ANDROGENESIS—ITS USE FOR
TRANSFER OF MAIZE CYTOPLASM
SHERRET S. CHASE*
NDROGENESIS in higher observed was the only successful plant
A plants, i.e., that aberration of
sexual reproduction leading to
the development of offspring with pater-
resulting from a cross of triploid Nico-
tiana tabacum var. macrophylla polli-
nated by normal N. Langsdorffii. It
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nal chromosomes only, affords breeders showed no trace of TV. tabacum, was like
and geneticists a means for direct trans- N. Langsdorffii in all respects save that
fer of cytoplasm from one strain to it had the gametic number of chromo-
another. This use of androgenesis was somes and the characteristic phenotypic
suggested by Chase4 in 1951 in connec- aspect of a monoploid. In 7the same year
tion with the special problem of con- Clausen and Lammerts recorded a
verting normal inbred lines of maize to paternal 'haploid' obtained in the pro-
forms characterized by cytoplasmically geny of the cross of a synthetic amphi-
conditioned male sterility. In maize, as ploid, N. digluta, pollinated by N.
in many other plants, the cytoplasm is of tabacum. This individual was diploid,
maternal descent; that is, in formation of the N. tabacum parent being a tetra-
the zygote, the sperm contributes nu- ploid. In 1934 Kostoff14 recorded
clear material only, whereas the egg con- another androgenetic monoploid in the
tributes both nucleus and cytoplasm. Tn progeny of the cross of the hybrid N.
androgenesis in higher plants, it is tabacum X N. sylveslris as female by N.9
probable that the male nucleus utilizes sylvestris as male. In 1936 Gerassimova
the cytoplasm of the egg or a synergid,
without participation of the female reported obtaining an androgenetic
nucleus, in forming the androgenetic monoploid from Crepis tectorum. In
'zygote'. In 1961 GoodselP1 demon- maize, paternal monoploids 20
have been
strated that the androgenetic sporophyte reported10 by Randolph , Seaney22,
in maize does indeed carry the cytoplasm Gerrish , Rhoades , and Goodsell11,
21
of the female parent. and have been observed regularly by the
author in the course of routine extraction
In this paper additional information of maternal monoploids. Rhoades21 ob-
on the frequency of occurrence of andro- tained two androgenetic monoploids,
genetic individuals in maize and prelimi- and an Fi diploid, from a maize kernel
nary data concerning effectiveness of a with three independent embryos. The
tetraploid cytoplasmic donor stock will same extraordinary occurrence has also
be presented. been observed by the author (unpub-
Literature Review lished) in another set of triplets.
Paternal individuals with the full
Sporophytes of paternal origin have somatic chromosome complement of the
been reported13 from several genera. In male parent have been reported over a
1929 Kostoff claimed the first definitely longer period of time. In 1916 Collins
confirmed paternal monoploid. (For and Kempton8 obtained a plant of
clarity, the term 'monoploid' will be Euchlaena mexicana in the progeny of
used here to indicate haploids derived the cross of Tripsacum dactyloides as
from diploid parents.) The individual female bv E. mexicana as male. Clausen
*DeKalb Agricultural Association, Inc., DeKalb, Illinois. The author wishes to express his thanks to
the DeKalb Agricultural Association, Inc., for financial support during the leave of absence in which
this paper was prepared; and to Dr. Donald F. Jones and his associates at the Connecticut Agricultural
Experiment Station and to Dr. Francois Mergen of Yale University for their helpful advice and assis-
tance.
Chase: Androgenesis 153
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PARENTAL MONOPLOIDS AND Fi DIPLOID
Figure 7
A—Maternal monoploid; B—Fi diploid; C—paternal monoploid. The contrasts among the three
classes of individuals are more pronounced at maturity, the mature color of the F! being purple, the
maternal monoploid green and the paternal monoploid brown.
and Lammerts7 in 1929 reported a Observations
paternal 'diploid' of Nicotiana tabacum
and postulated androgenetic origin for In connection with our studies of
diploid N. sylvestris plants which occa- parthenogenesis in maize2"6, it has also
sionally appear in the progeny of back- been possible to develop estimates of the
crosses of N. sylvestris X N. tabacum frequency of androgenesis. Androgenet-
hybrids as females to A^. sylvestris as ic individuals occur in a ratio of approxi-
male. Other androgenetic individuals mately one to every 80 monoploids of
with the somatic chromosome comple- maternal origin. For example, in a set of
ment of the male1 parent have been re- 577 monoploids obtained in the 1962-63
ported by Bleier in Vicia saliva, by winter season from progenies in which
Ichijima12 20in Fragaria virginiana, and by the 'Emerson brown, liguleless' stock,
Randolph in maize. a B PI C Ro Ig, was used as male, seven
In his study of androgenesis as a tool of the individuals were paternal mono-
for cytoplasmic transfer, Goodsell11 ploids. With average parthenogenetic
crossed the Nebraska Experiment Sta- rates in the range of 1/1,000, the fre-
tion inbred line N6 (a non-restorer of quency of androgenesis in our studies is
cytoplasmically conditioned male sterili- very similar to the rates noted by Good-
ty) to a 'Texas-sterile' cytoplasm stock, sell11, who found five paternal mono-
using N6 as the male parent. Among ploids in progenies of 400,000 plants or
the progeny, monoploid individuals of 1/80,000.
N6 were observed These paternals were In certain parentages, androgenetic in-
pollinated by the inbred N6. The pro- dividuals are easily distinguished from
geny were diploid, male-sterile indivi- the Fi heterozygotes and maternal indi-
duals of N6, (see Figure 8/f). viduals, (see Figure 7). The plants
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BREEDING DIAGRAMS
Figure 8
^ v p s conversion of inbred by androgenesis; cross of diploid X diploid. (1) Cytosterile
diploid, as female, pollinated by diploid inbred. (2) Androgenesis. (3) Paternal monoploid pollinated by
inbred. (4) Syngamy. (5) Cytoplasmicaily converted diploid inbred (progapated by continued back-
crossing). B—Synthesis of 'Emerson-brown', cytosterile, tetraploid stock. (1) Cytosterile diploid, as
female, pollinated by normal-cytoplasm, tetraploid, 'Emerson-brown'. (2) Syngamy (« + 2»). (3) Cyto-
sterile triploid, hybrid pollinated by tetraploid (first backcross). (4) Syngamy (n+ + 2n, and 2n+ + 2n).
(5) Selected tetraploid, cytosterile, brown plants pollinated by tetraploid (second backcross). (6) Syn-
Chase: Androgenesis
photographed for Figure 7, are from the phenotype and diploid. This individual,
cross of the 'Emerson brown, liguleless' grown in the greenhouse, was partially
marker pollinator to a Corn Belt dent fertile and set some seed upon self polli-
stock. At maturity, the contrasts nation. The second generation was field
among these three classes of individuals grown. Most individuals were complete-
are more pronounced, the mature plant ly male sterile, with the barren tassels
•color of the Fi being purple, the maternal characteristic of male cytosterility; a few
monoploids green, and the paternal plants were partially fertile; all were
monoploids brown. phenotypically indistinguishable in other
A few paternal diploids have also been characteristics from normal H52 (Figure
observed. Though these paternal di- 9). Apparently, an unreduced gamete
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ploids could have resulted from early from the male functioned androgenet-
doubling of the chromosome complement ically and, as expected, acquired the
of a paternal monoploid, it is thought cytoplasm of the female parent. This
more likely that this class of individuals particular gamete was, it seems most
derives primarily from androgenesis of likely, heterozygous in factors for partial
unreduced male gametes. (In partheno- fertility.
genesis, the maternal diploids that occur
are' generally heterozygous when de- Discussion
rived from a heterozygous female stock, For cytoplasmic conversion of inbred
though in such cases homozygous di- lines, the androgenetic method promises
ploids are also occasionally found). both a saving in time and, since the chro-
The usefulness of androgenesis in con- mosomal system is not disturbed by the
version of inbred lines of maize to male conversion process, greater technical
cytosterility is limited by a) the rarity of efficiency than in the backcross method
androgenesis and b) the meiotic sterility of conversion. The method is not as yet
of monoploids. With these two problems recommended for routine conversion of
in mind an investigation of the effective- lines.
ness in androgenesis of tetraploid cyto- In respect to the choice of diploid and
plasmic donor, stocks was started. In and tetraploid cytoplasmic donors, the
crosses of tetraploids by diploids in hoped for advantages in use of a tetra-
maize many types of aberrant repro- ploid donor were: a) increased frequen-
duction are possible (see Figure 8C). cies of androgenetic individuals among
For this phase of the study a 'Texas- viable progeny and b) increased oppor-
sterile' version of a tetraploid 'Emerson tunity for functioning of unreduced male
brown, liguleless' tester stock was de- gametes in androgenesis. The prelimi-
veloped. The scheme used in developing nary results indicate only that diploid
this stock is outlined in Figure 85. cytosterile paternal individuals can occur
Several inbred lines were crossed to this in the offspring when a tetraploid cyto-
specially synthesized cytosterile tetra- plasmic donor is used. Because of the
ploid, using the inbreds as pollen parents, large number of progeny required to put
with the hope of recovering paternal practical fiducial limits on rare events, a
diploids among the offspring. In pro- substantial body of data will be required
geny from a cross using Purdue inbred to determine the frequencies of diploid
H52 as pollinator, one individual was androgenesis in crosses to both diploid
recognized that was both paternal in and tetraploid female parents.
gamy (2n + 2«). (7) Established tetraploid, cytosterile, 'Emerson-brown' stock: (propagated by con-
tinued backcrossing). C—Theoretical progeny classes of tetraploid cytosterile stock, as' female, polli-
nated by diploid. (1) Cross of tetraploid, as female, by diploid; possible progeny: (2) Triploid (2n + n);
(3) Tetraploid (2n + 2« unreduced male gamete); (4) Parthenogenetic diploid (2n + 0). (5) Partheno-
genetic tetraploid (4» + 0). (6) Androgenetic monoploid (0 + n). (7) Androgenetic diploid (0 + 2»).
(8, 9) Other possibilities.
Key—(a) Sporophyte with normal cytoplasm, (b) Sporophyte with 'male-sterile' cytoplasm, (c) Egg
with normal cytoplasm, (d) Egg with'male-sterile'cytoplasm, (e) Sperm. The size of cell diagrams is
proportional to genome complement.
The Journal of Heredity
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CYTOPLASMIC CONVERSION
Figure S
^—Ears of normal inbred H52; B—ears of H523' (cytoplasm exchanged by androgenesis, inT X 2n
cross).
The functioning of unreduced male that the associated endosperm derives
gametes in androgenesis is of major in- from the fusion of polar nuclei and
terest. Since androgenesis, formation of sperm. In parthenogenesis it is possible
unreduced gametes, and competitive that both sperm fuse with the polar
success of diploid gametophytes in the nuclei. In the potato this has been
presence of monoploid gametophytes are demonstrated cytologically23.
all individually unusual, the coincidence In the formation of the triplets re-
of the three events is most interesting. ported by Rhoades and the author, it is
Colchicine, heat, or other treatments likely that four male nuclei were in-
of the tassels during the meiotic stages volved, with one of these four fusing
designed to double the chromosome with the polar nuclei to form a normal
complement of microspores, might great- endosperm, one fusing with the egg to
ly increase the production of unreduced form a normal zygote, and each of the
gametes and thereby the frequency of two extra nuclei forming paternal indi-
diploid androgenesis, especially if one viduals with a female contribution of
first sorted out the diploid pollen for use cytoplasm only.
in these crosses. Since there are distinct In respect to the paternal individuals
size and weight differences between n observed by Clausen and Lammerts",
and In pollen grains, sorting is feasible. both those with reduced chromosome
It is to be expected that androgenetic numbers and those with unreduced com-
rates will vary with the particular male plements, these authors state ". . . in
and female parents used. This has been neither case was any influence of the
found to 2be the case in parthenogenesis maternal cytoplasm discernible." The
in maize '0 and also in parthenogenesis drastic effects of special cytoplasms as
in the potato19. evident now in male cytosterile strains of
What is the fate of the egg nucleus in many plants, e.g., maize, onion, flax,
androgenesis? Perhaps it fails to func- petunia, carrot, sugarbeet, evening prim-
tion because of inviability or imbalance; rose, sorghum and wheat, has intensified
perhaps it simply fails to fuse with the interest in inherited cytoplasmic differ-
male nucleus and is lost in the first divis- ences and the structural basis of cyto-
ion the 'zygote'. The same possibilities plasmic inheritance. Studies of Mazo-
can be considered in regard to the fate of ti15"18 of a single homozygous maize
the male nucleus i n parthenogenesis. In genotype on maize and Euchlaena mexi-
both phenomena it has been established cana cytoplasmic backgrounds, have
Chase: Androgenesis 157
demonstrated major differences in pollen fertile individual segregated for partial-fertiles and
sterility, nucleolar size, chromosome completely sterile individuals. Apparently an
unreduced gamete of H52, heterozygous for par-
knob size, expression of the gene iojap, tial-fertility factors, functioned androgenetically
precosity and yield, dependent on the utilizing the cytoplasm of the female.
cytoplasmic environment. Whether, for cytoplasmic conversion in maize,
For investigators considering cyto- the use of a tetraploid cytoplasm donor will prove
more efficient than use of a diploid cytoplasm donor
plasmic studies in isogenic material, the remains to be demonstrated. The next step in
androgenetic technique offers special these investigations will be to attempt to increase
advantages in the establishment of the rate of occurrence of diploid pollen in the
stocks. In the studies of Mazoti, four- inbred male parents and to separate such pollen
for use on the tetraploid cytoplasmic donor stock.
teen crosses were required to develop the
isogenic, cytoplasmically differentiated
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stocks used in the comparisons. Literature Cited
Parenthetically, considering the phe- 1. BLEIER,H. Karyologische Untersuchugen an
nomenon of androgenesis in general, one Lensen-Wicken-Bastarden. Genetica 11:111-118.
should not discount the possibility of 1929.
obtaining rye seedlings from wheat ker- 2. CHASE, S. S. Monoploid frequencies in a
nels and similar occurrences. To quote commercial double cross hybrid maize, and in its
Zirkle24 in "Genetics in the 20th component single cross hybrids and inbred lines.
Genetics 34:328-332. 1949.
Century": "Virgil and Pliny told how 5. . Spontaneous doubling of the chro-
the farmers planted wheat and barley mosome complement in monoploid sporophytes of
only to discover that some of the seed maize. Proc. Iowa Acad. Sci. 56:113-115. 1949.
suddenly produced wild oats. A grain of 4. . The monoploid method of develop-
ing inbred lines. Proc. 6th Annual Hybrid Corn
oats was reported occurring in an ear of Industry-Research Conj. 29-34. 1951.
wheat by Thomas Johnson in 1633 and a 5. . Monoploids in maize. In: Heterosis
grain of rye in an ear of barley by Ole (ed. by [. \V. Gowen) Iowa State College Press,
Worm in 1655. These records are offered Ames. 1952.
6. —. Production of homozygous diploids
here only for comparison with Lysenko's of maize from monoploids. Agron. "Jour. 44:
supposed discovery. . . of grains of rye 263-267. 1952.
occurring in an ear of wheat." Lysenko's 7. CLAUSEN, R. E. and VV. E. LAMMERTS. Inter-
theories of the origin of such individuals specific hybridization in Nicotiana. X. Haploid and
are nonsense, yet through androgenesis diploid m'erogony. Amer. Nat. 63:279-282. 1929.
8. COLLINS, G. N. and J. H. KEMPTOX. Patro-
all these events are within the realm of genesis. Jour. Hered. 7:106-118. 1916.
possibility. Indeed we have a good 9. GERASSIMOVA, H. Experimental erhaltene
parallel already in the plant of Euchlaena haploids Pflanze von Crepis tectorum L. Planta 25:
mexicana obtained by Collins and Kemp- 696-702. 1936.
10. GERRISH, E. E. Studies of the monoploid
ton8 from a Tripsacum dactyloides seed method of producing homozygous diploids in
parent. Zea mays. Ph. D. Thesis, University of Minnesota.
Minneapolis. (Diss. Abs. 16:2285-2286). 1956.
11. GOODSELL, S. F. Male sterility in corn by
Summary androgenesis. Crop Sci. 1:227-228. 1961.
12. ICHIJIMA, K. Studies on the genetics of
In maize, androgenesis has been observed in Fragaria. Zeilschr. ind. Abst. Vererb. 55:300-347.
both diploid by diploid crosses, and in tetraploid by 1930.
diploid crosses. The work of Goodsell (1961) and 13. KOSTOFF, D. An androgenic Nicotiana
the observations reported here indicate that andro- haploid. Zeitschr.f. Zellforsch- «. mikro. Anat. 9:
genesis may be used to effect cytoplasmic transfer 640-642. 1929.
from one source to another, without rearrange- 14. . A haploid plant of Nicotiana svl-
ment of the chromosomal system of the nuclear vestris. Nature 133:949. 1934.
donor. Data presented indicate that in diploid by 15. MAZOTI, L. R. Nuevos hallazgos acerca del
diploid crosses, paternal monoploids occur with a comportamiento de las unidades de la herencia,
frequency of the order of one per 80 maternal geney y plasmonic. Rev. Argent. Agron. 17:
monoploids, or one per 80,000 progeny. 145-162. 1950.
In a special study, a single diploid androgenetic 16. . Heritable cytoplasmic characters
individual was recovered from the progeny of the derived from the hybrid of Euchlaena with Z.ea.
cross of a specially devised 'Texas-sterile' cyto- Rev. Invest. Agric. B. Aires 8:175-183. 1954.
plasm, tetraploid version of the 'Emerson brown, 17. . Study of heritable cytoplasmic
liguleless' maize tester strain, pollinated by the differences in Z. mavs and E. mexicana. Rev.
Purbue inbred line H52. Progeny of this partially Argent. Agron. 25:12-44. 1958.
The Journal of Heredity
18. . Nucleo-cytoplasmic interactions in 22. SEANEY, R. R. Studies on monoploidy in
maize inheritance. Rev. Fac. Agron. La Plata 36: maize. Ph.D. Thesis, Cornell University {Plant
63-71. I960. Breed. Abstr. 25:3035). 1955.
19. PELOQUIN, S. J., R. W. HOUGAS and A. C. 23. v. WANCENHEIM, K. -H., S. J. PELOQUIN
GABERT. The frequency of haploids in Solarium and R. W. HOUGAS. Embryological investigations
tuberosum. Amer. Potato Jour. 37:350. 1960. on the formation of haploids in the potato {Sola-
rium tuberosum). Zeitschr. ind. Abst. Vererb. 91:
20. RANDOLPH, L. F. Some effects of high temp- 391-399. 1960.
erature on polyploidy and other variations in 24. ZIRKLE, C. The knowledge of heredity be-
maize. Proc. Nat. Acad. Set. 18:222-229. 1932. fore 1900. In: Genetics in the 20th Century (ed.
21. RHOADES, M. M. Androgenesis. Maize by L. C. Dunn). The MacMillan Company, New
Genetics Coop. Newsletter 22: 10. 1948. York. 1951.
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THE NORMAL HUMAN KARYOTYPE
The London Conference 28th-30th, August, 1963*
T HE present meeting was called to consider
developments since the Denver Conference!
and to assess the degree to which these may have
A comparison of data submitted by different
investigators revealed that surprisingly good
agreement on relative chromosomal length and
aided characterization of individual chromosomes arm ratios or centromeric indices can be obtained
or may have revealed inadequacies in the earlier in different laboratories provided that mitotic
identifications. There was general agreement that figures, clearly delineated and free from irregu-
the seven groups proposed in the Denver number- larities of condensation pattern, are selected for
ing system have stood the test of time, and that analysis. The dispersion of chromosomal measure-
little or no uncertainty exists in the group assign- ments appears to be due to both biological and
ment of any chromosome. However within certain technical factors and, while some of these varia-
groups identification of individual chromosomes is tions are random, others appear to be systemati-
questionable. The groups are sometimes designa- cally produced, as in the effect of exposure to
ted by letters, such as A to G, as well as by the colchicine which may contact some chromosomes
numbering scheme described in the Denver classi-
fication. {Continued on Page 182)
*Ciba Foundation Guest Symposium, sponsored by The Association for the Aid of Crippled Chridren,
New York. The participants were: N. A. Barnicot, Department of Anthropology, University College
London, London, W. C. 1., England; M. A. Bender, Biology Division, Oak Ridge National Laboratory,
Oak Ridge, Tennessee; D. H. Carr, Department of Microscopic Anatomy, University of Western Ontario,
London, Ontario, Canada; E. H. Y. Chu, Biology Division, Oak Ridge National Lab. Oak Ridge, Tennes-
see; M. A. Ferguson-Smith, Genetics Department, The University, Glasgow, W. 2. Scotland; C. E. Ford,
M. R. C. Radiobiological Research Unit, Harwell, Didcot, Berks, England; M. Fraccaro, Gruppo Eura-
tom i Universita di Pavia, Via Sant'Epifanio, 14, Pavia, Italy; J. German, The Rockefeller Institute,
New York, 21, N. Y.; J. L. Hamerton, Paediatric Research Unit, Cameron House, Guy's Hospital
Medical School, London, S. E. 1. England; K. Hirschhorn, Department of Medicine, New York Univer-
sity School of Medicine, 550 First Avenue, New York 16, N. Y.; P. A. Jacobs, M. R. C. Clinical Effects of
Radiation Research Unit, Western General Hospital, Crewe Road, Edinburgh 4, Scotland; J. Lejeune,
Institut de Progen£se, 15, rue de 1'Ecole de Medecine, Paris Vie., France; W. Lenz, Inst. fur Human-
genetik der Universitat, Universitats-Krankenhaus Eppendorf, Martinistrasse 52, 2 Hamburg 20,
Germany; J. Lindsten, Department of Endocrinology & Metabolism, Karolinska Sjukhuset, Stockholm
60, Sweden; O. J. Miller, Department of Obstetrics & Gynaecology, College of Physicians and Surgeons of
Columbia University, New York 32, N. Y.; P. S. Moorhead, Wistar Institute, Thirty-Sixth Street at
Spruce, Philadelphia 4, Pennsylvania; K. Patau, University of Wisconsin, Department of Medical
Genetics, Madison 6, Wisconsin; L. S. Penrose, (convener), Galton Laboratory, University College
London, London, W. C. 1, England; T. T. Puck, Department of Biophysics, University of Colorado,
Denver 20, Colorado; A. Robinson, Department of Biophysics, University of Colorado, Denver 20,
Colorado, W. Schmid, M. D. Anderson Hospital and Tumor Institute, Texas Medical Center, Houston
25, Texas, and Kinderspital, Zurich, Switzerland; C. Stern, Department of Zoology, University of Cali-
fornia, Berkeley 4, California; J. H. Tjio, National Institutes of Health, Bethesda 14, Maryland; M. J. D.
White, Department of Zoology, The University, Melbourne, Australia.
tA proposed standard system of nomenclature of human mitotic chromosomes. Jour. Hered. 51:214-
221. 1960.
