生物有机肥对小麦根腐病的防效及其机理初探

为研究生物有机肥对小麦根腐病的防效并初步分析其影响机理,采用盆栽试验法,分析微生物菌有机肥、复合生物有机肥、商品有机肥和发酵干鸡粪对小麦根腐病的防治效果以及对土壤酶活性、土壤养分和作物产量等的影响。结果表明,复合生物有机肥和微生物菌有机肥对小麦根腐病的防效分别为29.38%和27.91%,明显优于其他肥料;施用有机肥均能明显提高小麦根部土壤中脲酶、中性磷酸酶和蔗糖酶的活性,显著增加土壤速效养分含量,且以复合生物有机肥和微生物菌有机肥处理土壤的磷酸酶活性和有效磷含量增幅最大;复合生物有机肥能显著促进小麦生长,其株高、地上干重、根干重、穗粒数、千粒重、产量分别较对照增加8.08%、37.32%、37.50%、14.27%、17.16%和40.77%,差异均达到显著水平,微生物菌有机肥处理的地上干重也显著高于对照。总之,复合生物有机肥和微生物菌有机肥能有效提高土壤中性磷酸酶活性,促进磷素循环,增强拮抗和有益微生物对禾谷镰孢菌的抗性并促进小麦生长。     

不同抗性大麦品种对叶斑病的生理响应及主成分分析

为了解大麦对叶斑病响应的生理机制,以对该病害抗性不同的两个大麦品种蒙啤麦1号(感病)与蒙啤麦3号(抗病)为材料,接菌后测定了大麦苗期、拔节期和孕穗期叶片氧化还原酶等抗病相关酶活性、MDA及渗透调节物质含量等11个生理指标,比较了两个品种间的差异。结果表明,两个品种对叶斑病菌侵染的反应差异因生育时期和指标而异,其中与蒙啤麦1号相比,蒙啤麦3号苗期的脂氧合酶(LOX)、β-1,3-葡聚糖酶和苯丙氨酸解氨酶(PAL)活性及丙二醛(MDA)和可溶性糖(SS)含量的升高幅度较大,拔节期的超氧化物歧化酶(SOD)、过氧化物酶(POD)、过氧化氢酶(CAT)、多酚氧化酶(PPO)、LOX、β-1,3-葡聚糖酶和PAL活性及脯氨酸(Pro)和MDA含量的上升程度较大,孕穗期的SOD、POD、PPO、LOX和β-1,3-葡聚糖酶活性及Pro和SS含量增加幅度较大。通过对11个指标进行主成分分析,LOX、PPO、POD、PAL和β-1,3-葡聚糖酶活性及SS含量对大麦抗叶斑病能力贡献率较大。由此可见,大麦对叶斑病的生理响应因品种、生育时期而异;LOX、PPO、POD、PAL和β-1,3-葡聚糖酶活性及SS含量6个指标可作为评价大麦抗叶斑病的重要相关指标。     

玉米圆斑病病原的快速检测

玉米生平脐蠕孢菌(Bipolaris zeicola)是引起玉米圆斑病的病原菌。本研究通过对玉米生平脐蠕孢菌及其近似种的EF-1α基因(elongation factor 1α)部分序列进行比对,设计出玉米生平脐蠕孢菌的特异性引物Y-EF-F和Y-EF-R,利用该引物可以从B.zeicola中扩增出137 bp的特异片段,而其余的17个参试菌株扩增结果为阴性。灵敏度实验表明该对引物可以检测到目标DNA的浓度为1 pg·μL~(-1)。用B.zeicola接种玉米叶片、苞叶以及玉米粒,然后以接种发病的病组织DNA为模板,利用引物Y-EF-F和Y-EF-R进行PCR扩增,可以扩增出137 bp的特异性条带,而健康玉米组织DNA中未能扩增出任何条带。用B.zeicola孢子悬浮液接种大田玉米叶片,接种第3 d可以检测到未发病组织中有B.zeicola病原菌,第5 d可以看到明显的病斑。研究结果表明该方法可用于快速、准确和灵敏地检测玉米组织中的潜伏期玉米生平脐蠕孢菌,为玉米圆斑病的快速检测,进而及早采取防治措施提供积极的指导。     

玉米圆斑病菌的生物学特性研究

采用形态学和分子生物学方法,鉴定了玉米圆斑病菌并对其生物学特性进行了研究。结果表明,菌落生长最适培养基为PDA,最适温度为25℃;分生孢子萌发的最适碳源为蔗糖,浓度为0.5%,最适pH为7,最适温度为25℃,在不同培养基上分生孢子形态变化比较大。     

两种同时提取小麦根腐离蠕孢菌DNA和RNA的方法比较

获得足够数量和高质量的RNA和DNA是进行分子生物学研究工作的基础。以小麦根腐离蠕孢菌为原料,分别采用溶菌酶法和液氮研磨法破碎真菌细胞壁提取DNA,用琼脂糖凝胶电泳、紫外吸收及聚合酶链式反应（PCR）技术进行核酸完整性和纯度检测。结果表明：两种方法在提取DNA时都同时得到了高质量的RNA;PCR检测结果表明DNA完整性比较高,两种方法得到的DNA质量相当;溶菌酶法中得到的RNA质量优于液氮法。利用溶菌酶破壁法提取小麦根腐离蠕孢菌DNA的提取体系可用于RNA的提取,为小麦根腐离蠕孢菌核酸的DNA和RNA的同时提取提供了一种简便、安全、可行的方法。     

6种杀菌剂对小麦叶斑根腐病菌的毒力测定

筛选高效的防治药剂并研究出其配套的田间防治应用技术是防治小麦根腐病的重要措施,而通过不同杀菌剂对病原菌的毒力测定分析,可为田间防治药剂的筛选提供重要的参考依据。此研究采用生物测定的方法,研究了6种化学药剂对小麦叶斑根腐病菌菌丝生长的抑制作用。结果表明,不同药剂对病原菌生长的影响不同,世高对菌丝生长的抑制作用最显著,在质量浓度为40 mg/L时抑制率达94.9%;其次为唑醚?代森联、丙森锌、福美双、多菌灵和代森锰锌,在该浓度下对菌丝生长抑制率依次为89.5%、86.3%、48.2%、40%和37.8%。对病原菌菌丝生长抑制效果最好的世高对病原菌孢子萌发同样具有强烈的抑制作用,抑制萌发中浓度为6.45 mg/L,可在生产中继续合理使用。     

Characterizing and Estimating Fungal Disease Severity of Rice Brown Spot with Hyperspectral Reflectance Data

Large-scale farming of agriculture crops requires real-time detection of disease for field pest management.Hyperspectral remote sensing data generally have high spectral resolution, which could be very useful for detecting disease stress in green vegetation at the leaf and canopy levels. In this study, hyperspectral reflectances of rice in the laboratory and field were measured to characterize the spectral regions and wavebands, which were the most sensitive to rice brown spot infected by Bipolaris oryzae (Helminthosporium oryzae Breda. de Hann). Leaf reflectance increased at the ranges of 450 to 500 nm and 630 to 680 nm with the increasing percentage of infected leaf surface, and decreased at the ranges of 520 to 580 nm, 760 to 790 nm, 1550 to 1750 nm, and 2080 to 2350 nm with the increasing percentage of infected leaf surface respectively. The sensitivity analysis and derivative technique were used to select the sensitive wavebands for the detection of rice brown spot infected by B. oryzae. Ratios of rice leaf reflectance were evaluated as indicators of brown spot. R669/R746 (the reflectance at 669 nm divided by the reflectance at 746 nm, the following ratios may be deduced by analogy), R702/R718, R692/R530, R692/R732, R535/R746, R521/R718, and R569/R718 increased significantly as the incidence of rice brown spot increased regardless of whether it's at the leaf or canopy level. R702/R718, R692/R530, R692/R732 were the best three ratios for estimating the disease severity of rice brown spot at the leaf and canopy levels. This result not only confirms the capability of hyperspectral remote sensing data in characterizing crop disease for precision pest management in the real world, but also testifies that the ratios of crop reflectance is a useful method to estimate crop disease severity.     

QTL analysis for brown spot resistance in American rice cultivar ‘Dawn’

Rice brown spot (BS), caused by Bipolaris oryzae, causes yield loss and deterioration of grain quality. Using single-nucleotide polymorphism (SNP) markers, we conducted quantitative trait locus (QTL) analysis of BS resistance in backcross inbred lines (BILs) from a cross between an American rice cultivar, ‘Dawn’ (resistant), and ‘Koshihikari’ (susceptible). Four QTLs for BS resistance were detected in a three-year field evaluation, and ‘Dawn’ contributed the resistance alleles at all QTLs. The QTL with the greatest effect, qBSR6-kd, explained 15.1% to 20.3% of the total phenotypic variation. Although disease score and days to heading (DTH) were negatively correlated in all three years, qBSR6-kd was located near a QTL for DTH at which the ‘Dawn’ allele promoted heading. Another BS resistance QTL (qBSR3.1-kd) was unlinked to the QTLs for DTH. Therefore, these two QTLs are likely to be useful for breeding BS-resistant varieties without delaying heading. The other two BS resistance QTLs (qBSR3.2-kd and qBSR7-kd) were located near DTH QTLs at which the ‘Dawn’ alleles delayed heading. The QTLs reported here will be good candidates for developing BS-resistant cultivars.     

21种天然产物对小麦根腐病菌的抑菌活性

为探索小麦根腐病生防途径,筛选有效抑菌物质,达到农药减量的目的。以禾谷镰刀菌(Fusarium graminearum)、刺腐霉菌(Pythium spinosum)和小麦离蠕孢菌(Bipolaris sorokiniana)为供试材料,采用菌丝生长法测定了21种天然产物的抑菌活性。结果表明,反式细辛醚和顺式细辛醚对禾谷镰刀菌效果显著,EC₅₀值分别为69.99 mg/L和130.23 mg/L;白藜芦醇、顺式细辛醚和柠檬精油对刺腐霉菌有明显抑菌活性,EC₅₀值分别为24.4683 mg/L、71.02 mg/L和79.83 mg/L;丁香酚油和白藜芦醇对小麦根腐离蠕孢菌抑菌活性显著,EC₅₀值分别为14.06 mg/L和22.09 mg/L。因此,丁香酚、反式细辛醚、顺式细辛醚、白藜芦醇能有效抑制小麦根腐病菌的生长,在小麦根腐病生物防治方面具有很大应用潜力。