Effects of 2-iodobenzanilide on potassium uptake, H+ extrusion and K+-stimulated ATPase of corn roots

The carboxanilide systemic fungicide 2-iodobenzanilide (2-IB) after 2 h pretreatment at 0.25 m M inhibited K⁺ and SO₄^2- uptake by excised corn roots ( Zea mays L., cv. Dekalb 342) up to ca 70 and 40%, respectively. Proton extrusion from corn roots was also reduced by ca 50% after 1 h contact, and the microsomal K⁺-stimulated ATPase activity from corn roots and pea stems ( Pisum sativum L., cv. Alaska) inhibited by 50 and 72%, respectively. In contrast, the Mg²⁺-ATPase activities of microsomes and mitochondria at pH 6.0 and 8.7, respectively, were unaffected. After 2 h of preincubation with 0.25 m M 2-IB, O₂ consumption by corn roots and pea stems was inhibited by 12 and 18%, respectively. ATP content of corn roots was not altered by 2-IB treatment. Therefore, energy availability "in vivo" was unaffected and the primary effect on corn roots is suggested to be at the plasmalemma ATPase which forms the proton gradient.
With isolated pea stem mitochondria, 0.25 m M 2-IB inhibited O₂ consumption by ca 60% when NADH or malate plus pyruvate were added as substrates; when succinate was used O₂ consumption was unaffected. The mode of action on isolated mitochondria was different from that shown for carboxin and also formerly attributed to the whole class of carboxanilide fungicides.     

Comparison of trypsin inhibitors from several types of corn

Trypsin inhibitors were isolated from seeds of floury-2 corn, dent corn, and popcorn and found to be similar in physicochemical and immunological properties to the inhibitor that was previously isolated from seeds of opaque-2 corn. Thus, very similar and perhaps identical trypsin inhibitors occur in genetically diverse types of corn. Our results therefore suggest that the differences between the opaque-3 inhibitor and the amino acid sequence reported by Hochstrasser et al. for a trypsin inhibitor from an unspecified type of corn, do not stem from variations in source material.     

Characterization and genetic variability of Fusarium verticillioides strains isolated from corn and sorghum in Brazil based on fumonisins production, microsatellites, mating type locus, and mating crosses

Fusarium verticillioides (Gibberella fujikuroi mating population A) is a producer of fumonisins and one of the main contaminants of corn grains. In Brazil, some studies analyzing strains isolated from corn have demonstrated high levels of fumonisins, whereas the levels for strains isolated from sorghum have been found to be low. In the present study, we investigated the genetic diversity of 22 F. verticillioides strains isolated from corn and 21 strains isolated from sorghum cultivated in the State of S?o Paulo, Brazil. Differences in the genetic profile were observed between the strains isolated from the two substrates using single primer amplification reaction by polymerase chain reaction (SPAR-PCR). Fumonisins levels were higher in strains isolated from corn than in those isolated from sorghum. The MAT-1 and MAT-2 alleles were identified by PCR, and the isolates were subsequently crossed with Fusarium thapsinum (G. fujikuroi mating population F) reference strains because this species is morphologically similar to F. verticillioides and produces low levels of fumonisins. The SPAR haplotypes of some strains isolated from sorghum were similar to the F. thapsinum reference strain haplotypes, but there was no fertile mating between the strains isolated from the two substrates and the F. thapsinum references strains. The MAT-1:MAT-2 proportion was 5:17 and 14:7 for isolates from corn and sorghum, respectively.     

Pathogens associated with southwestern corn borers and southern corn stalk borers (Lepidoptera: Crambidae)

A study was undertaken to isolate entomopathogens of southwestern corn borer, Diatraea grandiosella Dyer, and southern corn stalk borer, Diatraea crambidoides (Grote). Field-collected diapausing larvae of southwestern corn borer (three sites in Mississippi) and southern corn stalk borer (one site in North Carolina), and a laboratory strain of D. grandiosella in the diapause state were maintained in a simulated winter followed by a simulated spring environnent. Few larvae (< or = 6%) collected from any of the field sites died in the winter environment, and most insect mortality (11-25%) occurred after transfer of the larvae to the simulated spring environment. Mortality during the simulated spring period differed among the collection sites, and the highest mortality was recorded for southwestern corn borers from Washington County (25%), followed by Marshall (16%) and Oktibbeha (11%) Counties. A high level of mortality was also observed in southern corn stalk borers during the simulated spring period (27%). No viruses were observed, but a number of bacteria, microsporidia, and fungi were isolated from both southwestern corn borer and southern corn stalk borer larvae and pupae. In most instances, numerous bacterial taxa were isolated from cadavers, but on some occasions a single taxon predominated. The most prevalent bacterial taxon from larval and pupal cadavers was Enterococcus faecalis (Andrewes & Horder) Schleifer & Kilppel-Balz, but Bacillus spp., Pseudomonas aeruginosa (Schroeter) Migula, and Serratia marcescens Bizio were frequently isolated as well. Few fungi (1-7%) were recovered from southwestern corn borer and southern corn stalk borer larvae and pupae. The most common entomopathogenic taxon isolated was Beauveria bassiana (Balsamo) Vuillemin from southern corn stalk borer larvae. Microsporidia were not isolated from southern corn stalk borers. However, Nosema spp. were isolated from southwestern corn borer cadavers from Washington (15%), Marshall (1%), and Oktibbeha (3%) Counties in Mississippi. In addition, we observed parasitism of southern corn stalk borer larvae by Macrocentrus cingulum Reinhard (Hymenoptera: Braconidae). No parasitism of southwestern corn borers was observed. Isolates of Bacillus, Beauveria, Entercoccus, Nosema, Pseudomonas and Serratia were all pathogenic to southwestern corn borer larvae under controlled environmental conditions, and with the exception of B. bassiana, these are novel pathogens of Diatraea corn borers.     

Actinomycetales from Corn

Mesophilic Actinomycetales were isolated from whole corn, brewer's grits, and break flour received from three different mills. In addition, strains were isolated from high-moisture (27%) field corn; high-moisture, silo-stored corn (untreated); and high-moisture corn treated with ammonia, ammonium isobutyrate, or propionic-acetic acid. According to standard techniques, 139 strains were extensively characterized and 207 additional strains were partially characterized. On the basis of these characterizations, the streptomycete strains were identified by both the systems of Pridham et al. and Hütter because these systems are rapid and accurate. In general, only Streptomyces griseus (Krainsky) Waksman and Henrici was isolated from high-moisture whole corn (treated or untreated) except from grain exposed to ammonium isobutyrate. Strains isolated from high-moisture corn subjected to that treatment represented both S. griseus and S. albus (Rossi Doria) Waksman and Henrici. The strains isolated from corn and corn products from the three mills were identified with a number of streptomycete species. Of all Actinomycetales isolated, only three were not streptomycetes—two from brewer's grits and one from break flour.     

Plant nucleases. I. Separation and purification of two ribonucleases and one nuclease from corn

Three enzymes with ribonuclease activity, one of which also had deoxyribonuclease activity, have been isolated and partially purified from corn seeds and seedlings. The purification of Ribonuclease I from mature seed was previously reported. This enzyme has a pH optimum near 5.0, is loosely adsorbed to carboxymethyl-cellulose, and has a molecular weight of 23,000, determined by gel filtration.Ribonuclease II was isolated from the microsomes of corn roots, and was partially purified by gel filtration. It has a pH optimum plateau from 5.4 to 7.0, and molecular weight of 17,000.Nuclease I hydrolyzes both RNA and DNA. It was isolated from the large particles of a corn root homogenate and was partially purified on a carboxymethyl-cellulose column. It has a pH optimum at 6.2 and a molecular weight of 31,000.The relative activities of the 3 enzymes for deoxyribonuclease and at pH 5 and pH 6.2 for ribonuclease may be used to characterize them during purification operations. Assays on homogenates of corn roots, and especially of the root tips, suggested that a fourth enzyme, which possesses deoxyribonuclease activity, is also present.     

Effect of climatic conditions on natural mycoflora and fumonisins in freshly harvested corn of the State of Paraná, Brazil

Natural mycoflora associated with fumonisins were analyzed in 150 samples of freshly harvested corn from Central-Southern, Central-Western and Northern regions of the State of Paraná, Brazil and correlated to climatic conditions. The corn samples were frequently contaminated with Fusarium sp.(98.7 to 100%) and Penicillium sp. (93 to 100%), when compared to Aspergillus sp. (not detected to 27.7%). The highest contamination with potentially mycotoxigenic fungi occurred in corn harvested in the Central-Western region, where total mould and yeast counts ranged from 5.5 × 10³ to 5.2 × 10⁶ CFU/g, with 98.7% contaminated byFusarium sp. and 93% by Penicillium sp. In this region F. moniliforme (F. verticillioides) was the predominant Fusariumsp., and was isolated in 85.9% of the samples. Aspergillus sp. was isolated from 27.7% samples. FB₁ was detected in 100% of the samples (mean of 2.39 g/g) and FB₂ in 97.7% (mean of 1.09 g/g). Fumonisins were also detected in all samples from Northern region, with mean of 4.56 g/g (FB₁) and 2.20 g/g (FB₂).Considering 1.0 g/g as the threshold, 72% of the corn samples from the Central-West and 92% from the North were contaminated with concentrations above this value, in contrast to a 18.5% contamination rate from Central-Southern samples. Between corn planting to harvesting season, the average maximum temperature and relative humidity were 26 °C and 77.1%(Central-Southern), 27 °C and 69% (Northern)and 29.9 °C and 89.1% (Central-Western).Therefore, the higher fumonisins contamination of corn from Northern region when compared to the Central-South were due to the differences in rainfall levels (92.8 mm in Central-Southern, 202 mm in Northern) during the month preceding harvest.This revised version was published online in October 2005 with corrections to the Cover Date.     

Plant Nucleases. II. Properties of Corn Ribonucleases I and II and Corn Nuclease I

Three enzymes with ribonuclease activity, one of which also had deoxyribonuclease activity, have been isolated and partially purified from corn seeds and seedlings. The purification of Ribonuclease I from mature seed was previously reported. This enzyme has a pH optimum near 5.0, is loosely adsorbed to carboxymethyl-cellulose, and has a molecular weight of 23,000, determined by gel filtration.

Ribonuclease II was isolated from the microsomes of corn roots, and was partially purified by gel filtration. It has a pH optimum plateau from 5.4 to 7.0, and molecular weight of 17,000.

Nuclease I hydrolyzes both RNA and DNA. It was isolated from the large particles of a corn root homogenate and was partially purified on a carboxymethyl-cellulose column. It has a pH optimum at 6.2 and a molecular weight of 31,000.

The relative activities of the 3 enzymes for deoxyribonuclease and at pH 5 and pH 6.2 for ribonuclease may be used to characterize them during purification operations. Assays on homogenates of corn roots, and especially of the root tips, suggested that a fourth enzyme, which possesses deoxyribonuclease activity, is also present.

     

Murine B-cell mitogenic properties of corn proteins.

Extracts of corn have been found to induce mitosis in human peripheral blood and mouse splenic lymphocytes. The present investigation was initiated to characterize the mitogenic components of corn. Various classes of proteins such as albumins, globulins, zeins, and glutelins were isolated from defatted corn meal. With the exception of corn zeins, all classes of corn proteins possessed mitogenic activity for murine spleen cells. Because of the extreme insolubility of corn glutelins the present investigation was carried out only with corn albumins and globulins. These two classes of proteins stimulated spleen lymphocytes from C3H/HeN, C3H/HeJ, and athymic nu/nu mice as well as nylon-wool fractionated mouse B lymphocytes. Both corn albumins and globulins consist of a complex mixture of proteins. By gel filtration on Sephadex G-100 a low-molecular-weight protein (MW 12,000), which possessed maximum mitogenic activity, has been isolated from corn albumins.     

β-Carotene inhibition of aflatoxin biosynthesis amongAspergillus flavus genotypes from Illinois corn

Thirty-nineAspergillus flavus genotypes (DNA fingerprinting) isolated from corn grown in a field near Kilbourne, Illinois were evaluated for their sensitivity to β-carotene (50 μg/ml) inhibition of aflatoxin B₁ biosynthesis. Inhibition of aflatoxin was greater than 90% for 28 of the genotypes and >70% for 38 of the 39 genotypes. FiveA. flavus strains (4 fingerprint groups) isolated from molded raw peanuts, NRRL 3239, NRRL 3357, NRRL 6514, NRRL 6515 and NRRL 13135, produced greater quantities of aflatoxin than all 39 genotypes isolated from corn, and were less sensitive to β-carotene inhibition.Aspergillus flavus NRRL 3357 is commonly used as inoculum in variety trials for aflatoxin resistance. Isolate identity and sensitivity to potential inhibitors in corn can be critical in assessing corn resistance to aflatoxin.     

Isolation and identification of xylitol-producing yeasts from agricultural residues

Selected yeast strains isolated from corn silage and viticulture residues were screened for their capacities to convert D-xylose into xylitol A conventional TLC was adapted for easy determination of xylose and xylitol in the culture supernatant solutions. This technique is suitable for the first steps of a screening program to select xylitol-producing yeasts from natural environments. Candida tropicalis ASM III (NRRL Y-27290), isolated from corn silage, appears to be a promising strain for xylitol production with a high yield (0.88 g xylitol per g of xylose consumed).     

Plant development from isolated microspores of Zea mays L.

This paper reports the development of plants from mechanically isolated microspores of corn (Zea mays). Large populations of corn microspores were isolated using technology previously developed for rapeseed. Embryos and callus were developed from microspores in the late uninucleate stage. Scutellar-type embryos developed after two weeks and these could be transferred and germinated on a hormone free medium. However, the large majority of plants recovered from embryos developed only upon transfer to a corn embryogenic callus medium. These embryos produced shoots through organogenesis, and subsequently could be induced to form roots. Plants were developed from these colonies and grown in the greenhouse. The frequency of mature plants developed from the embryos was approximately 5 %. Non embryogenic callus which developed from some microspores have thus far either failed to develop or have developed only roots. Seed set has been obtained on some of the regenerated plants.     

一株抗玉米纹枯病内生细菌的分离鉴定及其抗病促生作用

玉米纹枯病是玉米主要的真菌病害,从优良玉米品种川单13号中分离到一株对玉米纹枯病菌具有明显抑制作用并能够促进玉米生长的内生细菌.通过形态、生理生化特性分析以及16SrDNA序列同源性比较,鉴定该菌株为枯草芽孢杆菌.盆栽试验表明,菌株能够抑制玉米纹枯病的发生,抑菌率为 43.01%,促生试验表明,该菌能够显著促进玉米苗的生长,促生作用与植物生长刺激素(IAA)和效果相似,显示出良好的开发前景.     

Immunoadsorbent isolation of trypsin inhibitors from corn and teosinte seeds

An immunoadsorbent was constructed by covalently coupling 100 mg of affinity-purified antibodies against corn trypsin inhibitor to cyanogen bromide-activated agarose. With the immunoadsorbent, we could purify the trypsin inhibitor in aqueous extracts of corn (Zea mays) seeds in a single chromatographic step. The immunoadsorbent was found to have the capacity to bind about 6 mg of corn inhibitor. Recovery of exclusively virgin corn inhibitor required the addition of protease inhibitors to the corn extract. The virgin inhibitor isolated by use of the immunoadsorbent was found to be indistinguishable from the single-chain inhibitor recovered previously from chromatography of corn extracts on trypsin-agarose (Swartz, M.J., Mitchell, H.L., Cox, D.J. and Reeck, G.R. (1977) J. Biol. Chem. 252, 8105–8107). The immunoadsorbent was also used to isolate trypsin inhibitor seeds of teosinte (Zea mexicana), which is believed to be the progenitor of corn.     

Cloning and characterization of the bgxA gene from Erwinia chrysanthemi D1 which encodes a β-glucosidase/xylosidase enzyme

A -glucosidase/xylosidase gene from Erwinia chrysanthemi strain D1 was cloned and sequenced. This gene, named bgxA, encodes a ca. 71 kDa protein product which, following removal of the leader peptide, resulted in a ca. 69 kDa mature protein that accumulated in the periplasmic space of E. chrysanthemi strain D1 and Escherichia coli cells expressing the cloned gene. The protein exhibited both -glucosidase and -xylosidase activities but gave no detectable activity on xylan or carboxymethyl cellulose. The enzyme was classified as a type 3 glycosyl hydrolase, but was unusual in having a truncated B region at the carboxyl-terminus. Several E. chrysanthemi strains isolated from corn produced the glucosidase/xylosidase activity but not those isolated from dicot plants. However, bgxA marker exchange mutants of strain D1 were not detectably altered in virulence on corn leaves.     

Fumonisin occurrence in corn from high- and low-risk areas for human esophageal cancer in China.

Forty-seven corn samples were collected in 1989 from Linxian and Shangqiu Counties in Henan Province, the high- and low-risk areas, respectively, for human esophageal cancer in the People's Republic of China. The samples were analyzed for fumonisin (fumonisin B1 [FB1] and FB2) contamination. Of the fumonisin-positive samples, the mean levels in Linxian corn were found to be 872 ng/g for FB1 and 448 ng/g for FB2, while the Shangqiu corns had 890 ng of FB1 and 330 ng of FB2 per g. The incidence of fumonisin contamination of Linxian corn (48%) was about two times higher than that of Shangqiu corn (25%), and the former corn samples were frequently cocontaminated with trichothecenes. Fusarium species isolated from corn from Linxian County produced FB1 at levels ranging from 1,280 to 11,300 micrograms/g.     

木质素降解菌shu-0801降解玉米废弃物的研究

目的:研究分离到的木质素降解菌降解玉米废弃物效果.方法:利用苯胺蓝法从土壤中分离到木质素降解菌shu-0801.结果:研究表明shu-0801木质素降解菌能够显著提高纤维素的转化,提高纤维素酶的降解效率,还原糖的生成量明显提高,shu-0801菌与玉米秸秆粉共培养,可降解玉米废弃物干重约20%.结论:分离到木质素菌降解木质素效率较高,属巨大芽孢杆菌属.     

Systemic infection of stalks and ears of corn hybrids by <Emphasis Type="Italic">Aspergillus parasiticus</Emphasis>

This study was conducted to explore systemic infection by the Aspergillus flavus group into corn ears via the stalk. An A. parasiticus mutant which produces norsolorinic (NOR) acid (a visible orange intermediate of the aflatoxin biosynthetic pathway) was used in field studies to monitor systemic infection of corn stalk and ear tissues. Corn hybrids resistant and susceptible to aflatoxin contamination were grown in the field and inoculated prior to tasseling by inserting A. parasiticus infested toothpicks into stalks between the 5th and 6th node below the lowest ear shoot. Beginning 2 weeks after inoculation, systemic infection by the NOR mutant was assessed weekly by collecting ear shank tissue and stalk tissue from the nodes between the infection sites and the developing ears. Ears were collected at the end of the growing season to determine the level of kernel infection by the NOR mutant. In two separate studies, the A. parasiticus NOR mutant was isolated from stalk tissues at all of node positions and ear shank tissue from several susceptible corn hybrid plants at the first harvest date 2 weeks after inoculation. The NOR mutant was also isolated from stalk and ear tissue of a resistant hybrid. The NOR mutant was only isolated from kernels of susceptible hybrids in 2003 and 2004. Infection rates of kernels in infected ears were very low (<1%). In 2005, the fungus was found in only one kernel from an ear of the resistant hybrid. The NOR mutant was not isolated from stalks, ears, or kernels from control (uninoculated) plants grown in the plots with inoculated plants. Although infection levels of corn kernels were low, systemic movement of the A. parasiticus up the stalk appears to be another possible route to infection of developing corn ears.     

Differential Effects of Lithium Chloride on In Vitro Growth of Clavibacter michiganense subsp. nebraskense Depending upon Inoculum Source

The bacterium Clavibacter michiganense subsp. nebraskense (Corynebacterium michiganense subsp. nebraskense) was grown in broth cultures and inoculated into corn plants. The plating efficiency of cells from broth cultures was essentially the same on nutrient broth-yeast extract and the semiselective medium for this bacterium, CNS. However, when cells were isolated from Goss bacterial wilt- and blight-infected corn, very few were recovered on CNS compared with the amount recovered on nutrient broth-yeast extract agar. When lithium chloride was omitted from the CNS, recoveries from infected corn were nearly the same as on nutrient broth-yeast extract agar. No other ingredient of CNS was inhibitory, nor did substitution of other salts for lithium chloride cause equal inhibition. The amount of inhibition was proportional to lithium chloride concentration. The inhibition by lithium chloride occurred with several strains of the bacterium isolated from one corn cultivar and with one of the strains recovered from three different cultivars of infected corn.     

Isolation of Zeanic Acid,a Natural Plant Growth-regulator from Corn Steep Liquor and Its Chemical Structure

Corn steep liquor markedly inhibited the growth of the grass, alfalfa, as previously reported. The active principles from corn steep liquor were isolated. From the acidic fraction of corn steep liquor, white needles and yellow needles were isolated. The compound of white needles, which inhibited the germination of alfalfa seeds at the concentrations above 500 ppm, was found to be identical with ferulic acid known as germination inhibitor of plant seeds. The compound of yellow needles did not inhibit the germination of alfalfa seeds, but stimulated the growth of the plant. It has a structure related to β-acid which has been isolated from rice-bran and has been identified as 2,6-dihydroxycinchoninic acid. This active substance is a new quinoline, 2,8-dihydroxycinchoninic acid and was named zeanic acid.