Vol.2, No.3, 291-296 (2011)
Copyright © 2011 SciRes. Openly accessible at http://www.scirp.org/journal/AS/
Agricultural Sciences
Mango malformation: II. mangiferin changes associated
with fusarium pathogens
Wafaa M. Haggag1*, Mahmoud Hazza2, Mohamed E. Abd El-Wahab1
1Department of Plant Pathology, National Research Center, Dokki, Cairo, Egypt;
2Science Faculty, Botany Department, Banha University, Egypt; Corresponding Author: Wafaa_haggag@yahoo.com
Received 16 February 2011; revised 23 May 2011; accepted 25 July 2011.
Mangiferin (1,3,6,7-tetrahydroxy xanthone-C2-b-
D-glucoside) promoted vegetative growth and
exhibited inhibitory role on the occurrence of
malformation. Mangiferin changes associated
with mango malformation pathogens were fol-
lowed after inoculated mango seedlings (three
years) with malformation pathogens i.e. Fusa-
rium subglutinans, F. sterilihyphosum, F. ox-
ysporum and F. proliferatum. Mangiferin re-
mained at lower level in leaves of malformed
shoots as compared to healthy one. The floral
malformation was observed to be associated
with the reduction of mangiferin. S trong positive
correlations between mangiferin activity and
malformation incidence were observed. Mangiferin
level at panicle initiation may give a possible
estimate of malformation incidence in mango.
Keywords: Fusarium; Mangiferin; Mango
Mango (Mangifera indica L.) is the most important
fruit crop in Egypt. Mango ranks third in exports after
citrus and grapes. Egypt produces 232,000 tones of
mango annually and exports moderate amount (about
1500 tones) annually to 20 countries in near east and
Europe (FAO, 2007).
Mango Malformation is one of the most destructive
mango diseases [1]. Accumulation of mangiferin and
toxic metabolites of Fusarium moniliforme has been
suggested to be responsible for the malformation disease
of mango (Mangifera indica L.). Differences in the low-
and medium M, phenolic and steroidal compounds in
healthy and malformed florets of Mangifera indica, the
latter infested with Fusarium moniliforme var. subg luti-
nans are reported. Mangiferin, which are not normal
constituents of healthy florets were found in substantial
amounts in the diseased florets. Both mangiferin and 1,3,
6,7-tetrahydroxyxanthone were found to be potent anti-
fungal agents[2]. Accumulation of mangiferin, a natural
metabolite of Mangifera indica at the site of differenti-
ating buds, influences the changes from reproductive to
vegetative growth. Mangiferin in high concentration
suppressed the activity of peroxidase, catalase, α-amy-
lase and IAA-oxidase. Polyphenoloxidase and invertase
showed increased activity. Mangiferin accumulation
increased the rate of photosynthesis but lowered those of
transpiration and respiration. Mangiferin treatment in-
creased the contents of chlorophyll, carbohydrates, total
nitrogen, protein nitrogen, nucleic acids (RNA and
DNA) and indole-3yl-acaetic acid (IAA) [3]. Mangiferin
acts as a phytoalexin-like compound in M. indica. Non-
pathogenic strains of F. moniliforme isolated from maize
cob and banana fruits induced more mangiferin synthesis
than the pathogenic isolate from malformed mango
shoots in vivo. Activity of the mangiferin degrading en-
zymes, polyphenoloxidase of non-pathogenic strains of
F. moniliforme was higher than that of the pathogenic
strain. In vitro tests also confirmed rapid and massive
degradation of mangiferin by non-pathogenic strains.
Biochemical events associated with elicitation, degrada-
tion and accumulation of mangiferin determine the host
specificity of F. moniliforme in M. indica [4]. At the
stages of bud pre-emergence and initial differentiation,
mangiferin content in the leaves from the axils of which
healthy and malformed buds emerged did not show sig-
nificant differences although in the latter one mangiferin
content was higher. But in the fully developed panicles
at pre-blooming stage mangiferin content of the leaves
attached to the malformed panicle was in traces and sig-
nificantly lower than that of healthy one. Mangiferin
content in malformed panicles also did not differ sig-
nificantly at the initial stage. But the difference become
highly significant after influx of mangiferin from axil
leaves to the fully developed panicle at pre-blooming
W. M. Haggag et al. / Agricultural Science 2 (2011) 291-296
Copyright © 2011 SciRes. Openly accessible at http://www.scirp.org/journal/AS/
stage. Mangiferin due to its vegetative growth promoting
property tilted the hormonal balance of malformed pani-
cles in favour of vegetative growth resulting into trans-
formation of malformed florets into green leafy struc-
tures [5]. The mangiferin treated strains produced more
aerial hyphae but less pigment. Prolonged mangiferin
treatment affected the saprophytic ability of the strains
but improved its parasitism. The significance of mangiferin
induced changes in evolving the host-specific strain of
F. moniliforme of M. indicia lies in showing that an e-
cological disadvantage of survival in one niche (sapro-
phytic) may prove advantageous in another (parasite)
[8]. The fungal and mite populations were initially posi-
tively related to the mangiferin content and the disease
incidence. Further increase in mangiferin content re-
duced the fungal and mite populations [6]. The symp-
toms are the combined effects of aberrant host metabo-
lites produced in response to infection and phytotoxins
secreted by the pathogen. The pathogen has been identi-
fied as a physiological race of F. moniliforme (F.
moniliforme f. sp. mangifera) developed due to interac-
tion with the host metabolite, mangiferin for a prolonged
period. The disease cycle is greatly influenced with the
biochemical changes in the host tissues. Host metabo-
lites also effect the seasonal variation of population of
the pathogen Vis-a-Vis disease incidence. Proper bal-
ance of mangiferin and the Fusarium population is re-
quired for disease manifestation. Either by suppressing
or avoiding elicitation of hypersensitive reaction of the
host at the initial stage of infection, colonization by the
pathogen and subsequent symptom production could be
effected [3].Thus, objective of the present study is to
evaluate mangiferin change in mango plant due to infec-
tion with pathogenic fungi.
Mango (Mangifera indica L.) seedling cv. Sedekia
(three years old) was inoculated with 105 colony form-
ing units of Fusarium spp. i.e. F. subglutinans , F. ster-
ilihyphosum, F. oxysporum and F. proliferatum, the
causal organisms of mango malformation as inoculated
soil. Sterilized water was used as a control. Transplanted
seedlings were monitored for development of malforma-
tion. At the end of the experiment (120 days), all surviv-
ing seedlings were examined for apical disease symp-
toms. Samples for mangiferin study were taken from
leaves one-two cm below the tip of young seedlings.
2.1. Isolation of Mangiferin
The leaves were macerated with acetone in a high
speed blender. After 4 h, the mixture was filtered and the
solvent was removed under pressure. The extractions
were poured into 100 ml distilled water and the suspen-
sion was successfully extracted with ether and ethyl
acetate. At the interface of the aqueous ethyl acetate,
brown solid mass was precipitated and this was collected
by filtration. The identity of the brown residue was de-
termined as mangiferin according to Ghosal et al. [2].
Subsequently, a small volume of the extract was filtered
through 13 mm membrane filter (0.45 µm; polypropyl-
ene) directly into HPLC sample vial for injection with-
out further dilution.
2.2. Mangiferin Analysis
Extract were done using HPLC system (prominence
Lc, Shimaduz, Kyoto, Japan) equipped with a Lichro-
spher 100RP-18 (5 µm) column (250 mm × 4 mm,
Merck, Darmstadt, Germany), a C18 guard column and a
photoiode-array detector (Shimadzu, SPD-M20A). THE
elution system (0.8 mL·min–1) involved 2 mM phospho-
ric acid in water (eluent A) and MeOH (eluent B). The
gradient was as follows: 0 min, 25% eluent B,0 - 40 min,
80% eluent B, linear the retention time and spectral
characteristics of each sample were compared to a ref-
erence sample of mangiferin (Extrasynthese, Lyon,
France) [7].
2.3. HPLC Analysis and Method
The HPLC system consisted of a ternary solvent pump
(Gynkotek Model 480), autosampler (Gynkotek Gina
50), decade electrochemical detector with a glassy car-
bon electrode (Antec) and a diode array detector
(Gynkotek 340 S). Gynko soft software V5.60 was used
to control the HPLC system and for data acquisition and
analysis. The equipment was supplied by Dionex Softron
(Idstein, Germany). Three columns, i.e. Multosphere
C18 (3 μm; 125; 4 mm ID), Phenomenex Synergy
MAX-RP C12 80 A with TMS end-capping (4 μm; 150;
4.6 mm ID) and Phenomenex Synergi Polar RP (ether
linkedphenyl phase with polar end-capping) were tested
for the chromatographic separation of the above-men-
tioned substances. The Multosphere column was purchased
from CS, Langer-wehe, Germany and Phenomenex, As-
chaffenburg, Germany supplied the Phenomenex col-
umns. Peak identify was determined by means of reten-
tion time and UV spectra that were recorded for all sam-
ples 250 nm. During method development, three solvent
gradients were tested: program I: 0 - 6 min (12% B), 7
min (18% B), 14 min (25% B), 19 min (40% B), 24 min
(50% B), 29 min (12% B) (solvent A = 2% acetic acid in
aqueous solution (v/v) and solvent B = acetonitrile);
program II: 0 min (5% B, 5% C); 4,5 min (6,5% B, 5%
C), 7 min (18% B), 14 min (25% B),19 min (40% B), 24
W. M. Haggag et al. / Agricultural Science 2 (2011) 291-296
Copyright © 2011 SciRes. Openly accessible at http://www.scirp. o rg/journal/AS/
min (50% B), 30 min (5% B, 5% C) [solvent A = 2%
acetic acid in aqueous solution (v/v), solvent B = ace-
tonitrileand solvent C = tetrahydrofurane (THF)]; pro-
gram III: identical to Program I except that solvent A
was purified water buffered to pH 4 with citrate buffer.
This program, in combination with the Synergy
MAX-RP C12 column, was used exclusively for electro-
chemical detection. In all cases a flow rate of 1 ml/min
was used. The injection volume was 20 μl for each
analysis and separations were carried out at room tem-
perature. Linear calibration lines for mangiferin were
compiled using standard series of six dilutions between
4.7 and 98.3 µg/ml. The repeatability of HPLC method
was determined by ten injections of the same sample of
extracted. The reproductively of the complete assay was
tested by means of ten sample preparations [8].
Changes in mangiferin (1,3,6,7-tetrahydroxy xanthone-
C2-b-D-glucoside) C10H10O11 of healthy and malformed
shoots of mango cultivar Sedekia (three years old) was
followed after inoculated with mango malformation
pathogens i.e. F. subglutinans, F. sterilihyphosum, F.
oxysporum and F. proliferatum. Data pertaining to artifi-
cial inoculations revealed that, Fusarium subglutinans
proved to be the dominant fungus with 100% sample’s
infection in inoculated soil (Table 1). Fungi F. ox-
ysporum, F. sterilihyphosum and F. proliferatum showed
moderate infection in induced typical malformation
symptoms in inoculated mango seedling and were re-
isolated. Changes in mangiferin activity in mango seed-
lings cv. Sedekia as response to infection with mango
malformation pathogens i.e. F. subglutinans, F. sterili-
hyphosum, F. oxysporum and F. proliferatum grown
under greenhouse was determined. Mangiferin activity
varied widely amongst different inoculated pathogens.
From the data given in Ta ble 1 and Figure 1, it is clear
that accumulation of mangiferin was maximum (0.96
µg/g–1 FW) in leaves of healthy shoots. Mangiferin re-
duced in mango shoot in response to infection with F.
sterilihyphosum by 0.52 µg/g–1 FW. The amount of
mangiferin is 0.15 µg/g–1 FW due to infection with F.
subglutinis. Mangiferin is detected as 0.12 µg/g–1 FW as
result of infection with F. proliferatum. It is clear that
the least amount of mangiferin is detected as result of
infection with F. oxysporum (0.0039 µg/g–1 FW). Strong
positive correlations between mangiferin activity and
malformation incidence were observed. Mangiferin pro-
moted vegetative growth and exhibited inhibitory role on
the occurrence of malformation. It was also observed
that the healthy had highest activity of mangiferin as
compared to infected ones. Results of the present study
Table 1. Status of mangiferin in mango shoot explants as
influenced by different inoculation with pathogenic
Mangiferin (µg/g–1 FW)Malformation (%)
0.12 100
F. subglutinans
0. 52 50
F. sterilihyphosum
0.0039 50
F. oxysporum
0.15 50
F. proliferatum
clearly indicate that level of mangiferin could be consid
ered as a potential biochemical indicator for the malfor-
mation disease of mango. Interaction among Fusarium
moniliforme and colonizing malformed mango panicles,
and mangiferin (1,3,6,7-tetrahydroxyx-anthone C2-ß-D
glucoside), a defensive metabolite of the host plant in
relation to floral malformation, was in- vestigated. The
fungal populations were initially positively related to the
mangiferin content and the disease incidence. Further
increase in mangiferin content reduced the fungal and
mite populations; however, the increase in infection rate
was not affected until the Fusarium population was too
low. The fungal conidia remained adhered to the body
surface of the mites inhabiting malformed panicles, and
its plating on potato dextrose agar showed a trail of fun-
gal colonies along the pathway of its movement.
Tyrolicus casei facilitated the ingress of the fungus into
the host cells while F. moniliforme served as the feed of
T.casei and increased its multiplication [6,9]. Mangiferin
acts as a phytoalexinlike compound in M. indica. Activ-
ity of the mangiferin degrading enzymes, poly-
phenoloxidase of non-pathogenic strains of F. monili-
forme was higher than that of the pathogenic strain. In
vitro tests also confirmed rapid and massive degradation
of mangiferin by non-pathogenic strains. The disease
incidence could be minimized by removing stress factor
(the pathogen) through removal of malformed plant parts
and supplying the malformed plants necessary micronu-
trients by spraying with mangiferin metal chelates
[10,11]. The biochemical significance of the changes in
these constituents, resulting from the hypersensitive re-
sponses in the host specie, is appraised in respect to the
Fusarium pathogens, the causal organisms of mango
alformation. m
W. M. Haggag et al. / Agricultural Science 2 (2011) 291-296
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F. subglutinans
Openly accessible at
W. M. Haggag et al. / Agricultural Science 2 (2011) 291-296
Copyright © 2011 SciRes. Openly accessible at http://www.scirp. o rg/journal/AS/
F. oxysporum
F. sterilihyphosum
W. M. Haggag et al. / Agricultural Science 2 (2011) 291-296
Copyright © 2011 SciRes. http://www.scirp.org/journal/AS/
F. proliferatum
Figure 1. Changes in Mangiferin (1,3,6,7-tetrahydroxy xanthone-C2-b-D-glucoside) C10H10O11 of healthy and malformed shoots of
mango cultivar Seddek when inoculated with Fusarium pathogens.
This manuscript funded from the project “New applied approaches
to promote productivity and Quality of some fruit crops (Mango)” PI:
Prof. Wafaa M. Haggag. National Research Centre, 2007 to 2010.
Openly accessible at
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