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\begin{document}

\title{Chlorophytum kolliensis (Asparagaceae), a new species from the Eastern
Ghats of Tamil Nadu, India}



\author[1]{Sebastian Soosairaj}%
\author[1]{Prakasam Raja}%
\author[2]{Balakrishnan Balaguru}%
\author[3]{Narayanasamy Dhatchanamoorthy}%
\affil[1]{St.Joseph's College}%
\affil[2]{Jamal Mohamed College}%
\affil[3]{Foundation for Revitalisation of Local Health Traditions}%


\vspace{-1em}



  
  \date{June 17, 2024}


\begingroup
\let\center\flushleft
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\maketitle
\endgroup





\selectlanguage{english}
\begin{abstract}
A new species of Chlorophytum is described and illustrated in this
article. Morphological, root anatomical, pollen and molecular features
were compared with its allied species C. laxum R. Br. which has
apiculate anther, lanceolate bract, 16-18 nerved flat leaf and ellipsoid
pollen with contrast to obtuse anther, ovate bract, 11-13 nerved
conduplicate leaf and ovoid pollen. The molecular features too
compliment the differences.%
\end{abstract}%



\sloppy


\textbf{Introduction}

The genus \emph{Chlorophytum} Ker (Asparagaceae), a tuberous herb is
distributed in old world tropics of Africa, Asia and Australia with 202
species (POWO). In India, the genus is represented by 23 species
(Chandore and Yadav 2019) and most of them are distributed in Western
Ghats (Adsul 2014). Most of the species of Chlorophytum are ephemerals;
the size and colour of the flower closely resemble each other making it
difficult to differentiate one from the other. However, Chandore\emph{et
al} . (2012) propose that the features like roots, filament and anther
length are found to be more consistent and could be used taxonomically
to differentiate species of \emph{Chlorophytum} .\emph{Chlorophytum
comosum} ~(Thunb.) Jacques has been widely grown as ornamental plant in
various colours and forms such as indoor decoration, landscape designing
and hanging baskets.

During our field exploration in Kolli hills of the Eastern Ghats, we
collected some interesting specimens of \emph{Chlorophytum} from shola
borders in rocky grounds. Thorough examination of herbarium material and
relevant literature (Sardesai \emph{et al} . 2006; Malpure \& Yadav
2009; Chandore \emph{et al.} 2012; Prabhu kumar \emph{et al} . 2014;
Adsul\emph{et al} . 2014; Mathew \& George 2015; Shaikh \emph{et al.}
2017; Borah \emph{et al} . 2019; Lakshminarasimhan \emph{et al} . 2019)
revealed that it is an allied to \emph{Chlorophytum laxum} . A few
collections deposited at MH (MH00021359, MH00021371) and RHT (53495,
45549, 15077) resembles to \emph{C. kolliensis} and it was collected
from various hills of Eastern ghats (Sirumalai, Shevroys, Kalrayan).
Microscopic evaluation of root anatomy, seed, anther, pollen, leaf
epidermal and molecular features suggests that it is dissimilar to
\emph{C. laxum.}Hence it is described here as a distinct species of
\emph{Chlorophytum} .

\textbf{Material and methods}

\emph{Chlorophytum laxum} was collected from scrub forests of
Narthamalai, Pudukottai district of Tamil Nadu, India (Fig.
1).\emph{Chlorophytum kolliensis} was collected from Kolli hills,
Namakkal district, Tamil Nadu (Fig. 2). The collected specimens were
preserved following standard protocols. The morphological comparison was
done from the herbarium deposits at MH and RHT (for herbarium acronyms,
see Thiers 2017) along with relevant literature. In addition, digital
specimens from online resources, such as Kew Herbarium Catalogue
(http://apps.kew.org/herbcat/navigator.do) and the Plants of the World
online (http://www.plantsoftheworldonline.org/) were also referred.

Sequencing of \emph{rbcL} , \emph{matK} and trnL-trnF regions
of\emph{Chlorophytum} species were done at Rajiv Gandhi Centre for
Biotechnology, Thiruvananthapuram, Kerala, India. The sequences were
submitted to NCBI and the accession numbers are OP028638 and OR542640.
The sequences were aligned using Clustal W and phylogenetic tree was
constructed using the maximum likelihood (ML) method with the following
options: 1000 bootstrap replication phylogeny test, Tamura 3-Parameter
Model, Gamma distributed (G) using MEGA software (Zhu \emph{et al} .
2018). The T3P model was identified by jModeltest as the best fitting
substitution model based on AIC and BIC criteria (Posada 2008).

Scanning Electron Microscope (Carel Zeiss, EVO-18) image of seed, anther
lobe and pollen were obtained from ACIC, St.Joseph's College,
Tiruchirappalli, Tamil Nadu, India and free hand section of tuberous
roots of \emph{Chlorophytum} and safaranin staining were carried out at
Department of Botany, St.Joseph's College, Tiruchirappalli, Tamil Nadu,
India.

\textbf{Results}

Chlorophytum kolliensis S. Soosairaj, \emph{sp. nov.}(Fig. 2)

\emph{Chlorophytum kolliensis} is comparable to \emph{C. laxum} in
having a slender inflorescence, shape and size of perianth, tuberous
roots but differs by presence apiculate anther lobe, inflated filament,
ovoid pollen, ovate bract, conduplicate leaf with 11-13 nerves (Table
1).

\textbf{Type} : S. Soosairaj 3926, 27 Dec 2019, Kolli hills, Namakkal
district, Tamil Nadu, India, 1320 m, 11\selectlanguage{ngerman}° 19' 00'' N, 78° 21' 32'' E
(holotype: RHT!, isotypes: MH!).

Etymology

The specific epithet refers to Kollimalai, Namakkal district of Tamil
Nadu, from where the new species was collected and described.

Description

Herb up to 12 cm high. Root tubers 4 -- 6 per plant, sessile, filiform,
5 -- 14 cm long, ca. 4 mm thick with many fibrous root hairs. Leaves 6
-- 8, all radical, distichous, sessile; leaf lamina 8 -- 10 × 0.6 -- 0.8
cm, conduplicate, keeled, glabrous, 11 -- 13 veined, mid vein distinct,
base sheathing, apex subacute, margin hyaline, serrulate towards the
apex. Scape usually solitary, occasionally branched, 1 -- 2.5 cm long,
ca. 0.5 mm thick, terete, erect, originating in the axil of a leaf.
Inflorescence a raceme, simple or branched, 4 -- 10 cm long, 8 -- 12
flowers. Bracts triangular, ca. 0.4 × 0.2 cm, persistent until fruit
formation, apex acute, margin hyaline, main nerve ciliate on abaxial;
bracteole triangular, 0.3 x 0.1 cm; pedicel 0.3 -- 0.6 cm long,
glabrous, elongating in fruit up to 0.8 cm. Perianth lobes 6, in two
whorls, white, oblong, 0.45 -- 0.15 cm, glabrescent, 3 nerved. Stamens
6, filaments 1 -- 1.5 mm long, white, linear throughout, anthers 0.8 ×
0.35 mm, yellow, dehiscing by longitudinal slits. Ovary globose, 1 mm,
style 1.5 mm long; stigma minutely papillose at apex. Capsule
triquetrous to 3-sulcate, ca. 0.5 × 0.6 cm, obcordate. Seeds 1 -- 2 in
each cell, 2 x 2.2 mm, orbicular, discoid, black, shiny, testa echinate.

Phenology

Flowering and fruiting from November to March.

Distribution and Ecology

The species is found on the exposed rocks with shallow soil around the
ariyur shola of Kolli hills, Namakkal district, Tamil Nadu. In addition
it also has been collected from Sirumalai, Shevroys and Kalrayan hills
of the Eastern ghats. Chlorophytum \emph{kolliensis} grows at the shola
border which is an open places at an elevation range between 1250 and
1350 m. It is associated with \emph{Drosera burmanni} Vahl,\emph{Drosera
peltata} Thunb., \emph{Eriocaulon} sp., \emph{Setaria intermedia} Roem.
\& Schult., \emph{Impatiens fruticosa} Lesch. ex DC.,\emph{Arundinela
ciliata} Nees ex Miq., \emph{Heteropogon contortus}(L.) P. Beauv. ex
Roem. \& Schult. \emph{Utricularia graminifolia} Vahl, and
\emph{Utricularia scandens} Benj.

Specimen examined

\emph{Chlorophytum laxum} R. Br. INDIA. K000794564, K000794563,
K00079456, MH00021360, MH00021361, MH00021363, MH00021369, MH00021370,
E00179754; AUSTRALIA. BM000939294,

\emph{Chlorophytum kolliensis} S. Soosairaj. INDIA. MH00021359,
MH00021371; RHT 53495, 45549, 15077.

Notes

Leaf epidermal features such as the epidermal cell (length and width)
and stomata (size) of\emph{C. laxum} is similar in the present
collection as reported by Nalawade and Gurav (2017) i.e. 332±20 x 22±1.5
\selectlanguage{greek}µ\selectlanguage{english}m and 27.5\selectlanguage{ngerman}±1.3 x 25.6±0.5 \selectlanguage{greek}µ\selectlanguage{english}m respectively. The features of \emph{C.
kolliensis} are 228.7\selectlanguage{ngerman}±21 x 28.3±0.3 \selectlanguage{greek}µ\selectlanguage{english}m (epidermal cell) and 27.6\selectlanguage{ngerman}±1.0 x
26.5±0.7 \selectlanguage{greek}µ\selectlanguage{english}m (stomata). Hence it is observed that the epidermal cell is
longer and narrower in \emph{C. laxum} when compared to\emph{C.
kolliensis} .

Cross sections of \emph{Chlorophytum kolliensis} tuberous root display
an outer rhizodermis, middle cortex and inner stele (Fig. 3). The
rhizodermis is made up of one layer of radially elongated closely
arranged cells. The cortex consists of ovoid or spherical parenchyma
cells with intercellular spaces. Strikingly, a few calcium oxalate
needle like crystals is observed. The endodermis consists of a single
layer of polygonal cells that are uneven in size with prominent
casberian stripes. The vascular system is characterized by 13 xylem and
phloem bundles in a radial and alternative arrangement. Xylem consists
of 2-4 protoxylem vessels, one or two metaxylem vessels alternated with
phloem elements. Tuberous root anatomy of \emph{C. kolliensis} having
broader pith (approximately 14 cells across) and more number of vascular
elements (13) differs from that of \emph{C. laxum} that has narrowed
pith (3-4 cells across) and a fewer vascular elements (8).

Seed of Chlorophytum is discoid having reticulate grove and echinate
projection. The reticulate grove is deep in \emph{C. kolliensis} and
shallow in \emph{C. laxum} (Fig. 4). Pollen grains are ellipsoid,
monosulcate with reticulate exine in \emph{C. laxum} and ovoid,
monosulcate with reticulate exine in \emph{C. kolliensis} . Size of
pollen in \emph{C.laxum} is 27-28 x 16-17 \selectlanguage{greek}µ\selectlanguage{english}m, but that
of\emph{C.kolliensis} is 21-23 x 18-19.5 \selectlanguage{greek}µ\selectlanguage{english}m. P/E ratio is 1.6 and 1.2
respectively.

The combined data matrix of rbcL, matK and trnL-trnF has 3508 characters
(rbcL-1355, matK-1510, trnL-trnF-643) in length, in which 183 variable
characters are identified. The maximum likelihood tree showed two
distinct clades, \emph{C. comosum, C. malabaricum} and \emph{C.
tuberosum} as one clade and \emph{C. laxum} and \emph{C. kolliensis} as
another clade (Fig. 5). Molecular evidence from two chloroplast loci
showed \emph{C. laxum} is distinct from \emph{C. kolliensis} , and at
least as distinct as other accepted species in the genus. The scape
of\emph{C. laxum} and \emph{C. kolliensis} is slender whereas it is
stout in other species of Chlorophytum. Molecular markers have been used
to identify different species of Chlorophytum based on RAPD and rbcL
sequences (Meenu \emph{et al} . 2010).

\textbf{References}

Adsul, A. A. 2014. Taxonomic Revision of Genus Chlorophytum Ker Gawl.
for India, Ph. D. thesis submitted to Shivaji University, Kolhapur.

Adsul, A.A. et al. 2014. \emph{Chlorophytum sharmae} (Asparagaceae): a
new species from Kerala, India. - Kew Bulletin 69: 9503.

Borah, D. et al. 2019. \emph{Chlorophytum assamicum} (Asparagaceae), a
new species from Northeast India. - Phytotaxa 394 (1): 123--125.

Chandore, A.N. et al. 2012. \emph{Chlorophytum belgaumense} , a new
species of Asparagaceae from the Western Ghats of India. - Kew Bulletin
67: 527--531.

Chandore, A. N. \& Yadav. S. R. 2019. \emph{Chlorophytum
tillariense}(Asparagaceae), a new species from Western Ghats of India. -
Phytotaxa 427 (4): 279--284.

Lakshminarasimhan, P. et al. 2019. Flora of Karnataka, Botanical Survey
of India, Kolkata, 3: 304 -- 312.

Mabberley, D. J. 2017. \emph{The Plant Book} , 4\textsuperscript{th} ed.
Cambridge University Press, Cambridge.

Malpure, N.V. \& Yadav, S.R. 2009. \emph{Chlorophytum gothanense} , a
new species of Anthericaceae from the Western Ghats of India. - Kew
Bulletin 64: 739--741.

Mathew, J. \& George, K.V. 2015. A new \emph{Chlorophytum}(Asparagaceae)
from the southern Western Ghats. - Telopea 18: 379--382.

Meenu Katoch et al. 2010. Identification of Chlorophytum species
(\emph{C. borivilianum, C. arundinaceum, C. laxum, C. capense} \emph{and
C. comosum} ) using molecular markers. - Industrial Crops and Products,
32: 389--393.

Nalawade, A. S. \& Gurav, R. V. 2017. Stomatal Studies in the
Genus\emph{Chlorophytum} (Asparagaceae). - Bioscience Discovery, 8(3):
574-581.

Posada, D. 2008. jmodeltest: Phylogenetic Model Averaging. - Molecular
Biology and Evolution 25: 1253-1256.

Prabhu kumar, K. M. et al. 2014. A new species of
\emph{Chlorophytum}(Asparagaceae) from southern Western Ghats of India.
\textbf{-}Phytotaxa 188 (5): 282--286.

Sardesai, M. M. et al. 2006. A new species of
\emph{Chlorophytum}(Anthericaceae) from Western Ghats, India. - Kew
Bulletin 61: 269--271.

Shaikh, M. et al. 2017. A new species of
\emph{Chlorophytum}(Asparagaceae) from Madhya Pradesh, India. -
Phytotaxa 291 (2): 159--162.

Zhu, S. et al. 2018. Phylogenetic analysis of \emph{Uncaria} species
based on internal transcribed spacer (ITS) region and ITS2 secondary
structure. - Pharmaceutical Biology 56: 548-558.

Thiers, B. 2017. Index Herbariorum: A global directory of public
herbaria and associated staff. New York Botanical Garden's Virtual
Herbarium.

\textbf{Table 1. Comparison of characters between \emph{Chlorophytum
laxum} and \emph{C. kolliensis}}\selectlanguage{english}
\begin{longtable}[]{@{}lll@{}}
\toprule
\begin{minipage}[b]{0.30\columnwidth}\raggedright\strut
\strut
\end{minipage} & \begin{minipage}[b]{0.30\columnwidth}\raggedright\strut
C. laxum\strut
\end{minipage} & \begin{minipage}[b]{0.30\columnwidth}\raggedright\strut
\textbf{C. \emph{kolliensis}}\strut
\end{minipage}\tabularnewline
\midrule
\endhead
\begin{minipage}[t]{0.30\columnwidth}\raggedright\strut
Plant height\strut
\end{minipage} & \begin{minipage}[t]{0.30\columnwidth}\raggedright\strut
\textgreater{}20 cm\strut
\end{minipage} & \begin{minipage}[t]{0.30\columnwidth}\raggedright\strut
\textless{} 15 cm\strut
\end{minipage}\tabularnewline
\begin{minipage}[t]{0.30\columnwidth}\raggedright\strut
Leaf\strut
\end{minipage} & \begin{minipage}[t]{0.30\columnwidth}\raggedright\strut
Flat, \textgreater{}16 cm long, \textless{}5 mm wide, 16 - 18
nerved\strut
\end{minipage} & \begin{minipage}[t]{0.30\columnwidth}\raggedright\strut
Conduplicate, \textless{} 10 cm long, \textgreater{}5 mm wide, 11 - 13
nerved\strut
\end{minipage}\tabularnewline
\begin{minipage}[t]{0.30\columnwidth}\raggedright\strut
Inflorescence\strut
\end{minipage} & \begin{minipage}[t]{0.30\columnwidth}\raggedright\strut
\textgreater{} 20 cm\strut
\end{minipage} & \begin{minipage}[t]{0.30\columnwidth}\raggedright\strut
\textless{} 13 cm\strut
\end{minipage}\tabularnewline
\begin{minipage}[t]{0.30\columnwidth}\raggedright\strut
Bracts\strut
\end{minipage} & \begin{minipage}[t]{0.30\columnwidth}\raggedright\strut
Lanceolate, \textgreater{}10mm, glabrous, apex acuminate\strut
\end{minipage} & \begin{minipage}[t]{0.30\columnwidth}\raggedright\strut
Ovate, \textless{}4mm, ciliate along the midnerve, apex acute\strut
\end{minipage}\tabularnewline
\begin{minipage}[t]{0.30\columnwidth}\raggedright\strut
Bracteole\strut
\end{minipage} & \begin{minipage}[t]{0.30\columnwidth}\raggedright\strut
Ovate\strut
\end{minipage} & \begin{minipage}[t]{0.30\columnwidth}\raggedright\strut
Triangular\strut
\end{minipage}\tabularnewline
\begin{minipage}[t]{0.32\columnwidth}\raggedright\strut
Filaments\strut
\end{minipage} & \begin{minipage}[t]{0.32\columnwidth}\raggedright\strut
Filament \selectlanguage{ngerman}±2 mm long, uniform throughout, anther lobe apex
apiculate\strut
\end{minipage} & \begin{minipage}[t]{0.32\columnwidth}\raggedright\strut
Filament 1 - 1.5mm long, inflated at base, anther lobe apex obtuse\strut
\end{minipage}\tabularnewline
\begin{minipage}[t]{0.30\columnwidth}\raggedright\strut
Ovary \& style\strut
\end{minipage} & \begin{minipage}[t]{0.30\columnwidth}\raggedright\strut
1 x 1 mm, 1 mm long\strut
\end{minipage} & \begin{minipage}[t]{0.30\columnwidth}\raggedright\strut
1 x 1.5 mm, 1.2 mm long\strut
\end{minipage}\tabularnewline
\begin{minipage}[t]{0.30\columnwidth}\raggedright\strut
Seed\strut
\end{minipage} & \begin{minipage}[t]{0.30\columnwidth}\raggedright\strut
2 - 4 in each cell, 3 x 3mm\strut
\end{minipage} & \begin{minipage}[t]{0.30\columnwidth}\raggedright\strut
1 - 2 in each cell, 2 x 2,2mm\strut
\end{minipage}\tabularnewline
\begin{minipage}[t]{0.30\columnwidth}\raggedright\strut
No. of Tuberous root\strut
\end{minipage} & \begin{minipage}[t]{0.30\columnwidth}\raggedright\strut
\selectlanguage{ngerman}±10\strut
\end{minipage} & \begin{minipage}[t]{0.30\columnwidth}\raggedright\strut
\selectlanguage{ngerman}±5\strut
\end{minipage}\tabularnewline
\begin{minipage}[t]{0.30\columnwidth}\raggedright\strut
Altitude\strut
\end{minipage} & \begin{minipage}[t]{0.30\columnwidth}\raggedright\strut
\textless{} 200 m msl\strut
\end{minipage} & \begin{minipage}[t]{0.30\columnwidth}\raggedright\strut
\textgreater{}1200 m msl\strut
\end{minipage}\tabularnewline
\begin{minipage}[t]{0.30\columnwidth}\raggedright\strut
Pollen shape and P/E ratio\strut
\end{minipage} & \begin{minipage}[t]{0.30\columnwidth}\raggedright\strut
Ellipsoid, 1.6\strut
\end{minipage} & \begin{minipage}[t]{0.30\columnwidth}\raggedright\strut
Ovoid, 1.2\strut
\end{minipage}\tabularnewline
\begin{minipage}[t]{0.30\columnwidth}\raggedright\strut
Pith of root\strut
\end{minipage} & \begin{minipage}[t]{0.30\columnwidth}\raggedright\strut
3 - 4 cells across\strut
\end{minipage} & \begin{minipage}[t]{0.30\columnwidth}\raggedright\strut
13 - 14 cells across\strut
\end{minipage}\tabularnewline
\begin{minipage}[t]{0.30\columnwidth}\raggedright\strut
No. of vascular elements\strut
\end{minipage} & \begin{minipage}[t]{0.30\columnwidth}\raggedright\strut
ca.8\strut
\end{minipage} & \begin{minipage}[t]{0.30\columnwidth}\raggedright\strut
ca. 13\strut
\end{minipage}\tabularnewline
\bottomrule
\end{longtable}\selectlanguage{english}
\begin{figure}[H]
\begin{center}
\includegraphics[width=0.70\columnwidth]{figures/Fig-1/Fig-1}

\end{center}
\end{figure}\selectlanguage{english}
\begin{figure}[H]
\begin{center}
\includegraphics[width=0.70\columnwidth]{figures/Fig-2/Fig-2}

\end{center}
\end{figure}\selectlanguage{english}
\begin{figure}[H]
\begin{center}
\includegraphics[width=0.70\columnwidth]{figures/Fig-3/Fig-3}

\end{center}
\end{figure}\selectlanguage{english}
\begin{figure}[H]
\begin{center}
\includegraphics[width=0.70\columnwidth]{figures/Fig-4/Fig-4}

\end{center}
\end{figure}\selectlanguage{english}
\begin{figure}[H]
\begin{center}
\includegraphics[width=0.70\columnwidth]{figures/Fig-5/Fig-5}

\end{center}
\end{figure}

\selectlanguage{english}
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