The allotetraploidization of maize

Summary Allotetraploidization is the creation of artifical allotetraploids from a normally diploid species. The possible value of allotetraploid maize has been discussed in Section I of this series. Allotetraploidization of maize can be achieved by restructuring a maize genome so that its chromosomes will not pair with those of the standard maize genome. This restructuring can be done by concentrating differential pairing affinity (DPA) factors into a single line by a recurrent selection type of breeding program. Because the divergence of the maize genome is a gradual process, it is necessary to devise a model for chromosome pairing and gene segregation in segmental allotetraploids. This has been done by considering pairing in each arm separately and then combining paired arms to form pairing configurations for whole chromosomes. The chromosome disjunction patterns are hypothesized and genetic ratios in relation to different levels of DPA are suggested.Contribution from the Science and Education Administration, U.S. Department of Agriculture, and the Agronomy Department, University of Missouri, Columbia, Missouri, Agricultural Experiment Station Journal Series No. 8090     

Agrobacterium-mediated transformation of maize

Maize may be transformed very efficiently using Agrobacterium tumefaciens-mediated methods. The most critical factor in the transformation protocol is the co-cultivation of healthy immature embryos of the correct developmental stage with A. tumefaciens; the embryos should be collected only from vigorous plants grown in well-conditioned glasshouses. With the protocol described here, approximately 50% of immature embryos from the inbred line A188 and 15% from inbred lines A634, H99 and W117 will produce transformants. About half of the transformed plants are expected to carry one or two copies of the transgenes, which are inherited by the progeny in a mendelian fashion. More than 90% of transformants are expected to be normal in morphology. The protocol takes about 3 months from the start of co-cultivation to the planting of transformants into pots.     

Plagiogravitropism of maize roots

The direction of root growth can be studied by analyzing the trajectories of roots growing in soil. Both the primary seminal root and nodal roots of maize attain a preferred, or liminal, angle of growth that deviates from the vertical. These roots are said to be plagiogravitropic. Experiments using plants grown in soil-filled boxes revealed that the primary seminal root is truly plagiogravitropic. It shows both positive and negative gravitropism in response to gravity stimuli and tends to maintain its direction even after growing around obstacles. These are experimental results suggesting that plagiogravitropic growth is controlled by internal factors. The orientation of the grain affects the establishment of the liminal angle of the primary seminal root, and both the position of their node of origin and the root diameter are closely related to the plagiogravitropic behaviour of nodal roots. Several external factors are also known to influence plagiogravitropism. Low soil water content causes a decrease in the angle of growth and soil mechanical resistance suppresses the gravitropic curvature. Plagiogravitropic behaviour of both seminal and nodal roots plays a significant role in shaping the root system.     

Anther cultures of maize

The presence of synthetic auxins (2,4-D at a concentration as low as of 0.5 to 5.0 mg I^-1, NAA at least at 5 mg I^-1) in the cultivation medium was essential for the induction of callogenesis in anther cultures ofZea mays L. The application of IAA was ineffective. Kinetin induced bursting, darkening and a rapid anther necrosis, but at an appropriate concentration ratio with 2,4-D it stimulated pollen maturation at the same time.     

The allotetraploidization of maize

Summary Four barley telotrisomics (Triplo 3S, 5S, 6S, and 7S) were studied. No major qualitative differences in morphology between the telotrisomics and their diploid sibs were found. The pollen and seed fertility of these telotrisomics was comparable to their diploid sibs. The meiotic study showed that the average frequency of 6_II + 1_III at diakinesis and metaphase I was 84.2% and 71.7%, respectively. The normal chromosome separation ranged from 77.2% to 89.4% at anaphase I through telophase II. The transmission rate of the extra telocentric chromosomes averaged 28.4% upon selfing and 28.7% through the female. All four telotrisomics showed various degrees of pollen transmission, the average being 3.6%. Ditelotetrasomic plants (2n = 14 + 2 homologous telocentrics) were obtained in the progenies of selfed monotelotrisomic plants of all four types. These ditelotetrasomic plants were viable and showed various degrees of seed fertility.Contribution from the Department of Agronomy. Supported by USDA-CSU Cooperative Research Project No. 58-82HW-6-8 and CSU Hatch Project.     

The allotetraploidization of maize

Summary Allotetraploidization is the creation of artificial allotetraploids. Allotetraploidization of maize can be accomplished by concentrating differential pairing affinity (DPA) factors into lines by a recurrent selection breeding system. Selection will be based on changes in genetic ratios which are the result of changes in the relative frequencies of various pairing configurations caused by DPA. Part 1 of this series gave extensive data on gene segregation in trisomic and tetraploid heterozygotes. Some of these tetraploids behaved like segmental allotetraploids. Part 2 presented a model for gene segregation in segmental allotetraploids. This paper presents an analogous model for gene segregation in trisomic heterozygotes. The pairing configurations of trisomes are analyzed by considering pairing in single arms which then are combined to obtain pairing configurations for whole chromosomes. The chromosome disjunction patterns of the various pairing configurations are hypothesized and expected genetic ratios are given that result from different levels of DPA expressed in several hypothetical trisomes. The model analyzes the effect of random pairing in one arm and non-random pairing in the other arms. Also, the effect of crossing over is taken into account. Because crossing over rates are affected by the environment, part of the variability in the data (Part 1) is explained. In addition, an hypothesis is advanced to explain the frequent enhancement of pairing affinity following x-irradiation.Contribution from Agricultural Research/Science and Education Administration, US Department of Agriculture, University of Missouri, Columbia, Missouri, Missouri Agric. Exp. Sta. Jounal Series No. 8 670     

The allotetraploidization of maize

Summary Allotetraploidization is the creation of synthetic allotetraploids. The allotetraploidization of maize can be accomplished by concentrating DPA (differential pairing affinity) factors into stocks by a recurrent selection breeding system. Selection is based on pairing configuration frequencies and altered genetic ratios that reflect DPA. Both an observed decline in the quadrivalent frequency per meiocyte from 8.10 to 7.31 and genetic data disclosing a reduction in the average frequency of recessive waxy (wx wx) pollen from Wx Wx wx wx plants from 17.48% to 13.35%, indicate considerable progress has been made toward allotetraploidization. A simple model for the effect of DPA on chromosome pairing and genetic ratios is presented.Contribution from the Agricultural Research Service, U.S. Department of Agriculture, University of Missouri, Missouri Agricultural Experiment Station, Journal Series No. 9806     

Similarity of maize and sorghum genomes as revealed by maize RFLP probes

Summary Densely saturated genetic maps of neutral genetic markers are a prerequisite either for plant breeding programs to improve quantitative traits in crops or for evolutionary studies. cDNA and genomic clones from maize were utilized to initiate the construction of a RFLP linkage map in Sorghum bicolor. To this purpose, an F₂ population was produced from starting parental lines IS 18729 (USA) and IS 24756 (Nigeria) that were differentiated with regard to many morphological and agronomical traits. A total of 159 maize clones were hybridized to the genomic DNA of the two parents in order to detect polymorphism: 154 probes hybridized to sorghum and 58 out of these were polymorphic. In almost all of the cases hybridization patterns were similar between maize and sorghum. The analysis of the segregation of 35 polymorphic clones in an F₂ population of 149 individuals yielded five linkage groups. The three principal ones recall regions of maize chromosomes 1, 3 and 5: in general, colinearity was maintained. A possible inversion, involving a long region of maize chromosome 3, was detected. Simulations were also performed to empirically obtain a value for the lowest number of individuals of the F₂ population needed to obtain the same linkage data.Prof. E. Ottaviano, to whom this paper is dedicated, suddenly died on June 7^th, 1991     

Paramutation in maize

Paramutation is a heritable change in gene expression induced by allele interactions. This review summarizes key experiments on three maize loci, which undergo paramutation. Similarities and differences between the phenomenology at the three loci are described. In spite of many differences with respect to the stability of the reduced expression states at each locus or whether paramutation correlates with DNA methylation and repeated sequences within the loci, recent experiments are consistent with a common mechanism underlying paramutation at all three loci. Most strikingly, trans-acting mutants have been isolated that prevent paramutation at all three loci and lead to the activation of silenced Mutator transposable elements. Models consistent with the hypothesis that paramutation involves heritable changes in chromatin structure are presented. Several potential roles for paramutation are discussed. These include localizing recombination to low-copy sequences within the genome, establishing and maintaining chromatin domain boundaries, and providing a mechanism for plants to transmit an environmentally influenced expression state to progeny.     

Enhancement of maize transformation efficiency by the use of maize matrix attachment regions

Improving genetic transformation efficiency is a major concern in plant genetic engineering. While various strategies have been investigated, the enhancement of selectable marker gene expression has not been tried extensively. We used maize matrix attachment regions (MARs) to bracket an herbicide resistance transgene, bar. MARs have been reported to enhance transgene expression level and stability. We show here that MARs not only enhance transformation efficiency by 50%, but are also able to increase or decrease relative efficiencies of each step of the regeneration process depending on MAR sequence combinations. Furthermore, we assessed the trans-effect of MARs in co-bombardment experiments with two independent plasmids, one including the MAR sequences and the other one the bar gene. As for simple bombardment, MARs enhanced transformation efficiency by having a positive influence on organogenesis step in the regeneration process.     

Phyllodied panicles caused by head smut of maize and vegetative multiplication of maize

Young in vitro cultured panicles of maize(Zea mays L.) isolated from plants previously infected under controlled conditions by Sporisorium reilianum (Kühn) Landon and Fullerton, the agent of head smut, were maintained in culture up to the regeneration of healthy plantlets. Production of plants remained low and depended on a synchronization of the date of inoculation with the physiological condition of the mother-plant, which is linked to the season. Use of fungicide throughout the experiment prevented the colonization by the parasite. Such a regeneration takes advantage of a vegetative reversal naturally induced by S. reilianum on certain spikelets. Our method produced regenerants whose characters are different from those of mother-plant: the panicle was always replaced by a fertile ear. This revised version was published online in June 2006 with corrections to the Cover Date.