中国东北地区水稻纹枯病病原菌种类及融合群的分子鉴定

为明确中国东北地区水稻纹枯病病原菌种类及融合群的归属情况, 2015－2017年从黑龙江省、吉林省和辽宁省的17个水稻主产区采集水稻纹枯病标样, 分离获得水稻纹枯病菌214株, 运用水稻纹枯病菌的不同病原菌及融合群的特异性引物对214株水稻纹枯病菌进行病原菌种类和融合群鉴定, 并利用rDNA内转录间隔区(ITS)序列, 对供试水稻丝核菌的融合群归属进行了分析。结果表明:供试214株水稻纹枯病菌分属于茄丝核菌Rhizoctonia solani和水稻丝核菌Rhizoctonia oryzae-sativae, 菌株数分别为198株和16株, 占比分别为92.52%和7.48%。茄丝核菌菌株分属于2个融合群, 分别为AG1-IA和AG4, 菌株数分别为191株和7株, 占比分别为96.46%和3.54%。水稻丝核菌菌株均属于AG-Bb融合群, 菌株数为16株。不同年份水稻纹枯病的病原菌种类及融合群出现的频率和地域分布无明显变化, 而不同地域间水稻纹枯病病原菌的种类及融合群具有明显的分化特征, AG1-IA融合群在中国东北三省各个水稻产区均有分布且均为优势融合群, AG4融合群在辽宁省盘锦市出现频率最高, 水稻丝核菌AG-Bb融合群在吉林省吉林市、通化市和梅河口市出现频率最高。     

Whole-Cell Fatty Acid Composition to Characterize and Differentiate Isolates of Rhizoctonia Species Associated with Turfgrass Diseases in Japan

Cellular fatty acids were analyzed to characterize and differentiate 34 isolates of Rhizoctonia species representing binucleate Rhizoctonia AG-D (I), AG-D (II), R. solani AG 2-2 IIIB, AG 2-2 LP, R. circinata var. circinata and var. oryzae associated with turfgrass diseases in Japan. Myristic, pentadecanoic, palmitic, palmitoleic, stearic, oleic, linoleic and linolenic acids were consistently present in varying quantities in all isolates. Heptadecanoic and 9-heptadecenoic acids were present in isolates of Rhizoctonia AG-D (I), AG-D (II), R. solani AG 2-2 IIIB and AG 2-2 LP but not in isolates of R. circinata var. circinata and var. oryzae. Palmitic, oleic and linoleic acids were the major fatty acids found, constituting 88.30-98.37% of the whole-cell fatty acid content. The remaining fatty acids were present in smaller amounts. Isolates within a single group were closely clustered, whereas isolates from different groups were clearly distinguishable based on average linkage cluster analysis of cellular fatty acids. Principal component analysis, based on all fatty acids detected, confirmed the distinct separation of isolates representing the six groups of Rhizoctonia species obtained from turfgrasses. These results suggested that fatty acid analysis is useful for the characterization and differentiation of isolates of Rhizoctonia species associated with turfgrass diseases. Received 21 May 2001/ Accepted in revised form 28 September 2001     

镧素对镰刀菌Fusarium solani及其致病酶的影响

采用琼脂平板生长速率法及液体培养基培养测定了La对镰刀菌Fusarium solani的抑制作用和毒力,并测定了其对病菌胞外的果胶酶、蛋白酶和纤维素酶等几种致病酶活性的影响。结果表明,随着La2O3浓度升高,对病菌菌丝生长的抑制作用增强,对病菌的EC50和EC95分别为278.2和552.0 mg/L;在一定浓度范围内,La提高了单位量菌丝所产生3种致病酶的活性,但由于菌丝生长受到抑制,除蛋白酶外,病菌胞外致病酶果胶酶和纤维素酶的总量或总活性受到了抑制,降低了病菌的致病力。     

几种除草剂对水稻纹枯病菌的影响

研究了丁草胺、环庚草醚、草甘膦、苄嘧磺隆、吡嘧磺隆和环丙嘧磺隆6种除草剂对水稻纹枯病菌菌丝生长和菌核形成的影响。结果表明,在一定浓度范围内,6种除草剂均能抑制菌丝生长,减少菌核形成数量,且浓度越高,抑制能力越强,但在较高浓度范围内,除草剂可增加菌核体积;吡嘧磺隆在5μg/ml、环丙嘧磺隆在10μg/ml时可增加菌核形成数量;在含除草剂培养基上产生的菌核都可萌发。     

Molecular detection of Fusarium solani f. sp. glycines in soybean roots and soil

A polymerase chain reaction (PCR)-based method was developed to detect DNA of Fusarium solani f. sp. glycines , the cause of soybean sudden death syndrome. Two pairs of primers, Fsg1/Fsg2 designed from the mitochondrial small subunit ribosomal RNA gene, and FsgEF1/FsgEF2 designed from the translation elongation factor 1-α gene, produced PCR products of 438 and 237 bp, respectively. Primer specificity was tested with DNA from 82 F. solani f. sp. glycines , 55 F. solani non-SDS isolates, 43 isolates of 17 soybean fungal pathogens and the oomycete Phytophthora sojae , and soybean. The sensitivity of primer Fsg1/Fsg2 was 10 pg while that of FsgEF1/FsgEF2 was 1 ng when using F. solani f. sp. glycines total genomic DNA or down to 10³ macroconidia g⁻¹ soil. Nested PCR increased the sensitivity of the PCR assay 1000-fold to 10 fg using primers Fsg1/Fsg2, and 1 pg using primers FsgEF1/FsgEF2. F. solani f. sp. glycines DNA was detected in field-grown soybean roots and soil by PCR using either single pairs of primers or the combination of two pairs of primers. The occurrence of F. solani f. sp. glycines was determined using nested PCR for 47 soil samples collected from soybean fields in 20 counties of Illinois in 1999. F. solani f. sp. glycines was detected in soil samples from all five Illinois Agricultural Statistic Districts including 100, 89, 50, 92 and 50% of the samples from East, Central, North-east and West Districts, respectively.     

枯草芽孢杆菌YB5对菜豆根腐病菌的抑菌机制测定及应用

腐皮镰孢引起的菜豆根腐病是一种世界性病害,在菜豆生产中危害严重。为对其有效的生物防治,针对菜豆根腐病利用枯草芽孢杆菌进行了拮抗机制及应用的研究。结果表明:YB5能有效抑制菌丝生长、孢子产生和萌发。YB5的无菌滤液对高温敏感。YB5菌液对种子发芽没有抑制作用,且能促进根系的发育。YB5菌液及无菌滤液对菜豆腐皮镰孢根腐病的盆栽均具有较好的防效,YB5菌液防效可达94.6%。     

草莓丝核菌根腐病病原菌鉴定及7种杀菌剂的抑菌作用测定

为了解草莓丝核菌根腐病的病原种类及筛选防治丝核菌根腐病的有效杀菌剂,本研究基于形态学特征、细胞核荧光染色、菌丝融合群测定以及rDNA-ITS的序列分析,对北京和河北承德地区的草莓丝核菌根腐病的病原菌进行了鉴定,并利用菌丝生长速率法测定了7种杀菌剂对丝核菌的抑菌作用。结果发现,北京地区的丝核菌为双核丝核菌(binucleate Rhizoctonia,BNR),属于融合群AG-A;河北的丝核菌为立枯丝核菌Rhizoctonia solani,属于融合群AG-4。氟啶胺、吡唑醚菌酯、噻呋酰胺、戊唑醇、咯菌腈、氟硅唑对2种丝核菌均有很强的抑制作用,EC_(50)值为0.063 9～2.485 7μg/mL,抑霉唑的抑制作用较差,EC_(50)值为9.966 8～11.236 8μg/mL。同一种杀菌剂对不同丝核菌的抑制作用存在差异,噻呋酰胺、戊唑醇、氟硅唑、咯菌腈和抑霉唑对立枯丝核菌的抑制作用强于对双核丝核菌。试验结果为生产上合理选用杀菌剂防治草莓丝核菌根腐病提供了科学依据。     

东北油豆角腐皮镰孢菌根腐病拮抗芽胞杆菌的筛选与鉴定

以东北油豆角腐皮镰孢菌为靶标菌,采用室内平板对峙培养和温室防效测定方法,从发病油豆角根际(围)土壤中筛选拮抗能力强的芽胞杆菌菌株,以期获得腐皮镰孢菌根腐病的生防资源。结果表明:该试验筛选到一株拮抗芽胞杆菌WJ-126,初步鉴定为贝莱斯芽胞杆菌(Bacillus velezensis),在温室人工接种条件下,该菌株对油豆角腐皮镰孢菌根腐病的相对防效可达60.92%。     

温度对玉米纹枯病菌生长与发病的影响

室内控制性测定结果表明,玉米纹枯病菌(RhizoctoniasolaniAG1鄄IA)菌丝生长的温度10～35℃,最适温度25～30℃;菌核萌发温度18～35℃,最适温度25～30℃。在幼苗和离体叶鞘上由菌丝引起侵染和发病的温度15～35℃,以25～30℃最适宜,潜育期最短,病斑较大。田间分期播种观察分析,气温和地面温度与病害始见的历期(播种期至病害初见期)分别呈显著负相关和极显著负相关。在田间自然条件下,始病的温度为20℃左右。这些结果为了解玉米纹枯病的发生规律,开展预测预报提供了科学依据。     

立枯丝核菌侵染玉米的研究

 用获自水稻及玉米的立枯丝核菌(Rhizoctonia solani Khn) AG-11A接种玉米发生典型纹枯症状,其致病力显著强于AG-4。玉米拔节期,上位叶鞘抗性较强,抽雄及抽丝期抗性减弱,下位叶鞘无论在拔节期或抽雄、抽丝期,均较上位叶鞘感病。接种玉米后8小时,形成侵染垫及附着胞,从这些结构上形成侵入钉侵入,AG-4侵染上位叶鞘时,常以菌丝直接穿透表皮或从气孔侵入。在去掉菌体的叶鞘表面,发现有周边光滑或稍破损的侵入孔。接种后12小时,在叶鞘细胞中发现菌丝,它们在穿过细胞壁进入邻近细胞时,明显变细。接种后16小时,新生出的菌丝从气孔成丛出现。