河北省玉米小斑病菌优势生理小种鉴定及ITS序列分析

2007~2016年在河北各地采集玉米小斑病标样,对分离出的356株玉米小斑病菌菌株进行生理小种鉴定。结果表明,在鉴定的年度范围内,玉米小斑病菌T小种、C小种、S生理型、O小种的分离频率在年度间存有差异,O小种的平均分离频率94.94%,是河北省玉米小斑病菌的优势小种;O小种对主栽品种郑单958和自交系C103的致病力呈下降趋势,在C103上致病力下降幅度小于郑单958。对河北省石家庄、保定、衡水、沧州邯郸、邢台采集的45株菌株进行ITS序列分析构建UPGMA进化树,分析表明,河北省内玉米小斑病菌株的遗传进化与地域有一定关系,差异不明显。     

日本结缕草叶斑病病原鉴定及其生物学特性研究

从日本结缕草Zoysia japonica发病叶片上分离得到病原菌,对病原菌进行形态特征观察、致病性测定及其生物学特性研究.结果表明,该病原菌为Bipolaris hawaiiensis.该菌能利用多种碳源和氮源,其中以蔗糖为最适碳源,以KNO3为最适氮源;菌丝生长最适温度为25～30 ℃;最适pH值7～9.菌丝的致死温度为70℃/15 min.PDA平板药剂筛选结果表明,30%爱苗的抑菌效果最好,15%三唑酮的效果最差.     

Characterization,phylogenetic analyses and pathogenicity of Bipolaris setariae associated with brown stripe disease on sugarcane in Yunnan,China

Brown stripe disease is a severe foliar fungal disease of sugarcane worldwide and is widespread in all sugarcane planting areas in China. Brown stripe is a major disease that seriously affects the output and quality of the sugarcane industry in Yunnan Province, China's second-largest sugar base, while the pathogen of this disease remains not yet fully understood. To address this, we isolated and identified the fungi associated with 68 leaf samples showing typical symptoms of brown stripe from 22 sugarcane varieties in different areas of Yunnan Province. A total of 113 isolates were obtained, which were morphologically similar. Of these, 64 representative isolates were sequenced for the internal transcribed spacer region (ITS), GAPDH and EF-1α loci. All representative isolates grouped with the type strain of Bipolaris setariae in the phylogenetic trees inferred with individual and concatenated sequences of ITS, GAPDH and EF1-α. Pathogenicity test results showed that B. setariae strains were able to induce typical symptoms of brown stripe. The results obtained in this study clarify that only B. setariae is associated with sugarcane brown stripe in Yunnan, China. It is recorded here for the first time as a pathogen causing sugarcane brown stripe in Yunnan, and it is able to infect many major cultivars and new varieties, posing a new threat to the sugar industry in Yunnan Province. In addition, these results provide the scientific basis for the future breeding of disease-resistant varieties and effective prevention and control of sugarcane brown stripe disease.     

Phylogenetic analysis based on ITS sequences and conditions affecting the type of conidial germination of Bipolaris oryzae

One taxonomic characteristic of Bipolaris species is the bipolar germination of conidia, but conidia of Bipolaris oryzae , the causal pathogen of brown spot in rice, are regularly observed to show intercalary germination, a characteristic of Drechslera species. The effect of selection, culture media and culture age on type of conidial germination was determined for three brown spot isolates from Cavinti, San Pablo and Palawan in the Philippines, obtained from infected leaves showing typical disease symptoms. Based on the analyses of their ITS1, ITS2 and 5·8S rDNA nucleotide sequences, the local isolates were clearly identified as B. oryzae . Selection for colonies of the three isolates derived from single conidia with either bipolar or intercalary germination had no effect on the number of spores showing bipolar germination in subsequent cultures. Germination on seven different culture media was tested; of these, rabbit food agar and water agar increased the percentage of bipolar germination of conidia, although this varied between isolates. Incubation of the cultures of all three isolates for longer periods prior to harvesting conidia increased the percentage of bipolar-germinating conidia from c . 40 to c . 90% with 5-day-old and 30-day-old cultures, respectively.     

条斑型玉米圆斑病病原鉴定及其生物学特性研究

 本研究通过致病性测定、形态学和分子生物学鉴定方法,首次明确陕西省发生的玉米条斑型叶斑病是玉米圆斑病的一种症状类型,由玉米生离蠕孢(Bipolaris zeicola)3号生理小种引起。供试品种豫玉22、农大108、农大3138、东单7号、沈单10号、沈单16号、户单4号、新陕资1号、高农1号和陕单9号都对3号生理小种表现感病,品种间抗病性差异不显著。此菌在PDA、PSA、OMA、CMA、WA和Czapek上均可生长,最适生长培养基为PDA。菌落生长温度为5~35℃,最适菌落生长温度为25℃。分生孢子在5~40℃均可萌发,低于5℃高于40℃萌发率极低,萌发的最适温度为28℃。孢子在pH 3~11均可萌发,萌发最适pH为5。在自来水中分生孢子可以萌发,蔗糖和葡萄糖溶液可以明显促进孢子萌发。     

苏丹草小斑病病原鉴定

通过对苏丹草新病害苏丹草小斑病病原与玉米小斑病菌和苏丹草大斑病菌的比较研究，鉴定出苏丹草小斑病病原与玉米小斑病病菌是同一个种，不玉蜀黍平脐蠕孢〔Ｂｉｐｏｌａｒｉｓ ｍａｙｄｉｓ（Ｎｉｓｈｉｋａｄｏ＆Ｍｉｙａｂｅ）Ｓｈｏｅｍａｌｃｅｒ＝Ｈｅｌｍｉｎｔｈｏｓｐｏｒｉｕｍ ｍａｙｄｉｓ Ｎｉｓｈｉｋａｄｏ＆Ｍｉｙａｂｅ＝Ｃｏｃｈｌｉｏｂｏｌｕｓ ｈｅｔｅｒｏｓｔｒｏｐｈｕｓ（Ｄｒｅｃｈｓ．）Ｄｒｅｃｈｓｌｅｒ（有性世代）〕，苏丹草是一种新寄主。     

Bipolaris australiensis HD-1胆红素氧化酶纯化、鉴定及其酶学性质

[目的]为了获得B.bipolaris HD-1胆红素氧化酶纯品,确定酶的同源性及其常规酶学性质.[方法]将B.bipolaris HD-1发酵液依次通过硫酸铵盐析、阴离子交换层析和凝胶过滤层析确定蛋白的纯度、浓度和酶活力;通过基质辅助激光解析电离飞行时间质谱（MALDI-TOF）法获得氨基酸序列,并利用Mascot软件在NCBI蛋白库中的比对,以确定同源蛋白来源;用SDS-PAGE方法测其表观分子量;用等电聚焦方法测定等电点;用Linwerver-Burk双倒数作图法测定米氏常数;并对该酶的酶学性质进行了常规的测定.[结果]获得具有胆红素氧化酶酶活性的电泳纯单体蛋白,酶活为46 000 U/L,比活为1.15 U/mg;MALDI-TOF共获得135个氨基酸,该蛋白与来源于Hel-minthosporium tritici-vulgaris的漆酶前体蛋白序列同源性达100％;蛋白的表观分子量为68 kDa;pI为4.1;它对ABTS和胆红素的米氏常数分别为Km（ABTS） =1.8×10-5 mol/L（pH 3.0）和Km（bilirnbin）=2.7×10-5 mol/L（pH 7.5）;对胆红素的最适催化活性为36℃,酶在-40℃下可保持2年,酶活保持在90％以上,在pH 8 ～9.5下可保持相当高的催化胆红素活性,能够耐受高浓度的硝酸钠和尿素,低浓度的叠氮化钠对酶活有促进作用而高浓度则抑制,对氯化钠、溶解氧敏感.[结论]该蛋白具有典型的胆红素氧化酶特性,又存在明显的不同.     

Relationship between Frost Tolerance and Cold-Induced Resistance of Spring Barley, Meadow Fescue and Winter Oilseed Rape to Fungal Pathogens

Plants exposed to one stress factor may become more tolerant to another. Cold is the most often documented factor inducing plant resistance to pathogens. The aim of this work was to investigate whether resistance of spring barley and meadow fescue to Bipolaris sorokiniana and resistance of winter oilseed rape to Phoma lingam induced at 5 °C for 2, 4 or 6 weeks are associated with frost tolerance, water potential and soluble carbohydrate content. Cold‐acclimated plants of each species showed increased resistance to the studied pathogens. Barley, fescue and rape plants demonstrated higher frost tolerance after hardening, but only in the case of fescue a correlation between resistance to frost and resistance to B. sorokiniana was found. A significant decrease in the water potential of leaf cells was observed in cold‐acclimated barley and fescue. In these two species, water potential greatly affected resistance to B. sorokiniana. However, only in barley did accumulation of fructose, glucose and sucrose correlate as well with changes in water potential as with cold‐induced resistance to the pathogen. In the case of hardened rape, no correlation between the studied parameters was found. The results obtained indicated that the temperature of 5 °C used during cold acclimation was not favourable for hardening of this plant species.     

狗牙根平脐蠕孢菌生物学特性及药剂筛选

 对狗牙根平脐蠕孢菌的生物学特性及5种杀菌剂对该病原菌的抑制作用进行了研究。结果表明：狗牙根平脐蠕孢菌对多种碳、氮源都能利用,以蔗糖和硝酸钾、L-天冬酰胺最好,但以脲为氮源的培养基生长较差。菌丝最佳生长温度是25~30 ℃之间，在25 ℃时最利于产孢,菌丝生长和分生孢子的致死温度是56 ℃；该菌在pH 3~11范围内均能生长，产生分生孢子，菌丝生长的最适pH 5~7，产孢量最大的pH是 3，9，10；糖浓度在2%~8%菌丝能生长，在糖浓度为2%时菌丝生长最好，产孢量最高；光照对病原菌的菌丝生长和产孢有明显影响,在全光照的条件下有利于菌丝的生长，但不会产孢，黑暗12h光照12h既利于菌丝生长又利于产孢，而全黑暗的条件下不利于菌丝生长和产孢。对5种杀菌剂的研究表明：退菌特和百菌清对狗牙根平脐蠕孢菌菌丝生长的抑制效果较好，其它杀菌剂也能有效的抑制菌丝的生长。