Detection of the apomictic mode of reproduction in maize-Tripsacum hybrids using maize RFLP markers

Polyploid plants in the genus Tripsacum, a wild relative of maize, reproduce through gametophytic apomixis of the diplosporous type, an asexual mode of reproduction through seed. Moving gene(s) responsible for the apomictic trait into crop plants would open new areas in plant breeding and agriculture. Efforts to transfer apomixis from Tripsacum into maize at CIMMYT resulted in numerou intergeneric F₁ hybrids obtained from various Tripsacum species. A bulk-segregant analysis was carried out to identify molecular markers linked to diplospory in T. dactyloides. This was possible because of numerous genome similarities among related species in the Andropogoneae. On the basis of maize RFLP probes, three restriction fragments co-segregating with diplospory were identified in one maize-Tripsacum dactyloides F₁ population that segregated 1∶1 for the mode of reproduction. The markers were also found to be linked in the maize RFLP map, on the distal end of the long arm of chromosome 6. These results support a simple inheritance of diplospory in Tripsacum. Manipulation of the mode of reproduction in maize-Tripsacum backcross generations, and implications for the transfer of apomixis into maize, are discussed.     

Comparative isozyme polymorphisms of north American eastern gamagrass,Tripsacum dactyloides var.dactyloides and maize,Zea mays L.

Random samples, consisting of at least 100 individual seedlings, were taken from the diploid (2n=2x=36) eastern gamagrass (Tripsacum dactyloides var.dactyloides) and assayed to determine which of 12 enzyme marker loci and isozyme systems would be most informative in providing satisfactory resolution of both maize andTripsacum isozyme systems. For comparison, eight maize inbreds were included in the study to aid evaluation and comparison of the various isozyme systems. In addition, evaluations were conducted to identify if the identified optimum isozyme system could be used to detectTripsacum introgression in maize following a maize ×Tripsacum backcrossing scheme. Using the established isozyme techniques for maize (Zea mays L.), theAdh, Pgd, Cat, Est, B-Glu, Got, Idh, Tpi isozyme systems detected no polymorphism among theTripsacum individuals assayed. TheEst andB-Glu systems forTripsacum were unscorable due to poor staining and resolution. TheAcp, Mdh, Pgm, andPhi isozyme systems were found to be satisfactory markers for differentiating between eastern gamagrass individuals as well as detectingTripsacum introgression in maize. The availability of useful isozyme systems which can simultaneously provide significant isozyme resolution of maize,Tripsacum and maize-Tripsacum backcross hybrids, on a single gel system, will be useful for the detection of marker assistedTripsacum introgression into maize. In addition, the identification of a set of variable biochemical markers should also assist breeding, selection and genetic manipulations in eastern gamagrass.The use of company names in this publication does not imply endorsement by the USDA-ARS, or the product names of criticism of similar ones not mentioned. All programs and services of the U.S. Department of Agriculture are offered on a nondiscriminatory basis without regard to race, color, national origin, religion, sex, age, marital status, or handicap.     

MORPHOLOGY OF TEOSINTOID AND TRIPSACOID MAIZE (ZEA MAYS L.)

Modern races of maize (Zea mays L.) are characterized by indurated glume and rachis tissues. The archaeological record, as well as experimental studies indicate that in North America this induration is associated with hybridization between domesticated maize and its closest wild relative Z. mays subsp. mexicana (Schrad.) Iltis (teosinte). Similar induration can also be introduced into maize through introgression from Tripsacum. North and South American indurated races of maize are not all closely allied morphologically. They evolved independently under domestication. Teosinte is absent from South America, but Tripsacum is widely sympatric with maize from about 42 N to 42 S latitude. For these reasons it has been postulated that induration in South American races may be the result of Tripsacum introgression. However, barriers restricting gene exchange between Zea and Tripsacum are difficult to overcome in nature. It is maintained that indurated South American races of maize were derived from indurated Mexican races, and that the presence or absence of such induration is due to different degrees of expression by intermediate alleles of the tunicate locus.     

Malate Dehydrogenase, Alcohol Dehydrogenase, and 6-Phosphogluconate Dehydrogenase Isozymes of Zea mays L. × Tripsacum dactyloides L. Hybrids and Parents

Electrophoretic patterns of malate dehydrogenase (Mdh), alcohol dehydrogenase (Adh), and 6-phosphogluconate dehydrogenase (Pgd) of Zea mays L. × Tripsacum dactyloides L. hybrids and their parents were compared. The components of enzymes specific to T. dactyloides may be used as markers to identify the following T. dactyloides chromosomes in the hybrids: Tr 16 (Mdh 2 and Pdg 1), Tr 7, and/or Tr 13 (Adh 2). The isozymes of Mdh 2 are supposed as a possible biochemical marker to evaluate the introgression of genes, determining an apomictic mode of reproduction from T. dactyloides (localized on Tripsacum 16 chromosome) into Z. mays. The isozymes may be used as markers for the identification of maize chromosomes 1 and 6 in the hybrids as well. Chromosome count taken on the examined hybrids showed the addition of 9 to 13 chromosomes of T. dactyloides to maize chromosome complement.     

Tripsacum dactyloides (Poaceae): a natural model system to study parthenogenesis

Diplosporous apomeiosis, formation of unreduced embryo sacs primarily of the Antennaria type, followed by parthenogenetic embryo development and pseudogamy (fertilization of the central cell) describe gametophytic apomixis within the Tripsacum agamic complex. Tripsacum dactyloides (Eastern gamagrass) is a close relative of domesticated maize and was chosen as a natural model system to investigate gene expression patterns associated with parthenogenesis. The genome size of diploid sexual and polyploid apomictic T. dactyloides was estimated by flow cytometry to be 7.37 pg (2C), 14.74 pg (4C) and 22.39 pg (6C), respectively. The diploid genome size is thus approximately 1.352 larger than that of maize. The apomeiotic-pseudogamous pathway of seed formation was demonstrated at a rate of 92% by the flow cytometric seed screen (FCSS) with single mature seeds in tetraploid accessions. This number includes twin embryos which were detected in 13% of the seeds analyzed. Fertilization of unreduced egg cells (B_III hybrids) was measured in 10% of apomictic seeds. Autonomous (fertilization-independent) embryo development and fertilization-dependent endosperm formation were confirmed by pollination of tetraploid T. dactyloides with a diploid transgenic maize line carrying an actin::#-glucuronidase (GUS) reporter construct. GUS expression was detected after pollination in the developing endosperm, but not in the embryo. In similar intraspecific crossing experiments with maize, GUS expression was detected in both the embryo and endosperm. A protocol was established for microdissection of embryo sacs and early parthenogenetic embryos of T. dactyloides. Together, these techniques provide new tools for future studies aimed at comparing gene expression patterns between sexual maize and sexual or apomictic T. dactyloides.     

Inheritance of parthenogenesis in Poa pratensis L.: auxin test and AFLP linkage analyses support monogenic control

 Gametophytic apomixis in Kentucky bluegrass (Poa pratensis L.) involves the parthenogenetic development of unreduced eggs from aposporic embryo sacs. Attempts to transfer the apomictic trait beyond natural sexual barriers require further elucidation of its inheritance. Controlled crosses were made between sexual clones and apomictic genotypes, and the parthenogenetic capacity of (poly)diploid hybrids was ascertained by the auxin test. A bulked segregant analysis with RAPD and AFLP markers was then used to identify a genetic linkage group related to the apomictic mode of reproduction. This approach enabled us to detect both an AFLP marker located 6.6 cM from the gene that putatively controls parthenogenesis and a 15.4-cM genomic window surrounding the target locus. A map of the P. pratensis chromosome region carrying the gene of interest was constructed using additional RAPD and AFLP markers that co-segregated with the parthenogenesis locus. Highly significant linkage between parthenogenesis and a number of AFLP markers that also appeared to belong to a tight linkage block strengthens the hypothesis of monogenic inheritance of this trait. If a single gene is assumed, apomictic polyploid types of P. pratensis would be simplex for a dominant allele that confers parthenogenesis, and this genetic model would be further supported by the bimodal distribution of the degree of parthenogenesis exhibited in the (poly)diploid progenies from sexual x apomictic matings. The molecular tagging of apomixis in P. pratensis is an essential step towards marker-assisted breeding and map-based cloning strategies aimed at investigating and manipulating its mode of reproduction. Received: 13 January 1998 / Accepted: 19 January 1998     

Heterochronic features of the female germline among several sexual diploid <Emphasis Type="Italic">Tripsacum</Emphasis> L. (Andropogoneae,Poaceae)

One element of gametophytic apomixis is unreduced embryo sac (ES) formation, which often occurs precociously displacing or replacing meiosis and causing apospory or diplospory, respectively. This study evaluated a premise that apomixis may evolve in hybridogenous plants that contain duplicate sets of allelically divergent ovule development heterochrony genes. The duplicate sets of genes would belong to duplicate genomic regions that are recombinationally isolated from each other (no gene flow) by allopolyploidy or paleopolyploidy, and this isolation would genetically stabilize apomixis. For apomixis to evolve, the ancestral donors of the duplicate regions must have differed from each other in timing of megasporogenesis, ES formation and embryony such that epigenetic misexpressions, or competitions in expression, of the duplicate heterochrony genes in hybridogenous derivatives would cause apomixis. Herein, we report substantial heterochrony in onset timing of germline stages among several sexual diploid Tripsacum genotypes, which may have been progenitors of apomictic polyploid Tripsacum. Tripsacum floridanum and Tripsacum zopilotense genotypes entered meiosis early. The former advanced rapidly through ES formation, but the latter entered a lengthy lag phase prior to ES formation. In two Tripsacum dactyloides var. dactyloides genotypes, meiosis occurred late and was followed by a distinct lag phase prior to ES formation. Likewise, the T. dactyloides var. meridonale genotype entered meiosis late, but the lag phase was brief. These differences appear to reflect allelic diversity at loci responsible for onset timing of different germline development stages within and across species and possibly across the recombinationally isolated duplicate chromosome regions in the Tripsacum paleopolyploid haplome (x = 18). Unique combinations of divergent alleles in hybridogenous plants coupled with polyploidy induced gene misexpressions may be required for apomixis to evolve. Electronic supplementary material Supplementary material is available in the online version of this article at and is accessible for authorized users.     

Risk assessment of transgenic apomictic tetraploid bahiagrass,cytogenetics, breeding behavior and performance of intra-specific hybrids

Pollen-mediated gene transfer from stress tolerant or herbicide-resistant transgenic plants may cause environmental or agronomic problems. Apomictic seed production found in some bahiagrass cultivars may serve as a natural transgene containment system. Under greenhouse conditions, the average gene transfer frequency from an herbicide-resistant apomictic tetraploid to a population of sexual diploid bahiagrass genotypes or apomictic tetraploid bahiagrass was 0.16% when the transgenic pollen donor was placed at 0.5–1.5 m distance from the non-transgenic pollen receptors. The herbicide-resistant hybrids were characterized for transgene integration, expression and ploidy, by Southern blot analysis, immuno-chromatography and flow cytometry, respectively. Hybrids resulting from open pollination of non-transgenic diploid female plants with transgenic tetraploid male plants were triploids or near-triploids, with 2n = 26–34. These hybrids displayed a wide range of phenotypic variability, including some non-persistent or non-flowering dwarf-type hybrids with good vigor, or hybrids with vegetative growth similar to non-transgenic plants, but with significantly reduced seed set. Non-flowering aneu-triploids with good vigor/field performance will provide the highest level of transgene containment. Embryo sac analysis of pollinated spikelets confirmed a high proportion of aborted ovules. An apospory-linked RFLP marker was detected in 13 of the 15 near-triploid hybrids. All flowering aneuploid hybrids displayed significantly reduced seed set, and none of the sexual near-triploid hybrids produced any seeds. All tetraploid gene transfer events carried the apospory-linked RFLP marker, suggesting that despite the presence of the aposporus locus, a low degree of sexuality co-exists in apomictic tetraploid cultivars. Thus, tetraploid apomictic bahiagrass does not provide complete transgene containment, although intra-specific gene transfer is drastically reduced compared to sexually reproducing perennial grasses.     

Dosage effects in the endosperm of diplosporous apomictic Tripsacum (Poaceae)

 Imprinting in the endosperm of angiosperms, a phenomena by which expression of alleles differs depending on whether they originate from the male or female parent, has been shown to explain most failure of interploidy or interspecific crosses in plants. Because of imprinting, seeds develop normally only if a specific dosage is represented in the endosperm, with the relative contributions of genomes in the ratio of two maternal doses to one paternal dose (2m:1p). In Tripsacum, a wild relative of maize, all polyploids reproduce through the diplosporous type of apomixis. Diplospory results from meiotic failure in megasporocytes that develop into eight-nucleate unreduced female gametophytes. The male gametophytes remain unaffected. Flow cytometry was used to determine ploidy levels in the endosperm of both apomictic and sexual Tripsacum accessions. In both cases, fertilization appeared to involve only one sperm nucleus. Therefore, endosperm of apomictic Tripsacum develops normally even though the ratio of genomic contributions deviates from the normal 2m:1p ratio. Ratios of 2:1, 4:1, 4:2, 8:1 and 8:2 were observed, depending on both the ploidy level of the parents and the mode of reproduction. Thus, specific dosage effects are seemingly not required for endosperm development in Tripsacum. These findings suggest that evolution of diplosporous apomixis might have been restricted to species with few or no imprinting requirements, and the findings have strong implications regarding the transfer of apomixis to sexually reproducing crops. Received: 17 February 1997 / Revision accepted: 7 July 1997     

Apomixis and ploidy barrier suppress pollen-mediated gene flow in field grown transgenic turf and forage grass (Paspalum notatum Flüggé)

Bahiagrass (Paspalum notatum Flüggé) is the predominant forage grass in the southeastern US. The commercially important bahiagrass cultivar ‘Argentine’ is preferred for genetic transformation over sexual diploid cytotypes, since it produces uniform seed progeny through apomixis. Pseudogamous apomictic seed production in Argentine bahiagrass may contribute to transgene confinement. It is characterized by embryo development which is independent of fertilization of the egg cell, but requires fertilization with compatible pollen to produce the endosperm. Pollen-mediated gene transfer from transgenic, glufosinate-resistant apomictic bahiagrass as pollen donor at close proximity (0.5–3.5 m) with non-transgenic sexual or apomictic bahiagrass cultivars as pollen receptors was evaluated under field conditions. Hybridization frequency was evaluated by glufosinate herbicide resistance in >23,300 seedlings derived from open-pollinated (OP) pollen receptor plants. Average gene transfer between transgenic apomictic, tetraploid and sexual diploid bahiagrass was 0.03%. Herbicide-resistant hybrids confirmed by immuno-chromatographic detection of the PAT protein displayed a single copy bar gene identical to the pollen parent. Hybrids resulting from diploid pollen receptors were confirmed as triploids or aneu-triploids with significantly reduced vigor and seed set as compared to the parents. Transmission of transgenes to sexual bahiagrass is severely restricted by the ploidy difference between tetraploid apomicts and diploid sexual bahiagrass. Average gene transfer between transgenic apomictic tetraploid and non-transgenic, apomictic tetraploid bahiagrass was 0.17%, confirming a very low frequency of amphimixis in apomictic bahiagrass cultivars. While not providing complete transgene containment, gene transfer between transgenic apomictic and non-transgenic bahiagrass occurs at a much lower frequency than reported for other cross-pollinating or facultative apomictic grasses.     

OBSERVATIONS ON INTROGRESSION OF TRIPSACUM INTO MAIZE

Derivatives of a cross between diploid Zea mays L. and Tripsacum dactyloides (L.) L. (2n = 72) were compared cytologically and morphologically. The objective of this study was to detect introgression from Tripsacum to maize that might have occurred during seven backcross generations with maize. Thirty-three morphological characters were used to analyze variation among aneuploid (20Zm + 2Td), 20-chromosome recovered maize, and the recurrent maize parent plants. Aneuploid and maize checks were extreme types, with 20-chromosome hybrid derivatives being morphologically intermediate. Several recovered maizes clustered with aneuploid plants and these hybrid derivatives have the greatest chance of Tripsacum introgression. Many traits such as endosperm abnormalities, tassel seed, albinos, tunicate glumes, tassel-tipped ears, fasciated and branched ear, and male spikelets between rows of kernels were observed. Although the genetic basis of many traits is unknown, mutations, epistatic effects or expression of Tripsacum chromatin are possible causes. The number of abnormal and tripsacoid traits observed in 20-chromosome recovered maizes indicates genetic transfer from Tripsacum to the maize genome.     

Interspecific hybridization between Vigna radiata (L.) Wilczek and V. glabrescens

Summary Interspecific hybrids of the mungbean, Vigna radiata (L.) Wilczek (2n=22) and V. glabrescens (2n=44) were generated with the aid of embryo culture. V. glabrescens x V. radiata hybrids were recovered via germination of the immature embryos. Reciprocal hybrids were obtained via shoot formation from embryonic callus. The authenticity of the hybrids was determined by morphological characteristics, chromosome number, and isozyme patterns. The hybrids were highly sterile upon selfing, but backcrossing to the diploid parent yielded viable seeds. Some of the plants resembled the diploid parent morphologically while others resembled neither parent. The backcross plants were sufficiently fertile to give a large number of mature, selfed seeds. Plants obtained differed morphologically and in their isozyme patterns from either parent, indicating introgression. These progeny populations will be used as bridging materials to transfer pest resistance from the wild tetraploid to the cultivated mungbean.     

Chromosome control of apomixis in maize-gamagrass hybrids

The results of long-term studies on the transmission of the mode of asexual reproduction through seeds to maize from gamagrass, a closely related wild plant, performed in the Laboratory of Plant Cytology and Apomixis are summarized. The first apomictic hybrids between Zea mays and Tripsacum dactyloides were obtained in this laboratory more than 40 years ago and have been maintained until the present time. Cytogenetic studies on the hybrids have shown that at least nine chromosomes of the wild parent are necessary for the expression of asexual reproduction through seeds. In addition, the genes controlling two elements of apomixis (apomeiosis and parthenogenesis) have been found to be inherited independently from each other.     

Megasporogenesis and megagametogenesis in several Tripsacum species (Poaceae)

The Tripsacum agamic complex (x = 18) will provide valuable characters for maize breeding, provided that apomixis can be manipulated. Apomixis in Tripsacum was first reported 40 years ago, but its prevalence in the genus has not been established. Reproductive development was determined for eight Mexican and two South American Tripsacum species by microscopic analysis of ovaries cleared in a benzyl benzoate-dibutyl phthalate solution using interference contrast optics. The occurrence and distribution of callose deposition during megasporogenesis were determined by fluorescence microscopy of ovaries optically cleared in an aqueous sucrose solution containing aniline blue. Diploid genotypes were sexual. Polyploid forms reproduced apomictically following the Antennaria type (complete meiosis abortion) of diplospory. The Taraxacum type (unreduced megaspore production through meiotic restitution nuclei) of diplospory also occurred but rarely. The walls of diplosporic megasporocytes lacked callose whereas the walls of sexual megasporocytes contained a normal complement of callose. The absence of callose suggests that the diplosporic forms of reproduction result from mutations affecting the normal meiotic process. Apomixis in the Tripsacum genus is facultative, and the production of new polyploid genotypes through genetic exchanges involving both apomictic and sexual genotypes is possible.     

Allotriploid somatic hybrids of diploid tomato (Lycopersicon esculentum Mill.) and monoploid potato (Solanum tuberosum L.)

Allotriploid somatic hybrids were obtained from fusions between protoplasts of diploid tomato and monohaploid potato. The selection of fusion products was carried out in two different ways: (1) The fusion of nitrate reductase-deficient tomato with potato gave rise only to hybrid calli if selection was performed on media lacking ammonium. Parental microcalli were rarely obtained and did not regenerate. (2) The fusion of cytoplasmic albino tomato with potato gave rise to albino and green hybrid calli and plants. Allotriploids were identified from the two somatic hybrid populations by counting chloroplast numbers in leaf guard cells and by flow cytometry of leaf tissue. Although some pollen fertility of allotriploids and pollen-tube growth of tomato, potato andLycopersicon pennellii into the allotriploid style were observed, no progeny could be obtained. The relevance of allotriploid somatic hybrids in facilitating limited gene transfer from potato to tomato is discussed.     

Sexual tetraploid and apomictic pentaploid populations ofHieracium pilosella (Compositae)

Five species are recognized inHieracium subgen.Pilosella sect.Pilosellina Fries. Four are diploid (2x, 2n = 18), one (H. pilosella L.) is highly variable morphologically and cytologically (from 2x to 10x), in its mode of reproduction (self-incompatibility, agamospermy, amphimixis, apo-amphimixis) and in its hybridization pattern. A part of this huge agamic complex was analysed by comparing sexual 4x and apomictic 5x plants (crossing and germination experiments, measurements of vegetative reproduction by stolons etc.). In the experimental garden apomictic 5x produced more stolons than the sexual 4x plants and the total length of the stolons per rosette was greater. However, in nature, the competitive potential of the sexual plants seems to be higher, presumably as a result of the higher mortality of ramets in 5x. Sexual 4x plants often grow in dense and grazed grass vegetation, whereas 5x apomicts often occur in dunes with patchy vegetation. Apomicts produce more capitula per rosette, and sexual rosettes form only about 60% of the number of viable achenes as compared to apomictic ones. Therefore, apomicts appear to be characterized by a greater colonizing ability than sexual plants. Apomictic plants produce equal numbers of viable achenes under conditions of both open pollination and isolation. Sexual plants do not form any viable achenes after isolation and produce a somewhat lower percentage of achenes after open pollination than do apomictics. 5xreproduce exclusively apomictically. Apo-amphimixis was never observed in pentaploids and only very rarely in tetraploids. Addition hybrids are very rare. The cross sexual 4x × apomictic 5x failed in 70% of the attempts, but the recombination of genomes carrying genes for apomixis is possible and results in apomictic 4x and sexual 5x, both with a reduced number of viable achenes. In nature sexual and apomictic plants may occur in close proximity. In such cases the germination rate of the achenes of 4x and 5x is lower; this may indicate that apomictic plants fertilize sexual plants in nature (unidirectional gene-flow). 5x plants form euploid gametes carrying two or three genomes. The results of the crossing experiments can be explained in terms ofNogler's theory of monogenic inheritance of apospory.Variation and evolution inHieracium subg.Pilosella sect.Pilosellina I.     

Attack of the clones: reproductive interference between sexuals and asexuals in the Crepis agamic complex

Negative reproductive interactions are likely to be strongest between close relatives and may be important in limiting local coexistence. In plants, interspecific pollen flow is common between co‐occurring close relatives and may serve as the key mechanism of reproductive interference. Agamic complexes, systems in which some populations reproduce through asexual seeds (apomixis), while others reproduce sexually, provide an opportunity to examine effects of reproductive interference in limiting coexistence. Apomictic populations experience little or no reproductive interference, because apomictic ovules cannot receive pollen from nearby sexuals. Oppositely, apomicts produce some viable pollen and can exert reproductive interference on sexuals by siring hybrids. In the Crepis agamic complex, sexuals co‐occur less often with other members of the complex, but apomicts appear to freely co‐occur with one another. We identified a mixed population and conducted a crossing experiment between sexual diploid C. atribarba and apomictic polyploid C. barbigera using pollen from sexual diploids and apomictic polyploids. Seed set was high for all treatments, and as predicted, diploid–diploid crosses produced all diploid offspring. Diploid–polyploid crosses, however, produced mainly polyploidy offspring, suggesting that non‐diploid hybrids can be formed when the two taxa meet. Furthermore, a small proportion of seeds produced in open‐pollinated flowers was also polyploid, indicating that polyploid hybrids are produced under natural conditions. Our results provide evidence for asymmetric reproductive interference, with pollen from polyploid apomicts contributing to reduce the recruitment of sexual diploids in subsequent generations. Existing models suggest that these mixed sexual–asexual populations are likely to be transient, eventually leading to eradication of sexual individuals from the population.     

Genome accumulation in eastern gamagrass,Tripsacum dactyloides (L.) L. (Poaceae)

Intermatings between sexual diploids (2n=36) and facultative apomictic tetraploids (2n=4n=72) of Tripsacum dactyloides var. dactyloides, (L.), L., have been generated. Subsequent exchange and addition of genetic material between the two cytotypes, via a backcrossing program, provides one method for the manipulation of genetic elements involved in apomictic reproduction (diplospory) of this species.Identification and development of fertile triploids, and their innate tendency to exhibit a high frequency of unreduced egg cells which remain receptive to fertilization by a sperm nucleus, allows for the unique development of B _III (2n=3x+n) tetraploids by genome accumulation. The resultant B _III progeny provide new breeding stock for investigating the inheritance and generation of such materials also allows for the occurrence of genetic recombination between sexual diploids and apomictic tetraploids. Potential use of fertile triploids, their tendency for genome accumulation and their use in exploiting apomictic tetraploid germplasm resources are also discussed.All programs and services of the U.S. Department of Agriculture are offered on a nondiscriminatory basis without regard to race, color, national origin, religion, sex, age, marital status, or handicap.     

Crossing maize with sorghum,Tripsacum and millet: the products and their level of development following pollination

Summary Maize was crossed with sorghum, Tripsacum and millet with the aim of introgressing desirable alien characteristics into maize. The products of crosses were analyzed as to their level of differentiation following pollination; their further development on artificial culture medium was compared. In spite of a stimulation rate close to 5%, no evidence of hybridization between maize and sorghum or millet could be obtained. The plants recovered proved to be of maternal origin. However, with an appreciable frequency, stimulation leading to hypertrophic growth of nucellar tissue was observed. This phenomenon is bound to pollination, never occurring in non-pollinated ears. In crosses involving Tripsacum, more than 140 true hybrids were isolated. The influence of the genotypes used as well as factors such as climatic conditions or in vitro techniques are discussed. Except for one haploid maize plant, all the plants recovered proved to be classical hybrids, most of them showing the expected complement of chromosomes from each parent (10 + 36 chromosomes), a few others being slightly hyperploid (2n = 47 to 50 chromosomes). No non-classical hybrids constituted by a nonreduced female gamete and a reduced male gamete were obtained.     

CYTOLOGY OF MAIZE-TRIPSACUM INTROGRESSION

The American Maydinae genera Zea and Tripsacum cross readily when not isolated from each other by gametophytic barriers, and it has been suggested that intergeneric introgression played a role in the evolution of maize. Four Zea chromosomes pair with members of at least one basic genome of tetraploid Tripsacum, and in hybrids involving octaploid Tripsacum all 10 chromosomes of the basic maize genome frequently compete successfully in synapsis with Tripsacum chromosomes. Hybrids that combine 36 Tripsacum and 10 maize chromosomes are female fertile. When they are pollinated by maize their offspring have 36 Tripsacum and 20 maize chromosomes, or again have 36 Tripsacum and 10 maize chromosomes, but the 10 Zea chromosomes are contributed by the new pollen parent. Later backcross generations also include plants with 36 Tripsacum and 12, 14, 16, or 18 maize chromosomes. Individuals with 2n = 56 produce an abundance of offspring with 18 Tripsacum and 20 maize chromosomes when backcrossed with maize. Further backcrossing results in elimination of Tripsacum chromosomes, and eventually plants with 2n = 20 Tripsacum-contaminated maize chromosomes are obtained. Two generations of selfing restore full fertility to these 2n = 20 plants and eliminate all obvious traces of Tripsacum morphology.