Genome-wide association mapping and genomic prediction of stalk rot in two mid-altitude tropical maize populations

Maize stalk rot reduces grain yield and quality.Information about the genetics of resistance to maize stalk rot could help breeders design effective breeding strategies for the trait.Genomic prediction may be a more effective breeding strategy for stalk-rot resistance than marker-assisted selection.We performed a genome-wide association study(GWAS)and genomic prediction of resistance in testcross hybrids of 677 inbred lines from the Tuxpe?o and non-Tuxpe?o heterotic pools grown in three environments and genotyped with 200,681 single-nucleotide polymorphisms(SNPs).Eighteen SNPs associated with stalk rot shared genomic regions with gene families previously associated with plant biotic and abiotic responses.More favorable SNP haplotypes traced to tropical than to temperate progenitors of the inbred lines.Incorporating genotype-by-environment(G×E)interaction increased genomic prediction accuracy.

Screening of antagonistic Trichoderma strains and their application for controlling stalk rot in maize

Maize is one of the major crops in China, but maize stalk rot occurs nationwide and has become one of the major challenges in maize production in China. In order to find an environment-friendly and feasible technology to control this disease, a Trichoderma-based biocontrol agent was selected. Forty-eight strains with various inhibition activities to Fusarium graminearum, and Fusarium verticillioides were tested. A group of Trichoderma strains(DLY31, SG3403, DLY1303 and GDFS1009) were found to provide an inhibition rate to pathogen growth in vitro of over 70%. These strains also prevented pathogen infection over 65% and promoted the maize seedling growth for the main root in vivo by over 50%. Due to its advantage in antifungal activity against pathogens and promotion activity to maize, Trichoderma asperellum GDSF1009 was selected as the most promising strain of the biocontrol agent in the Trichoderma spectrum. Pot experiments showed that the Trichoderma agent at 2–3 g/pot could achieve the best control of seedling stalk rot and promotion of maize seedling growth. In the field experiments, 8–10 g/hole was able to achieve over 65% control to stalk rot, and yield increased by 2–11%. In the case of natural morbidity, the control efficiency ranged from 27.23 to 48.84%, and the rate of yield increase reached 11.70%, with a dosage of Trichoderma granules at 75 kg ha^-1. Based on these results, we concluded that the Trichoderma agent is a promising biocontrol approach to stalk rot in maize.

Effect of Potassium on Ultrastructure of Maize Stalk Pith and Young Root and Their Relation to Stalk Rot Resistance

To study the mechanism of potassium （K） application on improvement of maize resistance to stalk rot at cellular level, scanning electron microscope and transmission electron microscope were used to observe the effect of K on the ultrastructure of maize stalk pith tissue and young root tip cell influenced by K and pathogen. In K deficient treatment, parenchyma cells of stalk pith had abnormal structure, and the cell wall between upper and lower adjacent cell was damaged, resulting in the loss of connections between vascular cells and insufficient supporting capacity. However, an improved K nutrition helped to keep a quite tight arrangement of root cell with thick cell wall, and prevent the invasion of pathogen effectively. Moreover, K treated root cell had abundant golgi apparatus, which could excrete large amount of secretions to degrade mycelium. Papillary and highly electronic intensity dot were accumulated at the invading point to prevent the deveJopment of the mycelium. Improved K nutrition could increase the resistant ability of maize plant to stalk rot, through keeping cell structure stability, preventing the expansion of intracellular space to reduce the chances of pathogen invasions, and through reinforcing cell wall and formation of intercellular and intracellular material to restrict further development of pathogen in host cell.

The antioxidant protein ZmPrx5 contributes resistance to maize stalk rot

Maize(Zea mays L.)stalk rot is a devastating disease worldwide,causing severe yield losses.Although previous studies have focused on the genetic dissection of maize resistance to stalk rot,the mechanisms of resistance remain largely unknown.We used a comparative proteomics approach to identify candidate proteins associated with stalk rot resistance.Statistical analyses revealed 763 proteins differentially accumulated between Fusarium graminearum and mock-inoculated plants.Among them,the antioxidant protein ZmPrx5,which was up-accumulated in diseased plants,was selected for further study.ZmPrx5 transcripts were present in root,stalk,leaf,ear,and reproductive tissues.The expression of ZmPrx5 in three inbred lines increased significantly upon F.graminearum infection.ZmPrx5 was localized in the cytoplasm.Compared to control plants,maize plants overexpressing ZmPrx5 showed increased resistance to F.graminearum infection,and ZmPrx5 mutant plants were more susceptible than wild-type plants.Defense-associated pathways including plant–pathogen interactions,phenylalanine metabolism,and benzoxazinoid and flavonoid biosynthesis were suppressed in ZmPrx5 homozygous mutant plants compared with wild-type plants.We suggest that ZmPrx5 positively regulates resistance against stalk rot in maize,likely through defense-oriented transcriptome reprogramming.These results lay a foundation for further research on the roles of Prx5 subfamily proteins in resistance to plant fungal diseases,and provide a potential genetic resource for breeding disease-resistance maize lines.

Resistance Analysis of 25 Corn Varieties to Stalk Rot and Evaluation of Yield Loss

Due to the meteorological factors, Henan Province had a large-scale outbreak of corn stalk rot in 2014, and the incidence status of 25 varieties was investigated. According to the relevant resistance identification standards, Xundan.509 was classified as resistance variety; Yudan606, Jinsai38 and Xundan3136 were classified as moderate resistance ones, while the other 21 varieties were highly susceptible varieties. Using the yield loss rate of 5% as the critical value of disease tolerance standard, XundanS09, Yudan606, Jinsai38, Xundan3136, Huaiyu5288, QiaoyuS, XY046, Zhengyu10, Lile66, WeikeT02 and Xundan29 were classified into tolerance varieties of corn stalk rot. The concept between disease resistance and disease tolerance was distinguished. The diseased plant rate was used as the classification basis of disease resistance and the yield loss rate was used as the evaluation standard of disease tolerance. The relationship between both was showed by a Venn diagram.

Pyramiding the disease resistant genes to southern rust and stalk rot in maize(Zea mays L.) with marker-assisted selection

Southern corn rust(SCR) caused by Puccinia polysora Underw and maize stalk rot caused by Pythium inflatum Matthews(MSR-2) are two destructive diseases of maize(Zea mays L.) in China.Our previous studies indicated that maize inbred line Qi319 is highly resistant to SCR but susceptible to MSR-2,while inbred line 1145 is highly resistant to MSR-2 but susceptible to SCR.The SCR resistant gene(RppQ) in Qi319 and MSR-2 resistant gene(Rpi1) in 1145 have been mapped on chromosome 10 and 4 respectively.In this research,through marker-assisted selection(MAS) with the molecular markers,bnlg1937 tightly linked to Rpi1 and phi041 tightly linked to RppQ,pyramid breeding of the two kinds of disease resistant genes were carried out from the year of 2003 to 2007.Two homozygotic inbred lines of F5 generation,DR94-1-1-1 and DR36-1-1-1 were identified.MAS result suggested DR94-1-1-1 and DR36-1-1-1 contained the two resistance genes RppQ and Rpi1.Field inoculation tests confirmed their high resistance to the two diseases.In addition,field investigation indicated that the two selected inbred lines,particularly DR94-1-1-1,had excellent agronomic traits such as plant height,ear height and yield-relating traits including ear length,ear diameter,ear weight,kernels per ear,kernels per row and kernel weight per ear.The two selected inbred lines DR94-1-1-1 and DR36-1-1-1 can either be directly developed into commercial variety or used as immediate donors of SCR and MSR resistance breeding programs in maize.

Stalk Rot Resistance Analysis and Yield Loss Evaluation on 25 Maize Varieties and Utilization of Resistant Germplasm Jun M9

Stalk rot outbroke in Henan Province in 2014 on maize,and the disease conditions of 25 varieties were investigated.According to national maize variety disease resistance identification standards,Jundan 509 was a resistant variety; Yudan 606,Jinsai 38 and Jundan 3136 were moderately-resistant varieties; and other 21 varieties were all highly susceptible.Based on the yield loss rate of 5% determined as the critical value for division of disease tolerance,disease-tolerant varieties included Jundan 509,Yudan 606,Jinsai 38,Jundan 3136,Huaiyu 5288,Qiaoyu 8,XY046,Zhengyu 10,lile 66,weike 702 and Jundan 29.Varietal disease resistance and tolerance were mainly divided according to incidence and yield loss rate,respectively,and the correlation between them was showed by a Venn diagram.With the male parent Jun M9 of Jundan 509 as a backbone material,multiple maize combinations resistant to stalk rot were formed,and Yongyou 618 among them was approved by Henan Provincial Crop Variety Approval Committee.

木霉微生物菌肥的制备及对玉米茎腐病的应用评价

以前期分离鉴定的生防菌株非洲哈茨木霉Tr35为研究对象,通过载体的筛选,制备木霉菌剂和木霉微生物菌肥。通过测定不同含量的木霉菌剂和木霉有机肥的株高、根长和鲜重,筛选促生效果明显的处理,并通过室内盆栽试验,评价其对玉米苗期茎腐病的防效。结果显示,玉米秸秆粉具有吸水性强、对Tr35无毒和高释放率的优点,与Tr35共同发酵制备木霉菌剂,当土与菌剂为5∶1时,促生效果明显,并对玉米苗期茎腐病防效达51.77%。菌剂土与腐熟羊粪为7∶1时的木霉有机肥促生效果最好,以该比例制备木霉微生物菌肥,对玉米苗期茎腐病防效达61.31%,该研究结果为Tr35菌剂和菌肥的应用开发提供了一定的前期基础。

玉米镰孢茎腐病致病机制及其防治的研究进展

玉米作为主要的农作物,它的生产对保障粮食安全具有重要意义,而茎腐病多年来都是严重威胁世界玉米安全生产的主要病害。本文将针对玉米镰孢茎腐病侵染机制和致病机制进行总结,进一步阐述玉米致病后的抗性遗传机制和免疫机制,同时,结合近年来国内外的相关研究进展概述玉米镰孢茎腐病的科学防治措施,旨在为玉米镰孢茎腐病的有关研究提供参考思路。

Natural variation in maize gene ZmSBR1 confers seedling resistance to Fusarium verticillioides

Maize seedling blight caused by Fusarium verticillioides is a widely occurring maize disease,but the genetics and mechanisms of resistance are not well understood.In this study,GWAS performed by MLM and 3VmrMLM identified 40 and 20 QTNs,associated with seedling blight resistance.These methods identified 49 and 36 genes,respectively.Functional verification of candidate gene ZmSBR1 identified by both methods showed that the resistance of a mutant line to seedling blight decreased by 0.37 grade points after inoculation with F.verticillioides,compared with the WT.The length of the stem rot lesion caused by F.verticillioides increased by 86%in mutant seedlings,and the relative length of the adult plant stalk rot increased by 35%in mutant plants compared to the wild type after inoculation with Fusarium graminearum.Transcriptome analysis showed that expression of defense-related genes after inoculation was down-regulated in the mutant compared to the wild type,synthesis of secondary metabolites associated with resistance was reduced,and the immune response triggered by PAMP decreased,resulting in decreased resistance of mutant maize seedlings.Candidate gene association analysis showed that most maize inbred lines carried the susceptible haplotype.A functional PCR marker was developed.The results demonstrated that ZmSBR1 conferred resistance to multiple Fusarium diseases at the seedling and adult growth stages and had important application value in breeding.

木霉菌对玉米茎腐病的田间防治效果及玉米产量的影响

玉米是我国主要的粮食作物之一。近年来,玉米茎腐病的危害面积和危害程度逐年增长,造成玉米产量降低和品质下降,严重影响了玉米产业的可持续发展。为筛选出防控玉米镰孢茎腐病的优势生防菌株,为玉米茎腐病的绿色防控提供备选材料,以前期室内筛选到的6株木霉菌株为研究对象,将玉米种子分别通过木霉发酵液包衣处理和孢子悬浮液浸种处理后,采用田间随机区组试验测定不同木霉菌株对玉米茎腐病的田间防治效果,并分析其对玉米产量的影响。结果表明,木霉菌株种类、处理方式和病原菌种类不同,对玉米茎腐病的防治效果和玉米产量的影响也不同;其中绿色木霉T43和康氏木霉KS包衣处理对禾谷镰孢茎腐病的防治效果分别达到54.21%、32.71%;绿色木霉T43浸种处理后,接种拟轮枝镰孢菌的前提下,玉米产量为6 760.24 kg/hm^(2),促生率达48.63%;康氏木霉KS浸种处理后,接种禾谷镰孢菌的前提下,玉米产量为6 353.74 kg/hm^(2),促生率达61.26%。从供试的菌株中,筛选出绿色木霉T43和康氏木霉KS作为优势生防菌株,绿色木霉T43的包衣处理对禾谷镰孢茎腐病表现出较好的防治效果,绿色木霉T43和康氏木霉KS浸种处理对玉米产量具有较好的增产效果。

玉米镰孢菌复合病原茎腐病抗性种质资源鉴定及抗性基因挖掘

【目的】玉米茎腐病是我国玉米主产区普遍发生的重要病害之一,自然条件下,玉米茎腐病多由多种病原菌共同侵染造成。筛选抗禾谷镰孢菌(Fusarium graminearum)和拟轮枝镰孢菌(Fusarium verticillioides)复合侵染的优异玉米种质,鉴定与复合病原引起的茎腐病抗性相关数量性状核苷酸(quantitative trait nucleotide,QTN),挖掘抗性候选基因,为玉米抗复合病原抗性分子育种提供基因资源和理论借鉴。【方法】以一个玉米自然群体为试验材料,接种F.graminearum和F.verticillioides复合病原菌,鉴定茎腐病表型;利用全基因组关联分析,筛选显著抗性SNP位点,预测抗病候选基因。【结果】通过田间和室内2个环境下接种复合病原后的茎腐病表型鉴定试验,发现不同来源和亚群的自交系对复合病原侵染的表型差异显著。田间表型鉴定结果表明,收集于中国的自交系普遍抗性较高,而美国的自交系普遍感病性较高;热带及亚热带亚群的自交系抗性较高,混合型亚群的自交系感病性较高。室内表型鉴定结果表明,收集于美国的自交系材料抗性较高,国际玉米小麦改良中心的自交系材料感病性高;硬秆亚群的玉米种质表现较高抗性,混合型亚群的种质表现较高感病性。于田间和室内2个环境下分别筛选出29份和16份对复合病原侵染具有较高抗性水平的种质,2个环境共同筛选到6份抗病种质。基于田间表型GWAS分析,鉴定到18个与复合病原茎腐病抗性显著相关的QTN,挖掘出93个抗病候选基因;有4个基因表现出单倍型变异,且接种后基因表达水平在抗病材料中呈上调趋势。【结论】利用遗传背景丰富的玉米自然群体,在2个环境中共同鉴定出6份玉米镰孢菌复合病原茎腐病抗性材料,可作为潜在的玉米抗茎腐病育种种质资源;挖掘出4个可能参与复合病原菌抗性的候选基因,为玉米镰孢菌复合病原茎腐病抗性育种提供基因资源。

马铃薯细菌性茎腐病综合防治技术研究初探

榆林是中国马铃薯优生区之一,靖边是榆林马铃薯的核心产区,适宜马铃薯规模化生产的资源要素目前已发挥到极致,在马铃薯产业迅猛发展过程中,连年重茬种植在所难免,加之种薯地病原基数呈逐年上升的态势,导致马铃薯黑胫病、细菌性茎腐病、疮痂病、软腐病、环腐病和青枯病等细菌性病害同趋势加重。特别是马铃薯细菌性茎腐病(气生性茎腐病、茎秆腐烂病),目前己成为制约榆林马铃薯生产的主要细菌性病害之一,常年田间平均发病率为10%~20%,严重时发病率高达60%~70%,减产损失达10%~40%,对榆林马铃薯产业发展极具威胁。

玉米茎腐病接种方法的研究

本文利用黑龙江省玉米茎腐病的两种主要致病菌腐霉菌 (Pythiumspp .)和镰刀菌 (Fusariumspp) ,分别用土埋伤根法和茎基注射法进行接种效果方法研究。试验结果显示两种病原菌致病效果没有显著差异 ,两种接种方法差异不显著 ;两种调查方法 :全株调查与剖茎调查计算病情指数没有显著差异 。

玉米茎腐病研究进展

茎腐病是国内外玉米生产中的重要病害之一,近年来,随着机械收获和籽粒直收,茎腐病已成为玉米生产中最具威胁性的病害,利用抗病品种是防治玉米茎腐病最为经济、有效的措施。因此,本文从茎腐病的发生与为害、病原菌种类、抗茎腐病种质资源筛选、抗性遗传和抗病基因发掘与定位等几个方面阐述了国内外玉米茎腐病的研究进展,旨在为该病害的综合治理奠定重要的理论基础。

氯化钾对玉米木质素代谢的影响及其与茎腐病抗性的关系

【目的】阐明氯化钾(KCl)对玉米木质素代谢的影响,探讨和揭示KCl对提高玉米茎腐病抗性的机制。【方法】通过田间试验与室内分析相结合,研究了KCl对玉米不同抗性品种在接种禾谷镰刀菌(Fursarium graminearum)前后茎基部髓组织木质素含量的影响以及相关酶活性的变化。【结果】抗病品种吉单180茎基部髓组织原生木质素含量低于感病品种吉单327,但诱导木质素高于感病品种;而施用KCl则可降低原生木质素含量,增加诱导木质素含量。抗病品种茎基部髓组织苯丙氨酸解氨酶(PAL)和酪氨酸解氨酶(TAL)活性高峰出现时间均早于感病品种。虽然KCl降低了接种前玉米茎基部PAL、TAL和过氧化物酶(POD)活性的本底值,却提高了接种后PAL、TAL和POD的活性,尤其是高峰值。而且在接种前后,KCl处理的玉米茎基部髓组织肉桂醇脱氢酶(CAD)活性均保持较高水平。【结论】诱导玉米在接种后PAL、TAL、POD和CAD酶系的快速反应,提高诱导木质素含量,进而增强植株对病原菌的抗性,是KCl增强玉米茎腐病抗性的机制之一。

玉米茎腐病菌产生的细胞壁降解酶种类及其活性分析

玉米茎腐病菌 Pythium aphanidermatum、Fusarium graminearum能够产生一系列细胞壁降解酶 ,即果胶甲基酯酶 ( PE)、多聚半乳糖醛酸酶 ( PG)、果胶甲基半乳糖醛酸酶 ( PMG)、多聚半乳糖醛酸反式消除酶 ( PGTE)、果胶甲基反式消除酶 ( PMTE)和纤维素酶 ( Cx)。Fusarium graminearum产生的细胞壁降解酶活性明显高于 Pythium aphanidermatum。病菌在活体内和活体外产生的细胞壁降解酶活性明显不同。酶动力学研究表明 :Fusarium graminearum产生的各种细胞壁降解酶均有特定的最适反应条件。水解酶类的 PG、PMG、Cx最大酶活的 p H分别为 6.0、5.0、6.0 ,温度分别为 50、4 0、50℃ ;裂解酶类的 PGTE和 PMTE最大酶活的 p H均为 9.0 ,温度分别为 30、2 0℃ ,与其它病菌产生的细胞壁降解酶的特性基本相同。

玉米种质资源抗腐霉茎腐病和镰孢茎腐病精准鉴定

【目的】建立和完善玉米种质抗腐霉茎腐病和镰孢茎腐病精准鉴定的方法,对遗传背景丰富的玉米种质资源进行腐霉茎腐病和镰孢茎腐病的抗性精准鉴定,筛选出对2种茎腐病具有稳定抗性的种质资源,为玉米抗病育种奠定基础。【方法】对2 004份具有广泛遗传背景的玉米种质进行多年多点的自然发病抗性鉴定,初步筛选出抗茎腐病或综合性状优良的代表性种质,用于人工接种鉴定;利用建立的茎基部注射接种法,对690份代表性玉米种质进行3年共6个环境的抗腐霉茎腐病和镰孢茎腐病精准鉴定。【结果】2016—2018年,在田间自然发病条件下,通过多年多点鉴定出508份种质对茎腐病表现出较稳定的抗性,高抗、抗病和中抗的材料分别为79、106和323份。建立和完善了玉米种质抗茎腐病精准鉴定的方法-茎基部注射接种法,利用该方法与土壤根埋接种鉴定的玉米对腐霉茎腐病抗性的相关系数(r)为0.87,镰孢茎腐病抗性的r值为0.84,表明2种接种方法的评价结果具有高度的相关性,但注射接种的玉米发病程度更充分。2018—2020年,在北京昌平、河南长葛和河南原阳,利用茎基部注射法对690份玉米种质进行了腐霉茎腐病抗性精准鉴定,表明各鉴定点的材料均发病充分,大部分表现感病或高感,仅有3.5%的种质表现出稳定抗性。6个环境条件下玉米对腐霉茎腐病抗性的相关系数为0.46—0.72,同一年度(2018、2019和2020年)2个鉴定点之间的r值分别为0.66、0.60和0.65;3个年度间种质综合抗性的相关系数为0.67、0.84和0.87,表明3年各点的鉴定结果具有较好的一致性。精准鉴定出24份对腐霉茎腐病具有稳定抗性的种质(冀资H676、辽2235、冀资14L88、冀资14L101、丹337、M02N-23、Y1747、HRB16232、T628358等)。在辽宁沈阳、吉林公主岭、吉林梨树3个鉴定点共6个环境条件下,对690份种质进行了抗镰孢茎腐病精准鉴定,其中,辽宁沈阳的种质发病充分,绝大部分材料表现为感病或高感,吉林梨树和公主岭的材料发病明显偏轻。6个环境下玉米种质抗性的相关系数为0.00—0.76,2018、2019和2020年2个鉴定点之间的r为0.12、0.04和0.05,表明同一年度2个点的鉴定结果差异大;3个年度间镰孢茎腐病综合抗性的r值为0.40、0.74和0.72,显示年度间综合抗性较为一致。共鉴定出5份种质(冀资C32、辽785、辽2235、吉资1034和16SD088)对镰孢茎腐病具有稳定抗性。690份种质对2种茎腐病抗性的r值为0.44,表明其对2种茎腐病的抗性存在中等水平的相关性。【结论】明确了2 004份玉米种质在田间自然发病条件下对茎腐病的抗性水平;建立了玉米抗茎腐病精准鉴定的新方法-茎基部注射接种法;精准鉴定了690份玉米种质对腐霉茎腐病和镰孢茎腐病的抗性,筛选出具有稳定抗性的种质各24和5份,是开展玉米抗茎腐病育种或品种抗性改良的重要抗源。

玉米种质和新品种对腐霉茎腐病和镰孢穗腐病的抗性分析

玉米是我国最重要的农作物之一,腐霉茎腐病和镰孢穗腐病是玉米生产上的重要病害。2006-2012年期间,对1647份玉米种质进行了抗肿囊腐霉茎腐病和拟轮枝镰孢穗腐病鉴定,筛选出高抗茎腐病和穗腐病的种质分别为564份和27份,占鉴定总材料的34.2%和1.6%,抗性材料分别为209份和352份,占比为12.7%和21.4%,表明高抗肿囊腐霉茎腐病的资源较为丰富,高抗镰孢穗腐病的种质相对匮乏。其中,13份种质对2种病害均表现高抗,207份种质对2种病害均表现抗性或对其中一种表现高抗而另一种表现抗性。自交系中对肿囊腐霉茎腐病和拟轮枝镰孢穗腐病表现抗性以上(含HR和R)的种质分别占总鉴定种质的56.5%和23.6%,在农家种中分别为21.2%和21.4%,表明玉米自交系中的抗性资源较农家种丰富。2009-2013年期间参加国家玉米区试的品种中,对腐霉茎腐病表现高抗、抗性、中抗、感病和高感的品种分别占11.5%、11.9%、40.1%、17.6%和18.9%。2009-2011年间,中抗以上的育成品种所占比例呈现明显上升趋势,但2012-2013年间,中抗以上的品种所占比例呈下降趋势。

玉米茎腐病研究现状与防治对策

论述了玉米茎腐病的优势病原菌、发病规律及其防治对策,阐明了玉米茎腐病的侵染机制是腐霉菌在高温条件下主要的初侵染原,而镰刀菌因其超强的定殖能力而进行二次侵染,两种病原菌侵染致病的主要活性物质为糖蛋白毒素。筛选功能菌株研制生防制剂和采取农业综合栽培措施将成为取代化学药剂防治该病的主要措施。