Diversity, distribution and frequency based attributes of Phytoliths in ...

Bioscience Discovery, 8(2): 224-234, April - 2017 © RUT Printer and Publisher Print & Online, Open Access, Research Journal Available on http://jbsd.in ISSN: 2229-3469 (Print); ISSN: 2231-024X (Online)

Research Article

Diversity, distribution and frequency based attributes of Phytoliths in some Panicum spp. of Tripura, North East India Ashish Kumar Chowdhury* and B K Datta Plant Taxonomy and Biodiversity Laboratory, Department of Botany, Tripura University, Suryamaninagar799022, Tripura. * [email protected] Article Info Received: 14-03-2017, Revised: 27-03-2017, Accepted: 29-03-2017 Keywords: Grass, Opal, Panicum, Panicoideae, Phytoliths

Abstract The Phytoliths are micrometric particles protect the plant from various biotic and abiotic stresses. The present study is aimed to find out the morphological diversity, distribution and frequency of opal phytoliths in the leaves of Panicum notatum, Panicum repens and Panicum brevifolium by wet oxidation method. A lot of variation was observed in the morphotypes of phytolith present in the leaves of three grass species. A total thirteen phytoliths morphotype were observed during the course of study. Short cell bilobate and short cell cross are noted as the most common phytoliths morphotype present in all the three species and possess the characteristic features of the subclass Panicoideae. Short cell bilobate, Cuneiform bulliform and Bulliform parallepipedal phytoliths morphotype were observed in Panicum notatum and Panicum brevifolium but absent in Panicum repens. Based on the observations it can be conclude that the phytoliths produced by grass species posses definite taxonomic significance and serve as a useful tool for the identification and classification of grass species.

1960; Rover, 1983; Twiss, 1987; Piperno, 1988; INTRODUCTION Phytoliths (Greek, phyto= plant, lithos= stones) or Carter, 1999). Majority of archaeologist and plant stones are hydrated amorphous silica (SiO2, palaeoecologist have been using this kind of study H O) absorbed by roots from soil as polymerized for the reconstruction of past environment as well as n 2 form of monosilicic acid [Si(OH)4] and are past plant -people relationship (Wilding and Dress, deposited in or between cells of living plant tissues 1971; Mulholland, 1989; Mulholland and Rapp, of several plant families (Jones and Handreck, 1992). 1967; Pearsall, 2000 ). Phytoliths provide Some plants were found to produce distinguishable, mechanical support, stiffness to the plant and characteristic or “diagnostic” shaped phytoliths and protect them from various biotic and abiotic stresses considered as useful tools in archaeology, (Mazumdar, 2011). Because of their consistent angiosperm taxonomy (Piperno, 1988; Ghosh et al., shape within species, phytoliths provide significant 2011; Barboni and Bremond; Jattisha and Sabu, taxonomic information (Jones and Handreck, 1965; 2012; Hari Babu et al.,2015;) and recently in Blackman, 1969; Raven, 1983; Piperno, 1988). nanotechnology (Neethirajan et al., 2009). The Since, the structural hierarchy and resistant family Poaceae produces abundant, diverse and siliceous structures of this element in the form of distinctive opaline silica bodies with diagnostic phytoliths can remain stable in soil for hundreds and morphological features that permit identification even millions of years after plant tissues decay, they upto subfamily, or in some instances, lower are important in palaeoecological studies (Matcalfe taxonomic levels (Hodson et al., 2005). http://jbsd.in ISSN: 2229-3469 (Print) 224

Chowdhury and Datta Now a day’s phytoliths were used as an emerging taxonomic tool for identification of plants including grasses (Jattisha and Sabu, 2012; Shakoor et al., 2014; Sobhakumari and Kumarasamy, 2014, Shakoor and Bhat, 2014; Hari Babu et al., 2015). Grass phytoliths indices are also used for vegetation and climate study (Biswas et al, 2016). However, existing literature revealed that there is meager study confining the Phytolith morphotype from the Grasses. In this connection the present study is concerned with the study of diversity, distribution and the frequency of phytoliths in the leaves of three grass species (Panicum repens, Panicum notatum and Panicum brevifolium) which provide taxonomically significant features and strive to construct taxonomic tool for the identification of the grass species in the vegetative stage.

Phytoliths were extracted from fresh and mature leaves of three grass species by wet oxidation method (Mazumdar and Mukhopadhyay, 2009) with slight modification. All the collected samples were cleaned and take to an ultrasonic water bath followed by dried in hot air woven at 60oC for 2 hours. After that approximately 2g of oven dried sample were taken in test tubes containing 15 ml of saturated Nitric acid for overnight. In the next day the mixture were centrifuged at 2500 rpm for 10 min, decanted, and then boiled with 10% Hydrochloric acid and washed with distilled water for 2-3 times. The processed materials were dipped in acetone and again centrifuged at 2500 rpm for 10 minutes. The acetones were then evaporated from the samples. The sediments were transferred to storage vials. The residual subsamples were mounted with Canada balsam and observed under light microscope (Olympus CX21i). A minimum of 300 phytoliths grains were counted for each samples. The phytoliths morphotype were classified by using International Code for Phytolith Nomenclature, ICPN (Madella et al. 2005) and Phytolith Core (Phytolith database, GPEG-2014) classification.

MATERIALS AND METHODS Sample collection Healthy, fresh and mature leaves of three grass species (Panicum notatum Retz., Panicum repens L. and Panicum brevifolium L.) were harvested from Suryamaninagar (23o45′38.4′′ N and 91o15′54.0′′ E). The collected species were identified with the help of standard literature and local flora (Deb, 1983) and their herbaria prepared following conventional method (Jain and Rao, 1997). The herbaria were deposited in the Department of Botany, Tripura University. Isolation of phytoliths from leaves

RESULTS AND DISCUSSION In the present study a wide range of phytoliths morphotype were observed from the leaves of three grass species namely Panicum notatum, Panicum repens and Panicum brevifolium and were depicted in the table 1.

Table 1: Different types of observed phytoliths with their abbreviations. Sr.

No.

Morphotype

Abbreviations

1 2 3 4 5 6 7 8

Short cell bilobate Cylindrical polylobate Short cell rondel Cuneiform bullifom Bulliformparallepipedal Long cell echinate Short cell cross Globular cell

SCB CP SCR CB BP LCE SCC

9 10 11 12 13

Cylindric smooth Stomata Epidermal appendage hair cell Epidermal appendage trichome Trapeziform short cell

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Chowdhury and Datta The length and width of each observed phytoliths isolated from the leaf samples were measured (Table 2). The frequency assembled denoted in the table 3 & figure 1, 2, 3. The distributions of Phytolith morphotypes were tabulated in the table 4. Photographs of different types of phytoliths were depicted in figure 4-6. The data obtained from the frequency phytoliths assemblages as well as measurements were diverse in nature. The different phytoliths morphotype that were isolated from the leaves of selected grass species can be summarized as follows: P. notatum Retz.: The different morphotype isolated from the leaves of P. notatum are short cell bilobate, cylindrical polylobate, short cell rondel,

cuneiform bulliform, Bulliform parallepipedal, long cell echinate, cylindric smooth, stomata, short cell cross, epidermal appendage trichome and trapeziform short cell (Table 2, Fig. 4). Short cell bilobate (SCB) includes bilobate with convex end, bilobate with concave end, bilobate with flat end and bilobate with nodular shank. Short cell cross (SCC) includes cross with thick shank and cross with thin shank. Among the morphotypes observed short cell bilobate (SCB) is the most frequent (almost 60%) morphotype followed by cylindrical polylobate (CP) with almost 16 % frequency and stomata (ST) shows least frequent (almost 0.49%) phytoliths morphotype. Other morphotypes shows very less frequency (Table 3, fig 1).

Table 3: Frequency (%) of Phytolith morphotypes of eight grass species studied. Phytolith Morphotypes

Panicum notatum

Panicum repens

SCB 60.26±1.88 26.47±2.03 16.37±2.01 CP 2.05±0.72 22.74±2.21 SCR 0.96±0.39 CB 1.18±0.33 BP 7.31±1.65 10.16±1.53 LCE 3.92±0.32 5.56±1.15 SCC 1.47±0.12 GC 4.29±1.74 16.05±1.15 CS 0.49±0.17 1.86±0.66 ST 3.43±0.12 EAHC 2.24±0.39 1.27±0.23 EAT 1.09±0.45 11.22±1.10 TSC Values are (Mean±SD), (n=5) Values with (-) denotes absence of phytolith morphotype. P. repens L. : The different morphotype isolated from the leaves of P. repens are short cell bilobate, short cell rondel, long cell echinate, globular smooth, cylindric smooth, stomata, short cell cross, epidermal appendage trichome, epidermal appendage hair cell and trapeziform short cell (Table 2, Fig. 5). Short cell bilobate (SCB) includes bilobate with convex end, bilobate with concave end, bilobate with flat end and all these types are short shanked few types have thick shank. Short cell rondel (SCR) includes spool tower flat tower and ellipsoid rondel. Long cell echinate (LCE) includes LCE with concave end and convex end. Among the morphotypes observed short cell bilobate (SCB) is http://biosciencediscovery.com

Panicum brevifolium 60.53±2.14 13.43±0.9 1.50±0.25 3.45±0.44 2.19±0.07 3.32±0.29 8.36±0.85 0.61±0.06 3.04±0.32 0.77±0.1 1.05±0.17 1.90±0.15

the most frequent (almost 26%) morphotype followed by short cell rondel (SCR) with almost 22% frequency, cylindrical smooth(CS) with almost 16 % frequency, long cell echinate with almost 10 % frequency and epidermal appendage trichome (EAT) shows least frequent (almost 1%) phytoliths morphotype. Other morphotypes shows very less frequency (Table 3, fig 2). P. brevifolium L.: The different morphotype isolated from the leaves of P. brevifolium are short cell bilobate, cylindrical polylobate, short cell rondel, cuneiform bulliform, Bulliform parallepipedal, long cell echinate, short cell 227

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Bioscience Discovery, 8(2): 224-234, April - 2017 cross, globular cell, cylindric smooth, stomata, epidermal appendage trichome and trapeziform short cell (Table 2, Fig. 6). Short cell bilobate (SCB) includes bilobate with convex end, bilobate with concave end, bilobate with flat end, bilobate with long shank, bilobate with distinct side lines and bilobate with nodular shank. Cylindrical polylobate (CP) includes three lobate as well as polylobate morphotype. Among the morphotypes observed short cell bilobate (SCB) is the most frequent (almost 60%) morphotype followed by cylindrical polylobate (CP) with almost 13 % frequency, Short cell cross with almost 8 % frequency, globuler cell (GC) and stomata (ST) shows less than 1% frequeny. Other morphotypes also shows very less frequency (Table 3, fig 3). A total of thirteen different phytoliths morphotype were observed in the three grass species and short cell bilobate (SCB) noted as most abundant morphotype and considered as an

important characteristic feature of the subclass Panicoidae (Twiss et al, 1969). SCB, SCR, LCE, SCC, CS, EAT and TSC types of phytoliths are common among the three species. CP, CB and BP phytoliths morphotype were observed in both the species of P.notatum and P.brevifolium, but absent in P. repens. EAHC type of phytolith was observed in P. repens and absent in P. notatum and P. brevifolium. GC type of phytolith was recorded as the least abundant phytolith morphotype found in P. repens and P. brevifolium whereas absent in P. notatum. Over the last few decades plant physiologist, taxonomist and archaeologist have been investigating the occurrence of phytoliths in various plant families and make an attempt to find out their role in the archaeological, palaeobotanical, morphological, physiological and taxonomical aspects (Piperno and Pearsall. 1998;Portillo et al., 2005; Jattisha and Sabu, 2012).

Table 4: Occurrence of Phytolith morphotype in eight grass species Phytolith Morphotypes

Panicum notatum

Panicum repens

Panicum Brevifolium

SCB CP SCR CB BP LCE SCC GC CS ST EAHC EAT TSC

+ + + + + + + + + + +

+ + + + + + + + + +

+ + + + + + + + + + + +

(+ ) means presence, (- )means absent During this study 13 morphotypes of phytoliths were observed with huge morphological diversity in each type. These shapes were reported by earlier workers in different grass species (Piperno and Pearsall, 1998; Jattisha and Sabu, 2012; Sobhakumari and Kumarasamy, 2014). Fredlund and Tieszen, 1994, reported the occurance of short shanked and convex ended bilobate cells in some panicoidae grasses. Ghosh et al., 2011, reported the occurance of lobate, short cell cross and elongated phytoliths morphotype in P. repens http://jbsd.in

and other Panicoidae grass with varying frequency. The present study reveals the occurrence of different forms of bilobate and short cell cross phytoliths morphotype along with other morphotype showing variation in frequency percentage and also in structure among the three species of Panicoidae grass. Hari Babu et al., 2015, reported the occurance of different phytoliths morphotypes in twenty two grass species including P.repens that represent only bilobate short cell and trapeziform short cell. 228

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Fig. 2: Graphical representation of percent (%) frequency (Mean±SD) (n=5) of Phytolith Morphotypes Panicum repens


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Fig.3: Graphical representation of percent (%) frequency (Mean±SD) (n=5) of Phytolith Morphotypes Panicum brevifolium Present study reveals the occurance of bilobate short cell and trapeziform short cell along with short cell rondell, long cell echinate, short cell cross, Globular cell, cylindric smooth, stomata, epidermal appendage hair cell and epidermal appendage trichome phytoliths morphotype. Ghosh et al., 2011, also reported the occurance of lobate, cross and elongate type of phytoliths in P. repens and also in other few Panicoidae grasses that are also observed in this present study. Jattisha and Sabu, 2012, reported the occurance of short cell bilobate, bulliform fan, long cell echinate and other phytoliths morphotype of some Chloridoideae grasses in Kerala that are varied in structure, measurements and also in frequency as compared to the present study. The present study has also brought to light some novel types of phytolith such as globular cell, bilobate with nodular shank and bilobate with long shank that may be useful as taxonomic marker for identification of grasses at lower taxonomic level. The analysis of three grass species reveals that the key combination of phytolith morphotypes that are diagnostic up to the subfamily and sub-tribe level. Surface structure and the morphometric measurements also provide an additional parameter for the characterization and identification of grass species. The present study may indicate that the http://jbsd.in

phytolith assemblages study can be used as a marker to differentiate between the species of grasses. It can also be conclude that the frequency and different morphotypic structures may be utilized as a marker for identification of the grasses up to the species level even in the vegetative stage. Apart from that the phytoliths morphotype is also helpful in the reconstruction of the vegetation of the plants. ACKKOWLEDGEMENT Authors are thankful to the Head, Department of Botany for encouragement and Lab facilities. The first author is thankful to UGC for providing financial support under SAP-BSR. REFERENCES Blackman E, 1969. Observation on the development of the silica cells of the leaf sheath of wheat. Can. J. Bot., 47:827–838. Carter JA, 1999. Late Devonian, Permian and Triassic phytoliths from Antarctica. Micropaleontology, 45: 56–61. Barboni D and Bremond L, 2009. Phytoliths of East African grasses: an assessment of their environmental and taxonomic significance based on floristic data. Rev. Palaeobot. Palynol., 158 (1-2): 29–41. 230

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Fig 4: Phytolith morphotypes of Panicum notatum at 400x, Scale = 10 µm. ( A-B) CP, (C) One CP & one SCB, (D-F) SCB with long shank, (G) SCB with nodular shank, (H) Two SCB with shank, (I,J,L)SCC, (K) One SCR, (M-N)LCE, (O) One CS, (P) OneEAT, (Q) One TSC, (R) One BP, (S) One ST, (T) One CB.


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Fig 5: Phytolith morphotypes of Panicum repens at 400x, Scale = 10 µm. (A –E) SCB, (F-I) SCR, (J) Ellipsoid SCR, (K-L) LCE with flat end, (M-O) LCE with concave end, (P-Q) EAT, (S) One ST, (T) One SCC, (R &U)EAHC, (V-W) CS, (X) TSC, (Y) GC

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Fig 6: Phytolith morphotypes of Panicum brevifolium at 400x, Scale = 10 µm. (A-C) SCB, (D) One SCR & one SCB, (E) One SCB with long shank & one SCC, (F-H) SCB with nodular shank, (I) One CP & one SCB, (J-L) CP, (M) EAT, (N) SCC, (O) Two SCB, (P) One SCB, (Q-R) CS, (S-T) LCE, (U) One CB & one LCE, (V) Two TSC, (W) One CB, (X) One GC, (Y) One ST. Biswas O, Ghosh R, Paruya DK, Mukherjee B, Thapa KK and Bera S,2016. Can grass phytoliths and indices be relied on during vegetation and climate interpretations in the eastern Himalayas? Studies from Darjeeling and Arunachal Pradesh, India. Quat. Sci. Rev.114:100-115.


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How to Cite this Article: Ashish Kumar Chowdhury and B K Datta, 2017. Diversity, distribution and frequency based attributes of Phytoliths in some Panicum spp. of Tripura, North East India. Bioscience Discovery, 8(2):224-234.

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