ARTIFICIAL AND NATURAL HYBRIDS IN THE GRAMINEAE, TRIBE HORDEAE. VI. CHROMOSOME PAIRING IN SECALE CEREALE x AGROPYRON INTER- MEDIUM AND THE PROBLEM OF GENOME HOMOLOGIES IN THE TRITICINAE G. L. STEBBINS, JR. AND FUNG TING PUN University of California, Davis Received April 27, 1953 LARGE number of hybrids have now been made between cereal grain A species of the genera Triticum and Secale on the one hand and various species of wild grasses belonging to the tribe Hordeae on the other. Of par- ticular interest have been those hybrids involving perennial species of the genus Agropyron. The well-known hybrids between Triticum and Agropyron have been studied cytologically by many workers, and are known to exhibit consistently a certain amount of chromosome pairing at meiosis ( SAPEHIN 1935; PETO1936; OSTERGREN1940; LOVEand SUXESON1945; MATSUMURA 1949). The presence of this pairing has led these authors to the assumption that chromosomes derived from Agropyron are pairing with those from Triti- cum, and therefore that homology exists between genomes belonging to these separate genera. The possibility has often been overlooked that, since both parents of the hybrids are themselves polyploids, much or all of the pairing observed in the hybrids may be autosyndesis between chromosomes derived from the same parental species rather than allosyndesis between chromosomes belonging to the different genera. Hybrids involving Secale provide particularly favorable material for judging whether auto- or allospndesis is taking place. Secale chromosomes are con- siderably larger than those of Triticum or Agropyron, and can be easily recog- nized at meiosis, whi!e those of Triticum and Agropyron cannot be distin- guished from each other at this stage. Careful study of meiosis in such hybrids, therefore, should provide critical evidence concerning the amount of autosynde- sis which can take place between the different sets of any Agropyron species. From such evidence, the amount of allosyndesis found in Triticum x Agro- pyron hybrids could be estimated at least to a certain extent. The only records in the literature known to us of hybrids between Agro- pyron and Secale are those of VERUSHKIN(1935), LJUBIMOVA(1937), and TSITSIN(1937) on Secale cereale x Agropyron intervtzediicm, and of CRASNIUK (1935) and FAVORSKY(1935) on S. cereale x A. cristatum. The cytology of the hybrids concerned was studied by LJUBIMOVAand FAVORSKY.LJUBI- MOVA reported 0-5 bivalents in Secale cereale x Agropyron intermedium (" A. glaucumn "), which she believed to be due to allosyndesis. In S. cereale x A. GENETICS38: 600 November 1953. HYBRIDS IN THE TRIBE HORDEAE 60 1 cristatztm, FAVORSKYfound 7 bivalents more or less regularly present, and therefore assumed that the chromosomes of S. cereale are homologous with 7 of the 14 chromosomes derived from A. cristatztm. But since MYERSand HILL(1940) found that tetraploid A. cristatum has the cytological behavior of an autotetraploid, the pairing observed by FAVORSKYis best explained as autos)-ndesis, with little or no association between the chromosomes of A. cristatum and those of rye. In the present paper, evidence will be presented which strongly supports the same interpretation for the pairing found in the S. cereale x A.intermedium hybrid. MATERIAL AND METHODS The techniques of hybridization, fixation of cytological material, and making aceto-carmine preparations are the same as those previously employed ( STEB- BINS and TOBGY1944; STEBBINSet al. 1946). In the summer of 1949, sixty florets from one head of cereal rye, var. " Merced " were emasculated and pollinated with spikes of Agropyron intermedium culture no. 632. Seeds for this culture were obtained from the Carnegie Institution Laboratory, Stanford, and are descended from a collection received from Bolu Turkey by the U. S. Department of Agriculture under the number F.P.I. 109,219. From this plli- nation, seven seeds were harvested, and six vigorous F1 hybrids were obtained in 1950. Cytological fixations were made during the seasons of 1950 and 1951. MORPHOLOGICAL CHARACTERISTICS OF THE HYBRID The F1hybrid between Secale cereale and Agropyron intermedium is com- pletely perennial, although it lacks the rhizomes characteristic of A. inter- medium. In spikelet characters it is far more like Agropyron than Secale, and in fact any systematist not acquainted with its origin would unquestionably call it an unusual form of Agropyron. It differs from A. intermedium in the shorter internodes of the rachis of the spike, the slight ciliation along the edges of these internodes, the smaller number of florets per spikelet, the smaller number of nerves on the glumes and lemmas, and the distinct though rather short awns. In addition, the glumes and lemmas are more strongly nerved than they are in A. intermedium or any of its relatives. The morphological charac- teristics of our hybrid agree very well with those reported by LJUBIMOVA (1937). The hybrid has proved completely sterile, although 30 clonal divisions of it have flowered in the garden for three successive seasons. The first attempt to double the chromosome number by colchicine treatment failed, and this attempt has not been repeated. CYTOLOGY OF THE HYBRID Meiotic chromosome behavior in the two parents was the same as has been described by previous authors for the species concerned. Plants of the S. cere- ale parent regularly form 7 bivalents at I metaphase, while in A. intermedium there are commonly 21 bivalents. The cytological material available of this 602 G. L. STEBBINS, JR. AND FUNG TING PUN species was not good enough for careful analysis, but in occasional sporocytes quadrivalents as well as univalents were seen. There was some lagging of chromosomes in the second anaphase, and in a small number of sporocytes a single bridge fragment configuration was seen. The tetrads, however, were entirely regular, with no micronuclei evident. Meiosis in the hybrid was very irregular, but a considerable amount of pairing was observed in every sporocyte (table 1). With a few exceptions, this pairing was autosyndesis of chromosomes derived from Agropyron, and the seven larger Secale chromosomes appeared as unpaired univalents (fig. 2). In two of the fifty cells recorded in table 1, a chromosome of Secale was paired TABLE 1 Chromosome behavior at metaphase 1 in 50 sporocytes of Secale cereale x Agropyron intermedium. Chromosomes from Number of Percent Agropyron Secale cells Cells with 10 I1 + 1 I 71 1 2 19 YD 9II+3I 71 1 2 Y? YD 7II+7I 71 8 16 YD YD 1 111 + 8 I1 + 2 I 71 5 10 YY PY 1 I11 + 7 I1 + 4 I 71 G 12 #Y YD 1 III+ 6 I1 + 6 I 71 9 18 1s YY 1 I11 + 5 11+ 8 I 71 9 18 7D YD 2 111 + 6 I1 + 2 I AS+6I' 2 4 79 2, 2 111 + 6 I1 + 3 I 71 4 8 DD YD 2 I11 + 5 I1 + 5 I 71 1 2 YD YY 2 111 + 3 I1 + 9 I 71 1 2 73 DY 3 111 + 5 I1 + 2 I 71 1 2 1s ss 3111+511+21 1II+51 1 2 YY s, 3 111 + 4 I1 + 4 I 71 1 2 Number of chromosomes synapsed per nucleus. Range: 12-21; mean 15.83 Number of chromosomes involved in multiple associations. Range: 0-9; mean 3.24 'The symbol -4s indicates a bivalenr corrpssed of one chromosome from Agropyron and one from Secale. with one from Agropyron by means of a single terminal connection (fig. 1). This same configuration was seen in another cell in which the other configura- tions could not he fully anal>zed. In one sporocyte two of the large chromo- somes from Secale were similarly connected with each other (fig. 3). The autos] ndesis among the chromosomes derived from Agropyron inter- medium consisted most often of seven bivalents and seven univalents, or of one trivalent with five or six bivalents and eight or six univalents. Most of the remaining configurations were those involving two or three trivalents, or with nine or ten bivalents. More than eight unpaired Agropyron chromosomes were found in only one cell. The bivalents were niostly of the open type, containing but one chiasma, but closed bivalents were not infrequent. The later stages of meiosis were all very irregular, and contained many 2 3 4 6 FlCVREs 1-5.--hfriotic configurations in Scra/r rcrca/c X lfgropyron ifitcrnlcdili!n, all X 900. 1. Photograph of I metaphase, showing synapsis of Secale and Agropyron chromosomes. 2 and 3. Drawings of I metaphase; 2, showing seven Secale univalents (at left), two trivalents antl five bivalents of Agropyron (center), antl five univalents of Agropyron (right) ; 3, showing five univalents and one bivalent of Secale (left), three trivalents and five hivalcnts of Agropyron (center), and t\vo univalents of Agropyron (right). 4 and 5. Anaphase, sho\ving lagging chrommomes antl hridge fragment configurations. lagging chroniosonies. IZritlge-fragment configurations were seen in 34 percent of 82 cells o1)served at first anaphase (figs. 4, 5). The pollen grains of the hybrid are conipletely a1,ortive. DISCI.SSIOS The most notalde feature of this hy1)rid is the high amount of autosyntlesis among the chronioson1es derived from .4qrop~ronintcritteditcirr. The most fre- quent pairing behavior was that which woultl he espcctetl if the haploid coin- pletnent of A. irtfrrnredirrur consists of two genonies which are strongly though 604 G. L. STEBBINS, JR. AND FUNG TING PUN not completely homologous with each other, plus a third genome which shows only a slight degree of homology with the other two. This pairing is considerably greater than that reported by LJUBIMOVA (1937) in the same hybrid, but differences of this magnitude have been found by other authors between different hybrid plants of the same combination. In particular, of the two sister plants of Triticum durum var.