Cytochemical investigation of long-term stored maize pollen

Summary Maize pollen quality was investigated after long-term storage both in a refrigerator and in liquid nitrogen by a combination of viability tests and cytochemical methods. Determination of the activities of a number of enzymes involved in important metabolic pathways was carried out. Quinone formation was also studied, as some products of secondary metabolism affect pollen grain viability. One year of pollen storage in liquid nitrogen had little effect on the activities of oxidoreductases and hydrolases and had no significant effect on pollen grain viability evaluated by acetocarmine, neutral red and acridine organe. Only the FCR test showed slightly decreased viability. After one and two years of storage in a refrigerator, pollen grain viability, tested using acetocarmine, neutral red and acridine orange, did not change substantially. Simultaneously the FCR test showed a considerable decrease in pollen grain viability. Long-term storage in a refrigerator resulted in the loss of cytochrome oxidase activity and rise of alcohol dehydrogenase, lactate dehydrogenase, peroxidase and polyphenoloxidase activities as well as of quinone formation.Abbreviations ADH Alcohol dehydrogenase - DOPA L-3,4-Dihydroxyphenylalanine - FCR Fluorochromatic reaction - FDA Fluorescein diacetate - GDH Glutamate dehydrogenase - IDH Isocitrate dehydrogenase - LDH Lactate dehydrogenase - NADI reaction with -NAphthol and DImethyl-p-phenylenediamine hydrochloride - 6-PGDH 6-phosphogluconate dehydrogenase     

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.     

Expression of mature plant resistance to Striga hermonthica in maize

Twelve maize genotypes with different degrees of resistance were evaluated in plots inoculated with seeds of the phytoparasite Striga hermonthica (Del.) Benth. and in Striga-free (control) plots for three seasons between 1991 and 1993. Resistant genotypes, although showing similar levels of underground infection as susceptible 9 weeks after maize planting (WAP), had significantly fewer emerged parasitic plants and sustained lower damage. Host damage was not determined by the severity of infection. Percent height and dry matter reductions increased from 3 to 6 WAP for resistant and susceptible genotypes. While percent height and dry matter reductions for resistant genotypes declined at 9 and 12 WAP, susceptible genotypes either maintained or increased their levels of damage. Correlation between the severity of height and dry matter reductions at the vegetative (3 and 6 WAP) and reproductive (9 and 12 WAP) stages of maize growth were not significant, indicating that damage at early stages of growth cannot be used to predict mature plant response. Maize damage score (1–9) at 8 WAP, a non destructive and composite shoot indicator of host performance under Striga infestation, was significantly correlated (r = 0.88, r = 0.82; p < 0.01) to the level of shoot reduction at mature plant stages. Emerged Striga count at 8 WAP was significantly correlated (r = 0.98, p < 0.01) to the count at 10 WAP, the time when parasite emergence was highest. Under high and uniform infestation, mature plant resistance can be detected at 8 WAP, the onset of flowering in maize. This revised version was published online in July 2006 with corrections to the Cover Date.     

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.     

Populations of maize with more rapid and reliable seedling emergence than cornbelt dents at low temperatures

Summary Selected races, populations and genotypes of maize (Zea mays L.) from regions where maize is sown under cool conditions were evaluated in controlled-environment rooms for time to seedling emergence and percentage emergence at low temperatures. The objective of the study was to identify populations with the ability to emerge more rapidly and more reliably than Cornbelt Dent, the race most widely used to produce cultivars for temperate regions.Several populations emerged markedly faster and more reliably than Cornbelt Dent populations. All of these populations contained germplasm of highland Mexican origin, except for Gaspé Flint, and the fastest population in each experiment always contained a high proportion of germplasm from the Cónico race. The advantage of populations containing Cónico germplasm was greatest in environments where emergence was slowest.     

Mass selection for improved cold and density tolerance of two maize populations

Summary Data of planting and plant density are two cultural practices influencing grain yield of maize (Zea mays L.). Our study was designed to evaluate the usefulness of a mass selection scheme to improve cold and density tolerance of the BS2 and BS3 maize populations. Populations were planted at an early planting date and a high plant density, and three cycles of mass selection for well-filled ears on erect plants were conducted at earch of three Corn Belt locations (i.e., Waseca, MN; Ames, IA; Portageville, MO).Results showed that selection improved cold tolerance traits of BS3, but not of BS2. Mass selection did not increase density tolerance of either population at any location. Selection did not improve response to planting dates, although the early planting date did improve agronomic performance and grain yield of all entries. We concluded that mass selection at high plant densities and early planting dates at diverse geographical locations did not produce cycles adapted to specific environmental conditions.     

Variation among Andean races of maize for cold tolerance during heterotrophic and early autotrophic growth

Cold in the initial growth stages is an important stressfactor for maize grown in regions with a temperate climate,particularly in case of early sowing. Sources of tolerancehave been identified in adapted genotypes, but promisinggenes for cold tolerance should also be found in materialdeveloped under the lower-temperature margins of the cropdistribution. This research was conducted in order to testAndean maize accessions for cold tolerance expressed duringboth the heterotrophic and early autotrophic growth stages.Experiments were conducted in controlled environments tostudy cold tolerance traits (germination %, germinationindex and plant growth rate) at continuous 10^°C (heterotrophic growth) and at varying 10–16^°C (autotrophic growth). An experiment was also performed inthe field with early sowing (both heterotrophic and autotrophic growth). In each experiment, a control trialwas conducted in more favourable conditions (i.e. continuous25^°C in a controlled environment or late planting inthe field) so that cold tolerance traits could also beexamined as the ratio between the stress and the controltrial. None of the accessions was superior for all coldtolerance traits. However, several Andean maize accessionsoutperformed the US Corn-belt hybrid checks for one or moretraits, both in heterotrophic and autotrophic growth. Overall, BOZM 855, PMS 636, Poblacion D, Poblacion E andBOZM 696 were the best accessions, suggesting that they canbe a promising source of genes for improving cold toleranceof adapted maize genotypes.     

Evaluation of maize plant introductions for cold tolerance

Summary We evaluated cold-tolerance responses of 144 plant introductions (PI) of maize (Zea mays L.), attempting to include in our sample at least one PI adapted to each of the countries or ecological zones respresented in the maize germplasm collection at the North Central Regional Plant Introduction Station at Ames, Iowa, USA. Plant introductions were grown for 42 days in plastic boxes (26 cm long × 19 cm wide × 10 cm deep) in a growth chamber maintained at 10±1 °C. Cold-tolerance responses of each PI were evaluated by three traits: 1) percentage emergence (recorded 30 days after planting), 2) emergence index, an estimate of rate of emergence, and 3) seedling dry weight (sampled 42 days after planting). Estimates of variances and heritabilities for these three traits were large, suggesting that genotypic variation for cold tolerance in the maize germplasm collection would be sufficient to permit selection advance. Furthermore, genotypic correlations among the three traits were high: therefore, improvement by index selection should be possible. Correlations between the cold-tolerance traits and days from planting to 50% silk emergence (an estimate of maturity obtained at Ames. Iowa, USA) were low. Days from planting to 50% silking emergence for the 25 most cold-tolerant PI's ranged from 46 for PI 214279 from Canada to 106 for PI 331440 from Ethiopia. It should be possible, therefore, to develop cold-tolerant genotypes adapted to all latitudes.Contribution from the Iowa Agriculture and Home Economics Experiment Station, Ames, Iowa 50011, and the Agricultural Research Service, U.S. Department of Agriculture, cooperating. Journal Paper No. J-8780 of the Iowa Agriculture and Home Economics Experiment Station, Ames, Iowa. Projects No. 1018 and 2152     

Variation and inheritance of response to acetochlor among maize inbred lines and hybrids

Summary The variation of response to acetochlor was studied in a two-year experiment carried out by subjecting 18 maize (Zea mays L.) inbred lines to three herbicide rates (0, 2.5 and 5 l a.i./ha). In both years some inbred lines consistently exhibited an evident susceptibility, with symptoms consisting of the seedling curling up below the soil surface and causing impaired field emergence. The results were poor plant density and lower grain yield in comparison to control. In contrast, other lines showed a satisfactory level of tolerance.Then, to gather data on the inheritance of response to acetochlor, four tolerant inbreds (T) and four susceptible inbreds (S) were crossed to obtain four T×T, four S×S, four S×T and the corresponding four T×S two-way hybrids. These hybrids were studied together with parental lines by applying the same herbicide rates used in the previous trial. The S×S hybrids showed susceptibility to the herbicide and the T×T were tolerant, whereas the S×T and the T× S hybrids showed a tolerance very close to that of the T×T hybrids. No difference was found between S×T and the corresponding T×S hybrids as to herbicide response. On average, the 16 hybrids exhibited greater tolerance than the eight parental lines, with each hybrid group being more tolerant than its parental line group. These results indicate that tolerance to acetochlor is prevailingly dominant, that action of extranuclear genes should be ruled out, and that the level of plant vigour can affect herbicide reactiveness.     

Effects of parents and planting dates on R-nj expression in maize

Summary Two sets of diallel crosses and a study using the cultivar Cudu were conducted to determine the extent to which the expression of the R-nj allele for purple aleurone pigmentation was influenced by the color level of the pollen parent, planting date, and the maternal genotype. Neither planting date nor color level of the pollen parent substantially modified relative color expression among hybrids. Maternal parent genotype primarily determined color expression. The data indicated that R-nj color differences did not differ among reciprocal crosses used as female parents. Expression of R-nj appeared to be modified by many loci.Journal Article No. 1295 of the North Dakota Agri. Exp. Stn., Fargo. This material is based upon work supported by the U.S. Department of Agriculture under Agreement No. 5901-0410-9-03337-0.     

Effects of recurrent selection for grain yield on plant and ear traits of five maize populations

Summary Maize (Zea mays L.) breeders are interested in the effects of recurrent selection for grain yield on other traits. Changes in plant traits could alter agronomic acceptability of the populations under selection, and observed improvements in grain yield could be explained by changes in ear traits. We evaluated changes in combining ability for plant and ear traits of BS10(FR), BS11(FR), BSSS(R), BSCB1(R), and Lancaster Surecrop associated with recurrent selection for grain yield.Recurrent selection procedures generally did not change plant and ear heights or date of silking of testcrosses of populations or of the population crosses, BS10(FR)×BSS11(FR) and BSSS(R)×BSCB1(R). Grain yield improvements, however, were associated with increases in ear-sink size.Journal Paper No. J 9517 of the Iowa Agriculture and Home Economics Exp. Stn., Ames, Iowa. Project 2152.     

A selection procedure for ear drying-rates in maize

Summary A selection procedure to change the drying rates of maize (Zea mays L.) ears was developed and tested. Results indicated that drying rates were affected by hybrid genotype, ear maturity at harvest, number of kernels per row, ear diameter, and moisture content at harvest.Mass selection was initiated in an early synthetic, NDSG, in an attempt to produce both fast and slow drying strains. After two cycles resultant substrains were evaluated in the laboratory for direct and in the field for correlated selection responses. Laboratory results indicated that selection effectively changed moisture loss rates in NDSG, and confirmed earlier observations that an ear's moisture content at harvest affects its drying rate.Data from field experiments grown at five locations in 1980 revealed that each of two selection cycles for slow laboratory drying rate significantly reduced ear moisture content at harvest, in the field. This lower harvest moisture content appeared to result from a lower moisture content at physiological maturity rather than a faster drying rate. Other correlated selection responses included lowered yield, plant height, and ear weight due to selection for fast laboratory drying, and lowered plant height and ear weight from selection for slow drying.Data indicated that this selection procedure can be used to change moisture loss rate and ear moisture content at harvest. Results also contributed to understanding of factors involved in ear drying rates which may lead to more effective selection procedures.Journal Article No. 1385 of the North Dakota Agri. Exp. Stn., Fargo.     

Controlling the release of pollen from genetically modified maize and increasing its grain yield by growing mixtures of male-sterile and male-fertile plants

There is public concern about the consequences of pollen dispersal from genetically modified (GM) crops. There lease of viable pollen from GM maize can be controlled by growing mixtures of cytoplasmic male-sterile plants and male-fertile non-transformed pollinator plants. Our experiments indicate that such associations can bring about grain yields as high or even higher than those produced by pure male-fertile maize crops, especially when the male-sterile component is pollinated non-isogenically. The grain yield benefits from cytoplasmic malesterility and xenia as well as the fact that seed of male-sterile varieties can be produced cheaply and reliably in large quantities would facilitate the implementation of the proposed system in agricultural practice. This revised version was published online in July 2006 with corrections to the Cover Date.     

Inheritance of tillering and flowering-time in early maturing maize

Summary A diallel cross and F₂ populations derived from eight early maturing maize inbreds were used to investigate the inheritance of tillering and flowering-time (anthesis), and the possible relationship between tillering and flowering-time. Incomplete dominance for increased tillering was observed; potence ratios, representing the overall degree of dominance, ranged from 0.26 to 0.52. Dominance for early flowering ranged from incomplete with a potence ratio of 0.55 to overdominance with a potence ratio of 1.40. Broad-sense heritabilities were low for both characters. The genetic component of variation for tillering was due to general combining ability effects; specific combining ability effects were not significant. A significant negative linear relationship between tillering and flowering-time was found. Lack of independent assortment of tillering and flowering-time in F₂ populations indicated that the two characters are genetically related.     

Hybrid reconstruction in maize

Summary The production of seed of the maize single cross hybrid F68^*NE2 is uneconomic because of the low grain yield of the maternal line. Therefore the aim was to produce it from newly developed inbred lines obtained by reshuffling the genes in the hybrid, accompanied by selection. Thus in open pollinating generations derived from this hybrid, i.e. in C0, C1, C2, C3 and C4, honeycomb selection for grain yield improvement was applied. Selfing of one ear and open pollination of another ear of selected prolific C4 plants yielded 20 pairs of S1/half sib progenies. Plants grown from remnant S1 seed corresponding to superior progeny pairs were selfed. In each S2-line a single plant was selected and selfed. The S3-lines were evaluated for yield. Two S3-lines, i.e. 6D and 2B, attracted attention because they yielded two and a half times as much as the best commercial inbred line B73.The S1-and S2-lines were tested for combining ability with the related inbred lines NE2 and F68 by means of honeycomb design experiments and for combining ability with unrelated, freely available inbred lines by means of randomized complete block designs. Two S2-lines, i.e. 5C and 6E, were selected for their good combining ability. The six single cross hybrids produced by crossing the four S3-lines 6D, 2B, 5C, and 6E were compared with the original hybrid F68^*NE2 in a honeycomb design at two sites. The grain yields of the single cross hybrids 6D^*6E and 5C^*6E were similar to that of F68^*NE2. However, these two reconstructed hybrids can be produced in a cheaper way because the new maternal inbred lines yield as good as B73 (line 5C) or much better (line 6D).     

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     

Genetic and environmental effects on abscisic acid accumulation in leaves of field-grown maize

Summary This study analyzes the components of phenotypic variation for abscisic acid (ABA) content in maize (Zea mays L.) leaves and the correlations with drought sensitivity index (DSI) and silk delay (SD), involved in the reaction to water deficit. Eight early- and seven medium-maturity inbreds were examined in field trials: in 1990 with low irrigation volume and in 1991 with low and high irrigation volumes. ABA concentration and DSI were investigated at growth stages (S) corresponding to stem elongation (S3), appearance of the first husks (S4), and mid-end of silking (S5). The ABA concentration was significantly higher in conditions of water deficit and in the later growth stage. The genetic component for ABA concentration attained higher relative values than those shown by DSI in the same growth stages and by SD; moreover, it increased from growth stage 3 to stage 5. The genotype × year and genotype × irrigation volume interactions were smaller for ABA concentration than for DSI and SD. The broad sense heritability on a plant basis, estimated in drought conditions, for ABA concentration ranged from 21.4 to 55.1% according to maturity group and growth stage. A wide variation was observed among lines for ABA concentration: the medium-maturity group showed a three-fold range (from 219 to 605 ng ABA g^–1 dry weight). No clear relationships between ABA concentration, DSI and SD were found. These results indicate the feasibility of a selection for ABA concentration within segregating populations derived from crosses between the inbred lines herein tested.Abbreviations ABA - abscisic acid - DSI - drought sensitivity index - DW - dry weight - SD - silk delay     

Relationships among American and Spanish populations of maize

Summary Two experiments were carried out with two objectives. First, to establish the phenetic relationships among the maize (Zea mays L.) landraces from Galicia (Northwestern Spain) maintained at the Misión Biológica de Galicia. Second, to assess the resemblance between a collection of Spanish populations (including the landraces from Galicia) and a set of US Corn Belt varieties. For the first objective 73 varieties from Galicia, along with 9 hybrid checks, were grown in 9×9 simple lattices at two locations for two years. For the second objective 131 populations from the US Corn Belt and Spain, along with 9 hybrid checks, were grown for three years in unreplicated experiments. Cluster analyses were carried out with the first principal components that accounted for a significant amount of the total variation. Four groups were found among the landraces from Galicia. The populations from Spain and America were classified as belonging to nine main groups. The replicated experiment was more accurate than the unreplicated one. However, it is concluded that an unreplicated test grown in several environments is accurate enough to detect the main groups, although some inaccuracies should be expected.     

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     

Properties of maternal haploid maize plants and potential application to maize breeding

Summary Presented are the results of a two-year study of haploid maize plants in the field. The haploids were produced with the aid of inducer line ZMS. In total, 604 and 1030 haploids were obtained and studied in the first and second years, respectively. Tassels of haploid plants were found to be almost completley sterile. Fertility of ears was studied by pollinating them with the pollen from diploid inbred lines, the cross resulting in almost all of the haploid ears carrying kernels. On average 27.4 kernels per ear of haploid plant were obtained in the first year of study and 26.3 in the second. These gave rise to normal diploid plants. This property allows genotypes selected at the level of haploid plants to be involved in breeding process. Unusual plants were found among haploids, phenotypically resembling homozygous lines. It was assumed that the plants had resulted from spontaneous chromosome doubling in haploids. The results of comparative studies of progenies of unusual plants and inbred lines derived from the same synthetic population are presented.