569

Breeding of Horticultural Crops Vol. 1 -Part B: l'lantation Crops (2017) : 569-589

Editors: V A Parthasarathy, C. Aswath, K. Nirma! Babu & R. Senthi! Kumar
Tod ay & Tomorrow's Printes and Publishers, New Delhi - 110 002, India

IS based
3nd the

nts a nd
risbane.
ARECANUT
K.S. Ananda, Anitha Karun and V. A. Parthasarathy
Jngress,

Arecanut (Areca catechu 1.) or betelnut is an extensively cultivated

tropical palm and the dry kernel is used mainly for masticatory purpose in
India and West and South East Asia. The major arecanut growing countries
are: India, Sri Lanka, Bangladesh, Nepal , China, Myanmar, Bhutan,
Thailand, Malaysia, Indonesia and Vietnam. The present production of
arecanut in the world is 1.22 million tonnes from an area of 0.90 million
hectares. In India, arecanut is cultivated in about 4.51 lakh ha with a
production of 7.4 7 lakh tonnes of nuts with the productivity of 1659 kg ha­
l. Arecanut is predominantly cultivated in traditional states like Karnataka,

Assam, Kerala, Maharashtra, West Bengal andAndaman and Nicobar group

ofIslands and it's cultivation is spread to non-traditional areas in Kamataka
and Tamil Nadu . Arecanut is providing livelihood security for millions of
people in rural and urban areas.
1. History, origin and distribution
The origin of arecanut is not exactly known. There are no fossil
remains of the genus Areca, but the fossil records of closely related genera
indicate its presence during tertiary period. The maximum species diversity
(24 species) and other indicators suggest its original habitat in Philippines,
Malaysia, and Indonesia (Raghavan, 1957; Bavappa, 1963). According to
Watt (1889) it is a native of Cochin China and Malay Peninsula. Beccari
(1919) described four cultivars and nine species of areca from Philippines.
Although it is not precisely known as to when areca nut found its
way to Indian subcontinent, several evidences about its antiquity exist
(Mohan Rao, 1982). Arecanut is mentioned in various ancient Sanskrit
scriptures (650-1300 BC); and its medicinal properties were known to a
famous Scholar Vagbatta (41h century A.D.). One of the well-known Ajanta
caves in central India (200 B.c. to 900 A.D.) has one exquisitely painted
arecanut palm providing a backdrop to Padmapani Budha. According to
Furtado (I 960) one of the earliest references to arecanut was in 1510 A.D.
570

The abundant use of arecanut in chewing and religious functions was Java

indicated even during the times of Aryans in India. Celebes

Australia
2. Botany
Solomon T~
Martius (1832-1850) was the first to attempt defining the limits of
New Guine
the genus, Areca. But the attempts were 110t satisfactory, as they were 110t
based on real affinities. Later various species, grouped under Areca, were
Philippines
separated into different genera, based on nature of albumen, position of
ovule, distribution of flowers, and limited the genus to close relatives of
the type of the genus, Areca catechu (Blume, 1836). Furtado (1933)
3. Specie
described the limits of the genus Areca and its sections. A list of Areca
species and their geographical distribution is given in Table 1. Blatter S
(1926) described the generic characters of genus Areca. Deta iled and Ph ilil
morphology, floral biology and embryology of arecanut have been described be separa
(Murthy and Piliai, 1982). The arecanut palm is a graceful, erect and un­ female f1(
branched palm reaching a height of upto 18-20m. The stem has scars of collection
falJen leaves in regular annulated forms. The girth of the stem depends on norman b)
genetic constitution, soil condition and plant vigour. The arecanut palm Pinanga (
has an adventitious root system. The crown of an adult palm contains 7­ 1999). T
12 leaves. The leaves are pinnatisect and consist ofa sheath, a rachis and 1999). A
leaflets. The Jeaf sheath completely encircles the stem. It is about 54 cm were intn
in length and 15 cm in breadth. The average length of Jeaf is 1.65 m, Indonesia
which bears about 70 leaflets. The leaflets are 30.0 to 70.0 cm in length Besides, a
and 5.8 to 7.0 cm in breadth depending on the position of the leaf. in differer
been desc
Arecanut is monoecious with both male and female flowers
occurring on the same spadix. It is cross-pollinated (Bavappa and T
Ramachander, 1967). The male phase lasts for 25-46 days. Female flowers internode
are cream coloured and turning green within a week. The female phase Shimoga
extends up to 10 days. The stigma remains receptive up to 6 days (Murthy Kanara ar
and Bavappa, 1960). Pollen is generally carried by wind. are also w
Table 1. Distributioll of Species of Areca
quality, ar

COUll try Species TI

1957 for I
India A. catechu, A. trianciro
Bavappa (
Andamans, India A catechu, A. taxa high yield
Sumatra A. catechu, A. Iriandra, A. laliloba T he chan
Sri Lanka A. catechu, A. concinna Thampan.
Malaysia A. catechu, A. triandra, A. laliloba, A. montana, A. ridleyana collection
named as
Borneo A. catechu, A. borneensis, A. kinabaluensis, A. arulldinacea.
A. bongayensis, A amojahl; A. mullettii, A. minuta, A.jurcata more nur
 571

Java A. catechu, A. lati/oba

tions was
Celebes A. eelebica, A. oxicarpa, A. paniculata, A. henrici
Au stralia A. catechu, A. alieae
Solomon Islands A.niga-so/u, A. reehingeriana, A. tondo, A. guppya no
,e limits of
New Guinea A. congesto, A j ob iensis, A. laderrnaniona, A. rn acrocaly x,
y were not A. nannospadix, A. warburgiana
reca, were
Philippines A. catechu , A. hutchinsonian a, A. vidaliana, A. costulata,
JOsition of A. rna cro corpa, A. parens, A. caliso , A. whitfordii, A.
datives of camariensis, A. ipot
do (1933)
3. Species and cultivars
~ of Areca
l. Blatter Several cultivars have been recognized in Karnataka (Rau, 1915)
Detailed and Philippines (Beccari, 19] 9) based on fruit and kernel. Cultivars can
1 described be separated on the basis of stomatal characters, size of nuts, leaves shed,
:ct and un­ female flowers, nut size (Bavappa, 1966; Bavappa and Pillai, 1976). A
IS scars of collection of five species viz., A. catechu, A. triandra, A. macrocalyx, A.
iepends on normanbyii and A. concinna and two genera Actinorhytis calapparia and
anut palm Pinanga dicksoni are available at the Regional Station, Virtal (Anuradha,
:ontains 7­ 1999). The gennplasm holding now consists of 113 accessions (Ananda,
rachis and 1999). Among these, 23 exotic accessions representing different species
lout 54 cm were introduced from countries viz ., Fiji, Mauritius, China, Sri Lanka,
is 1.65 m, Indonesia, Saigon, Singapore, British Solomon Islands and Australia.
n in length Besides, about 90 collections represent gennplasm from explorations made
eaf. in different areca nut growing states of India. About 39 accessions have
been described based on descriptors.
Ie flow ers
lappa ancl There is wide range of variations in fruit characters, stem height,
ale flowers internode length and leaf size and shape. The nuts in Malnad parts of
nale phase Shimoga and Chikmagalur districts are small in size whereas in North
( S (Murthy Kanara and Ratnagiri they are bigger (Murthy and Bavappa, 1962). There
are also wide variations in yield, earliness in bearing, fruit number/bunch,
quality, and dwarfness.
The exotic and indigenous collections are under evaluation since
1957 for morphology, nut characters and yield attributes (Ananda, 1999;
8avappa and Nair, 1982). Yield evaluation has resulted in release of four
high yielding cultivars of which three are selections from exotic colleytion.
The characteristics of these varieties have been described (Ananda and
Thampan, 1999 ; Nampoothiri et ai., 1999; Table 3). Among the exotic
collection, cultivar 'VTL - 3 ' introduced from China was released and
, A. rid /eya/la
named as 'Mangala' (Bavappa, 1977). This cultivar had earliness in bearing,
am nd i nacea,
more number of female flowers, high yield and lesser stem height as
{la, A.jurcala
572

compared to other accessions. Other varieties released for cultivation are Achrol1~
'Sumangala' and'S reemangala' that are introductions from Indonesia and (1945)/
Singapore respectively. High yield potential was observed in one of the (1956) :
indigenous collection from West Bengal and it was released as 'Mohitnagar' determir
variety. A comparative yield trial involving five high yielding varieties of 11=]6. S
arecanut viz., Mangala, Sumangala, Sreemangala, Mohitnagar and non-disj
Thirthahalli local was studied at Thirthahalli to identify suitable variety reported
for cultivation in the Malnad (hills) region of Karnataka (Ananda et of., four eco
2000). 'Mangala' registered significantly lower height with high percentage and met
of flowering palms. 'Mangala' performed better than other varieties with disj unct
highest yields in the initial years of bearing closely followed by 'Thirthahalli chromos
local' while 'Sreemangala' recorded comparatively lower yields. Five
arecanut cvs (VTL-3, VTL-ll, VTL-12, VTL-13 and VTL-17) were
quite nc
assessed for increasing production/unit area. The mean tree height, number
chromos
of leaves producedltree, girth at collar 45 months after planting, time taken also rep(
for first flowering, nut weight and processing out-turn were recorded.
cultivar'
Cultivars VTL-3, VTL-12 and VTL-13 were considered the most productive and Nair
(Thangaraj et of., 1980). Data on nut weight and nuts/tree were obtained
palms, 0
from two experiments, one involving 13 varieties of Areca catechu over even dec
eight years and the other 11 varieties over nine years. The three highest­
of chrom
yielding varieties proved suitable when planted in favourable enviro1U11ents
species.
only. Two others, with fairly high yields, were considered stable in all
mother CI
environments (Natarajan et of., 1982).
extent of
Bavappa (1974) recorded morphological, anatomical and yield high degJ
characters for 13 cultivars of A. catechu and four ecotypes of A. triandra The poss
during the years 1963, 1966 and 1972. The analysis of variance of the being une
results obtained in 1963 showed that the differences between cultivars are (Bavappc
highly significant for all the six morphological characters. A combined diakinesi~
analysis ofthe data for two years for 24 common characters recorded during 1974). D
1967 and 1972 also revealed significant interaction between cultivars for pamng a
all characters. A significant interaction between years and cultivars was triandra
seen for height, girth, internodal distance, number of bunches and distributi
inflorescences on the palm, length and breadth of leaf sheath, length and ensuring
volume of nut and length, breadth, weight and volume of kernel. was hight
A. triandl
4. Genetics and Breeding
may be d
The chromosome number of Areca catechu L. was first determined as compa
and reported by Venkatasubban (1945) as 2n=32. The chromosome number of the par
ofthe species was later confirmed by Sharma and Sarkar (1956), Raghavan satelJited
and Baruah (1958), Abraham et of., (1961) and Bavappa and Raman (1965). catechu
 573

:)11are A chromosome number of2n=32 reported by Darlington and Janaki Ammal

ia and (1945) for A. triandra Roxb was later confirmed by Sharma and Sarkar
of the (1956) and Bavappa and Raman (1965). Nair and Ratnambal (1978)
lagar' determined the meiotic chromosome number of A. macrocalyx Becc as
ties of n= 16. Sharma and Sarkar (1956) described meiotic abnormalities such as
rand non-disjunction, lagging chromosomes, univalents and pentads were
'ariety reported in A. catechu. Bavappa and Raman (1965) stuclied meiosis of
et al., four ecotypes of A. catechu. Abnormalities like univalents at diakinesis
~ntage and metaphase I, non-synchronisation of orientation, clumping, delayed
s with disjunction, chromosome bridges and laggards at anaphase r and II,
1ahalli chromosome mosaics and supernumerary spores were observed.
. Five Sharma and Sarkar (1956) found that the meiotic division was
I were
quite normal in A. triandra except for the presence of 14 and 18
umber chromosomes occasionally at metaphase II. Bavappa and Raman (1965)
: taken also reported regular meiotic division in A. triandra. There was intra­
orded. cultivar variation in meiotic behaviour of Areca (Bavappa, 1974; Bavappa
luctivc and Nair, 1978). While normal bivalent formation was observed in some
,tained palms, others had maximum association of hexavalents, octovalent and
u over even decavalent. Abnormalities like bridges and laggards, disorientation
ighest­ of chromosomes at anaphase I and anaphase II were also reported in this
unents species. Intra-palm variation in chromosome numbers exist in the pollen
in al1 mother cells of A. catechu, A. triandra and their hybrids . Cytomixis to an
extent of 39% seemed to have contributed to this abnormally. In spite of
i yield high degree of multivalents in A. catechu, pollen fertility was very high.
iandra The possibility of the frequency of multivalent formation and disjunction
of the being under genotypic control and being subject to selection was suggested
3rs arc (Bavappa and Nair, 1978). Partial desynapsis of chromosomes occurs at
nbined diakinesis in A. triandra and A. catechu x A. triandra hybrids (Bavappa,
during 1974). Desynapsis observed at diakinesis was followed by an increase in
ars for pairing at metaphase I as reflected by the frequency of bivalents in A.
rs was triandra and A. catechu x A. triandra hybrids. This was attributed to
~s and distributive pairing, a mechanism that has possibly been adopted for
~h and ensuring regular segregation of chromosomes. The extent of desynapsis
was higher in the F I hybrids of A. catechu and A. triandra as compared to
A. triandra, suggesting that the genetic mechanism controlling this character
may be dominant. The large number of univalents observed in the hybrid,
rmined as compared to A. triandra parent, has been attributed to reduce homology
lumber of the parental chromosomes (Bavappa and Nair, 1978). Two pairs of short
?;havan satellited chromosomes in the somatic chromosome complement of A.
1965). catechu were observed (Venkatasubban, 1945). Three pairs of long
574

chromosomes, six pairs of medium sized chromosomes and seven pairs of

short chromosomes were observed (Sharma and Sarkar, 1956) in A. catechu.
They categorized the chromosomes into seven groups based on their
morphology and relative length. Two pairs of long chromosomes next to
the longest were found to have secondary constrictions. They also observed
that the chromosomes of A. triandra were longer than those of A. catechu.
Bavappa and Raman (1965) found the chromosomes of A. catechu and A.
triandra to differ in size, total chromatin length, position of primary and
secondary constrictions and number and position of satellites. Based on 11
the assumption of Sharma and SarkaI' (1956) that gradual reduction in (
chromatin matter had taken place in the evolution from primitive to n
advanced forms of different genera and tribe of Palmae. Bavappa and y
Raman (1965) considered A. catechu as more advanced than A. triandra.
C
The chromosome morphology ofa few cultivars ofA. catechu from 1f

Assam was reported (Raghavan, 1957). Minor variation in structure and tv

length of individual chromosomes, total length of the complement and tv
position of constrictions among the types was noted. On the basis of al
morphology, he recognised nine groups in the somatic chromosomes of fl .
the cultivars. 5.
Studies on the karyotypes of eight cultivars of A. catechu and four
ecotypes of A. triandra (Bavappa , 1974; Bavappa et al., 1975) revealed
th
considerable differences in their gross morphological characteristics. The
ar
karyotypes of the A triandra ecotypes showed a higher frequency of sub
median and median chromosomes as compared to A. catechu. A 5.
classification of the karyotype of the two species according to the degree
of their asymmetry which recognizes three grades of size differences and
four grades of asymmetry in centromere position (Stebbins, 1958), showed
J111
that karyotypes 1B, 2A, 2B and 3B are represented in A. catechu cultivars
to
and only lA, 2A and 2B are represented in the ecotypes of A. triandra.
eX j
Even within the same cultivar of A. catechu, two different types of
int
asymmetry in karyotypes were observed. There was no such variation in
dif
A. triandra ecotypes. Evidently A. triandra has a more symmetrical
thE
karyotype than A. catechu. It was concluded that delineating the cultivars
cal
of A. catechu on the basis of standard karyotype is rather difficult. The
Se
fact that A. catechu has lesser chromatin matter and an asymmetrical
es~
karyotype compared to A. triandra shows that the latter is more primitive.
So
The hybridization programme was initiated with the objectives of So
exploiting the existing variability in the Areca germplasm (Bavappa and cy
Nair, 1982). The main concerns were to evolve high yielding, regular fOI
bearing, high quality and semi-tall ideotypes. Interspecific hybrids of A.
 575

)f catechu x A. triandra had only one stem as in A. catechu indicating

i. dominance of this character (Bavappa, 1974). The hybrids mostly equaled
lr the parents in internodal length. They also exhibited hybrid vigour for a
o number of characters like number of male flowers, female flowers , spadix
:d length and stem girth.
i. The tall nature of the palm hinders various operations like spraying
1 and harvesting and is quite labour intensive. One of the major thrusts in
ld the research on breeding in arecanut has been to induce dwarfness. A
10 natural dwarf mutant was identified and was named 'Hirehalli Dwarf'
In (Naidu, 1963). This however has low yields coupled with very poor quality
o nuts not suitable for chewing. An attempt was made to cross the high
ld yielding varieties with Hirehalli Dwarf to exploit the dwarfing nature
7. (Ananda, 1999; Nampoothiri et al., 1999). Maximum dwarfs and
:n intermediates were recovered from crosses Sumangala x HirehalliDwarf,
ld Mohitnagar x Hirehalli Dwarf and Mangala x Hirehalli Dwarf among the
ld twelve hybrids crosses made. Hirehalli Dwarf x Sumangala hybrid was
)f also found to be promising with a yield of 2.65 kg! palm dry nuts during
)f first year of yielding.
5. BIOTECHNOLOGY
II'
Biotechnological research in arecanut is still in infancy. During
:d the last ten years, research has been initiated in certain laboratories which
Ie
are reported below.
Ib

A 5.1. Tissue culture

~e
Research on tissue culture ofarecanut has been reported only during
ld the last ten years. Karun et aL., (2004) reported a protocol using leaf and
:d immature inflorescence of adult palms as a explant. They found Picloram
's to be the suitable callogenic agent. Initial standardization was done on leaf
1.
explants excised from one year old seedling and later modified for immature
)f inflorescence sampled from adult palm. The protocol was also tested with
,n different arecanut varieties viz., Mangala, Sumangala and Mohit Nagar;
11 the basal medium used was MS. Picloram was found to be the most suitable
's callogenic agent for both types of explants as well as for varieties tried
Ie Serial transfer of explants from high to low auxin concentration was
11 essenti~l for sustained growth of callus and somatic embryo induction.
..,.
Q

Somatic embryogenesis was achieved in hormone-free MS medium .

)f Somatic embryo was germinated in MS medium supplemented with
Id cytokininsand 20 11M BA was found to be best. No variation was noticed
1r for callus initiation and somatic embryogenesis and plantlet development
1.
576

in different varieties except for period of culturing. For rapid germination wounde
of somatic embryo, MS liquid medium supplemented with 5 iJM BA was structu!
used. Plantlet with 2-4 leaves and fairly good root system were weaned dicamb,
using sand: soil (5:1) potting mixture. These pl antlets were field planted callus ~
during 2006 at CPCRI (RS) Vittal for field evaluation. This protocol has subcultt
been applied for mass multiplication of field resistant Yellow Leaf Disease callus ti
(YLD) palms (Karun et af., 2005). Direct and indirect so matic after 10
embryogenesis from inflorescence explants of areca palms was reported After su
by Radha et af., (2006). However the numbers of direct somatic embryos to conta
formed were very less compared to indirect somatic embryos. The effect 24%.
of various cytokinins viz., TDZ, BA, Kinetin, 2i-P and Zeatin on growth 5.1.1. A
and maturation of direct somatic embryos were studied and it was found
that TDZ was essential for maturation and conversion of somatic embryos
into complete healthy plantlets (Radha et aI., 2008). Mass mUltiplication devastat
was also achi eved through indirect secondary somatic embryogenesis. in Keral:
susceptil
Mathew and Philip (2000) first reported the protocol for in vitro to be fie
propagation via direct adventitious shoot bud differentiation from embryo that heal
explants. Results obtained with excised embryos of A. catechu (cv. disease [
'Kasargodan' and 'Mangala') grown on Murashige & Skoog's medium, breeding
White's medium and Branton & Blake's (BB) medium with permutations used for
and combinations of auxins and cytokinins showed that activated charcoal, palms. T
2,4-D and high levels of phosphate in BB medium were critical for the villages v
differentiation of additional shoots from the cotyledon. Wang et af., (2003a) Kannada
developed a protocol for plantlet formation through shoot formation from plantlets
callus of arecanut. Greenish soft callus was formed from shoot tip explants compara1
within four weeks, when cultured on Gelrite-gelled MS basal medium
supplemented with BA in combination with TDZ. The highest percentage c
of callus formation (100%) was found on the medium supplemented with has also
0.2 mg 1. 1 BA and 0.2 mg 1. 1 TDZ. During subculture on the same medium 'Hi rehalli
for callus induction, most of calli proliferated and 50-60% formed shoots. in 1963
About 90% of shoots formed roots on BM containing 0.1 mg 1-1 NAA after Improven
four weeks in culture. Regeneration ofplantlets from shoot tips via primary of the int(
callus production and a two-step process of organogenesis, required about all the m:
20 weeks. hybrids v
way of in
Wang et af., (2003 b) obtained plant regeneration through somatic The breec
embryogenesis from zygotic embryo-derived callus of arecanut. An in Regional
vitro culture procedure was established for somatic embryogenesis and involving
plant regeneration from callus cultures. Segments of zygotic embryos were varieties
cultured on Murashige and Skoog basal medium supplemented with vitro plan
dicamba (9.05, 18.1,27.15, and 36.2 )lM). After 7-8 week in darkness, of these h
 577

,.on wounded regions of explants formed callus with yellow, soft, glutinous
las structures. Proliferation and maintenance of callus was on the same
led dicamba-containing medium. With regular subculture every 8 weeks, the
ted callus showed pale yellow, compact and nodular structures. During
1as subculture, somatic embryos were formed spontaneously from nodular
lse callus tissues within 2-4 months. The embryos developed into plantlets
tic after 10 weeks of culture on basal medium free of plant growth regulators.
ted After subculturing every month for 3 months, the plantlets were transferred
(OS to containers for acclimatization in the greenhouse. The survival rate was
eet 24% .
vth 5.1.1. Applications
md
yos One of the major production constraints of arecanut is the
IOn devastating Yellow Leaf Disease (YLD). This serious malady is prevalent
in Kerala and Karnataka states. All the cultivars have been reported to be
susceptible for this disease but few South Kanara Local cultivars are found
'tro to be field resistant. Surveys conducted in the hot spot regions revealed
ryo that healthy palms with good yields are available in the midst of the severely
cv. disease affected trees. These healthy palms are being used for resistant
1m, breeding programme. The tissue culture protocol developed at CPCRI was
Dns used for mass multiplication of field resistant Yellow Leaf Disease (YLD)
)al, palms. The field evaluation of these plants have been taken up in three
the villages viz. Gunadka, Balambi, and Nadubettu at Suliya Taluk ofDakshina
3a) Kannada District ofKarnataka State. The performance of the tissue cultured
om plantlets in the YLD hot spot region was found to be satisfactory and
mts comparable with seeding raised palms (Karun et al., 2005) .
urn
age Currently the arecanut tissue culture protocol developed at CPCRI
vith has also been applied for mass mUltiplication of dwarf arecanut (variety
'Hirehalli') and its hybrids. 'Hirehalli Dwarf', a natural mutant identified
urn
ots. in 1963 for its short stature, is a good genetic source for arecanut
fter improvement. The main features of the dwarf are the complete suppression
of the internodal space and erect crown shape. This dwarf has been used in
ary
lout all the major hybridization programmes of CPCRI. The identification of
hybrids with dwarfness and high yielding potential will benefit areca by
way of increased returns and reduced cost of various cultural operations.
atic The breeding programme at the Central Plantation Crops Research Institute
1 in Regional Station at Vittal, Karnataka have carried out crossing programme
and involving various combinations of a local dwarf with five high yielding '
,ere varieties and have succeeded in developing three promising hybrids. In
vith vitro plantlet development could be achieved from inflorescence explants
ess, of these hybrids.
578

5.1.2. Embryo rescue collection

for in vi!
In vitro germination of excised mature embryos of arecanut was
fertility. T
reported by Ganapathi ef af., 1997. Mathew and Philip (2000) reported
that are ('
the protocol for in vitro propagation via direct adventitious shoot bud
collected ,
differentiation from cotyledon explants. Karun ef af., (2002) repOlied an
collected
in vitro germination teclmique for rapid germination of arecanut embryos.
pollen ger
Seven month old embryos collected from four varieties 'Mangala', 'Sree
along witt
Mangala"Sumangala' and 'South Kanara Local' were cultured in vitro on
Fresh as
three agar gelled hormone free medium viz., Eeuwen's Y3 medium, 1"2
germinati(
strength MS and full MS. Embryos cultured in Y3 medium supplemented
pollen obs
with sucrose 3 % showed maximum germination (93.3 %). In vitro plantlets
59.5% in
after eight weeks in germination medium were transferred to liquid medium
'Sumanga
containing reduced level of sucrose (J.5 %) for proper development of
11.22% in
roots and expansion of leaves. Fully developed plantlets with minimum
three diffe
varietal difference with respect to germination was also recorded and
30Ve ,32{
showed that the varieties 'Sumangala' and 'South Kanara local' were more
germ inatic
responsIve.
pollen was
At present, arecanut germplasm is conserved mainly in field gene
Pc
banks which are exposed to climatic and biological hazards and are costly
in maximu
to maintain over a large area. Being a highly cross pollinated and
studies al!
recalcitrant species, cryopreservation of somatic embryo is the only option
germinatio
for long term conservation of arecanut genetic resources.
germinatio
5.1. 3 Cryopreservation of arecanut pollen was after t
observed"
Areca is a genus of about 76 species of palms in which Areca
cryopreser'
catechu L. is the only cultivable species. It is an allotetraploid palm with
and 63.98
chromosome number, 2n =32 and highly cross pollinated. Being a high
hybrids (H
valued commercial crop, its contribution in terms oflivelihood, employment
to be 78.3<i
and income to the national economy is significant. As a perennial tree crop
arecanut germplasm is maintained in the field gene bank. The arecanut Vic
palm is monoecious and its pollen remains viable for 8-9 hr under normal for differef
conditions. Increased longevity of pollen, from 15 to 21 days by storing in be 83.4%,
desiccators at room temperature was reported by Bhat et af., (1962). Since pollen for:
pollen is a useful source of diverse alleles within genepool, storage of
Pn
pollen is highly essential germplasm conservation as well as for hybrid
indicated it
seed production and also for assisted pollination. Pollen cryopreservation
feasible in
is the only larger scale, long term option for the ex situ conservation of
programm
areca nut pollen.
conducted:
Studies were conducted for standardization of procedure for pollen years cryos
 579

collection, its desiccation, germination media, and effect of temperature

for in vitro germination, pollen cryopreservation and its viability and
fertility. The fully matured male flowers, which had just opened as well as
that are expected to open (white colored perianth) the next day, were
collected and dried in at ambient temperature for 24 hours. The pollen was
colIected by sieving the dried male flowers. For selecting the media for
pollen germination, sucrose at various concentrations (2.5 %, 3 %, 4 %)
along with agar (l %), gelatin (1 %) and boric acid (0.01 %) were studied.
Fresh as well as desiccated pollen samples were incubated in pollen
germination media for a period of 90 minutes. Percent germination of
pollen observed are 82.65% in 2.5% sucrose, 51.43% in 3% sucrose and
59.5% in 4% sucrose in 'Hirahalli Dwarf' . In the case of tall cultivar
'Sumangala', it was 47.03% in 2.5% sucrose, 13% in 3 % sucrose and
11.22% in 4% sucrose. The desiccated pollen samples were incubated at
three different temperatures (room temperature: 28 to 30Ve, incubator:
30Ve, 32VC) for a period of90 minutes to optimize temperature for pollen
germination. Maximum pollen germination (75 %) was observed when
pollen was incubated at room temperature (28 to 30lJC).
Pollen germination medium containing 2.5% sucrose, that resulted
in maximum germination, was selected as standard for cryopreservation
studies also. The desiccated pollen samples were cryopreserved and
germination as well as pollen tube length measured. In cryopreserved pollen,
germination commenced after 90 minutes whereas in desiccated pollen, it
was after 60 minutes. In 'Hirehalli Dwarf', the germination percentages
observed were 91 % in fresh pollen, 72.2% in dried pollen and 70.1 % in
cryopreserved pollen whereas in 'Sumangala', it was 72.06 %, 47.03 %
and 63.98 % in fresh, dried and cryoprescrved pollen respectively. In
hybrids (Hirehalli dwarf x Sumangala), germination percentage was found
to be 78.3% in fTesh, 66.2 % in dried and 61.43 % in cryopreserved pollen.
Viability of cryopreserved arecanut pollen from Hirehalli dwarf
for different durations was studied. Germination percentage was found to
be 83.4%, 73.8%, 48%, 45% and 42% in fresh, oven dried, cryostored
pollen for 24 hours, one month and two months respectively.
Production of normal nut set with the use of cryopreserved pollen
indicated its fertility. The study concludes that pollen clyopreservation is
feasible in arecanut and can be utilized in large scale hybridization
programmes and also for germplasm conservation. Nut set studies
conducted at epeRI showed that normal nut set was observed with two
years cryostored pollen from Hirehalli dwarf palms.
 ------

580

5.2 Molecular Markers

confirrr
Arecanut germplasm have been characterized and evaluated based
polymo
on morphological and yield parameters (Ananda et aI., 2000; Rajesh, 2007).
of plant
However, the genetic diversity information provided by morphological
palms (
characters is limited and these parameters can be influenced by
et at. , ;
environmental, genetic and physiological factors. DNA-based molecular
coeffici
markers are an important tool for evaluating levels and patterns of genetic
and its I
diversity and have been utilized in a range of plant species and are available
were sh
in unlimited numbers.
and its I
Limited work has been conducted in arecanut for characterization shown!
of germplasm. Ananda and Rajesh (2002) utilized morphological and yield the larg
parameters and protein profiles for characterization of arecanut germplasm. Table 2.
Cluster analysis, based on protein profiles, did not show any association
between geographic and genetic affinities (Rajesh, 2007), highlighting the E "pla nts

limitations in using biochemical markers in arecanut. Bagindo (2011) used Zygo ti c E

RAPD for genetic diversity analysis of 30 Areca catechu individual s
collected in Indonesia (Papua, Sulawesi and Sumatra). Sets ofSSR markers Zygo tic e
were developed by Hu et af., (2009) and Zhan et af., (2012), while Ren
and Tang (20 I 0) optimized protocol for ISSR technique in arecanut, but
these markers have not been used for detailed genetic diversity analysis of
arecanut germplasm. Immature
Recently, genetic relationship among 60 arecanut germplasm, (S eedling

consisting of both indigenous and exotic accessions, were assessed using

14 polymorphic RAPD primers ( Bharatb et aZ., 2015). The average Immature
(adult pal l
polymorphism was 6.64 markers per primer. The PIC values among the 14
primers ranged from 0.19 to 0.49. Simjlarity values among the accessions
In fl o resce
ranged between 0.68 and 0.93. Cluster analysis revealed two major clusters.
The Indian collections Konkan 1, Konkan 11 and Maidhan formed a separate
cluster. Collections from Jndonesia, Sri Lanka, Vietnam, Fiji, Solomon R oot
Islands, Singapore China and some Indian collections ( Andaman and
Nicobar Islands and N0l1h East germplasm collections) formed a second
cluster. The clustering pattern was, in general, in accordance with the Leaf (Seel
geographical origin of the collections. The results obtained from this study
are crucial for developing effective management strategies for genetic
improvement of arecanut. The results of the study thus provides the first R oo t (See
basic information on the genetic relationship amongst arecanut accessions
from diverse geographical regions and can form tbe basis of advanced
Stem-(Sel
studies on germplasm characterization and conservation, breeding programs
and selection of possible parents to generate mapping populations of
arecanut accessions.
 581

Molecular markers have been successfully applied for the

confirmation of fidelity of tissue cultured plantlets. Random amplified
ased polymorphic DNA (RAPD) markers were used to evaluate clonal fidelity
107). of plantlets derived through direct somatic embryogenesis from two mother
gical palms (H and G) of Yellow Leaf Disease (YLD) resistant arecanut (Karun
i by et aZ., 2008). Pair wise genetic similarities were generated by Jaccard's
~ular
coefficient using the RAPD banding pattern between each mother palm
netic and its (eight plantlets /palm) progenies. Mother palm H and its progenies
lable were showing maximum similarity (99 %) where as, the mother palm G
and its progenies were showed 98 % similarity. The low level of variability
ation shown by plantlets of direct somatic embryogenesis can be exploited for
yield the large-scale mUltiplication of elite arecanut palms.
asm. Table 2. Summary of the research done on arecanut tissue culture
ation
Explants Basal Medium Response Reference
g the
used Zygotic Embryo Murashige and Skoog Adventitiou s shoot Mathew and
iuals formation Philip (2000)

rkers Zygotic embryo Eeuwens Y3 Varied with varieties Karun el ai.

- Sreemangala and (2002)
Ren
South Kanara Local
, but gave 93 %
;is of germ ination
Immature leaf Murashige and Skoog Plantlet regeneration Karun et al.
(Seedling) with picloram and through somatic (2004)
asm,
addition of BAP em bryogenesis
lSlllg
Immature Leaf ­ Murashige and Skoog Plantlet regeneration Karun et al.
;rage
(adult palm) with picloram and through somatic (2004)
1e 14 addition of BAP em bryogenesis
S10ns
I n florescence Murashige and Skoog Plantlet regeneration Karun et al.
sters. with picloram and through somatic (2004)
arate addition of BAP embryogenesis
)mon Root Murashige and Skoog Plantlet regeneration Radh a el al.
I and with picloram and through somatic (2009)
·cond addition of BAP embryogenesis
1 the Leaf (Seedling) Murashige and Skoog Regeneratio n of Wang et al.
;tudy with BA (0.2 mg/I plantlets through (2003a)
and TDZ ( 0.2 mg/I) organogenesis
netic
first Root (Seedling) Murashige and Skoog Regeneration of Wang el al.
plantlets through (2003b)
Slons organogenesis
lI1ced
Stem-(Seedling) Murashige and Skoog Regeneration of Wang et al. (2006)
;rams plantlets through
1S of organogenesis
Table 3. VARllirrlliS OF ARECANUT RELEASED IN INDIA Ul
00
N

Variety Year of Breeding Method Parents Important traits Institute/

Release (In trod u ction, University
Mutation, hybrid
etc. with details)

Mangala 1972 Introduction, evaluation VTL-3 (China) Semi Tall palm with partially drooping CPCRI,
and selection crown, earliness in bearing, more number Kasaragod
of female flowersllnl1orescence, higher
nutset, quicker stabilization, round and
medium sized ye llow coloured nuts.
Average chalildry kernel yield is 3.00kg!
palm/year

Sumangala 1985 Introduction , evaluation VTL-il (Indonesia) Tall palm with partially drooping crown, CPCRI,
and selection oval to round shaped deep yellow Kasaragod
coloured nuts. High recovery of chali
(26.50%) from fresh fruits. Average
chalildry kernel yield is 3.28 kg/palm/year

Sreemangala 1985 Introduction, evaluation VTL-17 (Singapore) Tall palm with sturdy stem, partially CPCRl,
and selection drooping crown. Round and bold with Kasaragod
deep yellow coloured nuts. Average chalil
dry kernel yield is 3.18 kg/palm/year

Mohitnagar 1991 Introduction , evaluation VTL-60 (\V. Bengal, Tall palm with medium thick stem, CPCRI,
and sele ction India) partially drooping crown, orange Kasaragod
yellow coloured oval to round shaped
nuts. Higher level of uniformity in

performance. The bun che<; ~r" wp ll

 t" ....... L ........... LJ
ana seleCIIon iHUW) .......... """"t:" ... · O _.
~

yellow coloured oval to round shaped

nutS. Higher level of uniformity in

performance. The bunches are well

placed and nuts loosely arranged on
spikes which help in uniform development.
Average chaliJdry kernel yield is
3.67 kg/palm/year.

Cal-17 /Samrudhi 1995 Introduction, evaluation VTL-37(Andaman & Tall palm with longer internodes, CPCRI and
and selection Nicobar Islands, India) partially drooping crown. Elongated bold CARl
nuts with orange yellow in colour.
Average chali/dry kernel yie ld is 4.34
kg/palm/year and recommended for
Andaman & Nicobar group of Islands.

SAS-I 1995 Introduction, evaluation VTL-52 (Sirs i Local, Tall palm with compact canopy, deep UAS, Dharwad
and selection India) orange colour with round and even
sized nuts. It is suitable for both tender
nut and ripe nut processing. Average chali/
dry kern el yield is 4.60 kg/palm /year and
recommended for Sirsi hill zone of
Karnataka.
Swarnamangala 2006 Introd uction, evaluation VTL-12 (Saigon) Tall palm with medium thick stem and CPCRI,
an d selection comparatively shorter internodes, partially Kasaragod
drooping crown . Nuts are bold and heavier
with high recovery of chali (26.40 %).
Average chal iJdry kernel yield is 3.88 kg/
palm/year.

VTLAH-I 2006 H ybri dization and Hirehalli dwarf (VTL- D warf in nature. Sturdy stem with super CPCRI,
tJl
evaluation 56) and Sumangala imposed nodes, reduced canopy size, Kasaragod 00
W
 fJl

we ll spread leaves, medium sized oval

....
00

(VTL-ll )
to round shaped nuts and early
stabilizat ion and me dium yielder.
Average chali/dry kernel yield is
2.54 kg/pa lm /y ear. Advantages-reduced
cost of cultivation in terms of harvesting
and spraying.

Hybridi zation and Hireh aUi dwarf (VTL­ Dwarf in nature. Sturdy stem with
VTLAH-2 2006
eva luati on 56) and Mohitnagar super imposed nodes, redu ced canopy
(VTL-60) size, well spread leaves, medium sized
oval to round shaped nuts and early
stabilization and med ium yie lder. Average
chali/dry kern el yield is 2.54 kg/palmi
year. Advantages-reduced cost of
cu ltivation in terms of harve sting and
sp raying.

Introduction, evaluat ion VTL-64 (Ass am, India) Tall in natu re with mediu m thick stem, CPCR I,
Kahikuchi 2009
longer intern odes, regular bearer, Kasaragod
and selection
consistent in yield, bunches are well
placed on the stem, oran ge colour, bold
and round shaped nuts, high recovery
(25. 16 %) ofch ali from fresh nuts,
comes to bearing by 5th year. Average
yield is 3.70 Kg dry kernel/palmi
year under normal conditions.

Madhuramangala 2013 Introduction , evaluation VTL62 (Maharastra, In dia) Sem i tall type, medium thick stem, CPCRI,
and selection regular bearer, medium maturity and Kasa r agod
'- • _ __ : __ _ 1 _. .A ,n __ _ _
 year under normal conditions.

Madhuramangala 2013 Introduction , evaluation VTL-62 (Maharastra, India) Semi tall type, medium thick stem, CPCRI,
and select ion regular bearer, medium maturity and Kasaragod
bearing by 4th year, synchronized
maturity of nuts, orange to yellow
colour nuts, oval to round shaped
nuts, marble appearance in split kernel,
suitable for making both chali and
processed tender nut. The average yield
under normal conditions is 3.54 Kg
ChaliJpalrnlYear and 2.95 Kg Dry tender
processed nuts/palmlYear.

Nalbari 2013 Introduction, evaluation VTL-75(Assam, India) The yield performance of this variety CPCRI,
and selection is higher than the earlier released Kasaragod
varieties.TaU type with medium thick
stem, shorter internodes, hom ogenous
population, regular bearer, well placed
bunches, yellow coloured round shaped
nuts, and belong to medium maturity
group, suitable for making chaliJdry
kernel. The average yield under normal
condition is 4.15 Kg Chalil dry kernel
palm/year.

'Jl
00
'Jl
 -
-- --~- -- - ­
586

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