Stimulation of Yam (Dioscorea) Tuber Growth
by Gamma Irradiation1
Franklin W. Martin2, Francis K. S. Koo and Jose Cuevas 3
Abstract. Treatment of aerial tubers and underground tuber pieces of Dioscorea alata L.t the 10-month
yam, with about 1250 rads of gamma irradiation stimulated tuber germination, vegetative growth, and
tuber yield. Higher dosages reduced viability and yields. When different parts of the tuber were irradiated, a
slight yield stimulation occurred when the developing shoot was treated, but treatment of parts of the tuber
reduced yield. Effects of irradiation disappeared entirely in the second vegetative generation after
irradiation.
For three quarters of a century the phenomenon of low dose
radiation stimulation of growth in higher plants has attracted
attention from a number of investigators. Many experiments
have provided affirmative evidence of radiation stimulation ( 2 ,
4 , 5, 6); but negative results have also been reported ( 6).
The most commonly observed manifestations of radiation
stimulation include accelerated germination, early growth,
increased overall vigor and size, early flowering and maturity,
and increased yield. Many studies have been performed with
cereals, grasses, legumes, vegetables, and ornamentals, but only a
few experiments have dealt with root and tuber crops. Sprague
and Lenz in 1929 (cf. 6) reported a positive stimulation effect
in irradiated seed potatoes, which produced larger tubers, but
no significant increase in tuber yield. Kuzin (3) mentioned that
yield of radish was increased by 20-30% after seed irradiation at
500 to l,000r levels, and Bilquez (cf. 1) also observed dry wt
increase in radish. On the other hand, negative results were
obtained by Fischnick et al. (cf. 6) in potatoes and equivocal
results by Bowen et al. in radish (measured in whole-plant dry
wt).
The development of new high-yielding cultivars of edible
yams (Dioscorea sp.) by conventional plant breeding methods
is difficult. The principal species do not flower regularly; they
have high and sometimes irregular chromosome numbers; the
flowers are often sterile. Propagation by asexual means,
including use of aerial tubers, has been responsible for the
widespread distribution of yams, and has maintained the genetic
material more or less intact.
Yams are important crop plants that have never been
thoroughly studied, and an opportunity exists for improving the
primitive cultivars for more adequate use throughout the
tropics. However, because of the limitations of sexual methods,
it would appear desirable to make use of new and
unconventional techniques to increase tuber yields. For this
study we have chosen gamma radiation as a stimulating agent to
test its feasibility.
Materials and Methods
Three cultivars of yams (D. alata L.) were chosen for this
study (Table 1). Aerial tubers or pieces of underground tubers
were used as seed pieces. Either the entire seed piece or a part of
it was treated with various levels of gamma radiation in the
60co gamma pool at the Puerto Rico Nuclear Center,
Mayaguez.
In the first experiment (1970), the stem ends of aerial tubers
of the cv. Feo were cut away, removing the principal sources of
preformed buds and some emerging shoots. The tubers were
treated with dosages of gamma rays from 0 to 3,500 rads, at
500 rads intervals. The dose rate was 1,000 rads per minute. The
1Received for publication November 5, 1973.
^Federal Experiment Station, USDA, ARS, Mayaguez, Puerto Rico.
^Puerto Rico Nuclear Center, operated by the University of Puerto Rico,
Mayaguez, under contract No. AT-(40-l)-1883 for the U.S. Atomic
Energy Commission.
282
Table 1. Cultivars o f D. alata used for tests of low dose gamma radiation
stimulation effects.
Recent geographical Production o f
Cultivar source aerial tubers
Feo Puerto Rico High
Trinidad Very low
Farm Lisbon
Cuello Largo India High
tubers harvested from the Ri (irradiated first generation) plants
were cut into seed pieces and grown for an R 2 generation the
following year. The experiments were repeated in 1971 and
1972 with the cv. Farm Lisbon, except that the stem ends of
the tubers were not cut away. Because the levels of gamma
radiation appropriate for stimulating foliage growth and tuber
yields were observed in 1970, only 2 intermediate doses, 1,250
and 2,500 rads were applied to ‘Farm Lisbon’ in 1971 and 1
dose (1,250 rads) in 1972. R i plants in 1971 were divided into
seed pieces and grown for an R 2 generation in 1972.
In 1972, the cv. Cuello Largo was chosen for the study of the
effects of irradiating different regions of the tuber. Since ‘Cuello
Largo’ had narrow and long aerial tubers, with appropriate lead
shielding it was possible to irradiate accurately specific regions
of the tuber. Matched groups of aerial tubers were selected, in
which the whole tuber, the upper half of the tuber containing
the dominant bud, the lower half of the tuber, and the First
shoot from the tuber were irradiated, then compared in
replicated blocks to the controls.
All the irradiated materials were planted without further
treatment at the Federal Experiment Station, Mayaguez, Puerto
Rico, and were grown over trellises to maturity in about 9
months, following established practices. Plants were not treated
for protection from diseases and insects. They were observed
regularly for vigor of growth, and for the possibility of gross
morphological changes. At harvest the tubers were dug, cleaned,
weighed, and observed for changes.
Results
Portions of aerial tubers of the cv. Feo, from which the stem
end containing preformed buds had been eliminated, differed in
rate of germination (sprouting) according to dosage of gamma
rays received (Table 2). Tubers irradiated with light doses
(500-1,500 rads) germinated more rapidly than tubers irradiated
with heavier doses or control tubers. The optimum radiation
dose for stimulating germination was 500-1,000 rads. The doses
at or higher than 2,000 rads retarded as well as reduced the
germination of the aerial tubers.
The foliage of all irradiated tubers developed more rapidly
and demonstrated more vigor than that of the nonirradiated
controls. For the first 7 months of the life cycle, vigor of foliage
was directly associated with gamma dosage. Some dieback and
death of plants occurred during heavy rains. Dieback appeared
to be most severe in nonirradiated plants. Later, during the final
2 months of growth, the lightly irradiated (500-1,000 rads)
tubers showed more vigorous foliage than controls or than
J. A m er. Soc. H ort. Sci. 9 9 (3 ) :2 8 2 - 2 8 4 . 1974.
Table 2. Percent germination (sprouting) of aerial tubers of the cv. Feo a second year, yield differences disappeared. In the second test
at different times after gamma irradiation.
in 1972, the radiation stimulation effect was not significant.
Gamma dosage Percent germination after irradiation When the entire aerial tuber of ‘Cuello Largo’ or either its
(rads) (weeks after treatment and planting) upper or lower part was irradiated, the yield of the subsequent
8
plant was significantly reduced (Table 5). When only the
9 10 11 12
emerging shoot was irradiated, subsequent yields were higher
0 29 46 61 77 100 than those of the control. The statistical probability of no
500 51 75 84 100 100
1,000 51
difference between the control yield and the higher yield from
75 84 100 100
1,500 35 51 67 84 100 treatment of the emerging shoot is 0.06.
2,000 25 42 67 75 84
2,500 17 35 42 67 75 Discussion
3,000 17 25 42 59 67
3,500
Gamma radiation significantly stimulated the growth of yams
8 25 25 51 67
from seed pieces. This stimulation was expressed as an increased
rate of germination, increased vigor of the foliage, and increased
size of the tuber. Since no consistent differences were seen in
heavily irradiated tubers. They stayed green longer and died the incidence of diseases, it is assumed that vigor and yield
back later. changes were not associated with control by irradiation of virus
At harvest, tubers from lightly irradiated plants weighed diseases. However, the stimulation was closely related to dosage.
more than those from controls or from heavily irradiated plants After the optimum dosage was reached, further increases in
(Table 3). These tubers were noticeably larger, but were not radiation decreased yields. The optimum dosage of radiation
mishapen in any way. There was very little rot in the tubers, may not be the same for the different types of stimulation. For
regardless of treatment. example, radiation doses that reduce yield may nevertheless,
Table 3. Tuber yields o f cv. Feo, as affected by gamma irradiation.

Dosage Of tubers Plants Total Per Statistical

(rads) treated surviving yield (kg) plant (kg) significance*
0 11 5 3.45 0 .6 9 z b
500 12 12 9.35 0.78 ab
1,000 12 9 9.00 1.00 a
1,500 12 9 8.76 0.97 a
2,000 12 10 7.84 0.78 ab
2,500 12 12 7.63 0.64 be
3,000 12 9 4.71 0.52 be
3,500 12 12 4.33 0.36 c

zMeans followed by the same letter are not significantly different (p = 0.05).

When the tubers from the first generation were cut into seed stimulate tuber germination or foliage growth (see Tables).
pieces and were grown a second year, no significant differences Cultivar differences in response to radiation stimulation were
were found in yields. Tubers from the heavily irradiated suggested but were not definitely demonstrated. The treatment
treatments were as large as those from controls and from lighter that increased tuber yield in 2 cultivars, ‘Feo’ and ‘Farm
irradiation treatments. Data for tuber weight in respect to Lisbon’, (1250 rads), decreased yield in ‘Cuello Largo.’ There is
radiation dosage are given below: also a suggestion of response differences in years and/or
No radiation, 2.56 kg. locations for a single cultivar. Similar results have been observed
1.000 rads- 2.12 kg. by many investigators, including Kuzin (3).
1.500 rads - 2.79 kg.
Table 5. Responses o f different parts o f the tuber o f ‘Cuello Largo’ to
2.000 rads-2.31 kg. irradiation with 1,250 rads of gamma rays.
2.500 rads - 2.20 kg.
3.000 rads - 2.40 kg. Mean weight
3.500 rads - 2.22 kg. o f tuber per
Part of tuber irradiated plant (kg)
In the experiments with ‘Farm Lisbon’, yields of tubers from
lightly irradiated (1,250 rads) seed pieces in the first test were Whole tuber 1.07 cz
much higher than those of tubers from untreated controls, or Lower part of tuber 1.18 be
from more heavily irradiated seed pieces (Table 4). When the Upper part of tuber 1.21 b
Emerging shoot only 1.50 a
tubers from the control and lightly irradiated series were grown Control (no irradiation) 1.38 a
Table 4. Effects of gamma irradiation on tuber yields o f cv. Farm Lisbon.
zMeans followed by the same letter are not significantly different
Mean wt (p = 0.05).
Test Treatment No. o f per plant
and year Generation (rads) plant (kg) The fact that the stimulation of the first generation was not
First test passed to the second generation is not surprising because the
1971 0 19 1.78 bz changes observed in the first generation after irradiation are
ri 1,250 20 2.88 a probably not of a genetic nature. In Sax’s review paper ( 6)
ri 2,500 15 1.65 b several similar findings are mentioned.
1972 _ 0 52 1.83 a T h e resu lts o b ta in e d b y irradiating d iffe r e n t p arts o f th e
r2 1,250 52 1.98 a tuber of ‘Cuello Largo’ are very perplexing. Since significant
Second test _ 0 39 2.03 a yield increase was obtained in both ‘Feo’ and ‘Farm Lisbon’
R1 1,250 39 2.31 a when the tubers were treated with similar doses, it is difficult to
see why ‘Cuello Largo’ failed to respond similarly to irradiation
zMeans o f a set followed by the same letter are not statistically different
(p = 0.05).
of its tubers in part or whole. A difference in cultivar response is

J. A m er. Soc. H o rt. Sci. 9 9 (3 ) :2 8 2 - 2 8 4 . 1974. 283

a possible explanation. Even more intriguing is the fact that the
yield of plants from the irradiated emerging shoots was not
decreased, but possibly slightly increased over that of the
control. If the dose damaged the tubers, then why did it not
damage the emerging shoots? Further experimentation will be
necessary to clarify these results.
Literature Cited
1. Bowen, H. J. M., P. A. Cawse, and S. R. Smith. 1962. The effects of
low doses of gamma radiation on plant yields. Intern J. Applied Rad.
and Isotopes 13:487-492.
2. Fowler, D. B„ and K. F. MacQueen. 1972. Effect of low doses of
gamma radiation on yield and other agronomic characters of spring
wheat (Triticum aestivum). Rad. Botany 12:349-353.
3. Kuzin, A. M. 1955. The utilization of ionizing radiation in agriculture.
Proc. 1st. Intern. Conf. Peaceful Uses Atomic Energy, Vol. 12, pp.
149-156.
4. MacQueen, K. F., J. W. Ketcheson, K. O. Lapins, M. D. Proverbs, L. H.
Lyall, V. W. Nuttall, and D. A. Rennie. 1971. Canadian studies on
applications of isotopes and radiation in agriculture and food
preservation. 4th U.N. Intern. Conf. Peaceful Uses o f Atomic Energy,
Paper No. A)Conf. 49/AI16Z
5. Nuttall, V. W., L. H. Lyall, D. H. Lees, and H. A. Hamilton. 1968.
Response of garden crop plants to low dose gamma irradiation of
seeds. Can. J. Plant ScL 48:409-410.
6. Sax, K. 1963. The stimulation of plant growth by ionizing radiation.
Rad. Botany 3:179-186.

Endomycorrhizal Fungi and the Development

of Citrus Seedlings in Florida Fumigated Soils1
N. C. Schenck and D. P. H. Tucker
University o f Florida
Gainesville and Lake Alfred

Abstract. The addition of Endogone calospora to fumigated soil significantly increased shoot and root
growth of citrus seedlings in the greenhouse but not in the field. The addition of E. macrocarpa
significantly increased growth of transplants and seedlings in nonfumigated and in methyl bromide-, and
dichloropropene-fumigated field soil. This beneficial effect did not occur in soils aerated by disking after
fumigation. Within 100 days, endomycorrhizal fungi were reestablished on weed roots growing in fumigated
soil. Citrus stunting was greater after methyl bromide fumigation than after dichloropropene fumigation,
but seedling stunt was not as severe as that observed in some commercial nurseries. Thus our results may
not be applicable to correcting citrus stunt as found in commercial nurseries.

In 1969, a Premium Quality Nursery Tree Program was
initiated in Florida for citrus nurserymen by the Florida
Department of Agriculture and Consumer Services, Budwood
Registration Bureau. As a result of this program, most citrus
seedbeds were fumigated with methyl bromide (453 g/9.29 m 2)
prior to planting. Shortly after the initiation of this practice,
several citrus nurserymen reported seedling stunting, marginal
leaf necrosis, and leaf bronzing in their citrus nursery seedbeds.
Tucker and Anderson (11) described the symptoms and history
of this problem in Florida and use of foliar and soil phosphate
applications to correct it.
Kleinschmidt and Gerdemann (4) reviewed the literature on a
similar citrus seedling problem from California, and related its
similarity to the stunting of citrus seedlings in Florida. After
inoculation with Endogone mossae Nicol. and Gerd. they
obtained an increase of citrus seedling growth in steamed and
fumigated soil in the greenhouse.2 This was similar to the
response obtained by Marx et al. (5), using the same
endomycorrhizal fungus on citrus seedlings in steamed soil. In
addition, Kleinschmidt and Gerdemann (4) reported an in­
creased growth response of citrus seedlings to E. mossae after
fumigation with methyl bromide in the field in Illinois.
O ur p u rp o se was 1) to evaluate the effect of
endomycorrhizal fungi of the Endogonaceae on citrus seedling
growth in fumigated soil in the field, and 2) to evaluate the
1Received for publication October 10, 1973. Florida Agricultural
Experiment Stations Journal Series No. 5062.
^Gerdemann and Trappe will soon publish a revision of the
Endogonaceae (3). Of the endomycorrhizal fungi mentioned in this
paper, Endogone mossae and E. macrocarpa were placed in a new genus
Glomus while E. calorpora and E. gigantea were placed in a new genus
Gigaspora.
feasibility of correcting citrus seedling stunt in Florida by
infesting soil in the field with endomycorrhizal fungi.
Materials and Methods
To evaluate the effect of endomycorrhizal fungi on citrus
seedling growth, studies were conducted in fumigated soil in
both the greenhouse and in the field. In the greenhouse, a
species of Endogone was used to inoculate seedlings of sour
orange {Citrus aurantium L.), ‘Cleopatra’ mandarin {Citrus
reticulata Blanco), and rough lemon {Citrus jambhiri Lush). This
species was similar to E. gigantea Nicol. and Gerd., producing
clusters of external, echinulate vesicles, but it had cream to
white colored azygospores rather than the typical green-yellow
color described for E. gigantea (2). This same species was the
most common species of Endogone on soybeans in Florida (9)
and, because of its cream colored azygospores, it was referred to
as E. calospora Nicol. and Gerd. It will also be termed E.
calospora in this paper.
Approximately 100 spores of E. calospora, sieved from a pot
culture of sorghum {Sorghum vulgare Pers.) were added to the
top 5 cm of methyl bromide-treated soil (908 g/0.83 m3 soil) in
15-cm plastic pots. Six pots of sour orange and 3 pots each of
rough lemon and ‘Cleopatra’ mandarin were planted with 25
seeds each. An equal number of pots of each citrus rootstock
received no E. calospora spores. After 14 months in the
greenhouse (18-35°C), plants were removed and assayed for
fresh weight of tops and roots, spore numbers in rhizosphere
soil, and mycelial development on the roots.
In 1971, E. calospora was used in field experiments at
Gainesville and Winter Haven, Florida. At Winter Haven, the
experimental design was a split plot of 3 replications. Main plots
were fumigated (methyl bromide 453 g/9.29 m2), and non­
fumigated soil. Fumigated plots were covered for 7 days

284 J. A m er. Soc. H ort. Sei. 9 9 ( 3 ) :2 8 4 - 2 8 7 . 1974.