New Zealand Journal of Crop and Horticultural Science

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Delayed self-incompatibility causes morphological

alterations and crop reduction in ‘Ataúlfo’ mango
(Mangifera indica L.)

M Gehrke-Vélez, A Castillo-Vera, C Ruiz-Bello, JL Moreno-Martinez & G

Moreno-Basurto

To cite this article: M Gehrke-Vélez, A Castillo-Vera, C Ruiz-Bello, JL Moreno-Martinez & G

Moreno-Basurto (2012) Delayed self-incompatibility causes morphological alterations and
crop reduction in ‘Ataúlfo’ mango (Mangifera indica L.), New Zealand Journal of Crop and
Horticultural Science, 40:4, 215-227, DOI: 10.1080/01140671.2011.632423

To link to this article: https://doi.org/10.1080/01140671.2011.632423

Published online: 19 Mar 2012.

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New Zealand Journal of Crop and Horticultural Science
Vol. 40, No. 4, December 2012, 215
227

Delayed self-incompatibility causes morphological alterations and crop reduction

in ‘Ataúlfo’ mango (Mangifera indica L.)
M Gehrke-Véleza*, A Castillo-Verab, C Ruiz-Belloa, JL Moreno-Martineza and
G Moreno-Basurtoa
a
Facultad de Ciencias Agrı´colas, Universidad Autónoma de Chiapas, Huehuetan, Chiapas, Me´xico; bEl Colegio
de la Frontera Sur, Tapachula, Chiapas, Me´xico
(Received 11 December 2010; final version received 25 August 2011)

Poor fruit set, premature fruit drop and excessive nubbin production are causing crop reduction
in ‘Ataúlfo’ mango orchards in southeastern Mexico. Fruit set was measured in self- and cross-
pollinated Ataúlfo mangoes in the field using houseflies (Musca domestica L.) as pollinators and
pollen from donor varieties. Pollen tube kinesis was monitored from pollen germination to
arrival at the ovule sac following manual self- and cross-pollination. Cross-pollinated
inflorescences produced more fruit set than selfed ones. ‘Joe Welch’ and ‘Criollo’ varieties were
the most efficient pollen donors tested. Seedling germination in Ataúlfo mangoes averaged 3%
during 2 consecutive years. Pollen tube development in vivo revealed positive contact with the
micropyle and normal zygote formation in cross-pollinated flowers. Selfed pollen tubes entered
the embryo sac but produced atrophied embryos and aborted or malformed fruit. Seedlessness
was not observed although selfed fruitlets contained malformed or necrotic embryos, suggesting
delayed self-incompatibility.
Keywords: nubbin; pollination; self-incompatibility; fruit set; embryonic atrophy; inbreeding
depression

Introduction 2000; Hanemann et al. 2008). From the mid-

In the Soconusco region of Mexico’s southern-
most state of Chiapas, ‘Ataúlfo’ mango has
become a profitable export product which
could be a solution to the land productivity
problem and ecological devastation that devel-
oped from the cotton and banana booms of the
past century (Richter 1993; Magallanes-Cedeño
2004). Previously abandoned cotton and
banana plantations and marginal soybean and
corn growing systems have been converted to
‘Ataúlfo’ orchards under a poorly planned,
non-systemised pattern of monocultivated
mango orchards, which have nonetheless pro-
vided sizeable prosperity to an otherwise deva-
stated land system in near bankruptcy (Richter
1960s to the turn of the century, mango
plantations grew from negligible to more than
15,000 hectares (IMPI 2003). In 2010, the
area planted with ‘Ataúlfo’ was estimated at
23,000 hectares in this region alone (Asociación
Agrı́cola Local de Fruticultores del Soconusco,
unpublished report, 2010). Nevertheless, ser-
ious physiological and morphological disorders
have been appearing during the reproductive
process and have resulted in unprecedented low
yields throughout mango plantations in the
region (Pérez-Quintanilla et al. 1998; Gehrke-
Vélez 2008). Foremost of these disorders are
lack of fruit set (despite abundant flowering),
fruitlet abortion and excessive nubbin or

*Corresponding author. Email: malc@unach.mx, malcrgehrke@hotmail.com

ISSN 0114-0671 print/ISSN 1175-8783 online
# 2012 The Royal Society of New Zealand
http://dx.doi.org/10.1080/01140671.2011.632423
http://www.tandfonline.com
216 M Gehrke-Ve´lez et al.
‘mango niño’ production. Although these phe-
nomena are not uncommon in a number of
mango varieties (Singh et al. 1962), in most
cases their incidence has not been reported as
drastic enough to cause serious crop reduction.
However, a high level of coincidence detected in
‘Ataúlfo’ mango between the occurrence of
these disorders and excessive crop reduction
leads to the suggestion that there is a cause-
and-effect relationship between these factors
(Gehrke-Vélez 2008). Causes of low fruit set
and fruitlet abortion have not been clearly
described although various investigators point
to exogenic causes or abiotic factors such as
excessively high or low temperatures during
anthesis or flower differentiation and at
fruit set (Shu et al. 1989; Gazit et al. 1992;
Issarakraisila et al. 1992).
Few investigators have investigated the
nubbin mango fruit and there is a general lack
of information regarding its morphological
characteristics and physiological causes.
Although Litz (1997) states that there is little
indication of self-incompatibility in most mango
cultivars, he cites diverse authors who have
reported complete or partial self-incompatibility
in some cultivars and supplies as evidence of this
phenomenon a degeneration of embryonic and
nucellar tissues and excessive loss of fruitlets.
Litz (1997) and Singh et al. (1962) describe
nubbins as stenospermocarpic fruits (Soule
1985)*products of embryonic atrophy*and
relate their incidence to exogenic factors or to
physiological restrictions of the fertilisation
process due to genetic incompatibility factors.
Pollination is essential in mango in order to
maintain genetic flow, especially in mono-
embryonic cultivars (Pimentel et al. 1984;
Robbertse et al. 1988). This occurs when pollen
is transported to the floral stigma, principally by
insect activity (Popenoe 1917; Peña 2003).
However, various authors (Bijhouwer 1937;
Mallik 1957, cited by Litz 1997) sustain
that pollination can occur by air movement or
gravity. Pollen transfer, with the participation of
external vectors, is known as cross-pollination
and can occur within or between flowers of the
same plant (self-pollination) and between flow-
ers of different plants (cross-pollination) (Peña
2003).
Mango inflorescences consist of a mixture
of small hermaphroditic (with a functional
ovary) and masculine (without a functional
ovary) flowers in variable proportions (Zee
1995). Participation of an external agent is
necessary for sexual reproduction in mangoes
because hermaphroditic flowers open during
the night and pollen grains are liberated in the
early morning, the stigma remaining receptive
for 3
72 hours after anthesis (Popenoe 1917;
Pimentel et al. 1984; Robbertse et al. 1988;
Diaz 2002), with optimum receptivity occurring
3 hours after anthesis. Furthermore, mango
pollen is most viable soon after dehiscence and
degrades rapidly within a few hours (Mallik
1957; Singh 1963). In mango, pollen is trans-
ferred mainly by diverse species of Hymenop-
tera (wasps, ants and bees) and Diptera
(houseflies and others) (Anderson et al. 1982).
The present investigation addresses the
compatibility issue in ‘Ataúlfo’ mango variety
under the hypothetical assumption that
this variety may be reproductively self-
incompatible, due to genetic (Sedgley & Griffin
1989) or environmental (Issarakraisila et al.
1992) factors. In order to evaluate the repro-
ductive behaviour of this variety, the pollina-
tion/fertilisation process and initial fruitlet
formation were observed, and fruit set was
measured after self- and cross-pollination with
other local varieties during three consecutive
floral flushes. Seed viability of ‘Ataúlfo’ man-
goes was also determined.
Materials and methods
Controlled pollination in the field
One location, the ‘La Norteña’ Federal Gov-
ernment Agricultural Experimental Station,
was chosen for all the experimental field treat-
ments due to its location in the centre of the
Soconusco mango growing area. ‘La Norteña’
experimental site is located 16 km due south of
Tapachula, Chiapas, at an altitude of 16 m
 Delayed self-incompatibility causes morphological alterations 217
above sea level. Coordinates: 14845?35??N
latitude, 92823?12??W longitude (INEGI 2009).
Mean annual temperature is 34.6 8C with a
mean night temperature of 24.9 8C and a mean
day temperature of 31.6 8C. Annual rainfall is
1354 mm and there is an average of 109 rainy
days per year. This facility contains an arbor-
etum with more than 30 different mango
varieties from which flowering panicles were
obtained for cross-pollination studies. Varieties
of mango that were flowering simultaneously
with the ‘Ataúlfo’ trees during each floral flush
were used as pollen donors.
Atmospheric temperature data generated at
a weather station close to the experimental area
were obtained from Instituto Nacional de
Investigaciones Forestales, Agricolas y Pecuar-
ias (INIFAP; National Institute of Agricultur-
al, Forestry and Animal Research, Mexico)
offices, although relative humidity data were
unavailable.
Pollination was achieved by placing one
branch of a cut panicle of each cultivar of a
different donor variety inside each cage con-
taining a previously selected ‘Ataúlfo’ inflores-
cence. Since different varieties flowered at each
‘Ataúlfo’ flush, pollen donors varied during
each specific flush of the receptor variety
(Table 1). Houseflies (Musca domestica L.
[Muscidae]) were used as pollinators in this
experiment because this species is known to be
a frequent visitor of mango inflorescences and
has been reported to be a preferential pollinator
of mangoes (Anderson et al. 1982; Litz 1997;
Dag & Gazit 2000). Furthermore, this species
was chosen due to its easy rearing and handling
Table 1 Pollen donors used for pollination of
‘Ataúlfo’ mango during different floral flushes.
Donor Local mango varieties
First flush Second flush Third flush
I ‘Platano’ Brooks Irwin M
II ‘Criollo’ Tommy Atkins Springfield
III ‘Joe Welch’ Irwin M None1
1
No other coincident flowering variety was available.
in the field for controlled pollination. House-
flies were collected from local sanitary landfills
and reproduced and reared in the laboratory
using a nutrient formula for larval development
containing wheat bran, yeast and powdered
milk (Castillo et al. 2007). Pupae were placed in
vermiculite and, upon emergence, adults were
placed in vials (100 adult flies per vial) for
transport to the field and liberation inside mesh
cages previously installed on selected inflores-
cences for controlled pollination.
The experimental design was completely
randomised with five treatments and 10 repeti-
tions. One row of 10, 18-year-old ‘Ataúlfo’ trees
was chosen for the experiment, each tree being
used as an experimental unit or repetition (R1
to R10). Selected trees were subjected to
normal cultural practices which consisted of
mechanical weed control, one irrigation and
fertilisation with a 17-17-17 NPK formula.
Experimental trees did not receive pruning or
floral acceleration with chemical inductors
(KNO3, paclobutrazol, etc.).
In each tree (or repetition), five ripe panicles
at the generative stage (Davenport & Nuñez-
Elisea 1997) were randomly chosen and covered
individually with fine mesh plastic mosquito
netting forming cages which served as treat-
ments as follows:
“ T1 Unpollinated control: panicle caged but
without flies or any donor inflorescence
“ T2 Self-pollinated control: cage containing
flies but no donor inflorescence
“ T3 Cross-compatibility I: cage with flies
and an inflorescence from donor number I
“ T4 Cross-compatibility II: cage with flies
and an inflorescence from donor number II
“ T5 Cross-compatibility III: cage with flies
and an inflorescence from donor number III.
In treatments T2 to T5, 100 mature
laboratory-reared houseflies were placed as
pollinators in each cage and in treatments T3,
T4, and T5, fresh panicles obtained from donor
varieties were placed in vials containing water
to avoid wilting and additionally introduced
218 M Gehrke-Ve´lez et al.
into each cage. These panicles were replaced
every third day in order to ensure their fresh-
ness and viability.
All the caged inflorescences were uncovered
25 days after initiating treatments and the
fruitlets that remained adhered were counted
as ‘set’ fruit. Three random samples were taken
from each of three trees in each of four
experimental sites. Set fruitlets were dissected
longitudinally along the stigma-style axis. Ma-
terial was then examined microscopically with a
VELAB Mod.VE-B3 microscope with 10
and 40 objectives in order to observe embryo
development of selfed and cross-pollinated
specimens at the early stages of fruit formation.
The experiment was repeated for each of
three flowering flushes (F1 to F3) which
occurred from 20 November to 10 December
2009 for the first flush, 25 December 2009 to
15 January 2010 for the second flush, and
20 February to 5 March 2010 for the third
flush. Data obtained were analysed with an
ANOVA of the completely randomised design
model using the Duncan (LSD) method.
Manual pollination under laboratory conditions
In order to complement findings at the field
level, laboratory experiments were conducted
to establish pollen tube behaviour within the
floral gynaecium.
Pollen tube kinesis
Ten panicles per experimental tree of the
‘Ataúlfo’ variety were randomly selected during
each of three floral flushes observed. From
these, 36 hermaphroditic flowers were picked,
emasculated and placed in vials containing a
Brewbaker-Kwack (1963) liquid nutrient solu-
tion for conservation. Pollen was then extracted
from the pollen sacs of ‘Ataúlfo’ and eight
other selected varieties (Table 1) and hand
pollinated on to the previously emasculated
flowers. Once pollinated, flowers were dissected
longitudinally along the stamen-pistil axis and
then fixed in a FAA solution (Kearns & Inouye
1993) at intervals of 8, 16, 24, 32, 40, 48 and
56 hours (four flowers for each interval).
Dissected material was then stained in a
0.1% aniline blue solution and placed on glass
slides for observation with a VELAB Mod.VE-
B3 contrast microscope with 10 and 40
objectives.
Pollen efficiency
Using plant materials collected at the field
experiment site described previously, 70 her-
maphroditic ‘Ataúlfo’ flowers were manually
self- and cross-pollinated with other common
mango varieties and the resulting pollination/
fertilisation process was microscopically ob-
served by tracing pollen tube displacement
from pollen adherence to the stigma to pene-
tration of the pollen tube into the ovule sac.
The experiment was conducted with material
obtained at the same site as the field experi-
ment. Pollen was obtained from the same
varieties used in the controlled field pollination
experiments (Table 1).
Pollen efficiency was determined as the
presence or absence of pollen tubes at the
micropyle entrance. Statistical design was com-
pletely randomised with seven treatments and
10 repetitions. Statistical analyses were con-
ducted using the Statgraphics 5.0 plus version
program. The ANOVA and LSD tests were run
in order to determine levels of significance.
Experimental seedling germination
In order to further establish varietal fecundity
under monocultivated field conditions,
‘Ataúlfo’ seeds were planted in a nursery and
observed for seedling production. The experi-
ment was conducted during two consecutive
crop cycles (May 2009 and May 2010). A
hundred ripe mango fruits were randomly
chosen from seven commercial orchards over a
146 km2 area of the Soconusco region. Fruits
were hand-peeled and depulped and the remain-
ing endocarps planted in plastic bags filled with
 Delayed self-incompatibility causes morphological alterations 219
Figure 1 Average hourly temperatures by calendar
month, measured at the ‘La Norteña’ Federal
Government Agricultural Experimental Station dur-
ing the 2009
10 mango flowering season.
soil/sand/potting compost, watered and allowed
to germinate.
Results
Controlled pollination in the field
Floral flushes
Three floral flushes were recorded during the
experiment. The first and most abundant flush
occurred during the second week of November
Figure 2 Fruit set in ‘Ataúlfo’ mango 25 days after pollination with different pollen donors in three floral
flushes. Means 91 SE.
2009, with an estimated average of 2000 to
2600 panicles per tree (PPT), followed by a
second flush during the first week of January
2010 which produced an average of 1000 to
1500 PPT. A third and relatively weak flush
(51000 PPT) occurred during the first week of
February 2010.
Figure 1 shows average hourly temperatures
recorded at the experimental site during the
2009
10 flowering season. No data were re-
corded regarding specific timing of floral an-
thesis in this experiment.
Fruit set
As shown in Figure 2, highest fruit set occurred
during the first floral flush in all treatments
except the unpollinated control. Cross-
pollinated inflorescences (T3, T4 and T5) pro-
duced higher levels of fruit set than self-
pollinated ones (T1 and T2) during the first
and second floral flushes. During the third
flush, fruit set was generally low showing no
significant differences (LSD P 50.05) between
treatments T3 and T4 and the control. In this
case only two donor varieties were tested
because only these produced flowers at the
time the ‘Ataúlfo’ variety was in bloom.
220 M Gehrke-Ve´lez et al.
Pollination under laboratory conditions
Pollen tube kinesis
Microscopic (100 ) observation of floral cut-
tings revealed that two to three pollen grains
normally adhered to the floral stigma after
manual pollination and at least one pollen tube
(PT) was generated in 85% of all pollen/ovule
varietal combinations. Pollen tube development
was traced along the style from the stigma to
the base of the style after which adequate
observation became impossible with the meth-
odology used due to the fact that the develop-
ing pollen tube travels within the ovary wall in
differing three-dimensional patterns, reaching
the ovule sac in an average of 48
56 hours after
pollen adherence (Fig. 3).
Pollen tube development
Actual fertilisation of the ovule was not ob-
served at the levels of microscopy used, but
posterior monitoring revealed embryo
Figure 3 Diagrammatic representation of a typical ‘Ataúlfo’ mango gynoecium 46
56 hours after cross-
pollination.
formation in 25% of all the material ob-
served. Pollen tubes developed normally at
similar rates in all cases within the stigma-style
tissue to the stigma base where it becomes part of
the ovary wall. Although PT movement within
the ovary could not be traced with the metho-
dology used due to the spherical form of the
ovary, flower dissection at 48
56 hours after
pollination revealed PT insertion into the micro-
pyle through the funiculum in ]60% of cross-
pollinated cases and 540% of self-pollinated
ones.
Varietal differences in pollen tube growth
Pollen tube growth from the stigma to the style
base occurred after an average of 36
40 hours.
Figure 4 shows the variation in growth rate
observed between pollen-donating varieties. No
significant differences (P ] 0.05) were observed
in growth rates between donor varieties during
the first and second floral flushes. ‘Irwin’
 Delayed self-incompatibility causes morphological alterations 221
showed the lowest growth rate during the third
floral flush.
Pollen efficiency
Statistical analysis revealed that there is sig-
nificant difference (P 50.05) in pollen effi-
ciency between certain donor varieties. LSD
test results showed significant differences fa-
vouring ‘Joe Welch’ and ‘Criollo’ over other
varieties tested (Table 2). Irwin showed the
lowest growth rate during the third floral flush.
Embryo formation
Embryo formation was similar in selfed and
cross-pollinated specimens (Fig. 5). At this
stage, however, embryos in the self-crossed
(Ataúlfo Ataúlfo) treatments showed an ab-
normal production of a colourless gum-like
substance and remained attached to the pla-
cental wall on the dorsal side of the embryo sac.
Initial fruitlet development
Fruitlet dissection at varying initial stages of
development revealed that in no case was
complete seedlessness observed. Presence of
rudimentary embryos within the developing
ovary sac was noted at fruit set in both self-
and cross-pollinated fruitlets, although defor-
mity was observed in all self-pollinated fruits
(Fig. 6A). Embryos in cross-pollinated ovaries
Table 2 Multiple range rests for pollen efficiency* of different mango donors on ‘Ataúlfo’ gynaecia (95%
LSD method).
Treatments Count
Control (ATA & ATA) 10
ATA & Irwin Mor 10
ATA & T. Atkins 10
ATA & Springfield 10
ATA & Platano 10
ATA & Criollo 10
ATA & Joe Welch 10
*Pollen efficiency is defined as the proportion of flowers for which pollen tubes were observed entering the ovary.
were free of the placenta and showed normal
development and growth with fruitlets elon-
gated normally (Fig. 6B). Selfed embryos were
shrivelled and necrotic or malformed and
dorsally adhered to the placenta (Fig. 7).
Experimental seedling germination
In 2009, seedling germination tests resulted in
emergence of a total of five seedlings and only
one in 2010. Hence ‘Ataúlfo’ mango seeds
showed only an average of 3% germination in
the two years.
Discussion
Under the conditions prevailing in the experi-
ment, the first floral flush proved to be the most
efficient for fruit set. It was also the most
intense and vigorous flush (]2000 inflores-
cences/tree) of the three flushes that were
studied, followed by the second (1300
1500
inflorescences/tree) and third (B1000 panicles/
tree) flushes which produced decreasing
amounts of flowers.
Climatic conditions occurring during floral
differentiation and development are important
factors affecting flush intensity and consequent
fruit set in ‘Ataúlfo’. Hourly temperatures
recorded at the experiment site (Fig. 1), show
differences in minimum and maximum daily
readings from November 2009 to April 2010.
Lower overall temperatures during November
Mean Homogeneous groups
0.3 A
0.3 A
0.4 AB
0.5 ABC
0.6 ABC
0.8 BC
0.9 C
222 M Gehrke-Ve´lez et al.

Figure 4 Pollen tube growth in ‘Ataúlfo’ mango pistils 36 hours after pollination using different pollen
donors during three consecutive floral flushes. Means 91 SE.

coincided with higher floral flush intensities, a
situation similar to that found by Ramirez &
Davenport (2010) who concluded that under
subtropical conditions, and, to a lesser degree,
in the low latitude tropics, cool temperatures
are important for floral induction in mangoes.
Further analysis of the data in Figure 1 shows a
gradual cooling trend from a high peak at
2:00pm to a daily low at 8:00am at which time
intense heating occurs during the following
6 hours. Such a pattern of slow cooling during
the night apparently coincides with floral
anthesis as observed by Pimentel et al. (1984)
in polyembryonic cultivars. Similarly, intense
heating with an expected lowering of relative
humidity enhances anther dehiscence creating
an apt scenario for natural pollination at mid-
day (Randhawa & Damodaran 1961, cited by
Davenport & Nuñez-Elisea 1997). However,
further research is needed in order to determine
which specific conditions are most important
and in what order and magnitude. Work by
Issarakraisila et al. (1992) on ‘Kensington
Pride’ mangoes indicates that fruit set and its
timing varies from year to year, apparently
depending on climatic conditions and physio-
logical constraints.
Inter-varietal differences in fruit retention
showed ‘Joe Welch’ and ‘Criollo’ to be the most
effective pollen donors of those tested (Fig. 2).
Although all donor varieties resulted in a high
number of aborted fruits in relation to fruitlet

Figure 5 A, Dissected self-pollinated ovary of ‘Ataúlfo’ mango 60 hours after pollination. B, Dissected cross-
pollinated ovary of ‘Ataúlfo’ mango 60 hours after pollination. Scale bars 100 micrometres *(mm).
 Delayed self-incompatibility causes morphological alterations 223

Figure 6 Dissected ‘Ataúlfo’ mango fruitlets 36 days after pollination. A, Self-pollinated, with embryo fused
to placental wall. B, Cross-pollinated, with embryo free of placental wall. Scale bar 10 mm.

production, these two produced the highest
number of net fruits per inflorescence at harvest
time (0.6 and 0.4, respectively). These results
Figure 7 Self-pollinated ‘Ataúlfo’ fruitlet 45 days
after anthesis. Note atrophied necrotic embryo and
empty placental sac (arrow). Scale bar 10 mm.
are similar to those reported by Issarakraisila &
Considine (1994) who observed an average fruit
set per flowering shoot of 0.45 and 0.37 in two
consecutive years in ‘Kensington Pride’. The
situation is common in most mango varieties
since a high rate of premature abortion and
‘set’ fruit drop occurs in this species as a result
of an extremely high flower:fruit ratio of up to
3000:1 in most mango varieties (Mukherjee
1997).
Our results shown in Figure 2 indicate that
in ‘Ataúlfo’ mango, out-crossing is favoured
over self-crossing, a situation reported by
Degani et al. (1997) who found ‘substantial’
out-crossing rates and an unexpectedly high
hybrid fruit production in ‘Tommy Atkins’ and
‘Maya’, suggesting the existence of self-incom-
patibility and a genetic selection favouring out-
crossed fruits. Dag et al. (1997) obtained
similar results using ‘Tommy Atkins’ pollen as
an out-crossing agent for pollinating ‘Lily’
224 M Gehrke-Ve´lez et al.
mangoes. These authors stated that the over-
whelming presence of hybrid fruits when trees
were caged with a pollinator was indicative of a
strong advantage for cross- vs self-pollination.
This would point to the presence of in-
breeding depression in ‘Ataúlfo’ and certain
other mango varieties, a characteristic de-
scribed by Charlesworth & Willis (2009) as a
fitness trait existent in natural populations,
directed toward increased heterosis favouring
species survival. It is defined as the decline in
vigour in the offspring of genetically closely
related organisms (Allaby 2006) and is often
considered the result of a ‘population bottle-
neck’ caused by the inbreeding of related
individuals (Mahy & Jacquemart 1999). Never-
theless, further and more detailed research is
required in this particular aspect of genetic
physiology before considering its causal role
within the problem investigated in ‘Ataúlfo’.
Our experimental results (Fig. 2) indicate
that adequate fruit set is dependent on
cross-pollination using other mango varieties
as suitable pollen donors. There is initial
compatibility and fertilisation occurs in the
self-breeding process in this variety, as is
evident from our observations and data. Self-
pollinated stigmas result in embryo formation,
but a delayed self-incompatibility becomes
evident at the final stage of the pollination
process leading to alterations in the fertilisation
process which evidently result in embryo atro-
phy. This results in immature fruitlet abortion
and, in advanced cases, to paranormal devel-
opment of disfigured fruit known as nubbins or
‘mango niño’.
True parthenocarpy was not evidenced
because seedlessness was not found in any of
the material sampled. This is substantiated by
findings by Chandler (1958), Singh (1961),
Singh (1964) and Ram et al. (1976), all cited
by Davenport & Nuñez-Elisea (1997), who
observed that seedlessness in mangoes is caused
by embryo abortion and not by a fertilisation
failure. It is attributed to stenospermocarpy as
defined by Soule (1985) rather than partheno-
carpy as described by Perez-Barraza et al.
(2009). This is consistent with findings reported
by Nuñez-Elisea & Davenport (1983) and by
Whiley et al. (1988) who found that stenosper-
mocarpic fruitlets are slower growing than
seeded fruit and result in misshapen fruit which
does not reach full size.
Observations at fruit set in this experi-
ment revealed normal embryo growth in
cross-pollinated samples but consistent ab-
normalities in embryo growth and development
of selfed material, such as stunted and shri-
velled or necrotic embryos (Fig. 6), that in-
dicate incomplete or abnormal zygote
formation probably due to an incomplete or
restricted ovule fertilisation. These observa-
tions are similar to those of Sharma & Singh
(1972) and Ram (1983) who associated these
phenomena with embryo abortion occurring in
diverse mango varieties in India.
Adequate cross-pollination is closely related
to varietal differences (Fig. 3). Clearly, for a
pollen donor to be successful as such, flowering
must be coincidental with the target receptor,
since pollen availability must occur for cross-
pollination to be possible. However, there is
clear evidence that certain specific varieties are
better donors than others, possibly due to
specific morphological and physiological char-
acteristics of each donor variety at the pollen/
ovule level and their affinity to ‘Ataúlfo’ traits,
although further investigation is necessary to
determine the conditions and extent to which
this occurs.
It is evident from our data shown in
Figure 3 that ‘Joe Welch’ is a superior pollen
donor for ‘Ataúlfo’ as far as fruit set is
concerned, followed by ‘Criollo’ and ‘Plátano’
in that order. Microscope observation of pollen
tube kinesis revealed that at the pollination
stage, ‘Ataúlfo’ gynoecia are receptive to pollen
from all varieties tested, including selfed mate-
rial. Self-pollinated pollen tubes penetrate the
ovule sac at the micropyle as do cross-polli-
nated PTs resulting in fertilisation evidently in
both cases. However, upon formation of the
zygote, the incipient embryo becomes adhered
to the placenta at a point similar to that
 Delayed self-incompatibility causes morphological alterations 225
described by Joel & Eisenstein (1980) as the
‘ponticulus’ on the dorsal side of the ovule. It
ceases to grow and becomes atrophied and, in
many cases, shrivelled and necrotic (Fig. 6), a
situation described by Ram et al. (1976).
However, Barrera & Gehrke-Vélez (2010)
reported that self-fertilisation initially induces
ovary enhancement and apparent fruit forma-
tion, but later results in the formation of
atrophied fruits or nubbins that are either
aborted by the mango plant at early stages of
development, or remain adhered to the panicle
until full ripeness is reached. This seems to be
the result of low self-compatibility at the post-
zygotic level combined with climatic conditions
adverse to the reproductive process (Gehrke-
Vélez 2008; Dag et al. 2009). However, addi-
tional research is required to substantiate this
hypothesis.
The remarkably poor germination of
‘Ataúlfo’ seeds from monocultivated orchards
during two consecutive seasons provides
further evidence that this variety is self-infertile
(Gehrke-Vélez 2008) since virtually no seedlings
were produced.
Conclusions
Floral flushes are erratic in ‘Ataúlfo’ and in
other varieties in the Soconusco region,
although three flushes generally occur during
the months of November to March. The timing
and intensity of these flushes seems to be
governed by numerous and diverse environ-
mental and intrinsic factors as has been sug-
gested by various investigators (Gazit et al.
1992; Davenport & Nuñez-Elisea 1997). How-
ever, when and how these flushes occur affects
the reproductive development of ‘Ataúlfo’
which appears to depend largely on floral
coincidence with other mango varieties in
addition to inter- and intra-varietal compat-
ibility. Self-incompatibility has been demon-
strated in ‘Ataúlfo’, despite apparent successful
pollination and pollen tube growth within the
gynaecium.
Incompatibility involves inadequate embryo
formation caused by abnormal adherence of
the embryo to the embryo sac, resulting in fruit
malformation (nubbins) and premature fruitlet
abortion. The mechanics causing abnormal
fruit set and nubbin production are likely to
be due to inbreeding depression or natural
defence mechanisms created to restrict endoga-
my (Mahy & Jacquemart 1999) similar those
reported in ‘Dashehari’ mangoes in India
(Singh et al. 1962) involving post-zygotic in-
compatibility, as described by Dag et al. (2009).
‘Ataúlfo’ has proved to be self-incompatible
as is evident from the very low frequency of
germination of seeds and the negligible fruit set
of self-pollinated flowers that we observed. It
appears that poor fruit set, premature fruit
abortion and embryo atrophy resulting in
nubbin production, is due to endogamic de-
pression resulting from the increased monocul-
tivation of this variety that is taking place in the
Soconusco region. This may be a consequence
of the progressive elimination of local varieties
that may have previously functioned as pollen
donors.
Acknowledgements
We thank Francisco Infante (ECOSUR) for labora-
tory facilities and instrumentation,Francisco Aguirre
(Rosario Izapa Experimental Station, CERI,
INIFAP), for providing field facilities at the ‘La
Norteña’ experimental site, Trevor Williams, for
help in revising this article, Eusebio Ortega (Asocia-
ción Agricola Local de Fruticultores del Soconusco)
for help with logistics and grower collaboration, and
Jaime Toledo for help in preparing computerised
images. The project was funded by Fundación
Produce Chiapas AC.
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