Mapping QTLs in breeding for drought tolerance in maize (Zea mays L.)

Summary Grain yield in the maize (Zea mays L) plant is sensitive to drought in the period three weeks either side of flowering. Maize is well-adapted to the use of restriction fragment length polymorphisms (RFLPs) to identify a tight linkage between gene(s) controlling the quantitative trait and a molecular marker. We have determined the chromosomal locations of quantitative trait loci (QTLs) affecting grain yield under drought, anthesis-silking interval, and number of ears per plant. The F₃ families derived from the cross SD34(tolerant) × SD35 (intolerant) were evaluated for these traits in a two replicated experiment. RFLP analysis of the maize genome included non-radioactive DNA-DNA hybridization detection using chemiluminescence. To identify QTLs underlying tolerance to drought, the mean phenotypic performances of F₃ families were compared based on genotypic classification at each of 70 RFLP marker loci. The genetic linkage map assembled from these markers was in good agreement with previously published maps. The phenotypic correlations between yield and other traits were highly significant. In the combined analyses, genomic regions significantly affecting tolerance to drought were found on chromosomes 1,3,5,6, and 8. For yield, a total of 50% of the phenotypic variance could be explained by five putative QTLs. Different types of gene action were found for the putative QTLs for the three traits.     

Storage of maize (Zea mays L.) pollen at −196°C in liquid nitrogen

Summary The water content of pollen has a decisive influence on its storability in liquid nitrogen. Pollen with an initial high water content cannot be stored successfully at extremely low temperatures, so a certain degree of drying must be carried out before storage. Provided the viability of the pollen is not significantly reduced during drying, the pollen remains viable and fertile when kept at –196°C.     

Maturity interaction and black layer occurrence in opaque-2 and normal hybrids in maize (Zea mays L.)

Summary Six opaque-2 lines and their normal counterparts were crossed in diallel crosses. The crosses were grown in 1970 in a split plot randomized complete block design. Harvests were made at 7-day intervals starting at 28 days after pollination and countinuing through 63 days.The average kernel weight of opaque-2 hybrids was inferior to that of the normal. Nevertheless, the opaque-2 gene performed differently in different hybrids. In B14 x B37 single cross the mutant had similar kernel weight as its normal counterpart in the first and second harvests. In contrast a wide difference was found between the opaque-2 and the normal, both at early and late stages of development in W64A x A545 background.The normal hybrids had greater cob weight, ranging from 9'7 to 11.8% more than the opaque-2. The difference in cob weight of the opaque-2 and the normal remained constant over the different harvest dates.At physiological maturity, the opaque-2 hybrids averaged 3.5% higher moisture content than the normal. In general, a slower accumulation of dry matter in the kernels was accompanied by a retention of more moisture.Shelling percentage was higher for the normal hybrids. Black layer, an indicator of physiological maturity, was formed at about the same time in the opaque-2 and normal.Journal paper No. 6094.     

Genetic variation in maize (Zea mays L.) for germination and emergence at 10°C

Summary Full-sib, paternal half-sib and S1 families were derived from a population of maize, Pool 5, with a wide germplasm base. These families were used to study genetic variation for germination percentage, emergence percentage and time to emergence at 10°C.Genotypic variances were large for all characters, and the best families from the population were markedly superior to hybrids used as checks. Maternal effects appeared to be of overwhelming importance for germination percentage, but variances due to males were significant for emergence percentage and time to emergence. Inbreeding depression at the seedling level was also important for these two emergence characters. Germination and emergence characters were not correlated with seed weight.We concluded that Pool 5 should be a valuable source of genes for improving germination and emergence of maize at low temperatures.     

Heterosis in relation to genetic divergence in maize (Zea mays L.)

Summary The magnitude of heterosis was examined in relation to genetic divergence among 10 parents in a 10 × 10 diallel cross of maize. The 10 parents (varieties) were selected on the basis of multivariate analysis of quantitative characters using Mahalanobis' D²-statistic from a lot of 64 varieties of maize belonging to different geograshical regions of the world. The magnitude of heterosis for grain yield and its components was found to be higher with moderate (intermediate) parential diversity than with extreme ones. The results indicated that not only the genetic diversity but the high per se performance with respect to specific characters of interest should be taken into consideration also while selecting parents for hybridization.     

Breeding for yield, in mixtures of common beans (Phaseolus vulgaris L.) and maize (Zea mays L.)

Summary Despite the growing industrialization, technification and transformation that is happening in the agriculture around the world, and despite that agricultural research has always concentrated its effort on sole crops, multiple cropping systems have historically been important for common bean production in tropical countries. The reasons for this fact, are economical and social, as well as biological. Bean breeders have always been questioned on their work, because the development of new varieties is usually done in sole crop, but the varieties are grown in either systems. This paper addresses a set of questions that are usually presented to the breeders, in light of the evidence obtained from many trials conducted in Brazil and in the U.S.A.: Will the genotypes bred for sole crop conditions, perform well when grown in intercrop; How different should a genotype be, for cultivation in intercropping compared to genotypes developed for sole crop conditions; Is there a need for special breeding programs for intercropping and How could a breeding program focus the question of multiple (associated) cropping?     

Effect of pollen storage by drying and deep-freezing on the expression of different agronomic traits in maize (Zea mays L.)

Summary Pollen from the inbred maize line HMv 1645 was used to study the effect of pollen treatments (drying and deep-freezing) on the phenotypic performance of the next generation. Fresh and artificially dried pollen samples with different water contents (56%, 18%, 13% and 10%) were used for sib pollinations immediately after collection or drying. Samples containing low amounts of water were then stored in liquid nitrogen for 7 days. Fertilization ability of the samples with 13% water was the highest after storage. Plant characteristics of the next generation originated from the seeds set by differently treated (fresh, dried to 13% water and deep-frozen) pollen were examined and statistically analysed. Pollen treatments due to the pollen storage procedure did not cause detectable changes in quantitative characters of the next generation.     

Genotype x environment interactions in a 10 × 10 diallel cross of quality protein maize (Zea mays L.)

Summary A 10×10 diallel cross experiment involving white modified opaque-2 maize inbred lines was grown at four sites. A stability analysis, based on both a combining ability and a heterotic pattern model, was developed. The stability analysis provided valuable information on the genotype x environment interaction properties of the 10 inbred lines. The Gail & Simon (1985) test for qualitative interactions provided a means of determining the nature of these interactions. The inbred, SO507W(M), is shown to have the best potential for use in a hybrid breeding programme, in terms of having the highest weighted general combining ability and line heterosis, and the best general adaptability to all four sites. The single cross, SO713W(P) x PO558W(F), has been identified as one of the genotypes to be used in a recurrent selection programme that favours specific combining ability.     

Genetic characterization of 218 elite CIMMYT maize inbred lines using RFLP markers

Characterization of genetic diversity among maize inbred lines can facilitate organization of germplasm and improve efficiency of breeding programs. A set of 218 phenotypically diverse inbred maize lines developed at CIMMYT for hybrid production was characterized using 32 RFLP markers to: (1) analyze the genetic diversity present; (2) define potential heterotic groups based on clusters formed with marker data; and (3) identify the most representative testers for each potential heterotic group. Lines were clustered using five different genetic distance measurements to find consensus non-hierarchical clusters. Dendrograms were produced to study hierarchical classification within smaller groups of lines. A very high average allelic diversity was seen in this germplasm. Lines did not cluster based on phenotype, environmental adaptation, grain color or type, maturity, or heterotic response (as determined based on hybrid performance with testers), but lines related by pedigree usually did cluster together. Previously defined testers from opposite heterotic groups were not genetically differentiated, and did not represent well their heterotic group. Discrete clusters were difficult to find; thus, potential heterotic groups will be difficult to suggest using RFLP markers alone. However, suggestions on how to use molecular markers and cross performance information to refine heterotic groups and select representative testers are presented.     

Glyphosate tolerance in maize (Zea mays L.). 1. Differential response among inbred lines

Summary Variation in susceptibility to the safe broad-spectrum herbicide glyphosate was investigated in maize. Eleven inbred lines, grown in a growth chamber, were evaluated for their tolerance to the herbicide at 2.4 mM (0.2 kg a.i. in 400 I ha^-1 of water). Following treatment with glyphosate at the three-leaf stage, significant variation in damage, expressed as visual injury ratings scored 7, 14 and 21 days after the application of the herbicide, was found. Effects on dry weight and shoot height were consistent with visual scores and the carbon-exchange rate was found to be a sensitive index of differential injury.Biochemical characterization of 5-enol-pyruvyl-shikimate-3-phosphate (EPSP) synthase, the main target of the herbicide, ruled out the possibility that this differential susceptibility was due to variations in the sensitivity of the enzyme. On the contrary, a positive correlation was found between in vivo tolerance and EPSP synthase levels, measured at different stages during seedling growth. This result suggests that a naturally occurring difference in EPSP synthase levels in the tissues may contribute to the differential response observed in vivo in maize inbreds.Abbreviations CER carbon-exchange rate - EPSP 5-enol-pyruvyl-shikimate-3-phosphate - ID₅₀ concentration causing 50% inhibition     

Identification of pedigrees of hybrid maize (Zea mays L.) cultivars by isozyme electrophoresis and reversed-phase high-performance liquid chromatography

Summary Determination of pedigree by laboratory means can provide an initial and rapid check of hybrid identity in studies designed to compare genotypes. In order to test the accuracy of such determinations, isozymic and chromatographic data were used in an attempt to ascribe pedigrees to 25 hybrids that were identified only by a letter code from a list of 30 inbred lines. The pedigrees of 9 single-cross hybrids were correctly deduced as was the direction in which each cross was made. The pedigrees of 2 hybrids could only be determined ambiguously, however these hybrids were reciprocals and the biochemical data were unable to distinguish between 2 related candidate inbred lines. Pedigrees of 3 hybrids were either incompletely determined or could not be determined from the list of 30 candidate inbred lines and were thus correctly identified as non-Pioneer^® brand hybrids. Pedigrees of 9 3-way hybrids could be determined accurately but the pedigrees of 2 non-Pioneer^® brand hybrids could not be deduced completely. The ability to correctly ascribe pedigrees of hybrids shows that these data provide a rapid means of determining whether additional field and laboratory tests would be warranted in tests designed to compare genotypic similarity of hybrids.     

The impact of genotype x soil texture interaction on the efficiency of selection for yield in maize (Zea mays L.)

Summary This study was undertaken to investigate the implications of genotype x soil texture interaction on response to selection in maize. Mass honeycomb selection for yield was applied for 11 cycles from the F₂ of the single cross maize hybrid F68×NE2 in a field B with silty-clay-loam soil texture. Response to selection compared to the original single cross hybrid was estimated both in absence of competition and under solid stand in the selection field B and in a nearby field A differing in soil texture (clay-loam).A strong crossover type of interaction occurred both under solid stand and in the absence of competition in the two tests the improved population outyielded the hybrid in field B in the two densities, but lagged behing the hybrid in field A. The results suggest that interaction between genotype and soil texture might affect efficiency of selection detrimentally unless provision is taken for parallel selection early in the crop improvement program in fields differing in soil texture.     

Glyphosate tolerance in maize (Zea mays L.). 2. Selection and characterization of a tolerant somaclone

Summary The progeny of 104 regenerated maize plants were screened for tolerance to the safe broad-spectrum herbicide glyphosate during seed germination and early growth. Seven somaclones showed varying degrees of resistance to the application of the herbicide at 1.2 mM (0.1 kg a.i. in 400 1 ha^-1 of water). Plants capable of a normal growth following treatment with 2.4 mM (0.2 kg ha^-1) glyphosate at the three leaf stage were selfed, and their progeny analyzed. A family able to tolerate the exposure to glyphosate at 2.4 mM was isolated and shown to maintain a photosynthetic rate comparable with control after the application of the herbicide.The selfed progeny of the tolerant somaclone was characterized as to the properties of two targets of glyphosate, the shikimate pathway enzymes 5-enol-pyruvyl-shikimate-3-phosphate (EPSP) synthase and 3-deoxy-D-arabino-heptulosonate-7-phosphate (DAHP) synthase. In vitro tests ruled out the possibility that the tolerance was due to altered forms of these enzymes. Families showed significant variability with regard to EPSP and DAHP synthase levels, measured at different stages during seedling growth; however, not even these traits were correlated with in vivo response to glyphosate. The possible role of other physiological processes in determining the increased tolerance to the herbicide is discussed.Abbreviations DAHP 3-deoxy-D-arabino-heptulosonate-7-phosphate - EPSP 5-enol-pyruvyl-shikimate-3-phosphate - CER carbon-exchange rate - ID₅₀ concentration causing 50% inhibition     

Variation among maize (Zea mays L.) accessions of Bendel State,Nigeria. Multivariate analysis of agronomic data

Summary Two multivariate techniques were used to characterize 30 maize accessions collected from three ecological zones of Bendel State, Nigeria. Differentiation of the 30 accessions into five distinct groups was achieved with the unweighted variable group method of the average linkage cluster analysis of 34 agronomic characters. Four of the taxonomic groups contained at least three accessions each, while a fifth group contained only one. The single accession contained in the fifth group was characterised by very early maturity, deeply pigmented leaves and ear husks and short statured plants. Clustering of the accessions into different phenetic groups followed substantially along geographical and traditionally trading routes. A few cases of overlapping of accessions from different geographical locations were obtained.Principal component analysis revealed that days to 50% tasseling and silking, number of nodes/plant, ear length, ear weight, leaf width, and kernel colour were the principal discriminatory characters that differentiated the accessions. Sixty-four percent of the total variation among the 34 characters were accounted for by the first five principal components while the first and second components accounted for 26 and 14 respectively.     

Genetic variation in digestibility of forage maize (Zea mays L.) and its estimation by near infrared reflectance spectroscopy (NIRS). An analysis

Summary Genetic variation is a major source of differences in digestibility of forage maize. Other sources are environmental conditions, location effects, and cultural practice. These factors initiate differences in cell-wall content and cell-wall digestibility. In vitro techniques with rumen microflora account efficiently for such differences. Cell-wall content can be determined easily and can also be predicted properly by Near Infrared Reflectance Spectroscopy (NIRS). However, no currently available chemical and physical laboratory techniques are able to discriminate between digestible and indigestible cell-wall fractions. Present NIRS functions do not predict cell-wall digestibility properly in maize. If further research on NIRS functioning overcomes this barrier, NIRS can become a useful breeding tool for digestibility in forage maize.     

Linkage of genes for anthocyanin pigmentation in rice (Oryza sativa L.)

Summary The inheritance of anthocyanin pigmentation and the interrelationship between genes governing anthocyanin pigmentation in different parts of the rice plant and genes governing panicle density were studied in the cross Suma × Wanar-1. A ratio of 45 pigmented: 19 non pigmented was realised for anthocyanin accumulation in sheath, nodal ring, internode, leaf tip, leaf margin and apiculus whereas a ratio of 3 normal:1 laxy was observed for panicle density. Pleiotropic action of a basic gene (A) and each of the two complementary duplicate genes (P _a or P _b) was detected. A linkage group of genes localising pigment in six plant parts was identified, the sequence of genes being P _nr-P _lm-P _la-P _a-P _sh-P _in. This may form part of the III S _p group of Misro et al. (1966), in which case the genes P _nr and P _la are proposed to be additions to this group.     

Reappraisal of Ashby's hypothesis on heterosis of physiological traits in maize,Zea mays L

Summary The present study was undertaken to reappraise the materials and methods used by various reports discussing Ashby's hypothesis on heterosis of physiological traits in maize. The review of 15 reports led to the conclusion that both Ashby's hypothesis and the subsequent refutations of the hypothesis were based on insufficient evidence. New experiments using suitable pollination techniques and an appropriate statistical analysis enabled us to confirm Ashby's hypothesis.This work is supported by the Green Energy Program' (GEP84-II-6-6) promoted by MAFF.     

Genetic improvement of cell-wall digestibility in forage maize (Zea mays L.). I. Performance of inbred lines and related hybrids

Summary The current study deals with genetic improvement of the nutritive value of forage maize. In separate field trials, maize inbred lines without the brown midrib trait and derived hybrids were evaluated for stalk quality as well as some other agronomic traits. The aim was to relate the performance of lines and hybrids. Quality traits studied were the contents of ash and cell walls expressed as percentage of dry matter and the digestibilities of organic matter and cell walls (stalk-dv% and stalk-dcw%, respectively). The performance of hybrids was established in a trial at two locations with three replicates per location and the performance of lines at one location in an unreplicated trial.The range for stalk-dcw% was about 10 percentage units between hybrids and 15 percentage units between inbred lines. Stalk-dcw% had of all quality traits of hybrids the highest broad-sense heritability (h ²=0.74), and determined about 80% of the variation in stalk-dv%. The only stalk quality trait where a significant correlation was found between the mean hybrid performance and the corresponding midparent value was stalk-dcw% (r=0.70, P<0.01).In conclusion, stalk-dcw% proved to be the only stalk quality trait worth evaluating at the inbred line level in a breeding programme aimed at producing commercial hybrid varieties of forage maize.     

Synchronization of flowering in maize (Zea mays L.) by the genetic system pg11 pg12

Summary The mutant genotype pg11pg11 pg12pg12-epistasis of double recessive factors-determines greater precocity of flowering. The possibility of using this mutant in the production of hybrid seed as a mean of synchronizing flowering in parent strains of different earliness is studied. The differences between mutant and green control genotypes in the characters of days to pollen shedding, pollen production, and height of plants are evaluated. Four mutant lines are considered together with their respective green near-isolines in two distinct localities and across four successive plantings.The mutant near-isolines presented lower values than those of the green near-isolines for all the characters. The maximum reductions in days to flowring were obtained in early plantings and late maturity strains. The reduction in pollen production was about 50%. The reduced height of the mutants is explained, at least in part, by the correlation of this factor with days to flowering. The mutants did not prove to be more sensitive to the environment than their green near-isolines. Seed production was not studied since the mutant is very deficient for this character and it could therefore only be utilized as the male parent. We concluded that the mutant, could be used to synchronize flowering in the industrial production of hybrid seed.     

The genetics of maize (Zea mays L.) growing at low temperatures II. Harvesting time,number of kernels and plant height at maturity

Summary A set of 11 inbred lines of grain maize and generations derived from them were grown at a latitude further north than the generally accepted limit of the crop in the U.K. Analyses of variance for harvesting time, number of kernels and final plant height showed significant differences between the inbred lines and between their F₁, F₂ and backcross generations.A- and C-scaling tests or the joint scaling test were used to analyse the means of parents, F₁, F₂ and the two backcross generations for 8 reciprocal crosses.These tests showed that for all the crosses the addivite-dominance models were adequate for the characters. i.e. there was little or no evidence of non-allelic interactions.There was some evidence for heterosis for number of kernels and plant height, that of number of kernels being particularly impressive.