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     

Selection of 5-methyltryptophan resistant rice mutants from irradiated calli derived from embryos

For increasing the contents of specificfree amino acids in rice (Oryzasativa L.) cultivar, Donganbyeo, mutantcell lines resistant to growth inhibitionby 5-methyltryptophan (5MT) were selectedfrom embryo cultured callus irradiated with50 Gy of gamma rays. Two lines, M₂-1and M₂-2, were obtained from theregenerated plants by selfing. Thesegregation ratios of resistance andsensitivity in these lines fitted 9:7 and3:1 ratios, respectively. Considering theagronomic traits, M₂-1 was about thesame as the original variety in culm lengthand a little longer or higher in paniclelength and number of tillers. However, thefertility of M₂-1 significantlyincreased compared to the originalvariety. M₂-2 had an extremely highculm length and tillering capacity. Four5MT-resistant homozygous M₃lines (MRclasses), M₃-1-40 and M₃-1-116from M₂-1 (MR I),and M₃-2-8 and M₃-2-12from M₂-2 (MR II), were obtained inthe same manner as with the M₂generation. Protein contents of brown ricewere increased about 19% and 32% and the total contents of 9 free essentialamino acids were 71% and 34% greater thanthe original variety in the MR I andMR II groups, respectively. However, forthe free essential amino acids of polishedrice, there was no difference from theoriginal variety.     

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.     

Detection of linkage between RFLP markers and genes affecting anthocyanin pigmentation in maize (Zea mays L.)

Summary Linkages between molecular markers and genes involved in the expression of agronomical traits have already been described in all of the major crops. In most cases, the genetic model underlying the Quantitative Traits Loci (QTL) is discussed. Here, Restriction Fragment Length Polymorphisms (RFLPs) and Mapmaker-QTL have been used to pinpoint seven regions of the genome significantly correlated with four pigmentation qualitative traits of maize (Zea mays L.). Two of these, located on chromosomes 2 and 10, explain most of the variation of these traits. The R and B gene loci known to be involved in the regulation of the anthocyanin pathway map to the same regions and we suggest that these loci could be the candidate genes involved in the correlations detected with RFLPs. This type of result is in accordance with the hypothesis of the candidate gene which supposes that, if we have a very high density map of randomly-selected cDNA clones, it should theoretically be possible to associate a cloned genic sequence with a phenotypic trait where correlations are found.     

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.     

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.     

Structural observations during androgenic microspore culture of the 4c1 genotype of Zea mays L.

Summary The capacity of the maize genotype 4c1 to regenerate microcalli and embryos from cultured microspores has been examined by comparing various cold pretreatments and culture media, using microspores and pollen at different stages of development. Viability of cultured cells was tested with FDA and their development was traced with light and fluorescence microscopy using DAPI as a nuclear dye.It was found that a pre-incubation of dissected flowers floating in a liquid nutrient medium at 8°C during 10–14 days was most successful for the induction of cell division. Among the developmental stages tested only the microspores appeared to regenerate. Subculture at 25°C in the same liquid medium, supplemented with 0.1 mg/l TIBA, gave highest rates of microspore division, i.e. up to 70% at 4 to 6 days of culture.All pathways described earlier for maize androgenic embryogenesis were observed within the 4c1 genotype. Symmetric divisions occurred in cultured microspores but most frequently asymmetric divisions lead to the formation of microcalli within 12 days of culture. In at least 60% of all dividing microspores cells were derived from the generative nucleus. Microcalli further developed either into loose or compact calli. Compact calli formed embryo-like structures.Abbreviations DAPI 4,6-diamidino-2-phenylindole - Dicamba 3,6-dichloro-2-methoxy benzoic acid - 2,4D 2,4 dichlorophenoxyacetic acid - FDA fluorescein diacetate - PAA phenylacetic acid - TIBA 2,3,5-triiodobenzoic acid - YP medium Yu-Pei basal salt medium     

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     

Cytoplasmic-nucleargenic effects in the C121 (Athens) Zea mays L. inbred with three cytoplasms

Summary Seeds of the long-time maize (Zea mays L.) inbred CI 21 (Athens) were prepared with the following cytoplasms: GA 199 and GT 112. Each source of cytoplasm as the female parent was corssed to the CI 21 (A) as the pollen parent. The F₁, as the female, was backcrossed successively to the CI 21 (A) genotype until the CI 21 (A) nucleus was substituted into each of the cytoplasms. Thus, each cytoplasmic source of seed assumed to have the same nuclear genotype.The cytoplasms were compared in all possible combinations for one year with 10 replications. Among the characters studied, significant differences were obtained for the following: effect of herbicide, reaction to Fusarium sp., silking on a given day, tasseling, plant height, ear height, lodging, number of ears, and yield.These results revealed a favorable interaction of GA 199 cytoplasm with the CI 21 (A) genotype for desirable characters. Apparently, plasmon-sensitive effects caused the unfavorable results. Results from these experiments also provide further support for cytoplasmic effects on agronomic characters in maize.Part of the dissertation submitted by the senior author to the Dep. of Agronomy, University of Georgia, Athens, Georgia 30602, USA.     

The characterization and assessment of genetic diversity among maize (Zea mays L.) hybrids that are widely grown in France: Chromatographic data and isozymic data

Summary Zein chromatographic and isozyme electrophoretic data were used to characterize 61 hybrids of maize (Zea mays L.) that are of current widespread usage in France. A similar study was also carried out for 88 hybrids that are widely available in the central corn belt of the U.S. Objectives were to 1) investigate the degree to which isozyme and chromatographic data could provide unique hybrid identification; 2) to evaluate the wealth of genetic diversity available among hybrids that are currently cultivated in France and in the U.S.; and 3) to compare the patterns and extent of genetic diversity found in both countries with respect to their registration, breeding, and seed production practices.Isozyme data showed that all French hybrids were unique germplasm. However, numerous hybrids had chromatographic profiles that were identical for 11 major peaks and these hybrids could thus have some common germplasm at least among their female parental lines. Although there was a broad base of isozymic diversity among French hybrids, the amount in cultivation was less since the few most widely used hybrids, which were planted on approximately half the cultivated area, were similar for their isozymic and zein chromatographic profiles. Approximately only 50% of the U.S. hybrids were shown to be unique with the remainder classified into groups; at least some of the hybrids within each group could be genetically identical products. Leading U.S. hybrids were unique germplasm but numerous hybrids that collectively covered a large acreage appeared to encompass a relatively narrow genetic base.For the continued success of agriculture, it will be important to improve breeding, registration, and marketing practices in order to ensure the continued supply of genetically different and improved elite hybrids.     

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.     

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 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.     

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.     

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.     

若干玉米胚乳突变体基因效应的研究

对５个玉米单隐性胚乳突变体、３个双隐性胚乳突变体的可溶性糖（还原糖、蔗糖）、水溶性多糖、淀粉、粗蛋白、赖氨酸含量测定，并对突变体种子结构扫描电镜观察。分析了玉米胚乳突主体的基因效应及其互作效应。     

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?     

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.     

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.