玉米大斑病菌ISSR反应体系的优化和遗传多样性分析

以玉米大斑病菌基因组DNA为模板,采用单因素水平优化的方法对DNA聚合酶的来源及浓度、引物浓度、dNTPs浓度、DNA模板浓度、Tm(退火温度)、PCR反应循环数等重要参数进行摸索和优化,建立了玉米大斑病菌ISSR-PCR优化反应体系,并从40条ISSR引物中筛选出9条多态性较好的ISSR引物。对来自河北、河南、辽宁等玉米主产区的44个菌株进行ISSR分析表明,ISSR标记在我国玉米大斑病菌中存在较高的多态性,多态性条带占40.3%。聚类分析显示,在阈值为0.8时菌株被分为7个类群。对ISSR揭示的玉米大斑病菌的遗传多样性与菌株交配型、地理来源之间的关系进行分析,结果显示菌株的遗传多样性与交配型间的关系密切,而与其地理来源无明显相关性。     

玉米自交系对大斑病菌的抗性鉴定及抗性来源分析

玉米大斑病是一种严重影响玉米产量的病害,在全国各地多有发生。通过筛选抗病玉米自交系,选育优良抗病品种可以有效防止大斑病的发生。试验于2017-2018年,用田间人工接种大斑病混合菌的方法,对185份玉米种质进行了玉米大斑病抗性鉴定与评价,筛选出高抗自交系16份,其中自选系7份,并对玉米自选系抗病性来源进行分析,发现Y6和J1577等品系抗病性强且可以稳定遗传。     

The Changes on Activity of Some Protective Enzymes in Maize Seedlings under Ht－toxin Stress

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玉米大斑病菌原生质体的制备

以玉米大斑病菌(Exserohilum turcium)菌株9948N为供试菌株，进行了原生质体制备条件的研究。主要探讨了酶、渗透压稳定剂、温度、时间等条件对原生质体产出率的影响。结果表明玉米大斑病菌原生质体制备的适宜条件是：28℃下以10mg／mL纤维素酶 10mg／mL蜗牛酶的混合酶处理经过研磨的菌体，酶解72h，以0．8mol/L甘露醇做渗透压稳定剂。     

Genes for resistance to northern corn leaf blight in diverse maize populations

Turcicum or northern corn leaf blight (NCLB) incited by the ascomycete Setosphaeria turcica, anamorph Exserohilum turcicum, is a ubiquitous foliar disease of maize. Diverse sources of qualitative and quantitative resistance are available but qualitative resistances (Ht genes) are often unstable. In the tropics especially, they are either overcome by new virulent races or they suffer from climatically sensitive expression. Quantitative resistance is expressed independently of the physical environment and has never succumbed to S. turcica pathotypes in the field. This review emphasizes the identification and mapping of genes related to quantitative NCLB resistance. We deal with the consistency of the genomic positions of quantitative trait loci (QTL) controlling resistance across different maize populations, and with the clustering of genes for resistance to S. turcica and other fungal pathogens or insect pests in the maize genome. Implications from these findings for further genomic research and resistance breeding are drawn. Incubation period (IP) and area under the disease progress curve (AUDPC), based on multiple disease ratings, are important component traits of quantitative NCLB resistance. They are generally tightly correlated (r_p? 0.8) and highly heritable (h²? 0.75). QTL for resistance to NCLB (IP and AUDPC) were identified and characterized in three mapping populations (A, B, C). Population A, a set of 121‐150 F₃ families of the cross B52×mo17, represented US Corn Belt germplasm with a moderate level of resistance. It was field‐tested in Iowa, USA, and Kenya, and genotyped at 112 restriction fragment length polymorphism (RFLP) loci. Population B consisted of 194‐256 F₃ families of the cross Lo951×CML202, the first parent being a Corn‐Belt‐derived European inbred line and the second parent being a highly resistant tropical African inbred line. The population was also tested in Kenya and genotyped with 110 RFLP markers. Population C was derived from a cross between two early‐maturing European inbred lines, D32 and D145, both having a moderate level of resistance. A total of 220 F₃ families were tested in Switzerland and characterized with 87 RFLP and seven SSR markers. In each of the three studies, 12‐13 QTL were detected by composite interval mapping at a signifcance threshold of LOD=2.5. The phenotypic and the genotypic variance were explained to an extent of 50‐70% and 60‐80%, respectively. Gene action was additive to partly dominant, as in previous generation means and combining ability analyses with other genetic material. In each population, gene effects of the QTL were of similar magnitude and no putative major genes were discovered. QTL for AUDPC were located on chromosomes 1 to 9. All three populations carried QTL in identical genomic regions on chromosomes 3 (bin 3.06/07), 5 (bin 3.06/07) and 8 (bin 8.05/06). The major genes Ht2 and Htn1 were also mapped to bins 8.05 and 8.06, suggesting the presence of a cluster of closely linked major and minor genes. The chromosomal bins 3.05, 5.04 and 8.05, or adjacent intervals, were further associated with QTL and major genes for resistance to eight other fungal diseases and insect pests of maize. Bins 1.05/07 and 9.05 were found to carry population‐specifc genes for resistance to S. turcica and other organisms. Several disease lesion mimic mutations, resistance gene analogues and genes encoding pathogenesis‐related proteins were mapped to regions harbouring NCLB resistance QTL.     

三空栽培模式对玉米大斑病流行的影响

为利用新型栽培模式解决黑龙江省玉米大斑病发生严重难以防治的问题,于2016年通过对三空栽培模式和常规栽培模式下不同时段玉米大斑病病情指数进行调查,采用DPS软件分析了两种栽培模式下病害发生情况。结果表明:三空栽培模式秋季的病情指数为59.78,极显著低于常规模式的68.58,同时,5个地点的试验平均病情指数三空栽培模式比常规模式低12.38%,产量提高了4.02%。说明三空栽培模式对大斑病的防治有一定的促进作用,同时也不减产。     

Epidemiology of Northern leaf blight on sweet corn

The epidemiology of northern leaf blight of corn, caused byExserohilum turcicum (Pass.) Leonard and Suggs, is reviewed. The minimal dew period required for infection is temperature-dependent. At 25°C, 1 h of dew is sufficient to cause infection and at this temperature the minimal dew period for sporulation is 14 h. Under natural conditions when one dew night is not long enough for conidia to develop, the dew period on the following night enables the completion of conidial formation. The amount of conidia formed is dependent on temperature, light, plant age, leaf position and plant susceptibility. Both qualitative and quantitative types of resistance were identified in several hybrids. Subsequently, there developed additional biotypes ofE. turcicum which are aggressive to plants containing qualitative monogenic resistance. Within the same physiological race, a significant variation in aggressiveness between isolates from various locations is observed. The pathogen overwinters as mycelia and conidia in infected leaves, husks and other plant parts, or onSorghum halepense L. Reduction in yield due to northern leaf blight is associated with the level of resistance of the host plant, with disease severity, plant age during infection, and position of infected leaves.     

中国玉米大斑病菌生理分化及新命名法的初步研究

 在分别含Ht1、Ht2、Ht3和HtN的单基因玉米鉴别寄主上测定了全国玉米各产区的百余份大斑病菌标样。结果表明,按传统方法命名的1号(或称0号)生理小种仍然是优势小种,占供试菌株的73.0%;2号(1号)生理小种占1.9%。虽然未出现已报道的3号(23号)、4号(23N号)和5号(2N号)小种,但特别值得注意的是出现了25.1%的未定名的新类群。为克服传统命名方法的局限性及指导抗病育种及品种布局,本文初步提出用毒力频率法分析玉米大斑病菌的生理分化状况,并进一步指明了不同致病类群在我国的分布频率及Ht基因的可用范围。     

Identification of RAPD and SCAR markers linked to northern leaf blight resistance in waxy corn (Zea mays var. ceratina)

Exserohilum turcicum causes northern corn leaf blight (NCLB), an important disease occurring in maize producing areas throughout the world. Currently, the development of cultivars resistant to E. turcicum seems to be the most efficient method to control NCLB damage. Marker-assisted selection (MAS) enables breeders to improve selection efficiency. The objective of this work was to identify random amplified polymorphic DNA (RAPD) and sequence characterized amplified region (SCAR) markers associated with NCLB resistance. Bulked segregant analysis (BSA) was used to search for RAPD markers linked to NCLB resistance genes, using F₂ segregating population obtained by crossing a susceptible inbred ‘209W’ line with a resistant inbred ‘241W’ line. Two hundred and twenty-two decamer primers were screened to identify four RAPD markers: OPA07₅₂₁, OPA16₄₅₇, OPB09₅₂₀, and OPE20₅₃₆ linked to NCLB resistance phenotype. These markers were converted into dominant SCAR markers: SCA07₄₉₆, SCA16₄₂₀, SCB09₄₆₄, and SCE20₄₂₉, respectively. The RAPD and SCAR markers were developed successfully to identify NCLB resistant genotypes in segregating progenies carrying NCLB resistant traits. Thus, the markers identified in this study should be applicable for MAS for the NCLB resistance in waxy corn breeding programs.     

油梨突脐蠕孢叶斑病病原菌鉴定及室内防治药剂筛选

【目的】为明确在广西崇左市油梨种植基地发现的一种叶斑病的病原菌及其生物学特性,以及该病害的防治药剂。【方法】通过形态学鉴定及分子生物学方法,对该病原菌进行鉴定,并对该病原菌进行了生物学特性测定及防治杀菌剂的室内筛选。【结果】该叶斑病的病原菌为嘴突脐蠕孢（Exserohilum rostratum）,该病原菌的最适生长温度为28℃,最适培养基为PDA培养基,最适pH为7,黑暗条件有利于该菌的生长。参试的250g/L吡唑醚菌酯乳油、10%苯醚甲环唑水分散粒剂、40%百菌清悬浮剂、430g/L戊唑醇悬浮剂和40%腈菌唑可湿性粉剂对嘴突脐蠕孢均有抑制效果,其中10%苯醚甲环唑水分散粒剂、430g/L戊唑醇悬浮剂和40%腈菌唑可湿性粉剂对该菌生长的抑制效果较好,其EC50分别为0.0163mg/L、0.0420mg/L、0.1109mg/L。