FLAVOUR AND FRAGRANCE JOURNAL

Flavour Fra`r[ J[ 1999^ 04] 026Ð039

First observation of little leaf disease and its impact on the

yield and composition of the essential oil of rose-scented
geranium (Pelargonium sp.)
B. R. Rajeswara Rao,1 P. N. Kaul,1 Gopal R. Mallavarapu2 and S. Ramesh2
1
Central Institute of Medicinal and Aromatic Plants Field Station, Boduppal, Uppal (PO), Hyderabad 500 039, India
2
Central Institute of Medicinal and Aromatic Plants Field Station, GKVK (PO), Bangalore 560 065, India

Received 07 Au`ust 0888

Revised 0 December 0888
Accepted 7 December 0888

ABSTRACT] Rose!scented geranium "Pelar`onium sp[# plants infected with little leaf disease\ probably caused
by mycoplasma\ were _rstly observed during 0885Ð0887[ The plants exhibited symptoms of reduced leaf:petiole
size "little leaf#\ yellowing and bunching of leaves and cessation of plant growth\ resulting in plant mortality[
Plant parameters\ viz[ plant height\ plant spread\ number of branches\ leaf size\ leaf weight\ etc[\ were adversely
a}ected by little leaf disease\ leading to signi_cant reductions in shoot "38Ð69)#\ root "54Ð73)# and oil "56Ð
80)# yields of diseased plants in comparison to healthy plants[ Healthy plants recorded higher concentrations
of geraniol\ while little leaf disease!a}ected plants had higher percentages of isomenthone\ citronellol\ citronellyl
formate\ 09!epi!g!eudesmol\ citronellyl tiglate and geranyl tiglate[ Copyright Þ 1999 John Wiley + Sons\ Ltd[

KEY WORDS] Pelar`onium sp^ Geraniaceae^ rose!scented geranium^ little leaf disease^ shoot yield^ root yield^
essential oil yield^ essential oil composition

Introduction the causal organisms for leaf blight:leaf spot diseases[

Rose!scented "Pelar`onium sp[\ family] Geraniaceae# is
a multi!harvest\ high!value\ aromatic crop\ the essential
oil of which is used in high!grade perfumes and some!
times for the isolation of commercial rhodinol[ The
crop is a}ected by a number of pests and diseases[ In
high!altitude locations of India\ nematodes "Mel!
oido`yne inco`nita\ Meloido`yne hapla\ Helicotylenchus
dihystera\ Scutellonema conicephalum#\ mites\ sucking
pests "species not mentioned#\ wilt "Pythium vexans#\
root rot "Fusarium sp[#\ leaf spot "Cercospora spp[#\ rust
"Puccinia spp[# and collar rot "Sclerotium rolfsii# a}ect
the crop\ resulting in plant mortality and biomass and
oil yield reductions[0Ð3 In Indian plains\ extensive dam!
age to plantations of rose!scented geranium is caused by
wilt and leaf blight:leaf spot diseases[ Wilt is reportedly
caused by a number of fungi\ namely Fusarium
redolens\4 Fusarium sp[\5 Botryodiplodia theobromae\6
Verticillium albo!atrum\7 Rhizoctonia solani\8 Pythium
sp[\09 Botryosphaeria obtusa00 and Phytophthora spp[\01
while Colletotrichum `loeosporioides\02 Colletotrichum
acutatum3 and Alternaria alternata3\01 were found to be
 Correspondence to] B[ R[ Rajeswara Rao\ Central Institute of Medicinal and
Aromatic Plants Field Station\ Boduppal\ Uppal "PO#\ Hyderabad 499 928\
India[
Other diseases\ namely root rot "Rhizoctonia solani#\8
basal rot of cuttings "Rhizoctonia solani\ Fusarium
moniliformae#\03 die!back of cuttings "Botryodiplodia
theobromae#04 and shoot tip burn and tip rot "Gleo!
sporium sp[#3\01 were also reported to cause damage to
the crop[ In this paper\ a possibly new disease of rose!
scented geranium is described and its e}ect on the crop
has been investigated[
Experimental
Field Observations and Isolation of Essential Oil
General plantation of rose!scented geranium cv[ Bour!
bon "Pelar`onium `raveolens L|Herit[ ex Aiton×P[
capitatum "L[# L|Herit[ ex Aiton# was maintained fol!
lowing recommended agricultural practices in the
Experimental Farm of Central Institute of Medicinal
and Aromatic Plants Field Station\ Hyderabad\ India[
The experimental location experiences semi!arid trop!
ical climate[ Two plants in February 0885\ one each in
April 0886 and February 0887 were observed to be
a}ected by little leaf disease[ Spraying of complete
Hoagland solution or broad!spectrum systemic fungi!
cides did not help in the recovery of the plants[ When the

Copyright Þ 1999 John Wiley + Sons\ Ltd[

027 B[ R[ RAJESWARA RAO ET AL[

Figure 1. Diseased (left) and healthy (right) leaves of rose-scented geranium (Pelargonium sp.)

plants showed symptoms of dying\ they were uprooted
along with root biomass and observed for insect pests[
In each year\ 09 healthy plants surrounding the diseased
ones were also dug out and examined for comparison[
Fresh seedlings were planted in the dug!out holes and
observed for their growth and development[ The shoot
biomass of both diseased "four plants# and healthy "09
plants:year# plants were submitted to distillation using
a Clevenger!type apparatus for 2 h for isolation of
essential oil[ The oil samples were dried over anhydrous
sodium sulphate and stored at 9>C until analysis[
GC Analysis
GC analyses of the above oil samples were carried out
using a Perkin!Elmer gas chromatograph 7499 _tted
with an FID and a BP!0 glass capillary column "14
m×9[4 mm i[d[# coated with dimethylsiloxane[ The car!

Table 1. Variation in plant parameters between healthy and diseased plants of rose-scented geranium (Pel-
argonium sp.)
Plant parameter Healthy Diseased Reduction in diseased
plant plant plant
"n  29# "n  3# ")#

Plant height "cm# 53 30 25

Plant spread
EastÐWest "cm# 52 23 35
NorthÐSouth "cm# 65 33 31
Number of branches per plant 15 09 51
Branch diameter "cm# 1[0 0[8 09
Number of leaves per branch 10 035 Ð
Leaf colour and thickness Green to dark green\ Pale yellow to yellow\ Ð
thick and hairy thin and less hairy
Leaf size
Length "cm# 5[3 0[5 64
Width "cm# 7[5 0[2 74
Leaf weight "mg# 0450[8 18[0 87
Petiole colour and appearance Green\ sturdy\ healthy\ hard to Pale green to yellow\ weak\ Ð
the touch and hairy thread!like\ soft to the touch and
less hairy
Petiole length "cm# 09[6 1[4 66

Copyright Þ 1999 John Wiley + Sons\ Ltd[ Flavour Fra`r[ J[ 1999^ 04] 026Ð039
 LITTLE LEAF DISEASE AND ESSENTIAL OIL OF GERANIUM 028

used as standards# with literature data\05Ð07 and by peak

enrichment on co!injection of authentic chemicals\
wherever possible[

Results and Discussion

Symptoms of the Disease
Plants a}ected by the disease exhibited symptoms of
reduction in leaf:petiole size "Figure 0# with pale green
to yellow\ soft\ thread!like petioles^ loss of green colour
of the leaves resulting in yellowing^ bunching of leaves
"Figure 1# and cessation of plant growth giving it a
dwarfed appearance[ In 0886\ when a young plant was
a}ected\ it did not develop tap root\ instead developed
thin\ hair!like\ brown coloured _brous roots[ The
infected plants ultimately died in all the 2 years[

Frequency, Occurrence and Transmission of the

Figure 2. Diseased (left) and healthy (right) branches of
rose-scented geranium (Pelargonium sp.). Symptoms of
little leaf, discolouration and bunching of leaves are
clearly seen in the diseased branch
rier gas used was nitrogen at 39 ml:min ~ow rate[ The
temperature programming was 59Ð119>C\ with a ramp
rate of 4>C:min and with a _nal hold time of 09 min[
Samples "9[0Ð9[1 ml# were injected with 0 ] 79 split ratio[
The essential oil constituents were identi_ed by com!
paring the retention times with those of reference com!
pounds run under similar conditions\ relative retention
indices of the peaks "computed from the gas chro!
matograms by logarithmic interpolation between
bracketing n!alkanes^ the homologous series of n!
alkanes C7ÐC11\ Poly Science Inc[\ Niles\ USA\ were
Disease
Diseased plants were observed during spring\ "Feb!
ruary\ March# and summer "AprilÐJune# seasons\ while
no occurrence of the disease was noticed in other
seasons[ Only one "0886\ 0887# or two plants "0885#
were a}ected by the disease in the 2 years[ However\
the causes for the low frequency or the susceptibility of
the infected plants to the disease are yet to be ascer!
tained[
Rose!scented geranium is propagated through rooted
stem cuttings[ Stem cuttings obtained from the infected
plants did not survive[ Similarly\ the disease could not
be transmitted by physical proximity or contact of
healthy plants with the diseased plants\ by rubbing
healthy leaves with sap from infected plant leaves\ by
grafting\ or by planting healthy seedlings at the same
place after uprooting the diseased plants[
Causal Organism of the Disease
No insect pests were found in the infected plants[ Dr R[
D[ V[ J[ Prasada Rao\ Senior Virologist at National

Table 2. Effect of little leaf disease on shoot, root and oil yields of rose-scented geranium (Pelargonium sp.)
Factor Shoot yield Root yield Oil yield
"g:plant# "g:plant# ")#

0885 0886 0887 0885 0886 0887 0885 0886 0887

Healthy plants 609 089 562 54 14 41 9[07 9[09 9[00

Diseased plants 254 47 297 10 3 07 9[95 9[91 9[90
Reduction in diseased plants ")# 38 69 43 57 73 54 56 79 80
 n  09:year of healthy plants\ 0 or 1:year of diseased plants[

Copyright Þ 1999 John Wiley + Sons\ Ltd[ Flavour Fra`r[ J[ 1999^ 04] 026Ð039
039 B[ R[ RAJESWARA RAO ET AL[

Table 3. Influence of little leaf disease on the chemical adverse e}ect of the disease on di}erent plant
composition (peak area percentage) of rose-scented ger- parameters\ the infected plants recorded 38Ð69) less
anium (Pelargonium sp.) essential oil shoot biomass yield\ 54Ð73) decreased root yield and
Compound RRI Healthy Diseased 56Ð80) reduced essential oil yield\ in comparison to
plant plant healthy plants "Table 1#[
a!Pinene 824 9[1 9[2 Little leaf disease induced some marked changes in
b!Pinene 865 9[1 9[3 the composition of the essential oil also[ Healthy plants
Limonene 0913 9[4 9[5 were richer in geraniol\ while the diseased plants rec!
cis!b!Ocimene 0917 9[0 9[0
trans!b!Ocimene 0939 9[1 9[1 orded higher concentrations of isomenthone\ citro!
cis!Linalol oxide "furanoid# 0952 9[4 9[4 nellol\ citronellyl formate\ 09!epi!g!eudesmol\
trans!Linalol oxide "furanoid# 0965 9[1 9[2 citronellyl tiglate and geranyl tiglate "Table 2#[
Linalol 0977 09[4 8[4
cis!Rose oxide 0986 9[4 9[7 This investigation has revealed that probably little
1!Phenylethyl alcohol 0091 tr tr leaf disease resulted in signi_cant yield losses and that
trans!Rose oxide 0005 9[3 9[4 it induced marked changes in the essential oil com!
Menthone 0025 9[7 9[8
Isomenthone 0036 6[6 7[8 position of rose!scented geranium[
Citronellol 0104 17[4 20[3
Acknowled`ements * The authors thank the Director\ CIMAP\
Geraniol 0131 13[2 01[3
Lucknow\ and the Scientist!in!Charge\ CIMAP FS\ Hyderabad\ for
Citronellyl formate 0150 4[9 5[1
facilities and encouragement\ and Dr R[ D[ V[ J[ Prasada Rao for
Geranyl formate 0173 1[0 0[8
examining the infected plants\ conducting laboratory tests and giving
Citronellyl acetate 0226 9[0 9[1
his opinion for the identi_cation of the causal organism[
Geranyl acetate 0251 9[2 9[2
b!Bourbonene 0275 9[4 9[7
b!Caryophyllene 0311 9[3 9[4
5\8!Guaiadiene 0333 9[0 9[0 References
Germacrene!D 0370 9[1 9[1
Citronellyl butyrate 0492 9[0 9[3
Geranyl butyrate 0423 9[3 9[5 0[ Doraswamy K\ Sundaram M\ Manian K[ Madras A`ric[ J[ 0868^
1!Phenylethyl tiglate 0444 9[8 0[3 55] 194[
09!epi!g!Eudesmol 0506 4[6 7[1 1[ Kumar S\ Nanjan K[ Indian Perfum[\ 0874^ 18] 010[
Citronellyl tiglate 0534 0[2 2[8 2[ Aravindakshan M\ Aslam SMA[ In Plantation Crops] Oppor!
Geranyl tiglate 0564 0[1 2[9 tunities and Constraints\ vol 0[ OxfordÐIBH Publishing Co] New
Delhi\ India\ 0875^ 218[
 Retention index relative to n!alkanes on BP!0 "methylsiloxane# column[ 3[ Annual Report[ Central Institute of Medicinal and Aromatic
tr  trace " ³ 9[94)#[ Plants] Lucknow\ India\ 0886Ð87^ 33[
4[ Sarwar M\ Parameswaran TN\ Shanmugam C\ Narayana MR[
Indian Perfum[\ 0871^ 15] 011[
5[ Staddard EMA[ Plant Dis[ Reptr[\ 0846^ 30] 425[
Bureau of Plant Genetic Resources Regional Station 6[ Manholay MN\ Naragund VR[ Indian Perfum[\ 0870^ 14] 60[
"Plant Quarantine#\ Hyderabad\ after observing the dis! 7[ Torgeson DC[ Plant Dis[ Reptr[\ 0841^ 25] 425[
8[ Kalra A\ Parameswaran TN[ Indian J[ Plant Pathol[\ 0877^ 5] 71[
eased plant in the _eld and examining the plant samples 09[ Singh R[ Proceedin`s of the Symposium on Essential Oils and
in his laboratory\ opined that the disease was probably Aromatic Chemicals\ Dehradun\ India\ 0844^ 051[
caused by mycoplasma "little leaf and bunching of 00[ Raghavendra Rao NN\ Chacko CI[ Annual Report[ Indian Insti!
tute of Horticultural Research] Bangalore\ India\ 0872^ 001[
leaves are characteristic symptoms of a mycoplasmal 01[ Annual Report[ Central Institute of Medicinal and Aromatic
disease#^ however\ this identi_cation is tentative and Plants] Lucknow\ India\ 0885Ð86^ 59[
needs further con_rmation[ 02[ Kalra A\ Parameswaran TN\ Ravindra NS[ Curr[ Sci[\ 0877^ 46]
0025[
03[ Ouf MF\ Sirry AR[ A`ric[ Res[ Rev[\ 0866^ 44] 026[
04[ Shehata MRA\ Tarabish AM\ Sania AF[ E`ypt[ J[ Phytopathol[\
Impact of the Disease on the Yield and 0866^ 14] 60[
Composition of the Essential Oil 05[ Davies NW[ J[ Chromato`r[\ 0889^ 492] 0[
06[ Fraisse D\ Schart C\ Vernin G\ Metzger J[ Proceedin`s of the
The disease adversely a}ected various plant parameters IXth International Essential Oil Con`ress\ Sin`apore\ Book 2[
of rose!scented geranium "Table 0#[ Except number of 0872^ 099Ð019[
07[ Jennings W\ Shibamoto T[ Qualitative Analysis of Flavor and
leaves per branch\ all other plant parameters were Fra`rance Volatiles by Glass Capillary Gas Chromato`raphy[ Aca!
markedly higher in healthy plants[ As a result of the demic Press] New York\ 0879[

Copyright Þ 1999 John Wiley + Sons\ Ltd[ Flavour Fra`r[ J[ 1999^ 04] 026Ð039