A. Delibes, I. López-Braña, M. Mena y F. García-Olmedo. 1988. Introgression of Aegüops triuncialis into Triticum aestivum. A progress report. An. Aula Dei 19 (1-2): 189-194. The wild grass Aegüops triuncialis (genomes CCUU) has been crossed with the tetraploid wheat Triticum turgidum (AABB) and the resulting ABCU sterile hybrid has been rescued with pollen from the hexaploid T. aestivum (AABBDD). Seven spikes from two hybrid plants were pollinated with pollen from T. aestivum cv. Almatense and 8 kernels were obtained. After five rounds of selfing, plants with 28 to 41 chromosomes have been derived. Low fertility (3-5 viable kernels per plant) was observed throughout the process. Further crosses to T. aestivum will be performed to obtain stable lines that will be screened for Ae. triuncialis genetic material with the aid of previously identified biochemical chromosome markers and specially developed DNA probes

Delibes Castro, Angeles

García Olmedo, Francisco

Lopez Braña, Isidoro

Mena, M.

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Introgression of Aegilops triuncialis into Triticum aestivum. A progress report.

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An. Aula Dei (1988), 19 (1-2): 189-194. 
Introgression of Aegüops triuncialis into 
Triticum aestivum. A progress report. 
by A. DELIBES, I. LOPEZ-BRAÑA, M. MENA AND F. GARCÍA-OLMEDO 
Department of Biochemistry. E.T.S. Ingenieros Agrónomos-UPM. E-28040 Madrid, Spain. 
Key Words: Triticum aestivum, Aegüops triuncialis, Introgression, Gene transfer. 
ABSTRACT 
A. Delibes, I. López-Braña, M. Mena y F. García-Olmedo. 1988. Introgression of Aegüops 
triuncialis into Triticum aestivum. A progress report. An. Aula Dei 19 (1-2): 189-194. 
The wild grass Aegüops triuncialis (genomes CCUU) has been crossed with the tetraploid 
wheat Triticum turgidum (AABB) and the resulting ABCU sterile hybrid has been rescued with 
pollen from the hexaploid T. aestivum (AABBDD). Seven spikes from two hybrid plants were 
pollinated with pollen from T. aestivum cv. Almatense and 8 kernels were obtained. After five 
rounds of selfing, plants with 28 to 41 chromosomes have been derived. Low fertility (3-5 viable 
kernels per plant) was observed throughout the process. Further crosses to T. aestivum will be 
performed to obtain stable lines that will be screened for Ae. triuncialis genetic material with 
the aid of previously identified biochemical chromosome markers and specially developed DNA 
probes. 
INTRODUCTION 
The transfer of genetic material from wild species to cultivated ones has been extensively 
exploited as a way to introduce certain agronomic traits, such as resistance to different diseases, 
into well adapted, highly productive cultivars. Previous work from this laboratory has demons-
trated the effectiveness of a particular strategy to transfer genes from the wild grass Aegüops 
ventricosa to the cultivated wheat Triticum aestivum (Delibes and García Olmedo, 1973; Delibes 
et al., 1977; Doussinault et al., 1983; Delibes et al., 1987). The strategy consists in crossing the 
donor species, Ae. ventricosa (genomes DVDVMVMV) with T. turgidum (AABB), which acts as 
a bridge, and rescuing the sterile ABDVMV hybrid with pollen from the recipient species T. aes-
tivum (AABBDD). Plants resulting from this cross were fertile and after repeated selfing, stable 
lines with 42 chromosomes were derived from them. A study of meiosis in hybrids of these lines 
and the recipient hexaploid wheat, together with an investigation of the distribution of bioche-
mical chromosome markers, indicated that genes from the donor had been incorporated into the 
190 A. DELIBES, I. LÓPEZ, M. MENA y F. GARCÍA 
transfer lines both by chromosonal substitution and by recombination (Delibes and García-
Olmedo, 1973; Delibes et al., 1977; García-Olmedo et al., 1984). The incorporation genes for 
resistance to eyespot disease, caused by the fungus Pseudocercosporella herpotrichoides, and to 
powdery mildew, caused by the fungus Erysiphe gramiñis, into recombinant wheat chromoso-
mes, following the above strategy, has been demonstrated (Doussinault et al., 1983; Delibes et 
al., 1987). 
We now report our present progress in the application of the described breeding scheme to 
Aegilops triuncialis (genomes CCUU) as donor species, using the same bridge and recipient spe-
cies as before. In this particular case, enhanced homeologous recombination should occur as a 
result of the known ability of the C genome to suppress the Ph diploidization mechanism of T. 
aestivum and T. turgidum (see Kimber and Feldman, 1987). 
MATERIALS AND METHODS 
Triticum aestivum cv. Almatense H-10-15, T. turgidum H-l-1, and Ae. triuncialis (Syn. T. 
triunciale) A-l were used in the described experiments. Seeds were germinated in agar and plan-
ted in pots. Crosses were carried out in the greenhouse. Chromosome staining for somatic chro-
mosome counts was carried out by the Feulgen reaction as described by Sharma and Sharma 
(1965), after allowing the chromosomes to contract in ice-water for 48 h. 
RESULTS AND DISCUSSION 
The breeding scheme is represented in Fig. 1. At least two other alternative strategies could 
have been followed: a) direct crossing with T. aestivum, and b) synthesis of an artificial amphi-
ploid AABBCCUU which would then be crossed with T. aestivum. Although natural and arti-
ficial hybrids of Ae. triuncialis with T. aestivum seem to yield plants that are sterile but viable, 
this type of hybrid would be a less adequate intermedíate in the gene transfer because it would 
carry all the hexaploid wheat genomes and, when rescued with ABD pollen, would have a grea-
ter tendency to eliminate the alien genetic material while selecting for the euploid chromosome 
number (2n = 42). On the other hand, the method involving prior synthesis of the AABBCCUU 
alloploid would be more time-consuming than the adopted strategy. 
Three kernels of the intermedíate hybrid were obtained, and two of them gave plants that 
were viable and vigorous, but m-sterile. Seven spikes from these plants were pollinated with 
pollen from T. aestivum cv. Almatense H-10-15, yielding 8 kernels. These were planted and sel-
fed, and their progeny subjected to four additional round of selfing. At all stages, cióse to 90% 
of the kernels germinated normally and although 20-40% of the plants died in the greenhouse, 
the surviving plants were notably vigorous (Fig. 2). These plants had high sterility, yielding 3-5 
kernels per viable plant throughout, with no discernible increase in fertility in the more advan-
ced generations. 
Somatic chromosome counts were carried out in mitotic roots of plants at the different sta-
ges. While in early generations plants with 28 to 41 chromosomes were observed, in the last two 
generations 95% of the plants had 40 chromosomes and only about 5% had 41 chromosomes 
(Fig. 3). 
The fertility observed in the progeny of the present transfer experiment seems to be signifi-
cantly lower that in the previous one, involving Ae. ventñcosa (DVDVMVMV) instead ofAe. triun-
cialis (CCUU). This might be due to the fact that the Dv genome of Ae. ventñcosa is closer to 
the D. genome of hexaploid wheat than either of the two different genomes (CU) present in Ae. 
triuncialis, thus possibly leading to a more regular meiosis in the former case and to a probable 
more extensive chromosomal rearrangement processs in the present case due to the P/z-suppres-
sing effect of the C genome. 
INTROGRESSION INTO TRITICUM 191 
Backcrosses to T. aestivum cv. Almatense H-10-15 are planned both to increase fertility and 
to obtain stable introgression lines with 42 chromosomes. Genetic transfer from Ae. triuncialis 
will be characterized with the aid of previously identified biochemical chromosome markers and 
recently developed DNA probes. 
ACKNOWLEDGEMENTS 
This work was supported by grant PR83-2046 from the Comisión Asesora de Investigación 
Científica y Técnica. Technical assistance from L. Lamoneda, J. García, and C. Rojas is grate-
fully acknowledged. 
REFERENCES 
Delibes, A.; F. García-Olmedo. (1973) Biochemical evidence of gene transfer from the Mv 
genome of Aegilops ventricosa to hexaploid wheat. In Sears, E.R.; Sears, L.M.S. (eds) Proc. 
4th Int. Wheat Genet. Symp. Mo. Agrie. Exp. Stn. Columbia Mo, pp. 161-166. 
Delibes, A.; R. Sánchez-Monge; F. García-Olmedo. (1977) Biochemical and citological studies 
of genetic transfer from the Mv genome oí Aegilops ventricosa into hexaploid wheat. A pro-
gress report. In Sánchez-Monge, E.; García-Olmedo, F. (eds). Interspecific hybridization in 
plant breeding. Proc. 8th Cngr. EUCARPIA. University Politechnica, Madrid, pp. 81-89. 
Delibes, A.; I. Lopez-Braña; M. Mena; F. García-Olmedo. (1987) Genetic transfer of resistance 
to powdery mildew and of an associated biochemical marker from Aegilops ventricosa to 
hexaploid wheat. Theor. Appl. Genet. 73: 603-608. 
Doussinault, G.; A. Delibes; R. Sánchez-Monge; F. García-Olmedo. (1983) Transfer of a domi-
nant gene for resistance to eyespot disease from a wild grass to hexaploid wheat. Nature 303: 
698-700. 
García-Olmedo F.; A. Delibes; R. Sánchez-Monge. (1984) Transfer of resistance to eyespot 
disease from Aegilops ventricosa to wheat. Vortr. Pflanzezuchtung 6: 156-168. 
Kimber, G.; M. Feldman. (1987) Wild wheat. An introduction special report 353 College of 
Agriculture University of Missouri-Columbia. 146 pp. 
Sharma, A.K.; A. Sharma. (1965) Chromosome techniques. Butteworth & Co. (Publishers) 116 
pp. 
RESUMEN 
Varias proteínas de endospermo pertenecientes a una familia de inhibidores de a-amilasas 
heterologas y de tripsina han sido purificadas y caracterizadas en trigo, cebada y centeno. La 
comparación de sus secuencias de aminoácidos y la localización cromosómica de sus genes 
estructurales indican que estos inhibidores están codificados por una familia multigénica dis-
persa que debió originarse por traslocaciones y duplicaciones intracromosómicas en una especie 
ancestral de trigo, cebada y centeno. La homología entre algunos miembros de esta familia de 
inhibidores y varias proteínas de maíz, ricino, mijo indio, etc., indica que la familia se extiende 
a otras especies filogenéticamente distantes de los tres cereales antes mencionados. 
192 A. DELIBES, I. LÓPEZ, M. MENA y F. GARCÍA 
? 
BRIDGE 
T. turgidum H-l-1 X 
2n = 28 
genomes AABB 
DONOR 
O Ae. triuncialis A-l 
2n = 28 
genomes CCUU 
Q m-sterile hybrid 
* 2n = 28 
genomes ABCU 
irregular meiosis: 
egg cells with up 
to 28 chromosomes 
RECIPIENT 
O T. aestivum cv. Almatense H-10-15 
2n = 42 
genomes AABBDD 
pollen ABD 21 chromosomes 
seed with up to 
49 chromosomes 
Selfing 5x 
AT lines 
2n = 28 to 41 
Figura 1.- Scheme followed for the genetic transfer from Aegilops triuncialis (syn. T. triunciale) 
to hexaploid wheat (T. aestivum). 
INTROGRESSION INTO TRITICUM 193 
Figura 2.- Plants obtained according to the breeding scheme of Fig. 1. 
194 A. DELIBES, I. LÓPEZ, M. MENA y F. GARCÍA 
-*í * * . í 
Ae. triuncialis 2n = 28 F. turgidum 2n = 28 7". aestivum 2n = 42 
4 
^ v j * » % TSb| 
* • « * * 
A* 
AT LINES 2n = 40 2n = 40 2n = 41 
Figura 3.- Cariotypes of parental material and representative 5 x selfed lines obtained as repre -
sented in Fig. 1. 


[EN] The wild grass Aegilops triuncialis (genomes CCUU) has been crossed with the tetraploid wheat Triticum turgidum (AABB) and the resulting ABCU sterile hybrid has been rescued with pollen from the hexaploid T. aestivum (AABBDD). Seven spikes from two hybrid plants were pollinated with pollen from T. aestivum cv. Almatense and 8 kernels were obtained. After five rounds of selfing, plants with 28 to 41 chromosomes have been derived. Low fertility (3-5 viable kernels per plant) was observed throughout the process. Further crosses to T. aestivum will be performed to obtain stable lines that will be screened for Ae. triuncialis genetic material with the aid of previously identified biochemical chromosome markers and specially developed DNA probes.
[ES] Varias proteínas de endospermo pertenecientes a una familia de inhibidores de @-amilasas heterologas y de tripsina han sido purificadas y caracterizadas en trigo, cebada y centeno. La comparación de sus secuencias de aminoácidos y la localización cromosómica de sus genes estructurales indican que estos inhibidores están codificados por una familia multigénica dispersa que debió originarse por traslocaciones y duplicaciones intracromosómicas en una especie ancestral de trigo, cebada y centeno. La homología entre algunos miembros de esta familia de inhibidores y varias proteínas de maíz, ricino, mijo indio, etc., indica que la familia se extiende a otras especies flogenéticamente distantes de los tres cereales antes mencionados.Peer reviewe

Introgression of Aegilops triuncialis into Triticum aestivum. A progress report.

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