Delaying corn rootworm resistance to Bt corn

Transgenic crops producing Bacillus thuringiensis (Bt) toxins for insect control have been successful, but their efficacy is reduced when pests evolve resistance. To delay pest resistance to Bt crops, the U.S. Environmental Protection Agency (EPA) has required refuges of host plants that do not produce Bt toxins to promote survival of susceptible pests. Such refuges are expected to be most effective if the Bt plants deliver a dose of toxin high enough to kill nearly all hybrid progeny produced by matings between resistant and susceptible pests. In 2003, the EPA first registered corn, Zea mays L., producing a Bt toxin (Cry3Bb1) that kills western corn rootworm, Diabrotica virgifera virgifera LeConte, one of the most economically important crop pests in the United States. The EPA requires minimum refuges of 20% for Cry3Bb1 corn and 5% for corn producing two Bt toxins active against corn rootworms. We conclude that the current refuge requirements are not adequate, because Bt corn hybrids active against corn rootworms do not meet the high-dose standard, and western corn rootworm has rapidly evolved resistance to Cry3Bb1 corn in the laboratory, greenhouse, and field. Accordingly, we recommend increasing the minimum refuge for Bt corn targeting corn rootworms to 50% for plants producing one toxin active against these pests and to 20% for plants producing two toxins active against these pests. Increasing the minimum refuge percentage can help to delay pest resistance, encourage integrated pest management, and promote more sustainable crop protection.     

Bt corn has a higher lignin content than non-Bt corn

Bt corn has been genetically modified to express the Cry1Ab protein of Bacillus thuringiensis to kill lepidopteran pests. Fluorescence microscopy and staining with toluidine blue indicated a higher content of lignin in the vascular bundle sheaths and in the sclerenchyma cells surrounding the vascular bundle in all ten Bt corn hybrids, representing three different transformation events, studied than of their respective non-Bt isolines. Chemical analysis confirmed that the lignin content of all hybrids of Bt corn, whether grown in a plant growth room or in the field, was significantly higher (33-97% higher) than that of their respective non-Bt isolines. As lignin is a major structural component of plant cells, modifications in lignin content may have ecological implications.     

Extruding foams from corn starch acetate and native corn starch

Because of the hydrophilic characteristics of native starch foams and the cost of modifying starch, the uses of starch and modified starch foams are hindered. To decrease hydrophilicity and cost of starch foams, native corn starch was blended with starch acetate and extruded. A twin-screw mixing extruder was used to produce the foams. Native starch content, screw speed, and barrel temperature had significant effects on molecular degradation of starches during extrusion. The melting temperature of extruded starch acetate/native starch foam was higher (216 degrees C) than that for starch acetate (193.4 degrees C). Strong peaks in the X-ray diffractograms of extruded starch acetate/native starch foam suggested new crystalline regions were formed. Optimum conditions for high radial expansion ratio, high compressibility, low specific mechanical energy requirement, and low water absorption index were 46.0% native starch content, 163 rpm screw speed, and 148 degrees C barrel temperature.     

Insecticidal proteins from Bacillus thuringiensis protect corn from corn rootworms.

Field tests of corn co-expressing two new delta-endotoxins from Bacillus thuringiensis (Bt) have demonstrated protection from root damage by western corn rootworm (Diabrotica virgifera virgifera LeConte). The level of protection exceeds that provided by chemical insecticides. In the bacterium, these proteins form crystals during the sporulation phase of the growth cycle, are encoded by a single operon, and have molecular masses of 14 kDa and 44 kDa. Corn rootworm larvae fed on corn roots expressing the proteins showed histopathological symptoms in the midgut epithelium.     

Management of corn leafhopper (Homoptera: Cicadellidae) and corn stunt disease in sweet corn using reflective mulch

Plastic reflective mulches significantly reduced populations of corn leafhopper, Dalbulus maidis (DeLong & Wolcott), adults and the incidence of corn stunt disease caused by Spiroplasma kunkelii (CSS) in late planted sweet corn (Zea mays L.). The reflective mulches were more effective than were either foliar or soil applied insecticides in managing both the leafhopper and the pathogen it transmits. Yields of marketable ears were 1.5 to 2 times greater in reflective mulch plots than from fallow plots. This was due to larger ears (individual ear weight and length) rather than an increase in the number of ears. The use of reflective mulches provides an alternative strategy to insecticides in the management of both D. maidis and corn stunt disease. Such a strategy may prove useful to growers in Latin America and to limited resource growers and organic growers in the United States who wish to grow corn without the use of insecticides.     

Incidence and levels of fumonisin contamination in Colombian corn and corn products

A survey was conducted to determine the levels of fumonisins B1 and B2 in corn and corn-based products available in Colombia for human and animal consumption. A total of 120 samples were analyzed by acetonitrile-water extraction, cleanup with a strong-anion-exchange column, and liquid chromatography with o-phthaldialdehyde-2-mercaptoethanol derivatization and fluorescence detection. The samples of corn and corn-based products for animal intake were taken at different feed manufacturing plants, whereas the samples used for human foods where purchased from local retail stores. The number of positive samples for fumonisin B1 was 20.0% higher in corn and corn-based products for animal intake (75.0%) than in corn and corn-based products for human consumption (55.0%). The levels of fumonisin B1 were also higher in corn and corn-based products for animal intake (mean = 694 μg/kg; range = 32–2964 μg/kg), than in corn and corn-based products for human intake (mean = 218 μg/kg; range = 24–2170 μg/ kg). The incidence and levels of fumonisin B2 were lower than those for fumonisin B1. Corn and corn-based products for animal consumption had an incidence of fumonisin B2 of 58.3%, with a mean value of 283 μg/kg, and a range of 44–987 μg/kg. The incidence of fumonisin B2 in corn-based products for human intake was 35.0%, with a mean value of 118 μg/kg and a range of 21–833 μg/kg. The highest incidence and levels of fumonisins were found in samples of hominy feed, with concentrations ranging from 86 to 2964 μg/kg fumonisin B1 and 57 to 987 μg/kg fumonisin B2.     

Patch clamping corn protoplasts

Summary Cell wall regeneration around protoplasts from Black Mexican Sweet corn suspension cells has been observed using scanning electron microscopy. A coherent array of cellulose microfibrils can be seen around protoplasts two hours after they have been isolated. This array does not form in the presence of 15 mg/l Congo Red. The frequency and electrical resistance of seals made between patch clamp pipettes and the plasmalemma around corn protoplasts is not significantly affected by the presence or absence of these fibrils (p₀=0.75); it remains relatively low. Some single channel records from BMS corn protoplasts are shown.     

采用玉米秸秆水解糖和玉米浆发酵生产丁二酸

研究了以玉米秸秆水解糖为碳源,不同氮源条件下琥珀酸放线杆菌Actinobacillus succinogenesSF-9的丁二酸发酵产酸能力。结果表明玉米浆可以替代酵母膏作为丁二酸发酵的廉价氮源。厌氧摇瓶丁二酸发酵单因素试验,得到在初糖浓度50 g/L时,玉米浆的较佳用量为20 g/L。在5 L搅拌罐上,考察了不同初始玉米秸秆水解糖浓度对A.succinogenes SF-9发酵生产丁二酸的影响,结果显示高初始秸秆糖浓度对琥珀酸放线杆菌的生长有抑制作用。采用补料分批发酵,发酵60 h丁二酸的产量达到42.7g/L,丁二酸产率82.7%,生产强度0.81 g/(L·h)。丁二酸的产量和生产强度较分批发酵有明显提高。     

RAPD-analysis of corn somaclones

The genetic difference between maize line A188 and A188-derived somaclones was assessed via analysis of randomly amplified polymorphic DNA (RAPD). In total, 15 out of 17 decanucleotide primers used each allowed amplification of 2-17 fragments ranging 200-2000 bp. The RAPD patterns did not differ between individual plants of line A188, which demonstrated again its high genetic homogeneity. The difference between the initial line and the somaclones was high, ranging 64-74%. On evidence of the genetic divergence, the somaclones formed two clusters. The distribution of somaclones between these clusters was consistent with their origin.     

Transgenic corn for control of the European corn borer and corn rootworms: a survey of Midwestern farmers' practices and perceptions

In 2001, a self-administered questionnaire was sent to 1000 corn, Zea mays L., farmers in each of five states (Illinois, Indiana, Iowa, Minnesota, and Nebraska) to evaluate their perceptions of transgenic corn designed to control the European corn borer, Ostrinia nubilalis (Hübner), and corn rootworms, Diabrotica spp. Respondents returned 1,313 surveys (26.2%). Farmers with small acreages planted a greater portion of their corn (54.5%) with transgenic corn for control of European corn borer than farmers with large farms (39.2%). The majority (75.2%) of farmers use crop rotation to control the corn rootworm. Nine insecticides comprised 92.2% of the commercial soil insecticides used for control of corn rootworm larvae. More than one-third of the farmers in Illinois (33.5%) and Indiana (39.4%) treated first-year corn for corn rootworm, primarily due to western corn rootworm egg laying in soybean, Glycine max (L.). When asked whether they would plant transgenic corn protected against the corn rootworm, 35.0% of farmers responded they would, whereas 40.5% said they were unsure. The two greatest farmer concerns about transgenic corn were the ability to sell harvested grain (59.3%) and additional technology fees (54.8%). Respondents indicated that less farmer exposure to insecticide (69.9%) and less insecticide in the environment (68.5%) were the primary benefits of transgenic corn. Farmers who had no concerns about transgenic corn for rootworm control were more likely to purchase the product (46.8%). The most common refuge-planting options farmers favored were adjacent fields (30.9%) and split fields (29.9%). Farmers (21.1%) observed a yield increase (23.7 bu/ha [9.6 bu/acre]) when using transgenic corn for European corn borer control compared with non-transgenic corn. These data can help in understanding farmers' knowledge and concerns regarding transgenic corn. This information may be of value to guide researchers, extension specialists, and policy makers in designing insect resistance management and integrated pest management programs.     

Insect-damaged corn stalks decompose at rates similar to Bt-protected, non-damaged corn stalks

The relative decomposability of corn (Zea mays L.) residues from insect (Bt)-protected hybrids and conventional hybrids cultivated under insect pressure was investigated in two studies. Above-ground biomass, residue macromolecular composition, and stalk physical strength were also measured. In the first decomposition study, chopped residues (stalks and leaves) were used from a corn rootworm-protected (Cry3Bb1) hybrid and its non-Bt near isoline that were grown in replicated plots infested with corn rootworms (Diabrotica spp.). In the second study, residue (intact stalk sections) was used from three European corn borer (ECB, Ostrinia nubilalis Hübner)-resistant (Cry1Ab) hybrids representing different seed manufacturer/maturity date series, their non-Bt near isolines, two Cry3Bb1-protected isolines, and three additional conventional hybrids, all cultivated in replicated plots under conditions of elevated ECB pressure. In both studies, insect-resistant residues decomposed at rates similar to their non-protected near isolines. No evidence was found that insect-protected hybrids produced more above-ground biomass or had distinct residue composition. While some measures of mechanical stalk strength indicated that ECB-damaged stalks were not as stiff as protected stalks, these physical differences did not translate into differences in residue decomposition. We conclude that while individual hybrids may vary in their production of biomass, residue composition or residue decomposability, these characteristics do not systematically vary with the presence of the Bt gene conferring insect resistance, even under conditions of insect pressure.     

Selection for resistance to mCry3A-expressing transgenic corn in western corn rootworm

To investigate the development of resistance to mCry3A, a laboratory colony of the western corn rootworm, Diabrotica virgifera virgifera LeConte, was established from field survivors of mCry3A-expressing (MIR604) corn, Zea mays L. Feral adults emerging from MIR604 (selected) and isoline (control) field plots were collected and returned to the laboratory. Progeny of each colony was reared one generation on isoline corn and then crossed reciprocally with a nondiapausing colony. The resulting nondiapausing progeny were then reared on greenhouse corn in accordance with the wild type parent's origin (on MIR604 or isoline corn). After four, seven, and 10 total generations of selection, the resistance ratio of the selected colony was 0.5, 4.3, and 15.4 in terms of lethal concentration (LC)50 values in toxicity assays, with the latter two LC50 values being significant. After seven generations of selection in total, selected and control colonies were screened on MIR604 and isoline corn under field conditions. There was a significant colony x corn pedigree interaction in terms of plant damage. There was no significant difference in damage between MIR604 and isoline corn, whereas this difference was significant for the control colony. After 14 generations of selection, a seedling bioassay was performed. Again, there was a significant colony x corn pedigree interaction, this time in terms of the number of larvae recovered. There was no significant difference in the number of larvae recovered from MIR604 and isoline corn for the selected colony, whereas this difference was significant for the control colony, although larval size was greater on isoline corn for both colonies. Resistance has developed in western corn rootworm laboratory colonies to all Bt proteins currently registered for corn rootworm management, which emphasizes the importance of adhering to resistance management plans for maintaining product efficacy.     

BT799玉米对亚洲玉米螟抗性研究

【目的】抗螟性鉴定是转基因抗虫玉米研发的重要一环。本文主要就转基因玉米BT799对亚洲玉米螟的抗性展开评价,同时测定了BT799植株组织中Cry1Ac蛋白的表达量。【方法】采用了酶联免疫吸附测定法(ELISA)、室内生测和田间人工接虫鉴定3种方法。【结果】转基因抗虫玉米BT799组织中Cry1Ac蛋白含量分别为768.0 ng/g(蛋白/鲜叶重)和1 452.8~2 978.5 ng/g(蛋白/花丝、苞叶或幼嫩籽粒干重)。田间心叶期接虫条件下转基因抗虫玉米BT799和CC-2XBT799表现高抗亚洲玉米螟。室内取食转Cry1Ac基因玉米不同组织的亚洲玉米螟敏感幼虫7 d后的存活率为0~37.5%,取食非转基因对照的存活率为89.9%~100.0%。亚洲玉米螟不同抗性品系取食郑单958K组织的存活率以Cry1Ie抗性品系最低,其次是Cry1F抗性品系,均显著低于取食对照郑单958,Cry1Ac抗性品系最高,与对照差异不显著。【结论】转Cry1Ac基因玉米BT799对亚洲玉米螟有很高的杀虫作用和良好的田间抗螟性。