temperature is very low (sub-zero to 15°C) which seems to disrupt the meiotic process by causing various meiotic abnormalities and consequently reduced viability of gametes. Ultimately these events hinder natural propagation of plants through sexual means.

Natural propagation observations revealed that majority of the species are perennial and propagate through asexual means such as rootstocks, offsets, root suckers, and stolons. Thus, it is assumed that in order to survive the harsh environmental conditions and gametic sterility caused by irregular meiotic events, the plants of the area have adopted alternate mechanism of reproduction through vegetative means. Vegetative means of propagation are important for species like Meconopsis aculeata Royle, Potentilla atrosanguinea Lodd. var. argyrophylla (Wall. ex Lehm.) Griers. and Long, Bergenia stracheyi Engl., Rosularia alpestris (Kar and Kir) Boriss., and so forth, where high pollen grain sterility have been observed. Occurrence of sterility on female side in the species may pose serious problem for species existence, especially, which are rare and engendered and are overexploited for medicinal purposes. Problem becomes more serious in species like Bergenia stracheyi, Engl. and Rosularia alpestris (Kar and Kir) Boriss. which propagate through vegetative means, and perennating parts are directly collected from the wild sources. Till now these species have not been included in any of the local cultivation programme.

To date numerous valuable herbal drugs have been discovered by following the knowledge of ancient folk healers [17]. The medicinal plants used to cure various diseases by the traditional healers are gradually becoming extinct. The two factors which have been identified as major threats to the floristic diversity are the overexploitation and habitat [18–21]. Majority of the plants (Aconitum heterophyllum Wall. ex Royle, Angelica glauca Edgew., Biebersteinia odora Steph. ex Fish., Bergenia stracheyi Engl., Delphinium denudatum Wall. ex Hook. f. and Thomson, Indigofera gerardiana Wall. ex Baker, Meconopsis aculeata Royle, Podophyllum hexandrum Royle, and Spiraea canescens D. Don) which have been studied presently are restricted to specific pockets with very low population density and belong to rare, critically endangered, endangered, vulnerable, threatened, and newly threatened category because of the fragile ecological conditions of this Himalayan region [20–22]. Some of the other factors which can also lead to the extinction of the local valuable herbs include agricultural expansion, heavy grazing, road building, increasing entry of tourists, and transport vehicles. So to study and examine different aspects of survival of the plants of the area in their natural habitats will be of colossal significance. Thus it becomes very urgent to collect information about the plants growing in such ecologically fragile region before it gets too late to respond. Immediate steps must be taken up on priority basis to preserve the ethnic knowledge and the plants of the region. The gradual exposure of younger generations to western civilization and limited employment opportunities are two important threats that may lead to serious losses of traditional knowledge, and it will not be surprising that after a decade or so nobody will be able to tell about the plants that were once used traditionally for ethnobotanical and medicinal purposes. In view of the above points it is suggested that an institute dealing with the conservation of plants and their traditional uses must be set up in the region. The benefits arising from ethnic knowledge must be shared among the local people and employment must be provided to local youth. Present investigations also form the basis for exploitation of intraspecific chromosomal variation/new cytotypes recorded in some of the presently studied species to detect biochemical diversity in these medicinally important plants.

Acknowledgments

The authors are thankful to the local people and medicine men for their help and cooperation during field study. The authors wish to thank the University Grants Commission, New Delhi, India, for providing financial assistance under DRS SAP I and II, ASIST programme, and Dr. D. S. Kothari Post-Doctoral Fellowship Award Letter no. F.4-2/2006(BSR)/13-427/2011(BSR) to Puneet Kumar. Thanks are also due to the Head of Department of Botany, Punjabi University, Patiala, India, for necessary laboratory and internet facilities. The authors declare that there is no conflict of interests regarding the publication of this paper.

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