International Research Journal of Biological Sciences ___________________________________ ISSN 2278-3202Vol. 4(6), 48-55, June (2015) Int. Res. J. Biological Sci. International Science Congress Association 48 Numerical Taxonomy of some members of Acanthoideae sensu Scotland and Vollesen (Acanthaceae) from North 24 Parganas District, West Bengal, India based on Gross Morphology and Pollen Characters Naskar SaikatPost Graduate Department of Botany, Barasat Govt. College, Barasat, Kolkata- 700124, West Bengal, INDIA Available online at: www.isca.in, www.isca.me Received 24th April 2015, revised 17th May 2015, accepted 3rd June 2015 Abstract Some members of Acanthoideae sensu Scotland and Vollesen from North 24 Parganas District, West Bengal, India have been classified following numerical taxonomic approach. Numerical taxonomic approach on Acanthoideae is rare. A total of 64 binary characters obtained from gross morphology and pollen morphology have been used for cluster analysis. Cluster analysis result shows remarkable differences of sub-tribal classification of Acanthoideae with that of previous phylogentic schemes. Besides Barleriinae no precise distinctions of Ruelliinae, Justiciinae and Andrographiinae are observed. Interestingly Phaulopsis-Rungia phenon has scored higher similarity value than the phenon of two species of same genus, i.e. Justicia adhatoda–J. gendarussa phenon. Finally two separate artificial keys are provided for identification purpose. Keywords: Acanthoideae, numerical taxonomy, gross morphology, pollen morphology.Introduction Acanthaceae is a large pan-tropic family of herbs and shrubs. This family comprises a large number of species ranges from 2500-26001,2 to 3175. The family divided into subfamilies Mendoncioideae, Thunbergioideae, Nelsonioideae and Acanthoideae. Among these subfamilies only Acanthoideae possesses retinaculate fruits. Acanthoideae again divided into Contortae and Imbricatae- based on corolla aestivation. Acanthoideae alone contains more than 95 percent species of Acanthaceae. Explosive and retinaculate fruits are generally accepted as synapomorphies of monophyletic Acanthoideae. Bremekamp retained only the genera with retinaculate fruits under Acanthaceae and excluded all others. He further subdivides Acanthaceae into Acanthoideae, based on monothecate anthers and colpate pollens, and Ruelloideae, based on presence of cystoliths and articulated stem. Molecular systematics on Acanthaceae7-10 do not support the separation of Nelsonioideae, Thunbergia and Mendoncia from Acanthaceae as proposed by Bremekamp. Scotland et al.placed Thunbergia (Thunbergioideae) and Elytraria(Nelsonioideae) as sister to Acanthoideae sensu Lindau. McDade10 placed Mendoncia (Mendoncioideae) as sister to Thunbergia. Pollens of Acanthaceae is tricolpate, triporate, diporate , pantoporate, or inaperturate11. Lindue classified pollens of Acanthaceae into 11 types. Different authors12-15 showed the importance of pollen characters for taxonomic treatments at generic and species levels. Phylogenetic relationships among the members of Acanthaceae were demonstrated by several authors, while phenetic relationships or overall similarities got less attention although several taxonomists still rely on phenetics. Since its resurgence by Sneath and Sokal16 it was widely applied in the field of biology to determine the natural relationships among taxa. Sneath and Sokal16 suggested that the groups of OTUs (Operational Taxonomic Units) established in numerical taxonomy by cluster analysis method are not fully synonymous with taxa. They used the term ‘Phenon’ for such groups which can be of any hierarchic rank. Phenons were represented with a number indicating the level of resemblance at which they are formed. Another important aspect of numerical taxonomy is consideration of large number of characters for better natural relationships among OTUs17. With this above background knowledge the present study is conducted to elucidate the natural relationships through numerical taxonomic method among some members of the subfamily Acanthoideae sensu Scotland and Vollesen14available in North 24 Parganas District of West Bengal, India. Gross morphological and pollen characters have been considered for study. Material and Methods Operational Taxonomic Units (OTUs): Characters for numerical analysis were obtained from freshly collected plants. A list of taxa (OTUs) is given in table-1. Identification of the OTUs was confirmed following standard taxonomic literature. Correct names of OTUs were determined following Scotland Research Journal of Biological Sciences ___________________________________________________________ ISSN 2278-3202 Vol. 4(6), 48-55, June (2015) Int. Res. J. Biological Sci. International Science Congress Association 49 and Vollesen14. Herbarium specimens of OTUs were prepared and deposited at Barasat Govt. College Herbarium for future reference. Gross morphology: Gross morphology was obtained from freshly collected worked out specimens. Freehand drawings were made for each specimen and descriptions were carried out following Harris and Harris18. Pollen morphology: Pollens were acetolyzed following Edtman’s technique. Descriptions were made following Hesse et al.19. Pollen pictures were captured under Carl Zeiss Axio Lab.A1 Microscope fitted with camera. Pollen photos are given in figure-1. Figure-1 Pollens of studied species of Acanthoideae. a – c: Andrographis paniculata; d – f: Asystasia gangetica; g: Barleria cristata; h – j: Barleria strigosa; k – l: Ecbolium viride; m – n: Eranthemum pulchellum; o – p: Hemigraphis hirta; q – r: Justicia adhatoda; s – t: Justicia gendarussa; u – v: Phaulopsis parviflora; w: Ruellia tuberose; x: Rungia pectinata. Bars- 10 m Research Journal of Biological Sciences ___________________________________________________________ ISSN 2278-3202 Vol. 4(6), 48-55, June (2015) Int. Res. J. Biological Sci. International Science Congress Association 50 Character coding: Characters were obtained from vegetative parts, floral parts and pollens. All total 64 characters were selected of which 57 from vegetative and floral parts and 7 from pollens. Binary character coding was employed for each character. A list of character and binary coding was given in table-2. Unavailable characters were presented as not assigned (NA). Cluster analysis: Data matrix (table-3) obtained from character coding were used for cluster analyses to draw phenetic relationship among OTUs. Cluster analysis of OTUs was conducted by applying Jaccard’s similarity coefficient or Jaccard Index (JI) and UPGMA methods in past software20. Results and Discussion Two primary phenons are formed by cluster analysis at lower JI (nearly 0.35), one contains two species of Barleria and rest OTUs fall under the second (figure-2). Near 0.4-Phenon line three phenons are observed. The 0.5- and 0.6-phenon lines show five and ten phenons respectively. Except two species of Barleria all OTUs are segregated at 0.7-phenon line. So, the highest similarity is observed between Barleria cristata and B. stigosa near JI= 0.75. The next highest similarity is appeared between Phaulopsis parviflora and Rungia pectinata near JI= 0.65 followed by similarity between Justicia adhatoda and Justicia gendarussa near JI= 0.60. Ruellia tuberosa and Hemigraphis hirta also show resemblance near JI= 0.60. Two major phenons are observed above 0.4-Phenon line. One ended with Ecbolium viride and Eranthemum pulchellum and another one ended with eight OTUs’ cluster, which further divides into three OTUs bearing Phenon and five OTUs bearing Phenon. Table-1 List of OTUs collected from North 24 Parganas District (23º15’ N to 22º11’ N & 89º5’ E to 88º20’ E) Sl. No. OTU Abbreviations Collection Number Collection places 1 Andrographis paniculata (Burm.f.) Nees Apn AC-003 Barasat, 24 Pgs (N), WB 2 Asystasia gangetica (L.) T.Anderson Agn AC-009 Barrackpore, 24 Pgs (N), WB 3 Barleria cristata L. Bcr AC-004 Barasat, 24 Pgs (N), WB 4 Barleria strigosa Willd. Bsg AC-014 Taki, 24 Pgs (N), WB 5 Ecbolium viride (Forssk.) Alston Evr AC-016 Taki, 24 Pgs (N), WB 6 Eranthemum pulchellum Andrews Epc AC-012 Duttapukur, 24 Pgs (N), WB 7 Hemigraphis hirta (Vahl) T.Anderson Hhr AC-006 Barasat, 24 Pgs (N), WB 8 Justicia adhatoda L. Jah AC-010 Barasat, 24 Pgs (N), WB 9 Justicia gendarussa Burm.f. Jgd AC-002 Barasat, 24 Pgs (N), WB 10 Phaulopsis parviflora Willd. Ppr AC-005 Barasat, 24 Pgs (N), WB 11 Ruellia tuberosa L. Rtu AC-001 Taki, 24 Pgs (N), WB 12 Rungia pectinata (L.) Nees Rpc AC-011 Birati, 24 Pgs (N), WB Table-2 List of Characters and binary character states used in cluster analysis Sl. No. Characters Character states (0/1) 1. Plant: Perennial (0), Annual (1) 2. Plant habit: Shrub (0), Herb (1) 3. Stem: Frutescent/Sub-frutescent (0), Herbaceous (1) 4. Stem surface: Glabrous (0), Pubescent (1) 5. Stem shape: Terete (0), Tetrangular (1) 6. Leaf symmetry: Symmetric (0), Asymmetric (1) 7. Leaf petiole: Sessile (0), Petiolate (1) 8. Lamina shape: Ovate/ Widely ovate (0), Lanceolate/ Elliptic (1) 9. Lamina apex: Narrowly acute/ Acute (0), Obtuse (1) 10. Lamina base: Narrowly cuneate/ Cuneate (0), Obtuse (1) 11. Lamina margin: Entire (0), Serrate (1) 12. Lamina surface: Glabrous (0), Pubescent (1) 13. Leaf vein type: Simple crespendodromous (0), Semi-crespendodromous (1) 14. 2º vein spacing of leaf: Uniform (0), Irregular/ Increasing toward base (1) 15. Inflorescence type: Thyrse (0), Other than thyrse (1) 16. Inflorescence: Receme/ Spike (0), Dichasial cyme (1) 17. Flowers position on inflorescence axis: One-sided (0), All sided (1) Research Journal of Biological Sciences ___________________________________________________________ ISSN 2278-3202 Vol. 4(6), 48-55, June (2015) Int. Res. J. Biological Sci. International Science Congress Association 51 Sl. No. Characters Character states (0/1) 18. Flowers density on inflorescence axis: Flowers loosely arrange on inflorescence axis (0), Flowers densely arrange on inflorescence axis (1) 19. Bract marginal growth: Armed (0), Unarmed (1) 20. Bract shape: Ovate/ Widely Ovate/ Elliptic (0), Linear/ Lanceolate (1) 21. Bract apex: Acuminate/ Caudate (0), Acute/ Narrowly acute/ Obtuse (1) 22. Bract margin Entire (0), Serrate (1) 23. Bract surface Pubescent (0), Tomentose (1) 24. Bract marginal special growth Non-scarious margin (0), Scarious margin (1) 25. Bractlets Absent (0), Present (1) 26. Flower symmetry Actinomorphic (0), Zygomorphic (1) 27. Flower stalk Sessile/ Sub-sessile (0), Pedicellate (1) 28. Calyx symmetry Actinomorphic (0), Zygomorphic (1) 29. Calyx lobes union Connate at base (0), Connate at middle (1) 30. Calyx lobes number 4 in number (0), 5 in number (1) 31. Calyx lobes surface Glabrous (0), Pubescent (1) 32. Calyx lobes margin Entire (0), Serrate (1) 33. Calyx lobes surface hair type Non-tomentose (0), Tomentose (1) 34. Calyx lobes shape Narrowly elliptic/ Linear (0), Lanceolate/ Ovate/ Lance-ovate (1) 35. Corolla symmetry Actinomorphic (0), Zygomorphic (1) 36. Corolla type Non 2-lipped corolla (1), 2-lipped corolla (0) 37. 2-lipped corolla type Upper lip non-galeate (0), Upper lip galeate (1) 38. Corolla aestivation Imbricate (0), Twisted (1) 39. Corolla tube length Short (0), Long (1) 40. Corolla tube bending Straight (0), Bent (1) 41. Corolla colour White (0), Colourful (1) 42. Corolla with purple spots Absent (0), Present (1) 43. Stamen Number 2 in number (0), 4 in number (1) 44. Stamens-corolla adhesion Adnate near the top or middle of the corolla tube (0), Adnate at the base of corolla tube (1) 45. Anther cell shape Oblong (0), Other than oblong (1) 46. Didynamous stamens (4 in number) character All fertile (0), Two rudimentary and two functional (1) 47. Anther attachment with filament Basifixed (0), Dorsifixed (1) 48. Anther lobes attachment Discrete (0), United (1) 49. Anther base Anther base without any appendage (0), Anther base with basal appendage (1) 50. Stamen insertion Cryptantherous (0), Phanerantherous (1) 51. Filament length Short (0), Long (1) 52. Filament surface Non-ciliated (0), Ciliated (1) 53. Ovary surface Glabrous (0), pubescent (1) 54. Ovule number per chamber 2 in each chamber (0), 3-6 in each chamber (1) 55. Style surface Non-ciliated (0), Ciliated (1) 56. Stigma character Entire (0), 2-lobed or 2-fid (1) 57. Capsule length Short (0), Long/ Linear (1) 58. Pollen type Homocolpate (0), Heterocolpate (1) 59. Pollen aperture number 2 to 3 in number (0), Many (1) 60. Pollen aperture type Colporate (0), Non-colporate (1) 61. Pollen aperture type Colporoidate (0), Colpoidorate (1) 62. Pollen shape Oblet-spheroidal (0), Other shapes (1) 63. Pollen shape Spheroidal (0), Prolate / Sub-prolate/ Prolate spheroidal (1) 64. Pollen wall sculpturing Reticulate/ Finely reticulate/ Coarsely reticulate (0), Lophoreticulate (1) Research Journal of Biological Sciences ___________________________________________________________ ISSN 2278-3202 Vol. 4(6), 48-55, June (2015) Int. Res. J. Biological Sci. International Science Congress Association 52 Table-3 Data matrix employed in the cluster analysis OTUs Characters 1 2 3 4 5 6 7 8 9 10 11 12 13 14 15 16 17 18 19 20 21 22 Apn 1 1 1 0 0 0 0 1 0 0 0 0 0 1 1 0 1 0 0 1 1 1 Agn 1 1 1 1 0 0 1 0 0 0 1 0 0 1 0 0 0 0 0 0 1 1 Bcr 0 0 0 1 1 0 1 0 0 0 0 1 0 1 1 0 1 1 1 0 1 NA Bsg 0 0 0 1 1 1 1 0 0 0 1 0 0 1 1 0 1 1 1 0 1 NA Evr 0 0 0 1 0 0 1 0 0 0 0 1 1 1 1 0 1 1 0 1 1 1 Epc 0 0 0 1 1 0 1 0 0 0 0 1 1 0 1 0 1 1 0 0 0 0 Hhr 1 1 1 1 0 0 1 0 1 1 1 1 0 0 1 0 1 1 0 0 1 1 Jah 0 0 0 1 0 0 1 0 0 0 0 1 0 0 0 NA 1 1 0 0 1 1 Jgd 0 0 0 1 0 0 0 1 0 0 0 0 0 0 1 0 1 0 0 1 0 1 Ppr 1 1 1 1 1 1 1 0 0 0 0 1 0 1 1 0 0 1 0 0 1 0 Rtu 1 1 1 1 1 0 1 0 1 0 0 0 0 0 1 1 1 0 0 1 1 1 Rpc 1 1 1 1 0 1 1 1 1 0 0 0 0 1 1 0 0 1 0 1 1 0 OTUs Characters 23 24 25 26 27 28 29 30 31 32 33 34 35 36 37 38 39 40 41 42 43 44 Apn 0 0 1 1 1 0 0 1 1 1 0 1 1 1 0 0 0 0 0 1 0 1 Agn 0 0 1 1 0 0 0 1 1 1 0 0 1 0 NA 0 0 1 0 0 1 0 Bcr 0 0 1 1 0 1 0 0 1 0 0 0 1 0 NA 0 1 0 0 0 1 0 Bsg 0 0 1 1 0 1 0 0 1 0 0 1 1 0 NA 0 1 0 1 0 1 0 Evr 0 0 1 1 0 0 0 1 1 0 0 1 1 1 0 0 1 0 1 0 0 0 Epc 0 0 1 0 1 0 1 1 0 0 0 1 0 0 NA 1 1 1 1 0 0 0 Hhr 1 0 1 1 0 1 0 1 1 0 1 1 1 0 NA 1 1 0 1 0 1 0 Jah 0 0 1 1 0 0 0 1 1 1 0 0 1 1 1 0 0 0 0 1 0 0 Jgd 0 0 1 1 0 0 0 1 1 1 0 0 1 1 1 0 0 0 0 1 0 0 Ppr 0 0 1 1 0 0 0 1 1 0 0 0 1 0 NA 1 0 0 0 0 1 0 Rtu 0 0 1 1 0 0 0 1 1 1 0 0 1 0 NA 1 1 0 1 0 1 0 Rpc 0 1 1 1 0 0 0 1 1 0 0 0 1 1 0 0 0 0 1 1 0 0 OTUs Characters 45 46 47 48 49 50 51 52 53 54 55 56 57 58 59 60 61 62 63 64 Apn 0 NA 0 1 0 1 1 1 1 1 1 1 1 0 0 0 NA 1 1 0 Agn 0 0 0 1 0 0 0 1 1 0 1 1 0 0 0 1 1 1 1 0 Bcr 0 1 1 1 0 0 1 1 0 0 0 1 0 0 0 0 NA 1 0 1 Bsg 0 1 1 1 0 0 1 1 0 0 1 0 0 0 0 0 NA 1 0 1 Evr 0 NA 0 1 0 1 0 0 1 0 1 1 0 1 1 1 1 0 NA 0 Epc 0 NA 0 1 0 1 0 0 1 0 1 1 0 0 0 1 0 1 0 1 Hhr 0 0 1 1 0 0 0 1 1 1 1 1 1 1 1 1 1 1 1 0 Jah 1 NA 0 1 0 0 1 0 1 0 1 0 0 1 0 1 1 1 1 0 Jgd 1 NA 0 0 1 0 0 1 1 0 1 1 0 0 0 1 1 1 1 0 Ppr 0 0 0 1 0 0 0 0 0 0 1 1 0 1 1 1 1 1 1 1 Rtu 0 0 0 1 0 0 0 0 0 1 1 0 1 0 0 1 1 1 1 1 Rpc 0 NA 1 1 0 0 0 0 0 0 0 1 0 1 1 1 1 1 1 0 Acanthoideae is a eurypalynous taxon. Pollen morphological variations are observed even within species of a genus. Carine and Scotlnad13 demonstrated that the genus Strobilanthesshowed diverse type of pollen morphology. Therefore pollen characters are important for taxonomic delimitation of the members of Acanthoideae and useful for taxonomic key preparation up to species. Both Lindau and Bremekamp used pollen characters for classification of Acanthoideae. Similarly the aestivation pattern within Acanthoideae got importance for taxonomic delimitation10. Besides, other gross morphological features have been utilized in the taxonomy of Acanthoideae Therefore in the present numerical taxonomic study the use of gross morphological characters with special emphasis on pollen characters is appropriate. Research Journal of Biological Sciences ___________________________________________________________ ISSN 2278-3202 Vol. 4(6), 48-55, June (2015) Int. Res. J. Biological Sci. International Science Congress Association 53 Figure-2 UPGMA dendrogram showing the resemblance among 12 species of Acanthoideae The selected OTUs represent 12 species under 10 genera of Acanthoideae. Two species of Barleria show the highest resemblance near Jaccard Index (JI) 0.75. In contrast, two species of Justicia show much lower level of resemblance near JI 0.6. Interestingly even being the members of two distinct genera, P. parviflora and R. pectinata, form a phenon with higher resemblance than J. adhatoda - J. gendarussa phenon. P. parviflora has been treated under sub-tribe Ruelliinae sensuScotnad and Vollesen and Rungia under Justiciinae sensuScotnad and Vollesen based on phylogenetic evaluations. The phenetic relationships among the taxa of Acanthoideae are varied with the result of cladistic analyses on Acanthaceae. Scotland and Vollesen 14 divided tribe Ruellieae into four sub-tribes, viz. Ruelliinae, Justiciinae, Andrographiinae and Barleriinae. Eranthemus, Hemigraphis, Paulopsis and Ruelliafall under Ruelliinae; Asystasia, Ecbolium, Justicia and Rungiafall under Justiciinae; Andrographis falls under Andrographiinae and Barleria falls under Barleriinae. But the dedrogram does not able to show such type of clear-cut distinction of tribes. Only near 0.4 Phenon line Barleriasegregated from rest of the studied taxa. So the sub-tribe Barleriinae is supported. The closeness of Ruellia and Hemigarphis supported the placement of these two genera under Ruelliinae, but the grouping of Eranthamus with Ecbolium(Justiciinae) and Phaulopsis with Rungia (Justiciinae) are ambiguous. Andrographis phenon is close to Justicia. Conclusion This study indicates that the phenetic relationship among members of Acanthoideae is distinct with that of the result of phylogenetic analysis. Now it is our discretion that which method should we follow. Two separate keys were prepared for identification purpose. Gross morphological characters key and pollen characters keys are given in table 4 and 5. Acknowledgement I would like to thank Principal, Barasat Govt. College and Head, PG Department of Botany, Barasat Govt. College for providing all necessary facilities to complete this research work. References 1.Long R.W., The genera of Acanthaceae in the southeastern United States, Journal of the Arnold Arboretum, 51, 257–309 (1970) 2.Willis J. C., A dictionary of flowering plants and ferns. Cambridge University Press, Cambridge (1980) 3.Mabberley D. J., Mabberley's Plant-Book- A portable dictionary of plants, their classification and uses, 3rd ed., Cambridge University Press, Cambridge, (2008) 4.Lindau G., Acanthaceae, In: Engler A. and Prantl, K. (eds.), Die Natfirlichen Pflanzenfamilien, 4 (3b), 274 - 353 (1895) 5.Olmstead R.G., Bremer B., Scott K.M. and Palmer J.D., A Parsimony Analysis of the Asteridae Sensu Lato Based on rbcL Sequences, Annals of the Missouri Botanical Garden, 80, 700-722 (1993) Research Journal of Biological Sciences ___________________________________________________________ ISSN 2278-3202 Vol. 4(6), 48-55, June (2015) Int. Res. J. Biological Sci. International Science Congress Association 54 Table-4 Artificial key to the specimens based on gross morphology 1 Herb; leaf asymmetric Phaulopsis parviflora 1 Shrub; leaf symmetric 2 2 Inflorescence one-sided spike or raceme 3 2 Inflorescence other than one sided spike 4 3 Rachis short, bract orbicular-lanceolate; corolla 2-lipped, stamens 2 Rungia parviflora 3 Rachis long, bract triangular; corolla 5-lobed, stamens 4 Asystasia gangetica 4 Shrub; inflorescence thyrse; Flower white with purplish spots, stamens 2 Justicia adhatoda 4 Herb or shrub; inflorescence spike; flower blue, stamens 4 5 5 Leaf margin serrate; ovule 3-8 in each cell Hemigraphis hirta 5 Leaf margin entire; ovule 2 in each cell 6 6 Corolla 2-lipped, corolla tube slender 7 6 Corolla 5 lobed, corolla tube funnel shaped 8 7 Upper lip and corolla tube narrow, flower light blue Ecbolium viride 7 Upper lip and corolla tube broad; flower purplish 9 8 Calyx-4, unequal; stamen exserted, 2 fertile and 2 rudimentary anthers 10 8 Calyx-5, equal; stamen inserted; 4 fertile anthers 11 9 Anther cells discrete Justicia gendarussa 9 Anther cells united Andrographis paniculata 10 Flower purplish in colour Berleria strigosa 10 Flower white in colour Barleria cristata 11 Stamens 4; ovule 8-10 in each cell Ruellia tuberose 11 Stamens 2; ovule 2 in each cell Eranthemum pulchellum Table-5 Artificial key to the specimens based on pollen morphology1 Pollen colporate 2 1 Pollen other than colporate 3 2 Pollen shape spheroidal 4 2 Pollen sub-prolate Andrographis paniculata 3 Pollen heterocolpate 5 3 Pollen homocolpate 6 4 Exine sculpturing finely lophoreticulate Barleria cristata 4 Exine sculpturing lophoreticulate Barleria strigosa 5 Aperture number 2-3 Justiciaadhatoda 5 Aperture number more than 5 7 6 Exine sculpturing reticulate 8 6 Exine sculpturing lophoreticulate 9 7 Pollen shape oblate-prolate Ecbolium viride 7 Pollen shape prolate or sub-prolate 10 8 Exine sculpturing coarsely reticulate, PA= 61.79 -83.5 m Asystasia gangetica 8 Exine sculpturing finely reticulate, PA= 30.06 – 41.75 m Justicia gendarussa 9 Pollen coporoidate, spheroidal in shape, exine c. 9.35 m thick Eranthemumpulchellum 9 Pollen colpoidorate, prolate-spheroidal in shape, Exine c. 5.84 m thick Ruellia tuberose 10 Aperture number 10 Hemigraphis hirta 10 Aperture number 10 11 11 PA= 41.75 – 50.10 m Phaulopsis parviflora 11 PA= 18.37 – 28.39 m Rungia pectinata Research Journal of Biological Sciences ___________________________________________________________ ISSN 2278-3202 Vol. 4(6), 48-55, June (2015) Int. Res. J. Biological Sci. International Science Congress Association 55 6.Bremekamp C.E.B., Delimitation and subdivision of the Acanthaceae, Bulletin of Botanical Survey of India, , 21-30 (1965) 7.Hedren M., Chase M.W. and Olmstead R.G., Relationships in the Acanthaceaea and related families as revealed by cladistic analysis of rbcL sequences, Plant Systematics and Evolution, 194, 93 – 109 (1995) 8.Scotland R.W., Sweere J.A., Reeves P.A. and Olmstead R.G. Higher-level systematics of Acanthaceae determined by chloroplast DNA sequences, American Journal of Botany, 82, 266-275 (1995) 9.McDade L.A. and Moody M.L., Phylogenetic relationships among Acanthaceae: evidence from noncoding trnL-trnF chloroplast DNA sequences, American Journal of Botany, 86, 70-80 (1999) 10.McDade L.A., Masta S.E., Moody M.L. and Waters E., Phylogenetic relationships among Acanthaceae: evidence from two genomes, Systematic Botany, 25, 106-121 2000) 11.Simpson M.G., Plant Systematics, Elsevier Academic Press (2006) 12.Furness C.A. and Grant M.C., Pollen morphology of some Ruellia species (Acanthaceae) from Africa and Madagascar, Grana, 35, 231 – 239 (1996) 13.Carine M.A. and Scotland R.W., Pollen morphology of Strobilanthes Blume (Acanthaceae) from southern India and Sri Lanka, Review of Palaeobotany and Palynology103, 143-165 (1998) 14.Scotland R.W. and Vollesen K., Classification of Acanthaceae, Kew Bulletin, 55, 513-589 (2000) 15.Rueangsawang K., Chantaranothai P. and Simpson D.A., Pollen morphology of Justicia L. (Acanthaceae) from Thailand and its taxonomic value, Grana52, 275–288 2013) 16.Sneath P.H.A. and Sokal R.R., Numerical taxonomy, Nature, 193, 855-860 (1962) 17.Sneath P.H.A and Sokal R.R., Numerical Taxonomy- The Principles and Practice of Numerical Classification, W.H. Feeman Company, USA (1973) 18.Harris J.G. and Harris M.W., Plant Identification Terminology- An Illustrated Glossary, 2nd ed. Spring Lake Publishing, Spring Lake, Utah (1999) 19.Hesse A.H., Halbritter R., Zetter M., Weber M., Buchner R., Frosch-Radivo A. and Ulrich S., Pollen terminology an illustrated handbook, Springer Wien, New York 2009) 20.Hammer Ø., PAST, ver. 3, Natural History Museum, University of Oslo (1999)