Available Online at http://www.recentscientific.com International Journal of CODEN: IJRSFP (USA) Recent Scientific

International Journal of Recent Scientific Research Research Vol. 10, Issue, 11(C), pp. 35918-35924, November, 2019 ISSN:0976-3031 DOI: 10.24327/IJRSR Research Article

SYSTEMATIC SIGNIFICANCE OF POLLEN MORPHOLOGY IN SOUTH INDIAN SPECIES OF SCHAEFFER ()

Remya Krishnan R.V1*, Santhoshkumar E.S2 and Radhamany P.M1

1Department of Botany, University of Kerala, Kariavattom P.O., Kariavattom, Kerala-695581, India 2Jawaharlal Nehru Tropical Botanic Garden and Research Institute, Karmankode P.O., Palode Thiruvananthapuram district, Kerala-695562, India

DOI: http://dx.doi.org/10.24327/ijrsr.2019.1011.4196

ARTICLE INFO ABSTRACT

Article History: Cinnamomum Schaeffer belongs to the family Lauraceae. Pollen morphological characters were studied in 25 taxa of Cinnamomum Schaeffer, which include 22 wild and 3 cultivated ones. All the Received 4th August, 2019 th taxa showed variation inpollen size, exine ornamentation, exine thickness and inter spinular Received in revised form 25 distance. The first Principal Component (PC) accounted for 97.6% of phenotypic variance followed September, 2019 by second for another 0.935%. Major pollen traits that accounted for more variability in PC1, PC2 Accepted 18th October, 2019 th and PC3 include polar length, equatorial diameter, exine thickness, nature of granules and Published online 28 November, 2019 arrangement of spinules. The unweighted pair- group method with mathematical averaging (UPGMA) clustering method revealed two principal clusters which separated all the accessions Key Words: between euclidean distances of 0.109 – 0.741. Both cluster analysis and principal co-ordinate

Cinnamomum, pollen morphology, PCA, analysis revealed that Cinnamomum moozhiyarense (CMz) collected from Pathanamthitta, UPGMA, PCoA. Cinnamomum travancoricum (CT), collected from Idukki and Cinnamomum walaiwarense (CWa) from Thrissur have morphologically distinct pollen grains. The phenogram and PCoA scatter plot showed the grouping of accessions with species delimitation, which emphasize the importance of pollen morphology as a key character in identifying Cinnamomum at species level.

Copyright © Remya Krishnan R.V, Santhoshkumar E.S and Radhamany P.M,2019, this is an open-access article

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INTRODUCTION of taxonomic uncertainty. The genus is usually recognised by trinerved and fragrant , paniculate inflorescence, The genus Cinnamomum Schaeffer belongs to the family with nine stamens and seated on a cupule. But there are a Lauraceae. The term ‘Cinnamomum’ is derived from the Greek lot of taxonomic disputes regarding the identification of word Kinnamon or kinnamomum meaning sweet wood. It at species level (Abeysinghe et al., 2009). Morphology of consists of about 350 species distributed in East Asia, South pollen is an efficient marker in identifying plants at all levels of East Asia, Australia, Samoa, Fiji and Tropical America taxa. But, the pollen morphology of Cinnamomum were (Mabberley, 2008).To date, The List includes 893 scarcely described and discussed. Therefore, in the present scientific plant names of species rank for the genus study, pollen morphology has been studied to delimit and Cinnamomum (http://www.theplantlist.org, last accessed 11 unravel the species complexities to solve the taxonomic April 2019) and 48 of these are attributed to species occurring disputes of Cinnamomum. in India, which are mainly distributed in the Western Ghats, Eastern Ghats and the Himalayas (Baruah & Nath, 2007; MATERIALS AND METHODS Geethakumary et al., 2015; Gangopadhyay, 2008). South India In the present study, twenty five taxa of Cinnamomum were houses about 26 species and they constitute 20 endemics, 4 collected from different localities of Kerala. Specimens were cultivated species (Cinnamomum verum, C. cassia, C. identified using authentic literature and comparing with the camphora & C. tamala) and two distributional records, like C. relevant type specimens deposited at various herbaria. Flower dubium and C. litseaefolium (Geethakumari et al., 2007; buds just before the anthesis were collected and fixed in glacial Geethakumari et al., 2012). acetic acid. The pollen grains were acetolysed according to the

following procedure proposed by Erdtman (1960). Cinnamomum is a highly complicated genus due to overlapping phenotypic characters in many species and become a complex Acetolysis

*Corresponding author:Remya Krishnan R.V Department of Botany, University of Kerala, Kariavattom P.O., Kariavattom, Kerala-695581, India International Journal of Recent Scientific Research Vol. 10, Issue, 11(C), pp. 35918-35924, November, 2019

The floral buds fixed in glacial acetic acid were crushed with a RESULTS AND DISCUSSION glass rod in centrifuge tube. Three milliliter of freshly prepared Palynological features have, however, attracted little attention acetolysis mixture (9 parts acetic anhydride to 1 part for defining the closely related species of Cinnamomum. The concentrated Sulphuric acid) was added to the content in the pollen grains of Cinnamomum were found to be spheroidal, tubes. The content was heated in a water bath from 70˚C to prolate-spheroidal or sub-prolate; apolar and inaperturate, with boiling point and stirred occasionally. The centrifuge tubes and highly reduced exine consisting of a thin coherent layer content were left in boiling water for 3 minutes and then ornamented with spinules. Spinules are usually broad at base centrifuged at 4000 r.p.m. for 5 minutes while still hot. The and with pointed, acuminate, acute or blunt tips. They are supernatant was decanted into acetolysis waste bottle before the characterised by the presence of a conspicuous, circular, addition of some water to the sediments in the tubes, and was cushion- like base which invariably protrudes proximally. Its shaken vigorously using a whirl mixer. Few drops of surface is studded with granular to globular elements (Figs. 1, methylated spirit were added to remove the foam formed and 2, 3 & 4). Out of the 25 taxa investigated, the largest pollen centrifuged again. The supernatant was decanted, washed with grains are those of C. moozhiyarense, followed by C. water, and centrifuged repeatedly four times. Fifty percent travancoricum and smallest are those of C. walaiwarense. The glycerin was added and left standing for two hours. The tubes pollen grains of the species differ in the exine ornamentation, were shaken vigorously using a whirl mixer and centrifuged at exine thickness, inter- spinular distance and size of pollen 4000 rpm. for 10 minutes. The supernatant was finally (Tables 1 & 2). Hence these characters can be used to support decanted off, and the tube was inverted over filter paper and the of Cinnamomum at species level. left overnight. 100% glycerol was added to the tubes, shaken and poured into labelled storage vials. The pollen grains were mounted in unstained glycerin jelly. The slides were examined with a Fisher scientific illumination microscope under (E 40; 0.65), oil immersion (E 100; 1.25) using 10× eye piece. The measurements were based on 20 readings from each specimen. Photomicrographs were taken using Leica CME with Digital Microscope. Light microscopic preparations were made by mounting the acetolysed pollen grains in glycerine jelly. The quantitative measurements were done at x100 magnification for 20 pollen grains of each taxon. Light microscopic studies were carried out by using Olympus CX21i.

Scanning Electron Microscopic (SEM) Analysis

Pollen grains for the SEM analysis were mounted directly on graphite tape and sputter coated for 45 seconds with gold in Fine Coater (JFC-1200, Joel, Japan), analysed and photomicrographed by SEM (JSM-5600 LV, Joel, Japan) at a voltage ranging 10-12 kV. The terminology for pollen description were adapted from Nair (1970) and Walker and Doyle (1975). Considering the longest diameter, the size classes of pollen have been determined following the classification proposed by Erdtman (1970). A total of 15 characters (9 qualitative and 6 quantitative) were selected for the study. Fig 1SEM Photographs of pollen of Cinnamomum. A & D- C. agasthyamalayanum, B & E- C. cassia, C & F- C. chemungianum, G & J- C. The pollens were observed, measured and recorded using 20 dubium, H & K- C. filipedicellatum, I & L- C. goaense. samples for each taxon. Observations on 15 morphological Analysis of variance traits were scored including both quantitative and qualitative characters. All data were processed using one way ANOVA Analysis of variance and Kruskal Wallis Test performed on and Duncan Multiple Range Test and the significance of quantitative and qualitative data respectively showed qualitative characters were tested using Kruskal Wallis Test significant (P <0.05) variation among the 25 taxa of using SPSS 16.0. A probability value of 0.05 was used as a Cinnamomum studied (Table 1). Polar length and equatorial bench mark for significant difference between parameters. diameter of pollen were significantly high (P <0.05) in C. Multivariate analysis was performed by numeric taxonomic moozhiyarense, while, C. walaiwarense has the lowest mean techniques using the procedure of Principal Component value for polar length and equatorial diameter of pollen. Analysis (PCA) (Sneath and Sokal, 1973) using MVSP Version Multivariate analysis was performed in order to find out the 3.1 (Kovach computing Services, Wales, U K). To bring out significant pollen characters and also to find out the species the patterns of similarity and dissimilarity, data was subjected relationships within the genus based on pollen characters. to cluster analysis based on UPGMA method to group the 25 taxa using DARwin 6. Principal Coordinate Analysis (PCoA) followed by construction of scatter plots based on ordination of accessions was performed using PAST 3.01.

35919 | P a g e Remya Krishnan R.V, Santhoshkumar E.S and Radhamany P.M., Systematic Significance of Pollen Morphology In South Indian Species of Cinnamomum Schaeffer (Lauraceae)

polar length, equatorial diameter, exine thickness, nature of granules and tip of spinules.

Fig 2SEM Photographs of pollen of Cinnamomum. A & D- C. heyneanum, B & E- C. keralaense, C & F- C. litsaeafolium, G & J- C. malabatrum, H & K- C. macrocarpum, I & L -C. mathewianum.

Fig 4SEM Photographs of pollen of Cinnamomum. A& D- C. sulphuratum, B & E- C. travancoricum, C & F- C. tamala, G & J- C. verum, H & K- C. wightii, I & L-C. walaiwarense, M & N- C.moozhiyarense

Cluster analysis

The unweighted pair group method with mathematical averaging (UPGMA) clustering method with 15 variables revealed two principal clusters which separated all the 25 taxa between euclidean distances of 0.109 – 0.741 (Fig. 5). Cinnamomum moozhiyarense was found to be most distantly placed among all the taxa followed by C. walaiwarense and C. travancoricum which have the smallest and second largest pollen grains respectively.

Fig 3SEM Photographs of pollen of Cinnamomum. A & D- C. mathewianum var. ponmudianum, B & E- C. mohananii, C & F- C. nicolsonianum, G & J- C. nilagiricum, H & K- C. perrottetii, I & L- C. riparium.

Principal Component Analysis

The first Principal Component (PC1) accounted maximum variation (Table 3). The PC1 had all the characters like polar length, equatorial diameter, exine thickness and tip of spinules as traits with the highest loadings. In addition to that, all the traits had positive values also. The second component accounted for 0.95% of the variability followed by third principal component with 0.5% variability. Major traits that Fig 5 UPGMA phenogram based on qualitative and quantitative pollen accounted for more variability in PC1, PC2 and PC3 include characters (DARwin 6).

35920 | P a g e International Journal of Recent Scientific Research Vol. 10, Issue, 11(C), pp. 35918-35924, November, 2019

Table 1 Quantitative pollen morphological characters of Cinnamomum

Equitorial Polar Length of Interspinular Taxa Abbreviations Diameter P/E X100 Exine Thickness(µm) length(P)µm Spinule(µm) Distance(µm) (E)µm

ijk jk c efgh def de C. agasthyamalayanum CAg 27.51 ± 0.29 27.11±0.323 101.6±10.07 0.94±0.023 0.83±0.090 1.32±0.090 de efg b a def a C. cassia CCa 31.41 ± 0.46 30.63 ±0.449 103.2±1.25 1.51±0.075 0.82±0.049 2.50±0.160

o op b fghi fghi efgh C. chemungianum CCh 24.04 ±1.17 23.27±1.120 103.5±0.342 0.89±0.052 0.67±0.048 1.00±0.100 jkl kl b cde fghi hi C. dubium CD 26.75 ±0.22 26.41±0.208 101.3±0.26 1.05±0.064 0.66±0.090 0.75±0.048

mno mno b k a hi C. filipedicellatum CF 25.12 ±0.19 24.42±0.325 103±0.667 0.61±0.058 1.61±0.136 0.72±0.029

lmn mn b cdef fghij b C. goaense CG 25.74 ±0.206 24.90±0.305 103.1±0.433 1.00±0.009 0.62±0.015 1.96±0.145 ghij jk b efgh ij efg C. heyneanum CH 27.93 ±1.08 27.02±0.898 103.2±0.742 0.96±0.013 0.50±0.032 1.22±0.079

cd de b cdef bcd cd C. keralaense CK 32.52 ±0.495 31.72±0.424 102.7±0.517 1.00±0.044 1.14±0.084 1.62±0.075 c cd b fghi cd efgh C. litseaefolium CL 33.35 ±0.598 32.72±0.620 102±0.558 0.88±0.011 1.01±0.073 1.00±0.098

fgh hi b cd efgh cd C. macrocarpum CMa 32.33 ±0.404 31.6±0.293 102.60±1.07 0.93±0.04 0.72±0.067 1.18±0.14 fghi ij b hi j ghi C. malabatrum CM 28.42 ±0.264 28.19±0.243 100.8±0.2 0.81±0.071 0.41±0.036 0.90±0.038 no nop b jk de i C. mathewianum CMt 24.5 ±0.539 24.09±0.630 102±0.516 0.71±0.024 0.92±0.123 0.66±0.038

C. mathewianum o p b defg defg def CMtP 23.66 ± 0.55 22.92±0.323 103.10±0.9 0.97±0.058 0.79±0.074 1.29±0.135 var.ponmudianum de ef b c b de C. mohananii CMo 31.51 ± 0.57 30.7±0.481 102.70±0.52 1.12±0.038 1.21±0.032 1.33±0.20 a a a cdef efgh efg

C. moozhiyarense CMz 42.2±0.40 36.32±0.216 116.1±0.24 0.95±0.05 0.720±0.041 1.30±0.35 C. nicolsonianum CNg 29.61 ± 0.09f 29.0±0.187hi 102.20±0.61 b 0.91±0.019efgh 0.49±0.077ij 1.10±0.07efg

fghi ij b fghi def bc C. nilagiricum CN 28.61 ±0.464 27.9±0.408 102.70±0.73 0.90±0.082 0.82±0.025 1.81±0.15 cd de b efgh efgh efg

C. perrottetii CP 29.42 ±0.319 29.0±0.192 101.50±0.48 1.11±0.029 0.72±0.052 1.56±0.09 klm klm b ghi j de C. riparium CR 26.23 ±0.516 25.8±0.575 102.00±0.49 0.85±0.039 0.41±0.022 1.34±0.12 d fgh b efgh hij cd C. sulphuratum CS 31.58 ±0.636 29.9±0.492 105.40±0.98 0.93±0.027 0.56±0.055 1.54±0.07 klm lmn b ghi efgh fghi C. tamala CTa 26.19 ±0.284 25.4±0.260 103.00±0.62 0.84±0.021 0.73±0.040 0.96±0.04

b b b cdefg de bc C. travancoricum CT 35.83 ±0.49 34.7±0.334 103.20±0.61 0.98±0.047 0.91±0.066 1.75±0.08 e ghi b b de efg C. verum CV 30.16 ±0.436 29.3±0.419 103.10±0.92 1.27±0.013 0.92±0.038 1.14±0.07

p q b ij efghi bc C. walaiwarense CWa 20.88 ±0.157 20.1±0.287 103.60±0.98 0.76±0.054 0.70±0.065 1.75±0.12 c c b fgh ghij efg C. wightii CW 33.76 ±0.445 33.2±0.457 101.40±0.37 0.91±0.02 0.59±0.054 1.11±0.11

Table 2 Qualitative pollen morphological characters of Cinnamomum

Size class of Tip of Ornamentation Nature of Arrangement of Nature of Ornamentation of

Taxa Shape class of Pollen Type of Granules Pollen Spinules of spinules Granules Spinules cushion base Cushion Base

Sparsely C. agasthyamalayanum Mediae Prolate spheroidal Acuminate Striated Monomorphic Closely arranged Prominent Absent

distributed

Acute- Compactly C. cassia Mediae Prolate spheroidal Striated Heteromorphic Prominent Prominent Present Acuminate arranged

Sparsely Compactly C. chemungianum Minutae Prolate spheroidal Acuminate Striated Monomorphic Prominent Not entirely distributed arranged

Acute- Sparsely C. dubium Mediae Prolate spheroidal Striated Heteromorphic Spatially arranged Not Prominent Absent Acuminate distributed

C. filipedicellatum Minutae Prolate spheroidal Acuminate Striated Monomorphic Prominent Spatially arranged Prominent Present

C. goaense Mediae Prolate spheroidal Acute Striated Heteromorphic Prominent Closely arranged Not Prominent Present

Acute- Sparsely Compactly C. heyneanum Mediae Prolate spheroidal Striated Monomorphic Prominent Not entirely Acuminate distributed arranged

Sparsely Compactly

C. keralaense Mediae Prolate spheroidal Acuminate Striated Monomorphic Prominent Not entirely distributed arranged

C. litseaefolium Mediae Prolate spheroidal Acute Striated Heteromorphic Prominent Closely arranged Prominent Not entirely

Compactly

C. macrocarpum Mediae Prolate spheroidal Acuminate Striated Heteromorphic Prominent Prominent Not entirely arranged

C. malabatrum Mediae Spheroidal Acuminate Striated Monomorphic Prominent Spatially arranged Prominent Absent

C. mathewianum Minutae Prolate spheroidal Pointed Striated Monomorphic Prominent Spatially arranged Not Prominent Not entirely

C. mathewianum Compactly Minutae Prolate spheroidal Acuminate Striated Monomorphic Prominent Prominent Not entirely var.ponmudianum arranged

Sparsely

C. mohananii Mediae Prolate spheroidal Blunt Not Striated Heteromorphic Spatially arranged Not Prominent Absent distributed

Sparsely C. moozhiyarense Mediae Sub-prolate Acute Not Striated Heteromorphic Spatially arranged Not Prominent Absent distributed

Sparsely Compactly C. nicolsonianum Mediae Prolate spheroidal Acuminate Striated Monomorphic Prominent Absent distributed arranged

Sparsely

C. nilagiricum Mediae Prolate spheroidal Acuminate Striated Monomorphic Closely arranged Prominent Absent distributed

Compactly C. perrottetii Mediae Prolate spheroidal Acute Striated Monomorphic Prominent Prominent Present arranged

Compactly C. riparium Mediae Prolate spheroidal Acute Striated Monomorphic Prominent Prominent Present arranged

Acute- Compactly

C. sulphuratum Mediae Prolate spheroidal Striated Heteromorphic Prominent Prominent Present Acuminate arranged

Compactly C. tamala Mediae Prolate spheroidal Acute Striated Monomorphic Prominent Prominent Present

arranged

C. travancoricum Mediae Prolate spheroidal Acuminate Striated Monomorphic Prominent Closely arranged Prominent Present Acute- C. verum Mediae Prolate spheroidal Striated Monomorphic Prominent Spatially arranged Prominent Absent Acuminate Acute- Sparsely C. walaiwarense Minutae Prolate spheroidal Striated Heteromorphic Spatially arranged Not Prominent Absent \ Acuminate distributed

C. wightii Mediae Prolate spheroidal Acute Striated Monomorphic Prominent Closely arranged Prominent Present

35921 | P a g e Remya Krishnan R.V, Santhoshkumar E.S and Radhamany P.M., Systematic Significance of Pollen Morphology In South Indian Species of Cinnamomum Schaeffer (Lauraceae)

Its pollen grains have usually been described as more or less Table 3 PCA variable loadings of Pollen morphological spheroidal, apolar and inaperturate, with the exine highly characters reduced and consisting only of a thin coherent layer Variables PC1 PC2 PC3 ornamented with spinules. In many taxa the spinules are Polar Length(P) 0.493 -0.105 0.028 formed by cable-like strands and surrounded by minute Equatorial Diameter(E) 0.489 -0.103 0.028 P/E X100 0.676 -0.067 0.015 granules at the base (Raj and Van der Werff, 1988). They Length of Spinules 0.097 -0.006 -0.037 considered the family eurypalynous and concluded that grains Interspinular Distance 0.083 0.034 0.068 of most genera are easily identifiable by their size and by the Exine Thickness 0.12 -0.069 -0.106 number and organization of the spinules. As discussed by Van Size class of Pollen 0.028 0.078 -0.421 Shape class of Pollen 0.077 0.143 0.086 der Merwe et al. (1988) pollen characters of Lauraceae are Ornamentation of Cushion Base 0.083 0.585 -0.326 taxonomically useful on the generic rather than specific level, Type of Granules 0.038 0.059 0.088 and they agree with the findings of Raj and Van der Werff Nature of Granules 0.038 0.278 -0.466 (1988). The pollen grains of Cryptocarya species have been Tip of Spinules 0.101 0.528 -0.019 Interspinular Space 0.063 -0.036 -0.519 described as inaperturate, spheroidal, with a characteristic Arrangement of Spinules 0.079 0.481 0.616 pollen wall, that was thin exine and a massive nature of the Nature of Cushion Base 0.017 0.184 0.036 intine (Raj and Van der Werff, 1988). Van der Merwe et al. Eigenvalues 9.273 0.089 0.049 (1988) reported that pollen grains of southern African Percentage 97.637 0.935 0.511 Cum. Percentage 97.637 98.572 99.083 Cryptocarya species are strongly flattened, peroblate/oblate. The exine surface of a pollen grain can be either more or less All the other taxa represent the second principal cluster. They smooth or variously verrucate (wrinkled). It is interesting to were again sub clustered. Cinnamomum agasthyamalayanum note that palynological studies can provide important spaces for and C. nilagiricum were found to be the closely associated understanding the evolutionary history of the taxon. From the species by having similar pollen grains. literature survey, it is clearly understood that the variations in floral and pollen morphology and their taxonomic importance Principal Co-ordinate Analysis have been employed in the taxonomy of a number of families, genera and tribes to solve their taxonomic disputes or to PCoA involving both qualitative and quantitative characters support the existing classification systems. considered all together (Fig. 6) gave true species co-ordination for all the taxa indicating them showing species delimitation. In Cinnamomum, pollen grains are apolar, inaperturate and Cinnamomum moozhiyarense collected from Pathanamthitta, their shape varies from spheroidal to prolate spheroidal except C. walaiwarense from Thrissur (CWa) and C. travancoricum the new Cinnamomum species having sub-prolate shape of (CT) from Idukki stood apart as it is distinguishable from all pollen. Exine ornamentation is spinulate with spinules having a the other taxa in the study due to the variability in pollen conspicuous cushion base. This cushion base is either morphology. Cinnamomum mohananii (CMo) also stood far prominent or not prominent and they are either ornamented or apart due to the non-striated spinules and their blunt tip. The not within the genus Cinnamomum. According to Sritharan findings are in tune with the plotted cluster diagram. et al. (1992), Cinnamomum is reported to have foveolate pollen grain. On contrary to this report, all the South Indian species of Cinnamomum are found to have excrescent type of ornamentation i.e., spinulate, during the present study, rather than depression type (foveolate) ornamentation. The exine surface (interspinular space) is ornamented with mono- or heteromorphic granules. The spinules are either striated or smooth in nature. The spinule tip is acute, acuminate, extremely pointed or blunt. Cinnamomum mohananii is the only member with blunt, non-striated spinules while C. mathewianum is the only species with extremely pointed spinules.

Based on size class, there are two groups within the genus-1) Fig 6 PCoA scatter plot based on qualitative and quantitative pollen characters Small 2) Medium. First group includes C. mathewianum, C. (PAST 3.01). mathewianum var. ponmudianum, C. filipedicellatum, C. Pollen characters have diagnostic value and comparative chemungianum and C. walaiwarense and the remaining species importance in taxonomy and evolutionary considerations at all constitute the second group. Among all the taxa, only one levels of taxa (Nair, 1991; 2004). Besides, there were several species i.e, C. moozhiyarense collected during the study have publications that address the history of palynology and its sub-prolate pollen and it is also the species with largest pollen application to disciplines like systematic biology and grain having wavy, ridges and furrow like interspinular space phylogeny (Blackmore, 2000; Ferguson, 2000). According to ornamentation. This new species also lacks a proper cushion Blackmore (2007), some inherent properties within the pollen base. grains make them so informative. Except C. verum and C. malabatrum, the pollen morphology of all the other taxa are reported for the first time. Even though, Palynological features have, however, attracted little attention Sritharan et al. (1992) reported the pollen grains of C. for defining groups in the closely related Cinnamomum. litseaefolium and C. dubium as foveolate type, the present Lauraceae is generally considered as a stenopalynous family. 35922 | P a g e International Journal of Recent Scientific Research Vol. 10, Issue, 11(C), pp. 35918-35924, November, 2019 study confirmed their pollens to be spinulate type, which is the varying between the species studied and thus useful in the characteristic pollen feature of the family Lauraceae itself. identification of Cinnamomum at species level. Out of the six quantitative characters selected, polar Length and Acknowledgements equatorial diameter of pollen were significantly high (P <0.05) The first author is grateful to the Department of Science and in C. moozhiyarense, indicating the large size of pollen in the Technology for the award of Inspire fellowship. The second species. It is followed by C. travancoricum having the second author is thankful to the Director, JNTBGRI for the facilities largest pollen grain. While C. walaiwarense has the lowest provided. mean value for polar length and equatorial diameter of pollen, indicating the fact of having the smallest pollen among the References species studied. Cinnamomum dubium showed some peculiar pollen grains by having unique dumbbell shaped projections on Abeysinghe, P. D., Wijesinghe, K. G. G., Tachida, H., exine in addition to spatially arranged small spinules. Yoshda, T., & Thihagoda, M. (2009). Molecular characterization of Cinnamon (Cinnamomum verum In PCA, the first Principal Component (PC) accounted Presl) accessions and evaluation of genetic relatedness maximum variation (97.637%). The PC1 had characters like of Cinnamon species in Sri Lanka based on trnL intron polar length, equatorial diameter, exine thickness and tip of region, intergenic spacers between trnT-trnL, trnL-trnF, spinules as traits with the highest loadings. In addition to that, trnH-psbA and nuclear ITS. Research Journal of all the traits had positive values also. The second component Agriculture and Biological Sciences, 5(6), 1079-1088. accounted for 0.935% of the variability followed by third Baruah, A. and Nath, S.C. (2007): Cinnamomum principal component with 0.5% variability. Major traits that champokianum sp. nov. (Lauraceae) from Assam, accounted for more variability in PC1, PC2 and PC3 include northeastern India. Nordic J. Bot. 25, 281-285. polar length, equatorial diameter, exine thickness, nature of Blackmore, S. (2000). The palynological compass: the granules and tip of spinules. Hence, it is understood that these contribution of palynology to systematics. In Plant pollen morphological traits very much contributed to the systematics for the 21st century: Proceedings from a variability between the species and are thus important in symposium held at the Wenner-Gren Centre, Stockholm, distinguishing them. Sweden, September 1998 (pp. 161-177). London, UK:

The UPGMA phenogram constructed using DARwin 6 based Portland Press. on pollen characters separated C. moozhiyarense alone to the Blackmore, S. (2007). Pollen and spores: Microscopic keys first principal cluster and the remaining taxa constitute the to understanding the earth's biodiversity. Plant second principal cluster. The separation of C. moozhiyarense Systematics and Evolution, 263(1-2), 3-12. may be due to their largest, sub-prolate pollen grain with non- Erdtman, G. (1960). The acetolysis method-a revised striated spinules lacking proper cushion base and wavy, ridges description. Svensk Botanisk Tidskrift, 54, 516-564. and furrow-like interspinular space ornamentation. The second Erdtman, G. (1970). Über Pollendimorphie distantly placed species was C. walaiwarense collected from inPlumbaginaceae-Plumbagineae (Unter besonderer Thrissur, due to the presence of the smallest pollen among the Berücksichtigung vonDyerophytum indicum). — taxa, not so prominent cushion base of spinules and sparsely Svensk. Bot. Tidskr.64(2), 184–188. distributed granules in the interspinular space, which is seen as Ferguson, I. K. (2000). Pollen-morphological data in only small protuberances. The third most distant species was C. systematics and evolution: past, present and future. In travancoricum due to the presence of second largest pollen Plant systematics for the 21st century: Proceedings from grain with closely arranged acute spinules and interspinular a symposium held at the Wenner-Gren Centre, space ornamented with heteromorphic granules. Cinnamomum Stockholm, Sweden, September 1998. London, UK: mohananii also stood apart from others because of their blunt, Portland Press. pp. 179-192. non-striated and spatially arranged spinules. The species which Gangopadhyay, M. (2008): Nine new taxa and a new is closely associated in the phenogram were C. combination in Lauraceae from India and Myanmar. agasthyamalayanum and C. nilagiricum inferring their Bangladesh J. Pl. Taxon. 15, 89-106. similarity in pollen morphology. Geethakumary M.P., Pandurangan, A.G., and Santhosh Kumar, E.S (2012): Cinnamomum PCoA involving both qualitative and quantitative characters litseaefolium(Lauraceae) – A new distributional record considered together gave true species co-ordination for all the for India. Rheedea. 22(2), 127-130. taxa indicating them showing species delimitation. The findings Geethakumary, M. P., Pandurangan, A. G., & Deepu, S. are in tune with the plotted cluster diagram. (2015). A new species of Cinnamomum (Lauraceae)

This study suggested that the pollen characters selected were from Nilgiris, southern Western Ghats, India. Phytotaxa, taxonomically important. The cluster and principal component 224(3), 283-290. analysis on pollen morphological characters revealed the Geethakumary, M.P., Santhosh Kumar, E.S., Pandurangan existence of variability among the investigated species. The A.G. and Shaju, T. (2007): Cinnamomum dubium Nees multivariate analysis recorded considerable variation among (Lauraceae) – a new record for India. Indian J. For. species, principally due to pollen size, exine ornamentation, 30(1), 73-74. exine thickness and inter spinular distance. The greater Mabberley, D. J. (2008). Mabberley's Plant‐book: A Portable importance of such characters was revealed in cluster and Dictionary of Plants, Their Classifications, and Uses. – PCoA. Hence, the distinguishing pollen morphological features Cambridge University Press, Cambridge, Uk, p. 1021. observed in this study are of systematic value because they are Nair, P. K. K. (1991). Pollen morphology, plant taxonomy and evolution. Rheedea, 1(1-2), 78-83. 35923 | P a g e Remya Krishnan R.V, Santhoshkumar E.S and Radhamany P.M., Systematic Significance of Pollen Morphology In South Indian Species of Cinnamomum Schaeffer (Lauraceae)

Nair, P. K. K. (2004). Plant taxonomy. Current Science, van der Merwe, J. J., & Van Wyk, A. E. (1988). 86(5), 665-667. Dahlgrenodendron, a remarkable new genus of Raj, B., & van der Werff, H. (1988). A contribution to the Lauraceae from Natal and Pondoland. South African pollen morphology of neotropical Lauraceae. Annals of Journal of Botany, 54(1), 80-88. the Missouri Botanical Garden, 75(1), 130-167. Walker, J. W., & Doyle, J. A. (1975). The bases of Sritharan, R., Jacob, V. J., Balasubramaniam, S., & Bavappa, angiosperm phylogeny: palynology. Annals of the K. V. A. (1992). Palynological and cytological studies of Missouri Botanical Garden, 62(3), 664723. the genus Cinnamomum. In WOCMAP I- Medicinal and Aromatic Plants Conference: part 4 of 4 330, pp. 107- 114.

How to cite this article:

Remya Krishnan R.V, Santhoshkumar E.S and Radhamany P.M.2019, Systematic Significance of Pollen Morphology In South Indian Species of Cinnamomum Schaeffer (Lauraceae). Int J Recent Sci Res. 10(11), pp. 35918-35924. DOI: http://dx.doi.org/10.24327/ijrsr.2019.1011.4196

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