Control of preharvest aflatoxin contamination in maize by pyramiding QTL involved in resistance to ear-feeding insects and invasion by Aspergillus spp.

Several resistance sources and resistance mechanisms to aflatoxin formation and corn earworm (Helicoverpa zea Boddie) damage to maize (Zea mays L.) have been identified. Based on this knowledge, experiments were initiated toward achievement of the following objectives: (1) to confirm earlier determinations on resistance traits of germplasm sources and to identify quantitative trait loci (QTL) associated with each of the traits, and (2) upon estimation of the degree of QTL effects on each trait, to generate a maize population, with chemical and physical resistance to Aspergillus spp. and ear-feeding insects, for inbred development. A 2-year field experiment to evaluate selected genotypes inoculated with A. flavus and infested with corn earworm revealed that significant variation exists among the genotypes for aflatoxin contamination and corn earworm damage. The protection of maize ears against aflatoxin contamination was primarily dependent on resistance to fungal infection and ear-feeding insects, and excellent husk coverage and tightness. A major QTL (p1) identified on chromosome 1S had effects of 54.0, 42.1, and 28.3% on the phenotypic variability for concentrations of silk maysin, 3′-methoxymaysin+apimaysin, and chlorogenic acid, respectively. Markers/QTLs for husk phenotypic traits and total aflatoxin concentrations have been determined, but more detailed mapping of these chromosomic regions will be necessary to locate precise markers/QTLs for husk traits and aflatoxin production. Realizing the complexity of the Aspergillus–aflatoxin-maize system and the factors affecting aflatoxin contamination, we are directing our program toward marker-assisted breeding to enhance or improve general genetic resistance to ear-feeding insects and invasion by Aspergillus spp.     

Heterotic affinity and combining ability of exotic maize inbred lines for resistance to aflatoxin accumulation

Aflatoxin accumulation in maize (Zea mays L.) kernels is a serious economic and health problem that reduces grain quality and nutritional values and causes death to livestock and humans. Understanding the genetic parameters and heterotic responses of exotic maize inbred lines can facilitate their use for developing aflatoxin resistant parents of hybrids in Africa. This study was designed to (1) determine the heterotic affinities of aflatoxin resistant exotic lines, (2) identify exotic inbreds with good combining ability, and (3) determine the mode of inheritance of resistance to aflatoxin contamination in these lines. A line?×?tester mating design was used to determine combining ability of 12 yellow and 13 white inbreds and classify them into heterotic groups. The inbreds were crossed to two adapted testers representing two African heterotic groups and the resulting testcrosses along with hybrid checks were evaluated in separate trials at two locations for 2 years in Nigeria. General combining ability (GCA) effects were more important than specific combining ability effects for aflatoxin and grain yield. Among 15 exotic inbred lines having negative GCA effects for aflatoxin and 13 with positive GCA effects for grain yield, six combined the two desired traits. Five white and six yellow endosperm testcrosses were found to be good specific combiners for the two desired traits. The exotic lines with negative GCA effects for aflatoxin accumulation will be used as donor parents to develop backcross populations for generating new inbred lines with much higher levels of resistance to aflatoxin accumulation.     

Development of a gene-based marker correlated to reduced aflatoxin accumulation in maize

Aflatoxins are carcinogenic and toxic metabolites produced by the fungus Aspergillus flavus during infection of maize (Zea mays L.) and other seed oil crops. Climatic conditions in the southeastern United States favor A. flavus infection and aflatoxin contamination in maize, making it a major issue for farmers in the region. One of the most promising avenues to combat aflatoxin contamination is the development of resistant maize lines. However, this has proven difficult due to a lack of reliable markers for resistance. Previous studies have identified candidate genes that were differentially expressed in response to A. flavus infection. One gene, encoding a chloroplast precursor, was found to contain multiple polymorphisms that were used to design a marker designated Mississippi Marker 1 (MpM1). The marker differentiates between the “resistance” and “susceptible” forms of the allele. This marker was used to screen three populations of F_2:3 mapping families, where it was found to map to chromosome 4 and was associated with a significant effect for resistance to aflatoxin accumulation in the Mp313E × B73 mapping population. Furthermore, the marker MpM1 identified a previously unknown quantitative trait loci for resistance to aflatoxin accumulation on maize chromosome 4. MpM1 is the first gene-based marker developed specifically for resistance to aflatoxin accumulation in maize and can now be integrated into existing marker assisted selection programs aimed at incorporating resistance into elite maize breeding lines.     

Stability of maize resistance to the ear rots caused by Fusarium graminearum and F. verticillioides in Argentinian and Canadian environments

Sources of resistance to Fusarium spp. are needed to develop maize hybrids resistant to the accumulation of fungal mycotoxins in the grain. In a search for resistant germplasm in 1999 and 2000, a set of Argentinian maize populations was evaluated in Ottawa, Canada, for resistance to ear rots after inoculation with local isolates of Fusarium verticillioides and F. graminearum. Sixteen of these populations, varying in observed resistance levels, were re-evaluated in 2003 and 2004 in Pergamino, Argentina, using local isolates of the same fungi. Conidial suspensions of each fungal species were inoculated into the silk channel of primary ears. Disease severity was assessed after physiological maturity using a scale based on the percentage of visibly infected kernels. Genotype effect was more important than genotype-by-fungal species or genotype-by-fungal species-by-environment interaction effects. In addition, disease severity levels associated with each fungal species were positively correlated (P < 0.05) (r = 0.90, r = 0.81, r = 0.87 and r = 0.53, in Ottawa 1999 and 2000, and Pergamino 2003 and 2004, respectively). Populations ARZM 01107, ARZM 07138, ARZM 10041, ARZM 13031, ARZM 16002 and Pora INTA exhibited the highest and most stable resistance to both species. Considering that disease resistance exhibited low specificity to the environment and to the fungal species in evaluations conducted in a wide range of environments and with fungal isolates collected from different hemispheres, the most resistant populations are potential sources of genes for stable resistance to these Fusarium spp.     

Pedigree selection for Gibberella ear rot resistance in maize

The pedigree method is often used for developing inbred lines in maize (Zea mays L.). This study was conducted to assess the effectiveness of pedigree selection for improving resistance to Gibberella ear rot in four maize populations. Selection was based on the severity of ear rot symptoms after inoculation with macroconidial suspensions of Fusarium graminearum (Schwabe) into the silk channel (for two populations) and into the developing kernels (for two other populations). Samples of the selfed families (S₁ to S₅), recovered from remnant seed from the selection programs, were evaluated for disease resistance during three years, using inoculation and evaluation protocols similar to those used during selection. Among-family selection was effective in both of the populations selected after silk inoculation and in one of the populations selected after kernel inoculation. Responses to selection were more evident in later than in earlier generations for both types of inoculation. Changes in the estimated genetic gain over generations were consistent with changes in the variances among families, which tended to increase in early generations and to decrease in later generations. Selection after kernel inoculation seemed to have been more effective than selection after silk inoculation in developing families with more stable resistance. Based on the results obtained here, it seems that responses to family selection could be accelerated without increasing operational costs by increasing selection intensity in later generations and inoculating fewer plants per family.     

Aflatoxin Accumulation and Associated Traits in QPM Maize Inbreds and their Testcrosses

Preharvest aflatoxin (AF) contamination by Aspergillus flavus Link:Fr is one of the main limitations for maize (Zea mays, L.) production in the southern USA, causing enormous economic losses and posing a risk to animal and human health. The objectives of this study were (1) to estimate aflatoxin accumulation and expression of associated traits in quality protein maize (QPM) inbreds and their testcrosses, (2) to compute their repeatabilities and correlations, and (3) to study the relationship between inbred lines and their testcrosses for aflatoxin accumulation. Forty-eight inbreds and their testcrosses plus checks were grown in one and three locations in south and central Texas, respectively. Aflatoxin concentration was evaluated in the plants following inoculation with A. flavus. Average aflatoxin concentration overall for inbreds was 286.3 ng g⁻¹, and for testcrosses 596.5 ng g⁻¹ at Corpus Christi, TX, 325.1 ng g⁻¹ at Weslaco, TX, and 105.1 ng g⁻¹ at College Station, TX. Flinty orange inbreds developed from CIMMYT Population 69 were the least susceptible to aflatoxin accumulation in both inbreds and testcrosses at all locations. Repeatability for aflatoxin was 0.67 in inbreds at Weslaco, TX and 0.54 in testcrosses across locations. Aflatoxin in testcrosses was positively correlated both phenotycally and genotypically with endosperm texture and kernel integrity, and negatively correlated with grain yield and silking date. Less aflatoxin accumulation was associated with flinty endosperm texture, better kernel integrity, and later maturities. Association between the expression of traits in inbreds and aflatoxin in testcrosses was relatively high for endosperm texture (R ² = 0.62), silking date (R ² = 0.44), kernel integrity (R ² = 0.39), and aflatoxin (R ² = 0.60 for log ng g⁻¹). It seems plausible to select for associated traits having high heritabilities and strong correlation with aflatoxin, in addition to low aflatoxin accumulation in inbreds and hybrids to reduce the risk of aflatoxin contamination.     

加拿大早熟群体杂交选系对玉米丝黑穗病抗性评价

为了了解加拿大早熟玉米群体杂交选系对黑龙江省主要玉米病害之一丝黑穗病的抗性水平,从而为中国北方玉米抗病育种提供优质种质资源,以80份加拿大早熟玉米群体杂交选系为材料,在哈尔滨、密山两地分别采用人工接种技术评价其对玉米丝黑穗病的抗性。结果表明,80份加拿大玉米群体杂交选系中没有高抗材料,发现SW1284等14份抗病材料,SW1084等13份中抗材料,SW1236等35份感病材料,SW1175等18个高感材料,中抗以上材料占全部鉴定材料的33.8%;杂交选系与供体双亲平均发病率相关极显著,相关系数r=0.30（r0.01,78=0.281）。     

Relationship between European corn borer resistance and concentration of mycotoxins produced by Fusarium spp. in grains of transgenic Bt maize hybrids, their isogenic counterparts, and commercial varieties

The European corn borer (ECB), Ostrinia nubilalis Hb., is a major pest of maize in central Europe and promotes the infection of maize with Fusarium spp. In this study, transgenic Bt maize hybrids were compared with their isogenic counterparts, and with commercial hybrids from the recommended list with regard to their level of ECB resistance and their concentration of deoxynivalenol (DON), its 15‐acetyl (15‐A‐DON) and 3‐acetyl (3‐A‐DON) derivatives, nivalenol (NIV), fusarenon‐X (FUS‐X), fumonisins (FUM), and zearalenon (ZEN) in harvested grains. The field experiments were performed in Germany at four locations in 1999 and at five locations in 2000. Transgenic Bt hybrids showed significantly lower means than their corresponding isogenic counterparts and than commercial hybrids for all resistance traits: damage rating of stalks, number of larvae per plant, number of larvae per ear, and percentage of damaged plants or ears under infestation. Among all mycotoxins analysed, DON consistently showed the highest concentration across all year × location combinations. Mycotoxin concentrations varied significantly between locations, years and genotypes, whereas mycotoxin concentrations were not significantly different between infested and protected plots. Associations between ECB resistance traits and mycotoxin concentrations were not consistent across years. It is concluded that under central European conditions, the use of Bt maize hybrids will only slightly reduce the contamination of maize kernels with mycotoxins produced by Fusarium spp.     

一个新的抗玉米矮花叶病基因的发现及初步定位

由SCMV引起的矮花叶病是我国的主要玉米病害之一, 鉴定和发掘新的抗病基因对于玉米抗病遗传育种具有重要意义。以抗病自交系海9-21和感病自交系掖478杂交的一个BC₂F₃群体为试验材料, 通过人工接种矮花叶病毒进行抗病性鉴定, 发现该分离群体中抗病植株与感病植株数符合1∶3的分离比例, 推测其抗病基因是由1对隐性基因控制。抗感池和SSR标记连锁分析表明, 存在一个新的玉米矮花叶病隐性抗病基因(或等位基因), 将该基因命名为scm3。scm3基因来源于抗病玉米自交系海9-21, 位于第3染色体短臂3.04~3.05区域, 在SSR标记umc1965和bnlg420之间, 遗传距离分别为45.7 cM和6.5 cM。连锁的标记还有umc1307、umc2265、bnlg2241和umc2166, 它们与scm3之间的遗传距离分别是8.3、13.3、15.5和19.7 cM, 这些SSR标记与scm3基因在染色体上的排列顺序为umc1965—scm3—bnlg420—umc1307—umc2265—bnlg2241—umc2166。     

Concentration of moniliformin produced by Fusarium species in grains of transgenic Bt maize hybrids compared to their isogenic counterparts and commercial varieties under European corn borer pressure

The European corn borer (ECB), Ostrinia nubilalis Hb., is a major pest of maize in Central Europe and is suspected to promote infection of maize with Fusarium species. The objectives of this study were to (1) determine moniliformin (MON) concentration in early maturing European Bt maize hybrids, their isogenic counterparts, commercial cultivars and experimental hybrids, (2) evaluate the association between MON concentration and ECB resistance and (3) correlate MON concentration with concentrations of other mycotoxins determined from the same plant materials. The field experiments were performed at five locations in Germany. MON concentration was significantly higher with manual infestation of ECB larvae (296 μg/kg) than under insecticide protection (66 μg/kg). Bt hybrids showed significantly lower MON concentrations and higher grain yields under manual ECB infestation than their corresponding isogenic counterparts, as well as commercial and experimental hybrids. All ECB resistance traits and grain yield under ECB infestation were significantly correlated with MON concentration. Correlations between concentrations of MON and other Fusarium mycotoxins were not significant. The use of Bt maize hybrids or insecticides to control ECB reduces the contamination of maize grains with MON in Central Europe. The presence of resistance genes against Fusarium species in the current elite maize germplasm was indicated by ECB susceptible non‐Bt hybrids with low‐MON concentrations.     

玉米种质资源大规模多年多点多病害的自然发病抗性鉴定

病害是影响玉米生产的重要因素。利用品种的抗性是控制玉米病害的经济、安全和有效措施。2016—2019年间,在黄淮海和东华北地区,首次对2000份来源广泛且遗传背景丰富的玉米种质资源进行了多年多点多病害的田间自然发生条件下抗病性鉴定,部分材料在西北地区也进行了田间鉴定,重点调查了小斑病、茎腐病、瘤黑粉病、弯孢叶斑病、南方锈病、粗缩病、大斑病、灰斑病和丝黑穗病的抗病性。综合4年共10个不同环境的自然发病下抗性鉴定数据表明,自然发病鉴定的结果受环境因素影响较大,表现为年度间和地域间的差异。在所有鉴定的病害中,小斑病在多个年份和多个鉴定点的发病均比较充分, 11份种质对该病害表现出稳定抗性;茎腐病、大斑病和灰斑病,在不同年份的部分鉴定点发病较为充分,对这3种病害表现抗病的种质分别为440、356和423份,综合抗性鉴定结果具有较大的参考价值;弯孢叶斑病、瘤黑粉病和粗缩病仅在1个鉴定点发病较为充分,南方锈病和丝黑穗病在所有鉴定点均发生较轻,鉴定结果有待进一步验证。本研究筛选出一批在不同环境条件下对多种病害均具有稳定抗性的材料,其中JN15、953、沈977、68122、K21、SC24-1、17MC7211、17MC7223、郑591、161191等种质的综合抗性突出,对种质的利用及后续人工接种精准鉴定的种质选择具重要参考价值。     

农作物收获前黄曲霉毒素污染与控制措施

黄曲霉毒素(aflatoxin,AFT)是曲霉属真菌产生的一大类生物毒素,是危及食品安全和人类健康的主要物质之一。农产品收获前黄曲霉毒素污染是热带、亚热带地区普遍存在的问题,其中在玉米、花生、棉籽、辣椒籽和一些木本坚果及其产品中尤为严重。国内外现有研究结果表明,多种因素可影响作物收获前黄曲霉毒素污染,其中干旱和高温的综合胁迫是最主要的环境因素。作物抗性对降低毒素污染具有重要作用。综合运用分子生物学及常规育种技术改良作物品种对黄曲霉菌侵染或产毒的抗性以及对其他病虫害及干旱的抗(耐)性,是解决黄曲霉毒素污染问题的重要途径。作物生产过程中病虫害的防治和合理的田间管理是作物收获前黄曲霉毒素污染的有效防控措施。     

Indirect response to selection for improving resistance to the Mediterranean corn borer (<Emphasis Type="Italic">Sesamia nonagrioides</Emphasis> Lef) in maize

Mediterranean corn borer (MCB) (Sesamia nonagrioides Lef) and European corn borer (ECB) (Ostrinia nubilalis Hbn) are the most important biotic stresses of maize in Europe. The first selection program to improve stalk resistance to MCB was carried out in the maize population EPS12. It has shown that selection was effective to improve stalk resistance to MCB and ECB, while yield was not significantly diminished. The objective of this research was to determine if correlated changes in EPS12 occurred due to selection for resistance to MCB. Cycles of selection per se and testcrosses to three testers were evaluated under MCB and ECB artificial infestation at two different Spanish locations during 2 years. Selection has significantly reduced cob damage, days to silking, plant and ear height, and 100-kernel weight; meanwhile early vigor was increased. These changes could rather be a consequence of unconscious selection and/or the genetic correlation of these traits with resistance than a consequence of genetic drift.     

Creation of new maize germplasm using alien introgression from <Emphasis Type="Italic">Zea mays</Emphasis> ssp. <Emphasis Type="Italic">mexicana</Emphasis>

Zea mays ssp. mexicana, an annual wild relative of maize, has many desirable characteristics for maize improvement. To transfer alien genetic germplasm into maize background, F₁ hybrids were generated by using Z. mays ssp. mexicana as the female parent and cultivated maize inbred line Ye515 as the male parent. Alien introgression lines, with a large range of genetic diversity, were produced by backcross and successive self-pollinations. A number of alien introgression lines with the predominant traits of cultivated maize were selected. Genomic in situ hybridization (GISH) proved that small chromosome segments of Z. mays ssp. mexicana had been integrated into the maize genome. Some outstanding alien introgression lines were evaluated in performance trials which showed 54.6% hybrids had grain yield greater than that of hybrid check Yedan12 which possessed 50% Ye515 parentage, and 17.1, 9.9% hybrids had grain yield competitive or greater than those of Nongda108 and Zheng958, which were elite commercial hybrids in China, respectively. The results indicated that some of the introgression lines had excellent agronomic traits and combining ability for maize cultivar, and demonstrated that Z. mays ssp. mexicana was a valuable source for maize breeding, and could be used to broaden and enrich the maize germplasm.     

A survey of soybean germplasm for resistance to <Emphasis Type="Italic">Phytophthora sojae</Emphasis>

Phytophthora root and stem rot of soybean caused by Phytophthora sojae is a destructive disease affecting soybean production regions throughout the world. The utilization of resistant cultivars is the most economical and environmentally safe method for controlling this disease. This work aims to screen the effective sources of special and partial resistance for the development of resistant cultivars. A total of 611 soybean germplasm lines from three ecological regions were evaluated for their responses to three P. sojae strains, namely, PNJ1, PNJ3, and PNJ4, using the hypocotyl inoculation technique. The soybean germplasm lines elicited eight different reaction types with three strains. Among these, 106 were resistant and 253 were susceptible to the three strains. A total of 123 soybean germplasm lines identified as susceptible to the three strains by the hypocotyl inoculation method were evaluated for partial resistance to PNJ1 using the slant board assay. Thirty-nine cultivars displayed high levels of partial resistance to PNJ1. The results of this study can be utilized to plant appropriately resistant cultivars in infected fields and to provide good breeding materials.     

Partial resistance to black spot disease in diploid and tetraploid roses: general combining ability and implications for breeding and selection

Black spot disease, incited by the fungus Diplocarpon rosae Wolf, is the most important disease of roses (Rosa hybrida L.) in the outdoor landscape. Though partial resistance exists in cultivated germplasm, the genetic basis of this trait has not yet been elucidated. Six diploid and six tetraploid rose cultivars were crossed in two factorial combining ability arrays. Whole plant and detached leaf inoculation methods were used to assess partial resistance under two different disease pressures using a characterized single-spore isolate. Parents from both arrays had significant general combining ability effects across multiple inoculation methods and environments. Specific combining ability was not significant for either array. Parent per se performance was highly correlated with progeny performance on a family mean basis. High positive correlations among whole plant and detached leaf inoculation methods indicate that detached leaf assays can substitute for whole plant assays. Based on these results, a breeding strategy including parental selection and early, among-family selection is proposed. To our knowledge, this is the first investigation of combining ability for disease resistance in rose.     

Genetic variation for resistance to Fusarium head blight in bread wheat

Summary During a four year period, a total of 258 winter and spring wheat genotypes were evaluated for resistance to head blight after inoculation with Fusarium culmorum strain IPO 39-01. It was concluded that genetic variation for resistance is very large. Spring wheat genotypes which had been reported to be resistant to head blight caused by Fusarium graminearum were also resistant to F. culmorum. The resistant germplasm was divided into three gene pools: winter wheats from Eastern Europe, spring wheats from China/Japan and spring wheats from Brazil. In 32 winter wheat genotypes in 1987, and 54 winter wheat genotypes in 1989, the percentage yield reduction depended on the square root of percentage head blight with an average regression coefficient of 6.6. Heritability estimates indicated that for selection for Fusarium head blight resistance, visually assessed head blight was a better selection criterion than yield reduction.     

The role of ear environment in postharvest susceptibility of maize to toxigenic Aspergillus flavus

A kernel screening assay (KSA) was used to assess the genetic and environmental effects on the vulnerability of maize to aflatoxin accumulation. Kernels of 26 inbred lines that had been grown in seven environments, and 190 lines of the Intermated B73xMo17 (IBM) population grown in one location in the United States, were inoculated with a toxigenic strain of A. flavus and incubated in the dark at 30°C for 6 days. Percent kernel colonization (PKC), sporulation and aflatoxin were influenced by the maize genotypes (G), the location (“ear environment” or E) and the GxE interactions. Overall, low broad‐sense heritabilities were observed for PKC, sporulation and aflatoxin. PKC was significantly correlated with sporulation in all environments. Aflatoxin was positively correlated with colonization for two and with sporulation for all ear environments. Higher grain sulphur or magnesium in IBM was associated with less colonization or aflatoxin. Postharvest susceptibility of maize to aflatoxin is thus influenced by factors that are modulated by the ear environment. In a KSA, sporulation could be a proxy test for aflatoxin accumulation.