Preliminary Genetic Studies of the Phenotype of Betaine Deficiency in Zea mays L

Glycinebetaine-deficient inbreds of Zea mays do not exhibit a general deficiency of nitrogenous solutes; the total free amino acid levels of betaine-deficient lines are not significantly less than those of inbreds which exhibit >100-fold higher betaine levels. Betaine-deficient inbreds are characterized by extremely low betaine: total free amino acid ratios (<0.0015). Highly significant correlations are demonstrated between the expected mid-parent and observed betaine:amino acid ratios of 30 hybrids of known pedigree. In 12 hybrids constructed from a betaine-deficient male parent (inbred 1506), the observed betaine:amino acid ratios of the hybrids are proportional to the betaine:amino acid ratios of the female parents (r = 0.83). Two hybrids, 1146 × 1074 and 1146 × 1506, were chosen for further genetic analysis. The common female parent (1146) and inbred 1074 both exhibit betaine:amino acid ratios of 0.090, a value which is approximately 90-fold greater than the betaine:amino acid ratio of inbred 1506. Hybrid 1146 × 1074 exhibits almost exactly twice the betaine:amino acid ratio of hybrid 1146 × 1506. If inbred 1506 is homozygous recessive for a single nuclear gene responsible for the phenotype of betaine deficiency, and if inbreds 1146 and 1074 are homozygous dominant for this allele, then this twofold difference in betaine:amino acid ratio must be associated with the homozygous dominant and heterozygous conditions, respectively, for 1146 × 1074 and 1146 × 1506. Evidence is presented from both greenhouse and field evaluations of F₂ populations of these hybrids that a single nuclear recessive gene is most likely responsible for the phenotype of betaine-deficiency in inbred 1506. Approximately 25% of the F₂ segregants from 1146 × 1506 exhibited extremely low betaine:amino acid ratios (<0.0015), whereas 0% of the F₂ segregants from 1146 × 1074 exhibited this phenotype. The segregation patterns with respect to betaine:amino acid ratio suggest a 1:2:1 segregation ratio for homozygous recessive:heterozygous:homozygous dominant individuals within the 1146 × 1506-F₂ population.     

Changes in the intracellular pool of free amino acids of Streptococcus lactis under different cultivation conditions]

The intracellular pool of free amino of Streptococcus lactis--lysine producer contains a good number of amino acids when cultivated on the corn medium. Glutamic acid, proline, alanine, lysine, leucine, histidine and arginine are in predominance. An almost complete amino acid pool develops at an early exponential phase of Str. lactis growth under stationary cultivation conditions. The content of free amino acids increases 4-fold during the transition from the early exponential phase to the stationary phases under submerged cultivation conditions. This can be attributed to a more intensive amino acid exchange during the medium stirring than during stationary cultivation.     

Penetration and Development of Meloidogyne incognita in Roots of Resistant and Susceptible Corn Genotypes

Rates of penetration and development ofMeloidogyne incognita race 4 in roots of resistant (inbred Mp307, and S4 lines derived from the open-pollinated varieties Tebeau and Old Raccoon) and susceptible (Pioneer 3110) corn genotypes were determined. Seedlings grown in styrofoam containers were inoculated with 5,000 eggs of M. incognita. Roots were harvested at 3-day intervals starting at 3 days after inoculation (DAI) to 27 DAI and stained with acid fuchsin. Penetration of roots by second-stage juveniles (J2) at 3 DAI was similar for the four corn genotypes. Meloidogyne incognita numbers in Tebeau, Old Raccoon, Mp307, and Pioneer 3110 peaked at 12, 12, 15, and 27 DAI, respectively. Nematode development in the resistant genotypes was greatly suppressed compared to Pioneer 3110. Resistance to M. incognita in these genotypes appears to be expressed primarily as slower nematode development rather than differences in J2 penetration.     

Molecular cloning,sequence and expression analysis of <Emphasis Type="Italic">ZmArf2</Emphasis>, a maize ADP-ribosylation factor

A full-length cDNA encoding a maize GTP-binding protein of the ADP-ribosylation factor family was cloned by suppression subtractive hybridization and an in silico cloning approach. The cDNA was 938 bp in length and contained a complete ORF of 612 bp, which encodes a protein of 203 amino acid residues. Its deduced amino acids sequence had an 83% identity with that of a GTP-binding protein in rice. The gene was designated ZmArf2. The ZmArf2 gene consists of G1, G2, G3, G4 and G5 boxes, and Switch I and Switch II regions. Eight nucleotides differed and five amino acids changed between the popcorn inbred N04 and the dent corn inbred Dan232. One changed amino acid was in the G1 box. RT-PCR analysis showed that ZmArf2 expression increased in the early stages of endosperm development and was not tissue-specific.     

Development of two isogenic sweet corn hybrids differing for glycinebetaine content

A hybrid of sweet corn, Zea mays L. (`1720'; Rogers Brothers Seed Co.), was found to be comprised of glycinebetaine-positive and glycinebetaine-deficient individuals in a 1:1 mixture. This phenomenon was traced to segregation for a single, nuclear, dominant gene determining leaf glycinebetaine content within the female inbred parent of this hybrid. Selection for homozygous recessive (glycinebetaine-deficient) and homozygous dominant (glycinebetaine-positive) genotypes of the female inbred parent enabled production of two isogenic versions of hybrid `1720' differing with respect to a single copy of the dominant allele, by mating these female parent selections with the common homozygous recessive (glycinebetaine-deficient) male parent. These two isogenic hybrids are shown to differ by a factor of 300- to 400-fold in glycinebetaine titer of young expanding leaves of salinized plants, but exhibit no striking differences in the levels of free amino acids or the level of N-methylnicotinic acid (nicotinic acid betaine; trigonelline). The only significant difference between the two hybrids in terms of amino acid composition was found to be in the level of alanine under nonsalinized conditions. The betaine-deficient hybrid exhibited a 14% lower alanine level than the betaine-positive hybrid. Betaine deficiency was not associated with altered stress-induced accumulation of amino acids such as proline, serine, and asparagine plus aspartate, attesting to the high specificity of the genetic difference between these isogenic hybrids with respect to betaine accumulation. This germplasm offers unique opportunities to test whether a single dominant allele determining stress-induced betaine accumulation capacity influences stress resistance in maize.     

Performance of transgenic corn hybrids in Missouri for insect control and yield

The efficacy of Bacillus thuringiensis-transformed corn (Zea mays L.) hybrids compared with comparable nontransformed corn hybrids for controlling first- and second-generation European corn borer, Ostrinia nubilalis (Hübner), and second-generation southwestern corn borer, Diatraea grandiosella Dyar, was determined. Yield comparisons were obtained from the same plots of corn hybrids. Both generations of European and the second-generation of southwestern corn borer were effectively controlled, but the Bt hybrids varied in degree of control. Hybrids from Ciba Seeds, DEKALB, and Mycogen had more European corn borer tunneling than those from Novartis or Cargill, and this was generally ascribed to different transgenic events. The Bt-transformed hybrids had virtually no leaf-feeding damage and less tunneling than the non-Bt corn hybrids. Some Bt corn hybrids had no tunneling, whereas other Bt hybrids had a small amount of tunneling. All of the non-Bt hybrids had significant leaf-feeding damage and stalk tunneling from both insects. Only three live European corn borer larvae (stunted) were found in the Bt corn hybrids while splitting stalks to assess tunnel length. When insect damage was significant, and in some evaluations where damage was not significant, differences in yields among hybrids were observed. No significant insect population differences were observed for five genera of beneficial insects for Bt versus non-Bt corn hybrids. Corn hybrids that have been transformed with the Bt gene provide an effective means of control for corn borers and efforts to reduce the likelihood of development of borer resistance are warranted.     

Meiotic behavior as a selection tool in silage corn breeding

In breeding programs, commercial hybrids are frequently used as a source of inbred lines to obtain new hybrids. Considering that maize production is dependent on viable gametes, the selection of populations to obtain inbred lines with high meiotic stability could contribute to the formation of new silage corn hybrids adapted to specific region. We evaluated the meiotic stability of five commercial hybrids of silage corn used in southern Brazil with conventional squashing methods. All of them showed meiotic abnormalities. Some abnormalities, such as abnormal chromosome segregation and absence of cytokinesis, occurred in all the genotypes, while others, including cytomixis and abnormal spindle orientation, were found only in some genotypes. The hybrid SG6010 had the lowest mean frequency of abnormal cells (21.27%); the highest frequency was found in the hybrid P30K64 (44.43%). However, the frequency of abnormal meiotic products was much lower in most genotypes, ranging from 7.63% in the hybrid CD304 to 43.86% in Garra. Taking into account the percentage of abnormal meiotic products and, hence, meiotic stability, only the hybrids CD304, P30K64, SG6010, and P30F53 are recommended to be retained in the breeding program to obtain inbred lines to create new hybrids.     

施氮水平对小麦籽粒发育过程中氨基酸含量的影响

施氮能提高小麦籽粒蛋白质氨基酸的含量,并与施氮水平呈正相关;但对普通小麦必需氨基酸与蛋白质氨基酸的比值没有影响,而硬粒小麦4286随施氮水平的提高,该比值下降。在开花后32d以前,籽粒发育过程中游离氨基酸与施氮水平呈正相关,以后,籽粒中游离氨基酸趋于相近,表明施氮增加了游离氨基酸的库源,不同基因型小麦对施氮水平的反应不同,在同等施氮水平和栽培条件下,籽粒中蛋白质氨基酸和游离氨基酸含量为硬粒小麦4286＞小偃6号＞小偃107,不同施氮水平下,籽粒中氨基酸含量为高氮＞中氮＞低氮。     

Nutritional suitability of corn pollen for the predator Coleomegilla maculata (Coleoptera: Coccinellidae)

The nutritional suitability of corn pollen for the facultatively phytophagous predator Coleomegilla maculata was studied in the laboratory. Dry matter, organic matter, ash, crude protein, amino acid, and quercetin contents of pollen from 10 hybrids of field corn were determined. C. maculata were reared on pollen or aphids + artificial diet for their entire lives; larval duration, post-mortem adult dry weights, fecundity within 7 days of mating, and mortality rates were compared among the treatments. In another experiment, C. maculata larvae were reared on pollen; weight gained, pollen ingested, and frass produced were compared among instars. Also, consumption relative to increases in larval biomass and the efficiency with which larvae converted corn pollen into biomass were compared among instars. Beetles reared on aphids had greater weights and fecundity and a shorter larval duration relative to the pollen-fed beetles. The percentages of organic matter and ash in corn pollen were significantly correlated with C. maculata mortality, and we hypothesize that some micronutrient or phytochemical is at sub-optimal levels for C. maculata development in some of the pollens. We observed an increase in the conversion efficiency of pollen and a decrease in the consumption relative to biomass of C. maculata as the larvae aged, which suggests a physiological or behavioral alteration in the feeding behavior of C. maculata during the larval stage.     

Field screening of experimental corn hybrids and inbred lines for multiple ear-feeding insect resistance

Identifying and using native insect resistance genes is the core of integrated pest management. In this study, 10 experimental corn, Zea mays L., hybrids and 10 inbred lines were screened for resistance to major ear-feeding insects in the southeastern Coastal Plain region of the United States during 2004 and 2005. Ear-feeding insect damage was assessed at harvest by visual damage rating for the corn earworm, Helicoverpa zea (Boddie), and by the percentage of kernels damaged by the maize weevil, Sitophilus zeamais Motschulsky, and stink bugs [combination of Euschistus servus (Say) and southern green stink bug, Nezara viridula (L.)]. Among the eight inbred lines and two control populations examined, C3S1B73-5b was resistant to corn earworm, maize weevil, and stink bugs. In contrast, C3S1B73-4 was resistant to corn earworm and stink bugs, but not to maize weevil. In a similar manner, the corn hybrid S1W*CML343 was resistant to all three ear-feeding insects, whereas hybrid C3S1B73-3*Tx205 was resistant to corn earworm and maize weevil in both growing seasons, but susceptible to stink bugs in 2005. The silk-feeding bioassay showed that corn earworm developed better on corn silk than did fall armyworm. Among all phenotypic traits examined (i.e., corn ear size, husk extension, and husk tightness), only corn ear size was negatively correlated to corn earworm damage in the inbred lines examined, whereas only husk extension (i.e., coverage) was negatively correlated to both corn earworm and maize weevil damage on the experimental hybrids examined. Such information could be used to establish a baseline for developing agronomically elite corn germplasm that confers multiple ear-feeding insect resistance.     

The constancy and similarity of the amounts of free amino acids in inbred strains of Drosophila and outbred Calliphora

The composition of the free amino acids are compared in adults of two inbred strains of Drosophila subobscura and their hybrids, three inbred lines of Drosophila melanogaster, and in flies from a heterogeneous population of Calliphora erythrocephala. It appears that not only do the amounts of amino acid vary very little from fly to fly, but also very little between inbred lines. Furthermore, the relative amounts of the amino acids in Drosophila are similar to the relative amounts in the blowfly, Calliphora. This characteristic of invariant amounts of the free amino acids in these Diptera occurs in spite of the probably large numbers of genes affecting them.     

Host Suitability of Selected Hybrids,Varieties and Inbreds of Corn to Populations of Meloidogyne spp.

Rates of reproduction of root-knot nematodes on corn varied with Meloidogyne species, with different populations of certain species, and with corn cultivars. M. arenaria, M. incognita and M. javanica reproduced at varying rates on all corn cultivars tested. None of the three selections of M. hapla reproduced on corn. Most of the Meloidogyne populations increased more rapidly on ''Coker'' and ''Pioneer'' hybrids than on ''McNair'' hybrids or on open-pollinated varieties or inbreds. Nematodes often reduced root growth, but the differences within given nematode-cultivar treatments were not usually significant. Root growth of ''Coker 911,'' which supported a high rate of reproduction, was affected less than ''Pioneer 309B'' which supported a low rate of nematode reproduction.     

Ammonium and amino acids as regulators of nitrate reductase in corn roots

When amino acids or ammonia are added to plant systems, the effects on the development of nitrate-dependent nitrate reductase activity are variable. In addition, amino acids added singly or as casein hydrolysate may not support a normal growth. A physiologically correct mixture of amino acids, one similar in composition to amino acids released by the endosperm, has been shown to support normal growth and protein synthesis in corn (Zea mays) embryos. In this investigation, we have used the mixture of corn amino acids to determine whether amino acids have an effect on the appearance or disappearance of nitrate reductase activity. The results show that these amino acids partially inhibit the induction of nitrate reductase in corn roots. The effect is more pronounced in mature root than in root tip sections. When glutamine and asparagine are included along with the "corn amino acid mixture," the inhibition is more severe. Amino acids or amino acid analogues added singly to the induction medium have a similar effect: i.e. when the induction of nitrate reductase is inhibited in the root tips (lysine, canavanine, azaserine, azetidine-2-carboxylic acid, dl-4-azaleucine, asparagine, and glutamine), that inhibition is more severe in mature root sections. Arginine enhanced the recovery of nitrate reductase in root tips but inhibited it in mature root sections. The effect of the amino acids is apparently on some phase of the induction processes (i.e. the uptake or distribution of nitrate or a direct effect on the synthesis of the enzyme) and not on the turnover of the enzyme.     

Evaluating Tripsacum‐introgressed maize germplasm after infestation with western corn rootworms (Coleoptera: Chrysomelidae)

Maize (Zea mays L.) is a valuable commodity throughout the world, but corn rootworms (Chrysomelidae: Diabrotica spp.) often cause economic damage and increase production costs. Current rootworm management strategies have limitations, and in order to create viable management alternatives, researchers have been developing novel maize lines using Eastern gamagrass (Tripsacum dactyloides L.) germplasm, a wild relative of maize that is resistant to rootworms. Ten maize Tripsacum‐introgressed inbred lines derived from recurrent selection of crosses with gamagrass and teosinte (Zea diploperennis Iltis) recombinants and two public inbred lines were assessed for susceptibility to western corn rootworm (Diabrotica virgifera virgifera LeConte) and yield in a two‐year field study. Two experimental maize inbred lines, SDG11 and SDG20, had mean root damage ratings that were significantly lower than the susceptible public line B73. Two other experimental maize inbred lines, SDG12 and SDG6, appeared tolerant to rootworm damage because they exhibited yield increases after rootworm infestation in both years. In the majority of cases, mean yield per plant of experimental maize lines used in yield analyses was equal to or exceeded that of the public inbred lines B73 and W64A. Our study indicates that there is potential to use Tripsacum‐introgressed maize germplasm in breeding programs to enhance plant resistance and/or tolerance to corn rootworms, although further research on insect resistance and agronomic potential of this germplasm needs to be conducted in F₁ hybrids.     

Breadfruit (<Emphasis Type="Italic">Artocarpus altilis</Emphasis>): a source of high-quality protein for food security and novel food products

Protein deficiency has been observed as a leading cause of malnutrition and child death in the tropics. The current study evaluated the protein quality of 49 important breadfruit cultivars (41 Artocarpus altilis and 8 hybrids of A. altilis × A. mariannensis). While significant differences were found between cultivars, all varieties contained a full spectrum of the essential amino acids and are especially rich in phenylalanine, leucine, isoleucine, and valine. The cultivar Ma’afala contained significantly higher total essential amino acid content than other varieties and higher-quality protein than staples such as corn, wheat, rice, soybean, potato, and pea.     

Corn pith weevil,Geraeus penicillus (Coleoptera: Curculionidae), in transgenic,insect‐resistant and conventional field corn hybrids

The corn pith weevil Geraeus penicillus (Herbst) is occasionally found boring in corn stalks throughout the eastern and Midwestern United States. Injury caused by G. penicillus is not typically economical, but may be confused with that of the European corn borer Ostrinia nubilalis (Hubner), an important economic pest throughout the United States. During efforts to assess European corn borer infestations, we discovered G. penicillus in field corn in south‐eastern Pennsylvania, including hybrids genetically modified (i.e. Bt hybrids) to control European corn borer among other herbivore species. Our analysis across sites indicated that tunnels of G. penicillus were significantly more abundant in transgenic Bt hybrids than non‐Bt hybrids, but comparisons of Bt hybrids and their near isolines revealed mostly similar numbers of G. penicillus tunnels, suggesting other hybrid features might be affecting the distribution of G. penicillus. Tunnels of G. penicillus were equally distributed among the three transgenic trait packages represented in our study. In plants where we found G. penicillus, tunnels were more abundant in stalks free of European corn borer damage. Our report appears to be the first to note G. penicillus feeding in Bt corn hybrids. These findings are notable because they document insect damage in Bt hybrids that may be mistaken for European corn borer damage and may provide evidence of an insect herbivore proliferating following a mild winter or possibly even moving into competitor‐free space.     

Determination of Inositol in Inbred and Hybrid Corn (Zea mays) Seedlings

Four inbred (W_f9, Hy₂, B₁₀, and 38–11) plus three hybrid(W_f9 x 38.11, US13, and B₁₀ x Hy2) corn (Zea mays) seedlingstrains (minus seed coat and endosperm) were assayed for totaland free myoinositol content, using a mutant inositol-less straino Neurospora crassa. Over a 120-h germination period, it wasfound that the concentration of total myoinositol decreasedin both of the respective varieties. The inbred corn seedling strains, assayed in this study, appearto demonstrate a lower decrease in total myoinositol than dothe hybrids, over the same 120-h germination period. Concomitantly,the hybrid strains have a higher percentage increase in freemyoinositol than do the inbred varieties, over the same germinationtime period. The hybrid corn seedling strains appear to havea greater capacity to mediate the metabolism of total to freemyoinositol, during the early stages of germination, comparedto the inbred varieties.     

Contibution of the X chromosme to the increased resistance of inbred Drosophila melanogaster hybrids to physical effects]

Effects of high sub-lethal temperature and UV-irradiation on surviving of inbred lines of Drosophila melanogaster and its F1 hybrids derived from reciprocal crosses between these lines are investigated. High resistance of F1 hybrids to these factors was observed as compared with that of parental inbred lines. D. melanogaster females in inbred lines and F1 hybrids were more resistant than males. Differences in the resistance between females and males were more pronounced in hybrids.     

Reproduction of Belonolaimus longicaudatus,Meloidogyne javanica,Paratrichodorus minor,and Pratylenchus brachyurus on Pearl Millet (Pennisetum glaucum)

Pearl millet (Pennisetum glaucum) has potential as a grain crop for dryland crop production in the southeastern United States. Whether or not pearl millet will be compatible in rotation with cotton (Gossypium hirsutum), corn (Zea mays), and peanut (Arachis hypogaea) will depend, in part, on its host status for important plant-parasitic nematodes of these crops. The pearl millet hybrid ''TifGrain 102'' is resistant to both Meloidogyne incognita race 3 and M. arenaria race 1; however, its host status for other plant-parasitic nematodes was unknown. In this study, the reproduction of Belonolaimus longicaudatus, Paratrichodorus minor, Pratylenchus brachyurus, and Meloidogyne javanica race 3 on pearl millet (''HGM-100'' and TifGrain 102) was compared relative to cotton, corn, and peanut. Separate greenhouse experiments were conducted for each nematode species. Reproduction of B. longicaudatus was lower on peanut and the two millet hybrids than on cotton and corn. Reproduction of P. minor was lower on peanut and TifGrain 102 than on cotton, corn, and HGM-100. Reproduction of P. brachyurus was lower on both millet hybrids than on cotton, corn, and peanut. Reproduction of M. javanica race 3 was greater on peanut than on the two millet hybrids and corn. Cotton was a nonhost. TifGrain 102 was more resistant than HGM-100 to reproduction of B. longicaudatus, P. minor, and M. javanica. Our results demonstrated that TifGrain 102 was a poor host for B. longicaudatus and P. brachyurus (Rf < 1) and, relative to other crops tested, was less likely to increase densities of P. minor and M. javanica.     

Manifestation of heterosis during early maize (Zea mays L.) root development

Heterosis is typically detected in adult hybrid plants as increased yield or vigor compared to their parental inbred lines. Only little is known about the manifestation of heterosis during early postembryonic development. Objective of this study was to identify heterotic traits during early maize root development. Four German inbred lines of the flint (UH002 and UH005) and dent (UH250 and UH301) pool and the 12 reciprocal hybrids generated from these inbred lines were subjected to a morphological and histological analysis during early root development. Primary root length and width were measured daily in a time course between 3 and 7 days after germination (DAG) and displayed average midparent heterosis (MPH) of 17–25% and 1–7%, respectively. Longitudinal size of cortical cells in primary roots was determined 5 DAG and displayed on average 24% MPH thus demonstrating that enlarged primary roots of hybrids can mainly be attributed to elongated cortical cells. The number of seminal roots determined 14 DAG showed on average 18% MPH. Lateral root density of all tested hybrids was determined 5 DAG. This root trait showed the highest degree of heterosis with an average MPH value of 51%. This study demonstrated that heterosis is already manifesting during the very early stages of root development a few days after germination. The young root system is therefore a suitable model for subsequent molecular studies of the early stages of heterosis manifestation during seedling development.