15%苯乙锡·井可湿性粉剂防治稻曲病和稻瘟病试验与研究

近几年来,稻曲病和稻瘟病有流行加剧趋势,长期使用稻瘟灵、三环唑等常规药剂防治药效有所下降。江苏金凤凰农化有限公司研制的15%苯乙锡·井可湿性粉剂(瘟曲必克)经试验能有效地控制穗期稻曲病和稻瘟病的危害。     

香蕉茎叶粉固态发酵条件优化及鹅对其养分利用率的研究

本试验旨在研究应用固态发酵技术改善香蕉茎叶粉养分组成的工艺参数及鹅对其养分的利用率。选择米曲霉和产朊假丝酵母作为发酵菌种,逐步探究5个因素(单菌种发酵、硫酸铵添加量、米曲霉和产朊假丝酵母接种比例、混合菌液接种量)对发酵后香蕉茎叶粉蛋白质含量的影响。利用正交设计筛选最优发酵温度、底物水分和发酵时间的组合。结果显示:米曲霉和产朊假丝酵母单独发酵均能显著或极显著提高发酵后香蕉茎叶粉的粗蛋白质含量(P0.05或P0.01)。添加硫酸铵能显著或极显著提高发酵后香蕉茎叶粉的真蛋白质含量(P0.05或P0.01),其中添加2%硫酸铵组的真蛋白质含量最高。米曲霉∶产朊假丝酵母接种比例为2∶1的处理发酵后香蕉茎叶粉的蛋白质净增加量显著或极显著高于比例为1∶1、1∶3和3∶2的处理(P0.05或P0.01)。正交试验结果显示,以4%的接种量、接种比例为2∶1(米曲霉∶产朊假丝酵母)、2%的硫酸铵添加量,在基质水分为50%,30℃的环境下发酵4 d效果最佳。经过该工艺发酵后的香蕉茎叶粉粗蛋白质含量提高了33.82%,氨基酸分析结果显示,除赖氨酸和精氨酸外其余的15种氨基酸含量均有不同程度的提高。马冈鹅的代谢试验结果显示,发酵后香蕉茎叶粉中的粗蛋白质利用率提高了52.66%,极显著高于发酵前(P0.01);此外,代谢能和能量利用率也都略有提高(P0.05)。由此可见,经过该发酵工艺发酵后的香蕉茎叶粉营养价值不仅得到了改善,也促进了鹅对其养分的消化吸收。     

辽宁省稻瘟病菌对肟菌酯敏感性监测

为了明确稻瘟病菌对肟菌酯的敏感性,利用菌丝生长速率法测定了 2018～2019年辽宁省11个水稻产区分离的220株稻瘟病菌对肟菌酯的敏感性,并在此基础上对供试菌株的抗药性水平进行了分析.结果表明:稻瘟病菌对肟菌酯的EC50值分布范围为0.0111～0.4983 μg·mL-1,敏感性差异达44.89倍.220株稻瘟病菌...     

稻瘿蚊综合防治新技术与示范

20世纪80年代后，稻瘿蚊发生危害明显回升，损失相当严重。以大田试验示范为主，系统研究了新耕作条件下稻瘿蚊综合防治，首次总结提出“喷药除草、压低虫源、抓住晚秧、做好测报、挑治本田”的综合防治新技术体系。室内外试验和大田示范表明，应用草甘膦杀死越冬寄主游草，可显著减少虫源基数；灭线磷(商品名益舒宝)、氯唑磷(商品名米乐尔)是防治稻瘿蚊的高效药剂，抗蚊青占品种高抗稻瘿蚊。此外，合理利用杀虫植物、性引诱剂和天敌等亦是防治稻瘿蚊的重要措施。在面积达12．46万hm^2的示范推广实践中，本田防治效果均达80％以上。     

中国水稻主产区白叶枯病菌致病型分析及近等基因系鉴别寄主的构建

【目的】分析中国不同稻区白叶枯病菌（Xanthomonas oryzae pv. oryzae）致病型,建立近等基因系鉴别寄主,为白叶枯病菌群体结构田间实时准确监测、抗性品种应用以及抗病育种提供科学依据。【方法】利用中国鉴别寄主、IR24以及15个抗白叶枯病近等基因系等共21个鉴别寄主,采用人工剪叶接种方法,对2018—2021年采自广东、广西、海南、浙江、湖南、辽宁、云南共7个省（自治区）的954个单菌落分离菌株进行致病型测定,探明白叶枯病菌致病型种类、分布及毒性分化;基于测试菌株与15个近等基因系及IR24的抗感互作,应用主成分因子分析法,开展近等基因系与病菌互作的变量因子分析,构建白叶枯病菌致病型近等基因系鉴别寄主;基于抗病基因与测试菌株的抗感反应,分析抗病基因聚合效应。【结果】954个测试菌株在中国鉴别寄主上鉴定出11个致病型,包括SRRRR（I）、SSRRR（Ⅱ）、SSSRR（Ⅲ）、SSSSR（Ⅳ）、SSRRS（V）、SRSRR（Ⅵ）、SSSSS（Ⅸ）、SSSRS（新型1）、SRSRS（新型2）、SRSSS（新型3）以及SSRSS（新型4）,占测试菌株的比率分别为11.53％、4.82％、7.34％、6.18％、7.23％、1.05%、59.96％、1.57%、0.10%、0.10%、0.10%。Ⅸ型菌作为致病性最广的强毒菌系已上升为华南和长江中下游湖南和浙江稻区的优势致病型,西南稻区的云南以Ⅳ型菌为主,东北稻区的辽宁以I型菌为主。15个水稻抗白叶枯病近等基因系对954个菌株的抗感性分析结果表明,测试的15个近等基因系可分为5种类型,第Ⅰ类为高感基因系,包括IRBB1、IRBB2、IRBB10、IRBB11、IRBB4;第Ⅱ类为中感基因系,包括IRBB3、IRBB203、IRBB14;第Ⅲ类为中抗基因系,包括IRBB8、IRBB13;第Ⅳ类为抗病基因系,有IRBB21;第Ⅴ类为高抗基因系,包括IRBB5、IRBB7、CBB23、GDBB23;测试菌株中,出现可侵染抗病基因xa5的有42个、Xa7的有34个、Xa23的有31个。对以白叶枯病近等基因系为主的16个品种（系）与954个菌株组成的抗感互作变量数据矩阵进行因子分析,以解释总变量>85.0%为界,提取出8个主成分因子,组建了以近等基因系为主的10个品种(系)组成白叶枯病菌近等基因系鉴别寄主,按其对变量方差贡献大小,这些寄主分别为IRBB10（Xa10）、IRBB4（Xa4）、GDBB23（Xa23）、IRBB5（xa5）、IRBB7（Xa7）、IRBB21（Xa21）、IR24（Xa18）、IRBB13（xa13）、IRBB3（Xa3）、金刚30;新鉴别寄主可将954个测试菌株划分为55个致病型,对测试稻区的白叶枯病菌菌株表现出较好的鉴别力。基因聚合联合抗性分析表明,不同抗病基因聚合对病菌的抗性频率有一定的提升,不同抗病基因对测试菌株的抗性具有一定的互补性。【结论】监测稻区的白叶枯病菌系趋向多样化,毒性分化明显,强毒菌系Ⅸ型菌在部分稻区已上升为优势致病型,侵染xa5、Xa7及Xa23等广谱抗性基因的菌株有上升趋势;抗病基因聚合可拓宽品种对病原菌系的抗性谱,是培育广谱抗性品种的有效途径;近等基因系鉴别寄主的建立与应用可为白叶枯病发生流行的精准监测以及田间实时预警提供技术支撑。     

白叶枯病菌xopXoo基因在致病性中的作用

辣椒斑点病菌(Xanthomonas campestris pv. vesicatoria,Xcv)在非寄主烟草上的过敏反应与xopX基因有关,但在白叶枯病菌(Xanthomonas oryzae pv. oryzae, Xoo)基因组中并未标注xopX同源基因的功能。为研究白叶枯病菌中xopX同源基因在致病性中的作用,对白叶枯病菌的xopXoo基因进行了克隆和突变,并对xopXoo突变体在非寄主烟草上的过敏反应、水稻苗期的水浸症状以及成株期的致病性和病菌生长能力进行了分析。测定结果显示,xopXoo基因突变并不改变白叶枯病菌在烟草上激发过敏反应和在水稻上产生水浸症状能力,但突变菌株在成株期水稻上的致病性和菌体生长能力显著下降。遗传互补能够恢复xopXoo突变体的致病性和菌体生长能力,表明xopXoo是白叶枯病菌的致病性基因。还对白叶枯病菌激发非寄主烟草产生过敏反应的可能基因进行了讨论。     

Detection and Analysis of Magnaporthe oryzae Avirulence Genes AVR-Pib,AVR-Pik and AvrPiz-t in Heilongjiang Province

【Objective】The objective of this study is to investigate the distribution and variation mechanism of avirulence genes AVR-Pib, AVR-Pik and AvrPiz-t in Magnaporthe oryzae strains from different regions and years in Heilongjiang Province, to understand the pathogenic phenotypes of different avirulence gene alleles, and to provide a reference for utilization and distribution of resistance cultivars in Heilongjiang Province.【Method】Based on the avirulence gene sequences published in NCBI, specific primers were designed to amplify full length and the coding sequence (CDS) regions of three genes, respectively. From 2016 to 2017, 335 M. oryzae strains in different regions of Heilongjiang Province were collected and isolated, and their DNA was PCR-amplified using avirulence genes primers and analyzed by agarose gel electrophoresis. The PCR products with different band patterns and from representative strains of different regions were selected for sequencing. The sequencing results were compared with the corresponding avirulence gene sequences for base and amino acid. The pathogenic phenotype of M. oryzae strains with different variant types was determined based on the rice resistance to single-gene lines.【Result】The specific bands of AVR-Pib, AVR-Pik and AvrPiz-t were detected in PCR detection and appeared in different distribution frequencies and mutation types, indicating that these 3 avirulence genes were all distributed in Heilongjiang Province. The average amplification frequency of the 3 avirulence genes was 75.52%, 87.16% and 85.67%, respectively. Among them, 4 types of band (bandless, high band, mid to high band and low band) of AVR-Pib were detected by electrophoresis analysis and 5 variant types AVR-Pib (1-1, 1-2, 2, 3, 3-1) were detected by PCR product sequencing. The genotypes AVR-Pib-1-1, AVR-Pib-1-2, AVR-Pib-2 and AVR-Pib-3-1 are newly discovered variant types, of which genotypes AVR-Pib-1-1 and AVR-Pib-1-2 are insertions of transposon Pot2 but with different insertion sites. The genotype AVR-Pib-2 has a small fragment insertion in the upstream of CDS region. The genotype AVR-Pib-3-1 base sequence has 4 differences from the original sequence, namely 32 (C/G) 35 (T/A) 36 (T/A) 38 (T/A), and the amino acid translation was terminated prematurely. Pathogenic analysis showed that except for the normal genotype AVR-Pib-3, the other alleles lost their avirulence functions. Seven AVR-Pik alleles (D, A, B, C, E, F, F2) were detected after PCR product sequencing, and the alterations in the nucleotide sequences of these alleles all resulted in amino acid missense mutations. The 7 AVR-Pik alleles have been reported previously. The avirulence gene AvrPiz-t was analyzed by electrophoresis and sequencing of PCR products, and 2 types of band (high band and normal band type) and 4 genotypes of AvrPiz-t (A, B, C, D) were revealed. Among them, AvrPiz-t-A is the original genotype, while AvrPiz-t-B has a base A insertion at position 191, causing premature termination of amino acid translation. Genotype AvrPiz-t-C is a newly discovered allelic type, characterized by the presence of a nucleotide variation at position 17 (T/C) and the insertion of base C at position 19 compared with type A, leading to the frameshift mutation and premature translation termination. The high band type avirulent genotype AvrPiz-t-D was sequenced and verified as having an insertion of the Pot3 transposon. Rice single-gene lines infection showed that the strains with AvrPiz-t-A were avirulent to Piz-t line due to Piz-t recognition, whereas the strains with AvrPiz-t (B, C, D) were virulent to Piz-t line due to lost the ability recognized by Piz-t.【Conclusion】The avirulence genes AVR-Pib, AVR-Pik and AvrPiz-t of M. oryzae in Heilongjiang Province are widely distributed and the types of variation are abundant. The results of this study can provide a reference for breeding and popularizing rice cultivars with corresponding disease-resistance genes.     

长江下游粳稻稻瘟病广谱抗性基因组合模式分析

[目的]基因聚合是实现水稻稻瘟病广谱抗性的有效途径之一.通过构建粳稻背景下不同双基因聚合系,利用长江下游粳型稻瘟病菌(Magnaporthe oryzea)菌株评价其抗性效应并解析其抗性效应产生的构成因子,为长江下游粳稻抗稻瘟病育种提供广谱抗性基因组合模式和种质资源.[方法]以粳稻07GY31为背景的Piz基因座不同复...     

Two‐component cultivar mixtures reduce rice blast epidemics in an upland agrosystem

The effect of two‐component rice cultivar mixtures on the control of rice blast disease was studied in three different experiments under rainfed upland conditions in the Madagascar Highlands. The mixtures involved a susceptible cultivar (either susceptible or very susceptible) and a resistant cultivar in different mixture arrangements (random or row mixtures) and with different proportions of the susceptible cultivar (50, 20 and 16·7%), which were compared to the susceptible cultivar grown in a pure stand. The effect of these mixtures on the incidence and severity of leaf and panicle blast was measured weekly, and on yield and yield components at harvest time. The mixture effect was more efficient in reducing disease with a proportion of 16·7% susceptible component than with a proportion of 50%. Blast epidemic was significantly reduced in all three experiments. However, under high blast pressure, there was no reduction in the disease by the end of the epidemic and yields of the susceptible cultivar were almost zero whatever the mixture. In two other experiments performed under lower blast pressure, disease incidence and severity were significantly lower in mixtures, and yields of the susceptible cultivars grown in mixtures were higher than those of their respective pure stands. Cultivar mixtures are a promising strategy that could contribute to a more sustainable cultivation of rice under upland conditions in the context of subsistence agriculture in Madagascar, where all cropping operations are manual.     

水稻白叶枯病菌hrcU基因缺失突变体构建及功能研究

 通过基因敲除方式,构建了水稻白叶枯病菌hrcU基因的缺失突变体。突变体丧失了在水稻上的致病性和在非寄主植物上的过敏反应。携带hrcU基因的黏粒完全互补了hrcU突变菌株的表型,互补菌在水稻组织中的生长能力也与野生型菌株相当。免疫杂交结果显示,hrcU基因突变后,水稻白叶枯病菌的Harpin蛋白Hpa1不能从细胞内分泌到胞外。这些结果表明,水稻白叶枯病菌hrcU基因的主要功能是形成Ⅲ型分泌系统,将效应分子分泌到病原细菌胞外,从而决定了在非寄主上的过敏反应和在水稻上的致病性。