THE IDENTITY AND OCCURRENCE OF PHYLLANTHUS HOOKERI MUELL.-ARG. AND P. NOZERANII ROSSIGNOL & HAICOUR (EUPHORBIACEAE) IN INDIA

There are 16 species of Phyllanthus subgenus Phyllanthus reported from India. The present paper adds two more species that are invasive weeds in the paddy fields and on forest floors, namely Phyllanthus hookeri Muell.-Arg. and P. nozeranii Rossignol & Haicour of sect. Urinaria subsect. Urinaria of Phyllanthus. The former is somewhat woody and perennial whereas the latter is slender and monsoonal. The presence of these two taxa in India was brought to light in 1987 by Rossignol et al. based on the herbarium specimens collected earlier to 1863 and deposited at Paris from northeastern and southeastern India. Whilst Phyllanthus hookeri is overlooked or underrated by the taxonomists, P. nozeranii is misidentified and considered conspecific with P. urinaria L. Introduction The biovulate Linnean Phyllanthus L. (Euphorbiaceae) is not only a well-known medicinal plant genus with its diverse biomolecules but also has given the name to the recently resurrected segregate family Phyllanthaceae Martinov, by molecular taxonomists. The first comprehensive taxonomic treatment of this genus for India was provided by Hooker (1887) in his Flora of British India. Earlier, Roxburgh (1832) described 25 species of Phyllanthus. Chiefly confined to humid tropics of the world, the genus comprises 833 species (Govaerts et al., 2000). In India, it is represented by 53 species (Gangopadhyay et al., 2007). Phyllanthus L. (s.l.) is often divided into a number of subgenera, namely, Cicca, Emblica, Eriococcus, Isocladus, Kirganelia, Phyllanthus, Xylophylla, etc. The subgenus Phyllanthus is characterized by herbs or low woody shrubs bearing colporate pollen grains, tricarpellary capsular fruits with six, striate and/or foveolate seeds. It is represented by 16 species in India (Gangopadhyay et al., 2007). The present paper reports Phyllanthus hookeri and P. nozeranii of subgenus Phyllanthus as additions to the Indian flora. These, in fact, were reported earlier but either overlooked by authors or often misidentified, or treated conspecific with Phyllanthus urinaria; the fourth of the six species described by Linnaeus in his Species Plantarum (1753). Later, Roxburgh (1832: 660) considered P. urinaria as the second species under Section II (Leaves pinnate; obviously, Roxburgh mistook the phyllanthoid branchelets as pinnate leaves). Phyllanthus urinaria L. was kept in section Urinaria by *Corresponding author. Email: satyavatsa@yahoo.co.in


Introduction
The biovulate Linnean Phyllanthus L. (Euphorbiaceae) is not only a well-known medicinal plant genus with its diverse biomolecules but also has given the name to the recently resurrected segregate family Phyllanthaceae Martinov, by molecular taxonomists.The first comprehensive taxonomic treatment of this genus for India was provided by Hooker (1887) in his Flora of British India.Earlier, Roxburgh (1832) described 25 species of Phyllanthus.Chiefly confined to humid tropics of the world, the genus comprises 833 species (Govaerts et al., 2000).In India, it is represented by 53 species (Gangopadhyay et al., 2007).
The present paper reports Phyllanthus hookeri and P. nozeranii of subgenus Phyllanthus as additions to the Indian flora.These, in fact, were reported earlier but either overlooked by authors or often misidentified, or treated conspecific with Phyllanthus urinaria; the fourth of the six species described by Linnaeus in his Species Plantarum (1753).Later, Roxburgh (1832: 660) considered P. urinaria as the second species under Section II (Leaves pinnate; obviously, Roxburgh mistook the phyllanthoid branchelets as pinnate leaves).Phyllanthus urinaria L. was kept in section Urinaria by Webster while Central Asia is conceived as the centre of origin of this section which is of interest due to hepatoprotective and other medicinal uses (Lee et al., 2006;Komuraiah et al., 2009).Chaudhary and Khan (2003) studied the SEM of seeds of nine herbaceous species of Phyllanthus from India and described these under three morphotypes, with Phyllanthus urinaria placed in P. urinaria type.
It is the experience of the taxonomists in general and of experts of the genus in India in particular that there exist specimens of Phyllanthus evincing affinity with P. urinaria but do not match wholly with the technical description of it in the regional floras.Rossignol et al. (1987) were the first to draw the attention to the discontinuities within the section Urinaria in the characteristics like pilosity, length of the internodes and of plagiotrophic (phyllanthoid) shoots, the number of leaves on it and the number of colpii of the pollen, the presence or absence of foveoles on the lateral sides of seeds, ploidy level, etc.On the basis of morphology, cytology, genetics and biometry, a new classification was presented by Rossignol et al. (1987) in which the allied species of P. urinaria or "urinaria complex", are placed in the subsection Urinaria Haicour & Rossignol, recognizing two subgroups within it on spermoderm ornamentation.Each of these lines is represented by two species which differ from either in somatic chromosome number: 50 (P.nozeranii) and 100 (P.embergeri) in the "spiraled" line, 50 (P.urinaria) and 100 (P.hookeri) in the "radiated" line.The members of the subsection are characterized by 4-5 colporate, prolate pollen with bi-reticulate ornamentation (Chen and Wu, 1997;Chen et al., 2009).
Certain of the herbarium specimens of Phyllanthus urinaria complex available at Madras Herbarium, Coimbatore (MH) and Kakatiya University Herbarium, Warangal (KUH) including the live plants in the botanical garden, Kakatiya University campus belong to the less-known but described species, viz.Phyllanthus hookeri Muell.-Arg.and P. nozeranii Rossignol & Haicour.Curiously, there was no mention of P. nozeranii in the recent account of Phyllanthus from India by Gangopadhyay et al. (2007).However, Phyllanthus hookeri Muell.-Arg., P. urinaria var.hookeri (Muell.-Arg.)Hook.f. and P. urinaria var.oblongifolia Muell.-Arg.were treated conspecific with P. urinaria L. Ramla (1995) described and segregated the seeds of P. urinaria complex in Kerala State as P. urinaria -'spiraled' (S) and 'radiated' (R) on seed coat ornamentation, as has been done by Rossignol et al. (1984Rossignol et al. ( , 1987)).But, she did not go further to distinguish them at the species level though well aware of the work of Rossignol et al. (1987).Later, Chaudhary and Rao (2002) treated P. urinaria, without commenting on the apparent morphological variation.Chaudhary and Khan (2003:118) stated that the seed of P. urinaria has 12-15 transverse ridges and 1-3 circular pits on the sides (a routine description copied from floras, e.g.Philcox, 1997) while their photographs show only single large pit.

Discussion
Among the eight secondary metabolites screened for Phyllanthus hookeri, P. nozeranii and P. urinaria, alkaloids are present while iridoids absent in all.Steroids are exclusive to P. urinaria, lignins, methylene-dioxy compounds and triterpenoids to P. hookeri and ellagic acid to P. nozeranii.However, tannins are shared by P. hookeri and P. nozeranii.The distribution of 21 known and 24 unknown amino acids, 10 known and 4 unknown phenolic acids and 8 secondary metabolites scored for 17 species of Phyllanthus revealed 25.0 paired affinity between P. nozeranii and P. urinaria, 23.0 between P. hookeri and P. nozeranii, and 15.3 between P. hookeri and P. urinaria.The isolation values are 50 for P. urinaria, 30.7 for P. hookeri and 22.2 for P. nozeranii (Komuraiah, 2009).
Leaf proteins in P. hookeri and P. nozeranii were studied using SDS-PAGE.Based on the mobility of the protein bands, the species of Phyllanthus were categorized into A (Slow: R f 0.4-1.5),B (Intermediate: R f 1.6-2.1),C (Fast: R f 2.8-3.9) and D (Very fast: R f 4.0-5.1).P. hookeri showed exclusive bands in A and B while P. nozeranii developed a band is C (Fast) while both shared bands in D (Very Fast).When the paired affinity and isolations were calculated for 11 species of Phyllanthus, P. hookeri and P. nozeranii showed zero paired affinity whereas P. hookeri evinced highest isolation value of 54.5.