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At least 19 recordsLinked to original sources

Tropical maize germplasm: what can we say about its genetic diversity in the light of molecular markers?

Knowledge about genetic variability of a crop allows for more efficient and effective use of resources in plant improvement programs. The genetic variation within temperate maize has been studied extensively, but the levels and patterns of diversity in tropical maize are still not well understood. Brazilian maize germplasm represents a very important pool of genetic diversity due to many past introductions of exotic material. To improve our knowledge of the genetic diversity in tropical maize inbred lines, we fingerprinted 85 lines with 569 AFLP bands and 50 microsatellite loci. These markers revealed substantial variability among lines, with high rates of polymorphism. Cluster analysis was used to identify groups of related lines. Well-defined groups were not observed, indicating that the tropical maize studied is not as well organized as temperate maize. Three types of genetic distance measurements were applied (Jaccard's coefficient, Modified Rogers' distance and molecular coefficient of coancestry), and the values obtained with all of them indicated that the genetic similarities were small among the lines. The different coefficients did not substantially affect the results of cluster analysis, but marker types had a large effect on genetic similarity estimates. Regardless of genetic similarity coefficient used, estimates based on AFLPs were poorly correlated with those based on SSRs. Analyses using AFLP and SSR data together do not seem to be the most efficient manner of assessing variability in highly diverse materials because the result was similar to using AFLPs alone. It was seen that molecular markers can help to organize the genetic variability and expose useful diversity for breeding purposes.

Brazil↗

Evaluating genetic relationships between tropical maize inbred lines by means of AFLP profiling.

Diversity among tropical maize inbred lines that compose breeding programs, is not well known. The lack of this information has made the arrangement of heterotic groups to be used for breeding purposes difficult. Methods of molecular analysis have been used as efficient alternatives for evaluating genetic diversity, aiming at heterotic group arrangement and acquisition of new hybrids. In this study, AFLP (amplified fragment length polymorphism) was used to investigate the genetic relationships among 96 tropical maize inbred lines from two different origins. The polymorphism level among the genotypes and the possibility of their allocation in heterotic groups were evaluated. Besides, correlations among genetic diversity and flowering time were analyzed. Nine primer combinations were used to obtain AFLP markers, producing 638 bands, 569 of which were polymorphic. Genetic similarities (GS), determined by Jaccard's similarity coefficient, varied from 0.345 to 0.891, with an average of 0.543. The dendrogram based on the GS and on the UPGMA cluster method did not separate the inbred lines in well-defined groups. Aiming at separating the lines into more accurate groups, Tocher's optimization procedure was carried out, 17 groups being identified. Association between flowering time and germplasm pools was detected. AFLP showed itself to be a robust assay, revealing a great power of detection of genetic variability in the tropical germplasm, and also demonstrated to be very useful for guiding breeding programs.

Brazil↗

Assessment of concentrations of iron and zinc and bioavailable iron in grains of early-maturing tropical maize varieties.

Twenty elite early-maturing (75-90 days) tropical maize varieties grown in three diverse agroecologies in West Africa were evaluated to identify varieties with high kernel-Fe and -Zn and bioavailable Fe levels. Bioavailable iron was assessed using an in vitro digestion/Caco-2 cell model. Significant (P < 0.001) varietal differences were observed in mean kernel-Fe and -Zn levels. The ranges were 15.5-19.1 mg kg(-)(1) for Fe and 16.5-20.5 mg kg(-)(1) for Zn. Genetic component accounted for 34% of the total variation in kernel-Zn and for 11% of the variation in kernel-Fe levels. Mean bioavailable Fe in varieties ranged between 4% below and 49% above the reference control variety. A significant negative relationship was detected between kernel-P concentration and bioavailable Fe (R = -0.36; P < 0.004; n = 60). Two varieties, ACR90POOL16-DT and ACR86TZESR-W, were identified as the most promising for further evaluation to determine their efficacy as improved sources of iron in target populations.

Biological Availability↗

Incorporation of tropical maize germplasm into inbred lines derived from temperate x temperate-adapted tropical line crosses: agronomic and molecular assessment.

Exotic maize ( Zea mays L.) germplasm may allow for increased flexibility and greater long-term progress from selection if it can be incorporated at high rates into U.S. breeding programs. Crosses were made between a temperate line, NC262A, and each of eight different lines consisting of 100% temperate-adapted tropical germplasm. Pedigree selection was used to generate a set of 148 F(5)S(2) lines that were evaluated in testcrosses with FR992/FR1064 in nine North Carolina environments. Several entries had grain yield, grain moisture content and standability that were comparable to three commercial checks. The best testcrosses outyielded the cross NC262A x FR992/FR1064 by 9.5 to 10.9%, suggesting that a significant amount of tropical germplasm was retained in these lines and that this germplasm combined well with the Stiff Stalk tester. Previous researchers had suggested that tropical alleles could be rapidly lost during inbreeding in populations derived from tropical x temperate bi-parental crosses, leading to the development of lines that possess significantly less than 50% tropical germplasm. F(5)S(5) sub-lines corresponding to the 14 best testcrosses were genotyped at 47 to 49 polymorphic simple sequence repeat (SSR) loci across all ten chromosomes to estimate the amount of tropical germplasm that was retained. The estimated genetic contribution from the tropical parent ranged from 32 to 70%, with the average being 49%. Only two of the 14 lines deviated significantly from a 50%-tropical/50%-temperate ratio, suggesting limited overall selection against germplasm from the tropical parents. These experiments collectively demonstrated that tropical maize germplasm can be incorporated at high rates into a temperate line via pedigree breeding methods in order to derive new inbred lines with acceptable agronomic performance.

Alleles↗

Molecular mapping in tropical maize (Zea mays L.) using microsatellite markers. 1. Map construction and localization of loci showing distorted segregation.

Microsatellites have become the most important class of markers for mapping procedures. Primarily based on restriction fragment length polymorphism (RFLP) markers, several molecular genetic maps of maize have been developed, mainly using temperate inbred maize lines. To characterize the level of polymorphism of microsatellite loci and construct a genetic map in tropical maize, two elite inbred lines, L-08-05F and L-14-4B, were crossed to produce 400 F(2) individuals that were used as a mapping population. A survey of 859 primer pair sequences of microsatellites was used. The polymorphism screens of each microsatellite and genotype assignment were performed using high-resolution agarose gels. About 54 % of the primer sets gave clearly scorable amplification products, 13 % did not amplify and 33 % could not be scored on agarose gels. A total of 213 polymorphic markers were identified and used to genotype the mapping population. Among the polymorphic markers, 40 showed loci deviating from expected Mendelian ratios and clusters of deviating markers were located in three chromosome regions. Non-Mendelian scoring was present in 19 markers. The final genetic map with 117 markers spanned 1634 cM in length with an average interval of 14 cM between adjacent markers.

Genetic Linkage↗

Accumulation, assembly, and digestibility of amarantin expressed in transgenic tropical maize.

An amaranth ( Amaranthus hypochondriacus) 11S globulin cDNA, encoding one of the most important storage proteins (amarantin) of the seed, with a high content of essential amino acids, was used in the transformation of CIMMYT tropical maize genotype. Constructs contained the amarantin cDNA under the control of a tissue-specific promoter from rice glutelin-1 ( osGT1) or a constitutive ( CaMV 35S) promoter with and without the first maize alcohol dehydrogenase intron ( AdH). Southern-blot analysis confirmed the integration of the amarantin cDNA, and copy number ranged from one to more than ten copies per maize genome. Western-blot and ultracentrifugation analyses of transgenic maize indicate that the expressed recombinant amarantin precursors were processed into the mature form, and accumulated stably in maize endosperm. Total protein and some essential amino acids of the best expressing maize augmented 32% and 8-44%, respectively, compared to non-transformed samples. The soluble expressed proteins were susceptible to digestion by simulated gastric and intestinal fluids, and it is suggested that they show no allergenic activity. These findings demonstrate the feasibility of using genetic engineering to improve the amino acid composition of grain crops.

Alcohol Dehydrogenase↗

Molecular mapping in tropical maize (Zea mays L.) using microsatellite markers. 2. Quantitative trait loci (QTL) for grain yield, plant height, ear height and grain moisture.

A previous genetic map containing 117 microsatellite loci and 400 F(2) plants was used for quantitative trait loci (QTL) mapping in tropical maize. QTL were characterized in a population of 400 F(2:3) lines, derived from selfing the F(2) plants, and were evaluated with two replications in five environments. QTL determinations were made from the mean of these five environments. Grain yield (GY), plant height (PH), ear height (EH) and grain moisture (GM) were measured. Variance components for genotypes (G), environments (E) and GxE interaction were highly significant for all traits. Heritability was 0.69 for GY, 0.66 for PH, 0.67 for EH and 0.23 for GM. Using composite interval mapping (CIM), a total of 13 distinct QTLs were identified: four for GY, four for PH and five for EH. No QTL was detected for GM. The QTL explained 32.73 % of the phenotypic variance of GY, 24.76 % of PH and 20.91 % of EH. The 13 QTLs displayed mostly partial dominance or overdominance gene action and mapped to chromosomes 1, 2, 7, 8 and 9. Most QTL alleles conferring high values for the traits came from line L-14-4B. Mapping analysis identified genomic regions associated with two or more traits in a manner that was consistent with correlation among traits, supporting either pleiotropy or tight linkage among QTL. The low number of QTLs found, can be due to the great variation that exists among tropical environments.

Microsatellite Repeats↗

Competition of Bacillus thuringiensis Cry1 toxins for midgut binding sites: a basis for the development and management of transgenic tropical maize resistant to several stemborers.

Binding and competition of five Bacillus thuringiensis toxins--Cry1Ab, Cry1Ac, Cry1Ba, Cry1Ca, and Cry1Ea--for midgut binding sites from three pests, Spodoptera frugiperda, Diatraea saccharalis, and Diatraea grandiosella, were investigated as part of a strategy to develop tropical transgenic maize resistant to several stemborers. On S. frugiperda, Cry1Ab and Cry1Ac compete for the same binding site; Cry1Ba and Cry1Ca compete for a second binding site. Cry1Ea recognizes a third specific binding site in S. frugiperda and does not compete with any of the other toxins. On D. grandiosella and D. saccharalis, Cry1Ac competes with Cry1Ab and not with Cry1Ba and Cry1Ca. Cry1Ba and Cry1Ca recognize each a specific binding site and do not compete with any of the other four toxins. Cry1Ea does not recognize any binding site on Diatraea species. Combinations of toxins are proposed to develop transgenic maize resistant to the three stemborers while allowing resistance management.

Animals↗

QTLs associated with phenolic acid accumulation and antioxidant activity in tropical maize.

Maize represents a significant source of phytochemicals, with phenolic acids standing out as one of the most extensively studied functional compound families. These bioactive molecules have gained attention for their potent antioxidant properties and potential contributions to human health improvement. To evaluate the segregation of phenolic compounds in maize and its genetic basis, this study was conducted to identify quantitative trait loci (QTLs) associated with major phenolic compounds and their antioxidant capacity. The mapping population comprised 100 recombinant inbred lines (RILs) derived from the cross between P84 and Kilima. Twelve traits were analyzed: free and cell wall-bound antioxidant capacity, total phenolic content, and contents of p-coumaric acid, ferulic acid, three isomers of di-ferulic acid, and three isomers of tri-ferulic acid. The RILs exhibited substantial diversity in phenolic compound profiles. In total, 19 QTLs were identified for nine traits, with the number of associated regions ranging from 1 to 5 and explaining between 2.95% and 37.48% of the phenotypic variation. This research provides substantial evidence for the co-localization of major QTLs for principal phenolic acids in maize with genomic regions harboring genes putatively related to their biosynthesis and biotic resistance. This is the first study to report QTLs associated with triferulic acids in maize. The identified regions co-localizing with biotic stress resistance genes represent targets for marker-assisted selection toward the improvement of phenolic acid accumulation in maize breeding programs.

QTLs↗

QTL for insect resistance and drought tolerance in tropical maize: prospects for marker assisted selection.

Insects and drought cause severe losses in the production of maize in many developing countries. Conventional breeding efforts to enhance the level of resistance to a number of insect pests and tolerance to drought have been successful, although only through large efforts of many breeders and over a large period of time. Continued improvements will only be possible through substantial investment of resources. Recently, success in identifying quantitative trait loci (QTL) in several plant species using various molecular marker systems offers alternative methods for accelerating conventional breeding programs. As the first step towards using molecular markers in CIMMYT's maize breeding program, restriction fragment length polymorphisms (RFLPs) have been used to understand the genetic basis of resistance to two corn borer species, southwestern corn borer and sugarcane borer, and to one major component of drought tolerance, anthesis-silking interval. A number of QTL with effects large enough to be regarded as significant in breeding were detected for each of these traits and many of them presented stable effects over environments. While variability in the number and location of QTL has been found when compared across populations, several loci were found to be quite consistent. Simple calculations can be made which estimate that the total genetic potential in maize for these traits is high. It is argued that to ultimately access and manipulate this potential, the use of linked molecular markers as indirect selectable markers is both feasible and necessary.

Adaptation, Physiological↗

Recovery of exotic alleles in semiexotic maize inbreds derived from crosses between Latin American accessions and a temperate line.

Genetic diversity of elite maize germplasm in the United States is narrow relative to the species worldwide. Tropical maize represents the most diverse source of germplasm. To incorporate germplasm from tropical maize landraces into the temperate gene pool, 23 Latin American maize accessions were crossed to temperate inbred line Mo44. During inbred line development, selection was practiced in temperate environments, potentially resulting in the loss of substantial proportions of tropical alleles. Genotyping 161 semiexotic inbreds at 51 simple sequence repeat (SSR) loci permitted the classification of their alleles as either Mo44 or tropical and allowed estimation of the proportion of detectable tropical alleles retained in these lines. On average, the percentage of detectable tropical alleles ranged among lines from 15% to 56%, with a mean of 31%. These are conservative, lower-bound estimates of the proportion of tropical germplasm within lines, because it is not known how frequently Mo44 and the tropical maize accession parental populations shared SSR alleles. These results suggest that substantial proportions of exotic germplasm were recovered in the semiexotic lines, despite their selection in temperate environments. The percent of tropical germplasm in semiexotic lines was not correlated to grain yield or moisture of lines testcrossed to a Corn Belt Dent tester, indicating that the incorporation of a substantial percentage of tropical germplasm in an inbred line does not necessarily negatively impact its combining ability. Thus, tropical maize accessions represent a good source of exotic germplasm to broaden the genetic base of temperate maize without hindering agronomic performance.

Agriculture↗

Light microscopy observations of tetrazolium-reducing bacteria in the endorhizosphere of maize and other grasses in Brazil.

Roots of field-grown tropical maize, Panicum maximum Jacq. and Digitaria decumbens Stent., and of sorghum and wheat grown in monoxenic culture with the diazotroph Spirillum lipoferum (syn. Azospirillum spp.) were examined for tetrazolium-reducing bacteria following incubation of roots in a malate-phosphate buffer-2,3,5-triphenyltetrazolium chloride medium. Bacteria were observed between and in cells of the cortex, in intercellular spaces between the cortex and endodermis, in xylem cells, and in and between pith cells. In maize, colonization of the inner cortex and stele appears to occur in the absence of significant bacterial colonization or collapse of outerlying tissues. Bacteria in the stele remained viable after a 6-h treatment of roots with chloramine-t, indicating that the endodermis was intact. Infection of the inner cortex and stele appears to occur initially in branches, and then to spread longitudinally into main roots. Inter- and intra-cellular infections of the cortex were observed in monoxenic systems. Tetrazolium reduction and prominent crystal formation was not specific for diazotrophic bacteria, but S. lipoferum was isolated from surface-sterilized roots, and S. lipoferum-like organisms were observed in the endorhizosphere. A correlation of inner cortex and stele infections with the presence of branches appears to explain previous observations that excised roots of grasses exhibiting high nitrogenase activity are characteristically branched roots with an intact cortex.

Bacteria↗

Selection for resistance to aflatoxin formation in maize through seed inoculation.

The efficiency of selection for resistance to aflatoxin development caused by Aspergillus flavus L. ex Fries, in maize (Zea mays L.) grain was studied in three CIMMYT lowland tropical maize populations. Seed samples of each population were inoculated with a spore suspension of cultures of Aspergillus spp. known to produce aflatoxins. Inoculated seeds were maintained at 85% RH in plastic boxes kept at room temperature. At 50% germination, seed samples were planted in the field. At flowering time, selected plants were bulk pollinated. After three cycles of selection, gains for tolerance to aflatoxin formation and improvement for agronomic characters were evaluated. The results indicate there are differences in aflatoxin accumulation between different genotypes and cycles of selection. Populations 22 (DMR) and 28 (DMR) were the most damaged and Population 31 DMR suffered the lowest infection. The study suggests that there are differences in aflatoxin production between the maize genotypes tested. However, our results failed to show that tolerance to aflatoxin development can be accumulated through a recurrent selection breeding scheme.

Aflatoxins↗

Molecular marker-based genetic diversity assessment of Striga-resistant maize inbred lines.

Striga-resistant maize inbred lines are of interest to maize breeding programs in the savannas of Africa where the parasitic weed is endemic and causes severe yield losses in tropical maize. Assessment of the genetic diversity of such inbred lines is useful for their systematic and efficient use in a breeding program. Diversity analysis of 41 Striga-resistant maize inbred lines was conducted using amplified fragment length polymorphism (AFLP) and simple sequence repeat (SSR) markers to examine the genetic relationships among these lines and to determine the level of genetic diversity that exists within and between their source populations. The two marker systems generated 262 and 101 polymorphic fragments, respectively. Genetic similarity (GS) values among all possible pairs of inbred lines varied from 0.45 to 0.95, with a mean of 0.61+/-0.002 for AFLPs, and from 0.21 to 0.92, with a mean of 0.48+/-0.003, for SSRs. The inbred lines from each source population exhibited a broad range of GS values with the two types of markers. Both AFLPs and SSRs revealed similar levels of within population genetic variation for all source populations. Cluster and principal component analysis of GS estimates with the two markers revealed clear differentiation of the Striga-resistant inbred lines into groups according to their source populations. There was clear separation between early- and late-maturing Striga-resistant inbred lines. Considering the paucity of germplasm with good levels of resistance to Striga in maize, the broad genetic diversity detected within and among source populations demonstrates the genetic potential that exists to improve maize for resistance to Striga.

Breeding↗

Screening and selection of maize to enhance associative bacterial nitrogen fixation.

The ability of maize (corn, Zea mays L.) to support bacterial nitrogen fixation in or on maize roots has been increased, through screening and selection. Isotopic N fixed from (15)N(2) was found on the roots. The nitrogen-fixing association was found in germplasm from tropical maize, but this activity can be transferred to maize currently used in midwestern United States agriculture.

Journal Article↗

Marker-assisted selection to improve drought adaptation in maize: the backcross approach, perspectives, limitations, and alternatives.

A number of different marker-assisted selection (MAS) approaches do exist for the improvement of polygenic traits. Results of a marker-assisted backcross (MABC) selection experiment aimed at improving grain yield under drought conditions in tropical maize are presented and compared with alternative MAS strategies. The introgression of favourable alleles at five target regions involved in the expression of yield components and flowering traits increased grain yield and reduced the asynchrony between male and female flowering under water-limited conditions. Eighty-five per cent of the recurrent parent's genotype at non-target loci was recovered in only four generations of MABC by screening large segregating populations (2200 individuals) for three of the four generations. Selected MABC-derived BC(2)F(3) families were crossed with two testers and evaluated under different water regimes. Mean grain yield of MABC-derived hybrids was consistently higher than that of control hybrids (crosses from the recurrent parent to the same two testers as the MABC-derived families) under severe water stress conditions. Under those conditions, the best five MABC-derived hybrids yielded, on average, at least 50% more than control hybrids. Under mild water stress, defined as resulting in <50% yield reduction, no difference was observed between MABC-derived hybrids and the control plants, thus confirming that the genetic regulation for drought tolerance is dependent on stress intensity. MABC conversions involving several target regions are likely to result in partial rather than complete line conversion. Simulations were conducted to assess the utility of such partial conversions, i.e. containing favourable donor alleles at non-target regions, for subsequent phenotypic selection. The results clearly showed that selecting several genotypes (10-20) at each MABC cycle was most efficient. In the light of these results, alternative approaches to MABC are discussed, including recurrent selection, illustrated by an example of improving the adaptation of maize to low temperatures. Given the current approaches for MAS and the choices of marker technologies available now and potential for future developments, the use of MAS techniques in further improving grain yield under abiotic stresses in maize appears very promising.

Crosses, Genetic↗

Change in apoplastic aluminum during the initial growth response to aluminum by roots of a tolerant maize variety

Root elongation, hematoxylin staining, and changes in the ultrastructure of root-tip cells of an Al-tolerant maize variety (Zea mays L. C 525 M) exposed to nutrient solutions with 20 &mgr;M Al (2.1 &mgr;M Al3+ activity) for 0, 4, and 24 h were investigated in relation to the subcellular distribution of Al using scanning transmission electron microscopy and energy-dispersive x-ray microanalysis on samples fixed by different methods. Inhibition of root-elongation rates, hematoxylin staining, cell wall thickening, and disturbance of the distribution of pyroantimoniate-stainable cations, mainly Ca, was observed only after 4 and not after 24 h of exposure to Al. The occurrence of these transient, toxic Al effects on root elongation and in cell walls was accompanied by the presence of solid Al-P deposits in the walls. Whereas no Al was detectable in cell walls after 24 h, an increase of vacuolar Al was observed after 4 h of exposure. After 24 h, a higher amount of electron-dense deposits containing Al and P or Si was observed in the vacuoles. These results indicate that in this tropical maize variety, tolerance mechanisms that cause a change in apoplastic Al must be active. Our data support the hypothesis that in Al-tolerant plants, Al can rapidly cross the plasma membrane; these data clearly contradict the former conclusions that Al mainly accumulates in the apoplast and enters the symplast only after severe cell damage has occurred.

Journal Article↗

Genetic relationships between resistance to stalk-tunneling by the European corn borer and cell-wall components in maize population B73xB52.

The objective of this study was to assess the relationships among quantitative trait loci (QTL) detected for European corn borer (ECB) tunneling and cell-wall components (CWC) neutral detergent fiber (NDF), acid detergent fiber (ADF), and acid detergent lignin (ADL) content in leaf-sheath and stalk tissues in a maize recombinant inbred line population derived from inbred lines B73 and B52. Most of the QTL for ECB resistance (10/13) were at QTL positions for one or more CWC. Of the 12 QTL for NDF and ADF in leaf-sheaths, five for each trait were at or near QTL for ECB tunneling. Four of these five QTL for NDF and ADF mapped to common locations. Four of the eight leaf-sheath ADL QTL were detected in the same genomic regions as ECB QTL. For stalk tissue, four regions contained common/overlapping QTL for ECB tunneling, NDF, and ADF. Six such regions were observed for stalk ADL and ECB tunneling. Seven of the ten QTL associated with both CWC and ECB tunneling contributed to the negative correlations observed between these traits, while relatively few QTL effects were positively correlated. This suggests that while CWC contribute to ECB resistance in this population, other mechanisms and other genes also are involved. Several QTL contributing to the negative correlations between ECB tunneling and CWC in the leaf-sheaths mapped to similar positions as QTL detected in tropical maize populations for resistance to leaf-feeding by Diatraea grandiosella Dyar and Diatraea saccharalis Fabricus. These regions may contain genes involved in the synthesis of cellulose, hemicellulose, and lignin in the leaf-blades and leaf-sheaths of maize.

Animals↗