玉米遗传转化与商业化转基因玉米开发*

玉米是世界上种植面积最大的粮食作物,为了提高玉米产量和满足人类需求,转基因育种已经成为改良玉米性状的有效手段。自1996年美国种植商业化转基因玉米以来,利用玉米遗传转化技术开发商业化转基因玉米已取得巨大成功。综述了玉米遗传转化体系优化的重要步骤,系统总结了已开发的商业化转基因玉米的种类,并对玉米遗传转化未来发展方向进行了展望。     

Tissue localization of maize acetylcholinesterase associated with heat tolerance in plants

Our recent study reported that maize acetylcholinesterase (AChE) activity in the coleoptile node is enhanced through a post-translational modification response to heat stress and transgenic plants overexpressing maize AChE gene had an elevated heat tolerance, which strongly suggests that maize AChE plays a positive, important role in maize heat tolerance. Here we present (1) maize AChE activity in the mesocotyl also enhances during heat stress and (2) maize AChE mainly localizes in vascular bundles including endodermis and epidermis in coleoptile nodes and mesocotyls of maize seedlings.     

Chemical and physical properties of ginkgo (Ginkgo biloba) starch

Starch isolated from mature Ginkgo biloba seeds and commercial normal maize starches were subjected to α-amylolysis and acid hydrolysis. Ginkgo starch was more resistant to pancreatic α-amylase hydrolysis than the normal maize starch. The chain length distribution of debranched amylopectin of the starches was analyzed by using high performance anion-exchange chromatography equipped with an amyloglucosidase reactor and a pulsed amperometric detector. The chain length distribution of ginkgo amylopectin showed higher amounts of both short and long chains compared to maize starch. Naegeli dextrins of the starches prepared by extensive acid hydrolysis over 12 days demonstrated that ginkgo starch was more susceptible than normal maize to acid hydrolysis. Ginkgo dextrins also demonstrate a lower concentration of singly branched chains than maize dextrins, and unlike maize dextrin, debranched ginkgo shows no multiple branched chains. The ginkgo starch displayed a C-type X-ray diffraction pattern, compared to an A-type pattern for maize. Ginkgo starch and maize starch contained 24.0 and 17.6% absolute amylose contents, respectively.     

两类不同玉米田蚜虫发生动态数学模型与空间分布型测定

蚜虫是我国各地玉米植株上的重要刺吸类害虫。为了进一步阐述该虫的发生与环境之间的关系,以及其在转基因抗性玉米上的空间分布,本文在田间系统调查蚜虫种群动态的基础上,使用多元回归分析法、Iwao回归分析法和5种常用指标参数法,探明了玉米田蚜虫发生与环境因素之间相关的数学模型,以及在常规玉米和抗性玉米植株上的空间分布。结果表明：江苏扬州地区两类不同玉米植株上发生的蚜虫以玉米蚜为主,玉米蚜虫的发生与当日最低气温关系最密切;玉米蚜虫在玉米植株上部的分布数量可达70%~80%,其全株玉米上的蚜量（y）与玉米植株上部1/3上的蚜量（x）关系为y=1.33x;玉米蚜虫在植株上的分布为聚集分布,其聚集由自身特性和环境因素共同确定。本研究为开展转基因玉米田蚜虫的测报防控提供了重要依据。     

Systematic analysis and comparison of nucleotide-binding site disease resistance genes in maize

Nucleotide-binding site (NBS) disease resistance genes play an integral role in defending plants from a range of pathogens and insect pests. Consequently, a number of recent studies have focused on NBS-encoding genes in molecular disease resistance breeding programmes for several important plant species. Little information, however, has been reported with an emphasis on systematic analysis and a comparison of NBS-encoding genes in maize. In the present study, 109 NBS-encoding genes were identified based on the complete genome sequence of maize (Zea mays cv. B73), classified as four different subgroups, and then characterized according to chromosomal locations, gene duplications, structural diversity and conserved protein motifs. Subsequent phylogenetic comparisons indicated that several maize NBS-encoding genes possessed high similarity to function-known NBS-encoding genes, and revealed the evolutionary relationships of NBS-encoding genes in maize comparede to those in other model plants. Analyses of the physical locations and duplications of NBS-encoding genes showed that gene duplication events of disease resistance genes were lower in maize than in other model plants, which may have led to an increase in the functional diversity of the maize NBS-encoding genes. Various expression patterns of maize NBS-encoding genes in different tissues were observed using an expressed-sequence tags database and, alternatively, after southern leaf blight infection or the application of exogenous salicylic acid. The results reported in the present study contribute to an improved understanding of the NBS-encoding gene family in maize.     

基于比较基因组学的玉米ESTs定位方法

描述了以水稻基因组数据和玉米与水稻的比较遗传图谱为桥梁,基于水稻和玉米间存在的标记和序列水平上的广泛的共线性,对大量的玉米ESTs初步定位于玉米连锁群上新方法,为对ESTs开展进一步的基因组学研究和基因克隆提供参考信息。对139条玉米ESTs的定位发现,96条玉米ESTs（69%）可在水稻基因组中找到同源序列,77条ESTs（55%）可使用该策略进行定位,证实了该方法的可行性和有效性。      

Promotive effect of jasmonates on the senescence of detached maize leaves

Promotion of senescence of detached maize leaves by jasmonates was investigated. Senescence of detached maize leaves was promoted by linolenic acid, the precursor of biosynthesis of jasmonic acid, and retarded by inhibitors of lipoxygenase, the first enzyme in the biosynthetic pathway of jasmonic acid. Results support a role of endogenous jasmonates in the regulation of senescence of detached maize leaves. Silver thiosulfate, an inhibitor of ethylene action, was found to inhibit methyl jasmonate, linolenic acid- and abscisic acid-promoted senescence of detached maize leaves. It seems that jasmonate-promoted senescence is mediated through an increase in ethylene sensitivity in detached maize leaves.Abbreviations ABA abscisic acid - MJ methyl jasmonate - STS silver thiosulfate     

Maize pollen foraging by honey bees in relation to crop area and landscape context

The increasing demand for insect pollinated crops and high recent losses of honey bee colonies raise concerns about food security. Systemic insecticides are recognized as one of the drivers of worldwide honey and wild bee declines. Particularly honey bees in agricultural environments are exposed to pesticides when they collect crop pollen and nectar. However, landscape scale studies which analyze pollen use and foraging distances of honey bees on mass-flowering crops like maize to evaluate potential exposure risks are currently lacking. In an experimental approach on a landscape scale we took advantage of intra-colonial dance communication to gather information about the location of utilized pollen resources. During maize flowering, four observation hives were placed in and rotated between 11 different landscapes which covered a gradient from low to high maize acreage. A higher frequency of dances for foraging locations on maize fields compared to other land use types shows that maize is an intensively used pollen resource for honey bee colonies. Mean foraging distances were significantly shorter for maize pollen than for other pollen origins. The percentage of maize pollen foragers did not increase with maize acreage in the landscape. The proportion of grassland area providing alternative pollen sources did not reduce the percentage of maize pollen foragers. Our findings allow estimating the distance-related exposure risk of honey bee colonies to pollen from surrounding maize fields treated with systemic insecticides. Similarly, the results can be used to estimate the exposure to transgenic maize pollen, which is relevant for honey production in European countries. Provision of alternative pollen resources within agri-environmental schemes could potentially reduce exposure risk to pesticide contaminated crop pollen.     

Flow cytometric sorting of maize chromosome 9 from an oat-maize chromosome addition line

Large numbers of maize chromosome 9 can be collected with high purity by flow cytometric sorting of chromosomes isolated from a disomic maize chromosome addition line of oat. Metaphase chromosome suspensions were prepared from highly synchronized seedling root-tips of an oat-maize chromosome-9 addition line (OM9) and its parental oat and maize lines. Chromosomes were stained with propidium iodide for flow cytometric analysis and sorting. Flow-karyotypes of the oat-maize addition line showed an extra peak not present in the parental oat line. This peak is due to the presence of a maize chromosome-9 pair within the genome of OM9. Separation of maize chromosome 9 by flow cytometric sorting of a chromosome preparation from a normal maize line was not possible because of its size similarity (DNA content) to maize chromosomes 6, 7 and 8. However, it is possible to separate maize chromosome 9 from oat chromosomes and chromatids. An average of about 6×10³ chromosomes of maize chromosome 9 can be collected by flow-sorting from chromosomes isolated from 30 root tips (ten seedlings) of the oat-maize addition line. Purity of the maize chromosome 9, sorted from the oat-maize chromosome addition line, was estimated to be more than 90% based on genomic in situ hybridization analysis. Sorting of individual chromosomes provides valuable genomic tools for physical mapping, library construction, and gene isolation. Received: 28 February 2000 / Accepted: 14 July 2000     

Comparative Recombination Distances among Zea Mays L. Inbreds, Wide Crosses and Interspecific Hybrids

Recombination distances and linkage heterogeneity were compared among a wide range of maize inbreds, wide crosses and maize X teosinte hybrids. Twelve maize and four teosinte races were backcrossed to stocks fixed for rare marker alleles on chromosome arm 1L. Recombination fraction estimates were higher for exotic germplasm than for either U.S. maize or maize X teosinte crosses. Serrano, Tuxpeno and a US-adapted inbred line of tropical origin, NC300, exhibited enhanced recombination. Three of the four maize X teosinte hybrids had little or no recombination between two loci. The observed recombination ``shrinkage' resulted from an apparent inversion in the vicinity of the Amp1 locus. Average recombination distances among common marker loci for composite maps were highly variable, even when map construction was restricted to maize germplasm of similar origins.     

SAD,a stearoyl-acyl carrier protein desaturase highly expressed in high-oil maize inbred lines

As special maize with more than 6% oil concentration in the grain, high-oil maize has received increased interest recently. To date, little is known about the expressions of genes involved in fatty acid metabolism of high-oil maize. Stearoyl-acyl carrier protein desaturase (SAD) is a key enzyme that converts stearic acid to oleic acid. In this study, two-dimensional electrophoresis, gas chromatography, and real-time PCR were used to determine the expressions of SAD at three seed development stages in high-oil and normal maize inbred lines. SAD was significantly more abundantly expressed in high-oil maize than in normal maize, not only at the protein and mRNA levels, but also at the product level. These results suggested that a high expression of SAD may play an important role in increasing oil concentration in high-oil maize.     

NO缓解玉米幼苗盐胁迫伤害的生理机制

以玉米幼苗为材料,通过NO的供体硝普纳(SNP)的合成抑制剂L-NAME和NaN3、清除剂cPTIO组合处理,分析外源NO和IAA对盐胁迫下玉米幼苗生长,以及NO对盐胁迫下玉米幼苗叶片和根尖IAA含量、IOD和POD活性的影响,以探讨NO与IAA在提高植物抗盐性中的关系.结果表明,盐胁迫下,SNP和IAA均能显著促进玉米幼苗株高、主根长和侧根数的增加;SNP能显著提高玉米幼苗叶片和根尖IAA含量,降低IOD和POD活性;L-NAME和NaN3及cPTIO均能有效减弱SNP诱导的IAA含量的增加.由此可见,在盐胁迫条件下,NO信号可能位于IAA信号的上游,它通过促进玉米幼苗內源IAA的积累缓解盐胁迫对其生长的抑制.     

Source of the soybean N credit in maize production

Nitrogen response trials throughout the United States Corn Belt show that economic optimum rates of N fertilization are usually less for maize (Zea mays L.) following soybean (Glycine max L.) than for maize following maize; however, the cause of this rotation effect is not fully understood. The objective of this study was to investigate the source of the apparent N contribution from soybean to maize (soybean N credit) by comparing soil N mineralization rates in field plots of unfertilized maize that had either nodulated soybean, non-nodulated soybean, or maize as the previous crop. Crop yields, plant N accumulation, soil inorganic N, and net soil mineralization were measured. Both grain yield (6.3 vs. 2.8 Mg ha^–1) and above-ground N accumulation (97 vs. 71 kg ha^–1) were greatly increased when maize followed nodulated soybean compared with maize following maize. A partial benefit to yield and N accumulation was also observed for maize following non-nodulated soybean. Cumulative net soil N mineralization following nodulated soybean, non-nodulated soybean, and maize was 112, 92 and 79 kg N ha^–1, respectively. Net mineralization of soil N appeared to be influenced by both quality (C:N ratio) and quantity of residue from the previous crop. In addition to an increase in plant available N from mineralization, the amount of soil inorganic N (especially in soil 5 cm from the row) was greater following nodulated soybean than non-nodulated soybean or maize. Based on these data, the soybean N credit appears to result from a combination of a decrease in net soil mineralization in continuous maize production and an increase in residual soil N from symbiotic fixation.     

Diabrotica virgifera virgifera (Coleoptera: Chrysomelidae) larval feeding behavior on transgenic maize (MON 863) and its isoline

Diabrotica species (Coleoptera: Chrysomelidae) larval behavior studies have posed a challenge to researchers because of the subterranean life cycle of this pest. To fully understand how the western corn rootworm, Diabrotica virgifera virgifera LeConte, injures the maize, Zea mays L., root system, its behavior must be studied. For example, larvae that can detect an area of the root that has a lower amount of toxin, whether from an insecticide or a transgenic maize plant, have an increased chance of survival. This study assessed D. v. virgifera larval feeding behavior on rootworm-susceptible maize and maize containing a biotechnology-derived trait (MON 863) with resistance to D. v. virgifera first instar feeding. Maize plants were grown in a medium that allowed for direct observation and measurements during feeding of larval stadia. Neonates were placed on maize seedlings, and data were taken at 3, 6, 9, and 12 d postinfestation on resistant and susceptible maize. On rootworm-susceptible maize, neonate larvae aggregated at the root tips and began actively feeding, and then they moved to older root tissue. Conversely, some larvae that ingested Cry 3Bb1 from the resistant maize exhibited no movement. Other larvae on the resistant maize moved continuously, sampling root hairs or root tissue but not actively feeding. The continuously moving larvae had visibly empty guts, suggesting possible nonpreference for the resistant root. This study contributes to our understanding of D. v. virgifera larval behavior and provides insight into questions surrounding the potential evolution of behavioral and biochemical resistance to Cry3Bb1.     

Field-evolved resistance to Bt maize by western corn rootworm: predictions from the laboratory and effects in the field

Crops engineered to produce insecticidal toxins derived from the bacterium Bacillus thuringiensis (Bt) provide an effective management tool for many key insect pests. However, pest species have repeatedly demonstrated their ability to adapt to management practices. Results from laboratory selection experiments illustrate the capacity of pest species to evolve Bt resistance. Furthermore, resistance has been documented to Bt sprays in the field and greenhouse, and more recently, by some pests to Bt crops in the field. In 2009, fields were discovered in Iowa (USA) with populations of western corn rootworm, Diabrotica virgifera virgifera LeConte, that had evolved resistance to maize that produces the Bt toxin Cry3Bb1. Fields with resistant insects in 2009 had been planted to Cry3Bb1 maize for at least three consecutive years and as many as 6years. Computer simulation models predicted that the western corn rootworm might evolve resistance to Bt maize in as few as 3years. Laboratory and field data for interactions between western corn rootworm and Bt maize indicate that currently commercialized products are not high-dose events, which increases the risk of resistance evolution because non-recessive resistance traits may enhance survival on Bt maize. Furthermore, genetic analysis of laboratory strains of western corn rootworm has found non-recessive inheritance of resistance. Field studies conducted in two fields identified as harboring Cry3Bb1-resistant western corn rootworm found that survival of western corn rootworm did not differ between Cry3Bb1 maize and non-Bt maize and that root injury to Cry3Bb1 maize was higher than injury to other types of Bt maize or to maize roots protected with a soil insecticide. These first cases of field-evolved resistance to Bt maize by western corn rootworm provide an early warning and point to the need to apply better integrated pest management practices when using Bt maize to manage western corn rootworm.     

The maize genome contains a helitron insertion

The maize mutation sh2-7527 was isolated in a conventional maize breeding program in the 1970s. Although the mutant contains foreign sequences within the gene, the mutation is not attributable to an interchromosomal exchange or to a chromosomal inversion. Hence, the mutation was caused by an insertion. Sequences at the two Sh2 borders have not been scrambled or mutated, suggesting that the insertion is not caused by a catastrophic reshuffling of the maize genome. The insertion is large, at least 12 kb, and is highly repetitive in maize. As judged by hybridization, sorghum contains only one or a few copies of the element, whereas no hybridization was seen to the Arabidopsis genome. The insertion acts from a distance to alter the splicing of the sh2 pre-mRNA. Three distinct intron-bearing maize genes were found in the insertion. Of most significance, the insertion bears striking similarity to the recently described DNA helicase-bearing transposable elements termed HELITRONS: Like Helitrons, the inserted sequence of sh2-7527 is large, lacks terminal repeats, does not duplicate host sequences, and was inserted between a host dinucleotide AT. Like Helitrons, the maize element contains 5' TC and 3' CTRR termini as well as two short palindromic sequences near the 3' terminus that potentially can form a 20-bp hairpin. Although the maize element lacks sequence information for a DNA helicase, it does contain four exons with similarity to a plant DEAD box RNA helicase. A second Helitron insertion was found in the maize genomic database. These data strongly suggest an active Helitron in the present-day maize genome.     

Maize pyruvate decarboxylase mRNA is induced anaerobically

A cDNA was identified using an oligonucleotide designed by comparing the sequences of bacterial and yeast pyruvate decarboxylase. The sequence of the cDNA identified by the oligonucleotide contained an open reading frame that encoded a protein of 65 kDa that was similar in sequence to bacterial and yeast pyruvate decarboxylase. This protein was selectively precipitated by an antiserum specific for maize PDC. Northern-blot analysis shows that PDC mRNA is anaerobically induced. Southern-blot analysis of maize genomic DNA indicated that the maize PDC gene has a single or low copy number.     

Traditional Maize Storage Methods of Mayan Farmers in Yucatan, Mexico: Implications for Seed Selection and Crop Diversity

For small-scale farmers who maintain genetically diverse crop populations, aspects of the storage of their seeds and harvest may be just as important for successful farming as those related to productivity. The community of Yaxcaba, Yucatan, Mexico was studied to understand how the conditions under which Maya farmers store their maize harvests influence their seed selection practices and the diversity of maize varieties grown. Most farmers select their maize seed based primarily on ear characteristics and secondarily on grain characters. Farmers incorporate storability concerns by selecting for an archetypal healthy ear and by conducting selection in several steps between harvest and planting of the subsequent crop generation. Most farmers store their maize harvest in the husk, initially in the field and then in rustic granaries constructed of logs and palm thatch, in which farmers conserve separately their different seed lots. All local landrace populations show morphological adaptation (principally husk characteristics) for local storage conditions, indicating that storability has been an important selection pressure on traditional maize. Storability also appears to be a key factor working against the straightforward adoption of improved maize seed in Yaxcaba. Local farmers value many qualities of improved maize varieties, but their principal complaint is that improved maize cannot be conserved reliably under local storage conditions. Nearly all farmers who utilize improved seed stock in Yaxcaba grow locally adapted or ‘creolized’ versions of improved varieties, displaying characteristics of local maize landraces that facilitate their storage.