Chromosome Botany (2011) 6: 85-90 © Copyright 2011 by the International Society of Chromosome Botany Exploration of cytomorphological diversity in the Scrophulariaceae from Kashmir Himalaya, India Reyaz Ahmad Malik1, Raghbir Chand Gupta and Santosh Kumari Department of Botany, Punjabi University, Patiala, Punjab 147002, India 1Author for correspondence: ([email protected]) Received May 31, 2011; accepted August 11, 2011 ABSTRACT. Cytomorphological exploration has been undertaken in 14 species of the Scrophulariaceae from Kashmir, Himalaya. The varied gametophtyic chromosome counts of n=23 for Veronica cana and diploid chromosome number i.e. n=7 for V. persica are reported here for the fi rst time on global basis. Besides this 0 to 2 B-chromosomes have been observed in Verbascum thapsus for the fi rst time. Likewise Digitalis grandifl ora (n=28) and Scrophularia scopolii (n=13) are fi rst chromosome reports from India. The meiotic course observed in these scrophs is normal except some irregularities like bridge formation in Mazus japonicus and Pedicularis rhinanthoides, interbivalent connections in P. pectinata and precocious disjunction in Euphrasia offi cinalis. Morphologically the different accessions within a species showed no conspicuous differences except Digitalis purpurea that has been seen to exist as three morphotypes on fl ower colour basis. KEYWORDS: Chromosome, Digitalis, Kashmir Himalaya, Veronica Scrophulariaceae is commonly called the fi gwort family. MATERIALS AND METHODS Most of the species included under this family are annual Appropriate sized fl ower buds of the plants growing in or perennial herbs with usually zygomorphic and rarely their natural habitats were collected and fi xed in Carnoy’s actinomorphic fl owers. It is a large family comprising fi xative. After 24 hrs. the buds were transferred in rectifi ed around 280 genera and 3000 species (Watson and Dalwitz alcohol for longer storage purpose. The meiotic studies 1992) which are mostly distributed to temperate regions were carried out in young fl oral buds by standard aceto- of the world. From Kashmir Himalaya the family is carmine smearing technique. Photomicrographs of pollen reported to cover 19 genera and 77 species (Dar et al. mother cells (PMC’s) and pollen grains were taken using 2002). There is ambiguity in the reports regarding the Nikon 80i Digital Imaging System. Pollen fertility was number of taxa under this family due to some taxonomic estimated by their stainability in 1% glycero-acetocarmine revisions made from time to time and thereby transferring (1:1). Well fi lled and well stained pollen grains were some taxa from one family to other e.g. Digitalis to the regarded as fertile while as the shrunken and unstained Plantaginaceae (Olmstead et al. 2001). pollen grains were considered as sterile. For morphological Kashmir Himalaya (1600 to 4500 m) is a land of hubs of studies simply metric scale and hand lenses were used. many important medicinal and aromatic plants including species from the Scrophulariaceae. The examples are RESULTS AND DISCUSSION Digitalis spp. reported to be benefi cial in heart diseases The meiotic studies in 14 species of the Scrophulariaceae (Goldthorp 2009), Picrorhiza kurroa in dysentry, stom- have been carried out on population basis covering achache and fever (Ballabh and Chaurasia 2009), different altitudinal ranges of Kashmir Himalaya. All the Verbascum thapsus for mental relaxation and wounds individuals worked out are either diploids (2x) or (Kumar et al. 2009), etc. This family which is in the list of tetraploids (4x) and reveal normal meiosis except some top ten families of Kashmir Himalaya as far as species irregularity in Euphrasia offi cinalis, Mazus japonicus, composition is concerned (Dar et al. 2002) has not been Pedicularis pectinata and P. rhinanthoides. The informa- comparatively explored on cytological basis. The reason tion regarding the observed chromosome number, location behind this is the combined hindrances posed by with altitude, ploidy level, pollen fertility and pollen size topography of far fl ung inaccessible areas and political of each species is presented in Table. challenges of the valley. Keeping in view the above said importance of the family along with its unsatisfactory Digitalis grandifl ora Mill. (Common name: Big-fl owered cytological exploration it is imperative to make more Foxglove) The plant is usually distributed to subalpine attempts for further cytological contributions. This will Kashmir Himalaya and growing from 2200-2900 m. It is help in framing chromosome number data base and a perennial plant, 1-1.5 m tall; leaves simple, lanceolate & understanding meiotic behaviour and meiotic irregularities serrate. Corolla tubular and yellow. Flowering period: if any caused to these species growing there in cold and June-July. harsh climatic conditions. The gametophytic chromosome number of n=28 (Fig. 1a) for this species is fi rst report from India. This is in accordance with the previous report of Dobes and Hahn 86 MALIK ET AL. Table 1. Data regarding the name of the species with accession number, collection site & its altitude, haploid chromosome number (n), ploidy level, pollen fertility and pollen size Taxa/ Locality Altitude Chromosome Ploidy level Pollen fertility Pollen size Accession (PUN) (metre) No. (n) (%age) (μm) Digitalis grandifl ora Mill.* 54378 Gulmarg 2,600 28 4x 99.5 21.19x16.65 D. lanata Ehrh. 54374 Gulmarg 2,600 28 4x 100 13.72x12.91 D. purpurea L. 54372x Gulmarg 2,600 28 4x 99.8 16.59x14.45 54373y Gulmarg 2,600 28 4x 99.6 55468z Gulmarg 2,600 28 4x 99.9 Euphrasia offi cinalis L. 55171 Aharbal 2,200 22 4x 46.6 28.40x23.69 55467 Thajwas 3,300 22 4x 48.5 Mazus japonicus (Thunb.) Kuntze 54350 Pulwama 1,750 20 4x 100 18.70x16.85 Pedicularis pectinata Wall. ex Benth. 55170 Aharbal 2,200 8 2x 79.2 26.55x24.90 55466 Dhara 2,400 8 2x 76.4 P. rhinanthoides Schrenk. 54366 Aharbal 2,250 8 2x 95.8 (37.21x30.80 55465 Gulmarg 2,400 8 2x 95.5 & 55464 Kellar 2,150 8 2x 97.0 28.82x24.31) Scrophularia himalensis Royle 55586 Aharbal 2,300 12 2x 92.8 19.85x17.37 S. scopolii Hoppe* 55192 Aharbal 2,300 13 2x 58.4 26.26x24.98 Verbascum thapsus L. 54376 Pahalgam 2,200 17+0-2B** 2x 89.5 17.46x15.92 55469 Pakharpora 1,900 17 2x 97.6 Veronica beccabunga L. 35.70x33.68 54377 Thajwas 3,100 9 2x 90.3 & 24.90x21.70 V. cana Wall. ex Benth. 55172 Aharbal 2,250 23˚ 2x 62.0 23.50x21.80 55462 Yusmarg 2,400 23˚ 2x 61.2 V. persica Poir. 55463 Pulwama 1750 7˚˚ 2x 81.2 07.92x07.38 55470 Srinagar 1650 7˚˚ 2x 82.3 V. serpyllifolia L. 54371 Thajwas 3100 7 2x 63.9 23.04x21.86 * First chromosome report for India; **First B-chromosome report for the species; °First aneuploid chromosome report for the species; °°First euploid (diploid) chromosome report for the species; X, Y and Z: White, lilac/light pink and rose/bright pink morphotypes respectively. PUN=Herbarium Code of Botany Dept. Punjabi Univ. Patiala. (1997). Flowering period: June-July. The present chromosome count of n=28 (Fig. 1b) further confi rms the base number Digitalis lanata Ehrh. (Common name: Wooly Foxglove/ 7 for the genus and is in accordance with its previous Grecian Foxglove) The species is mainly distributed to cytological reports of n=28 by Gill (1972) and Gupta and subalpine Kashmir Himalaya from 1900-2600 m. It is a Molnar (1979). perennial up to 0.8 m high, unbranched; leaves elongated, wooly and hairy on underside. Corolla tubular, white or Digitalis purpurea L. (Common name: Purple Foxglove/ lilac and fl owers in spikes. Flowers and stem also wooly. Common Foxglove/Deas Men’s Bells) CYTOMORPHOLOGY IN THE SCROPHULARIACEAE 87 The species is distributed to the Himalayan range from also Eastern Himalaya. In Kashmir it is found growing 2000 to 2600 m. It is a biennial herb; leaves spirally between 2400-3500 m. It is a perennial herb up to 25-80 arranger, simple, 10-35 cm long and 5-12 cm broad, cm tall with cauline pinnatifi d and opposite leaves. compound and hairy and forming a rosette at the ground Flowers are rose-pink. Flowering Period: June-August. level in the fi rst year. Flowering shoot typically 1-2 m tall. The present chromosome count of n=8 (Fig. 1h) in both The fl owers in a terminal cluster, pendant with beautiful the populations confi rms its diploid nature. Except some spots on the inner surface of corolla; corolla tubular. sort of interbivalent connections (Fig. 1i) in Aharbal Flowering period: June-July. population, the meiosis has been found to be normal. The three morphotypes have been found in this species from the same site growing in thin associations. One Pedicularis rhinanthoides Schrenk. (Common name: Rattle morphotype bearing white colored fl owers is shorter in Lousewort) The species is distributed to subalpine and height than the other bearing light pink. The third one that mostly alpine Western Himalaya and Eastern Himalaya. bears bright pink fl owers is tallest among all. It seems In Kashmir it is found between 2400 to 5000 m. It is 5-25 here that there might be a multiple allelic gene with cm high. Leaves linear-oblong; the radical leaves form a quantitative inheritance responsible for this variation. rosette and are petiolate. Flowers pedicellate, axillary and The chromosome report of n=28 (Fig. 1c) presented pink or purple. Flowering period: June-August Three here has been already assigned to this species by Verma populations of this species have been worked out. and Dhillon (1967) and Chandan and Bhavananda (1987). Specimens collected from Aharbal and Gulmarg show some irregularity in the form of bridges (Fig.