环腺苷酸对向日葵大丽轮枝菌生物学特性及致病力的影响

为探究环腺苷酸(cAMP,cyclic AMP)对向日葵大丽轮枝菌生物学特性及致病力的影响,在实验室条件下研究了外源添加cAMP(10mmol/L)后,向日葵大丽轮枝菌的菌落形态、生长速率、产孢量、分生孢子萌发率、微菌核数量、粗毒素分泌量以及致病力的变化。结果表明,外源cAMP使向日葵大丽轮枝菌分生孢子的产孢量提高了55.45%;而在含有cAMP的水琼胶培养基上培养24h后,大丽轮枝菌的分生孢子萌发率降低了69%;在含有cAMP的PDA上培养25d后,大丽轮枝菌微菌核形成的数量相比对照降低了46.8%;在含有cAMP的液体培养基中振荡培养15d后,大丽轮枝菌粗毒素的含量仅为0.156mg/mL,比对照培养液中毒素降低了35.27%。向日葵接种10mmol/L cAMP处理的大丽轮枝菌后,其病情指数为34.38,相比对照组降低了7.28%。因此,外源添加10mmol/L cAMP除了能够提高向日葵大丽轮枝菌的产孢量外,对其生长速率、分生孢子萌发率、微菌核形成的数量、粗毒素分泌量以及致病力均有不同程度的抑制作用,尤其是对微菌核形成的抑制作用最为明显。     

向日葵枯萎病菌的分离鉴定及其生物学特性

为明确向日葵枯萎病的病原菌种类,调查来自不同地区2014-2015年田间向日葵枯萎病株(50份),经过柯赫氏法则鉴定,将分离获得的31株分离物鉴定为镰刀属下的7个不同的种,即尖孢镰刀菌(F.oxysporum)、轮枝镰刀菌(F.verticilloides)、砖红镰刀菌(F.lateritium)、锐顶镰刀菌(F.acuminatum)、芬芳镰刀菌(F.redolens)、木贼镰刀菌(F.equiseti)和层出镰刀菌(F.proliferum)。以每个种随机选取的菌株为研究对象,对其生物学特性进行了比较研究。结果表明:轮枝镰刀菌BC012的生长速度最快,而砖红镰刀菌BC03生长速度最慢;产孢量以轮枝镰刀菌BC012最多,而木贼镰刀菌NMG1的产孢量最少。在不同的培养温度下,除锐顶镰刀菌TBB212可在5℃低温下生长外,其余菌株的最适生长温度均为25℃~30℃。所有供试的菌株在p H 4~11的培养条件下均可生长。不同种的镰刀菌株产毒能力的比较结果表明,尖孢镰刀菌BC102的产毒能力最强,粗毒素含量为0.651 mg/m L;砖红镰刀菌BC03的产毒能力最弱,粗毒素含量仅为0.075 6mg/m L。不同种的镰刀菌的致病力测定结果表明,接种21d后,轮枝镰刀BC012、芬芳镰刀菌WLJ1和锐顶镰刀菌TBB212的病情指数分别为65.0、55.0、55.0,症状表现为侵染快、发病早、病叶边缘变黄变褐且叶面上出现斑驳状病斑;木贼镰刀菌NMG1、层出镰刀菌T5、尖孢镰刀菌BC102和砖红镰刀菌BC03病情指数在30.0至40.0;症状表现为发病较晚,病叶少且症状轻。而对照菌株向日葵黄萎病菌株大丽轮枝菌的病情指数仅为25.0。     

棉花黄萎病菌毒素对棉花作用机制的初步探讨

为了阐明黄萎病菌毒素在致病中的作用,本试验采用该菌强、弱二种不同致病力类型的菌株毒素进行对棉苗气孔抗性、脱落酸(ABA)增加、叶片细胞膜透性和叶片原生质体的破坏率等研究,对该病菌的致病机制进行探讨,并比较菌株间对棉苗致萎作用的差异。所用材料,供试菌株:致病力强的     

华南3省（区）水稻纹枯病菌的生物学性状与致病力分化研究

 为了探明华南地区水稻纹枯病菌的生物学性状、致病力分化与菌株地理来源之间的关系,为该病害的防治提供依据,从我国华南3省（区）广东、广西和海南的33个县（市）采集水稻纹枯病标本,在分离到335个水稻纹枯病菌菌株的基础上,对它们的培养性状（菌丝生长速率、菌落颜色、菌核数量）、菌丝融合群、菌丝细胞核数目以及致病力分化进行了研究。结果表明,菌株菌丝生长速率可划分为慢、中、快3种类型,菌株数分别为3株（0.90%）、136株（40.60%）和196株（58.50%）;产生菌核的数量可划分为4种类型,即无菌核或极少形成菌核、菌核量少、菌核量中等和菌核量多,各有4株（1.19%）、59株（17.61%）、238株（71.04%）和34株（10.15%）。菌株融合群测定结果表明,所有供试菌株均属于立枯丝核菌（Rhizoctonia solani）AG-1融合群的ⅠA亚群,即AG-1 ⅠA。从中随机选取50个菌株进行菌丝细胞核染色,结果显示,水稻纹枯病菌菌丝的细胞核数目介于7～13个。对其中270个菌株的致病力测定结果表明,所有供试菌株可划分为中等致病力和强致病力2个致病型,分别为208株（77.04%）和62株（22.96%）,未发现弱致病力菌株。综合分析表明,所测菌株的致病型与菌丝生长速率及菌核产生数量各异,表现出较大的多样性。     

香蕉枯萎病菌菌株致病力分化及其原因研究

以田间表现出枯萎病症状的香蕉和粉蕉组织中分离获得的13株尖孢镰刀菌(Fusarium oxysporum)为试验材料,分别测定了不同菌株对香蕉(巴西蕉)和粉蕉(广粉一号)的致病力以及在不同培养液中产生的细胞壁降解酶的活性和毒素含量。结果表明:不同菌株对香蕉和粉蕉的致病力存在明显差异;相关性分析显示,不同菌株产细胞降解酶活性与相应菌株侵染引起的寄主病情指数无相关性,产镰刀菌酸的含量与寄主病情指数呈极显著正相关(r=0.9326)。认为香蕉枯萎病菌菌株间产镰刀菌酸能力的不同是导致其致病力出现分化的主要原因之一。     

向日葵黄萎病病原鉴定

从自黑龙江、内蒙古东、西部、河北、陕西、宁夏、新疆向日葵黄萎病发生严重的地块采集有典型症状的200多个病株进行病原物分离,通过常规组织分离法分离到120个菌株。在PDA培养基上,其菌落表现黑色、白色及灰白色。显微镜下观察其营养菌丝透明、有分隔,呈轮状分枝;分生孢子卵圆或椭圆形、单细胞;大多产生黑色微菌核。提取DNA并进行ITS序列分析（ITS1/ITS4）,证实该120个向日葵黄萎病菌株均为轮枝孢属大丽轮枝菌（Verticillium dahliae Kleb.）。     

温室条件下中棉所96B对新疆棉花黄萎病菌的抗性评价

利用8个分离自新疆不同区域的不同致病型黄萎病菌菌株和2个分离自河北、河南的菌株，在温室中对中棉所96B进行抗病性鉴定，分析了不同致病型菌株对其发病及生长情况的影响，以及病情指数与生物量指标间的相关性，以期为中棉所96B品种区划提供参考。结果表明：接种10个黄萎病菌菌株后，中棉所96B的病情指数为12.93～34.44，说明其对这10个菌株均有较好的抗性。此外，分离自新疆的8个不同致病型黄萎病菌菌株侵染后棉花生物量指标无显著差异。通过棉花黄萎病病情指数与生物量指标相结合可较好地评价品种的黄萎病抗性，进一步为中棉所96B品种区划提供参考。     

Cry2Ab 蛋白对棉花黄萎病菌、枯萎病菌和红腐病菌的影响

研究Cry2Ab蛋白对棉花黄萎病菌、枯萎病菌和红腐病菌生长速率及对黄萎病菌分生孢子产量、分生孢子大小和粗毒素分泌量的影响。结果发现,当Cry2Ab蛋白质量浓度为1.0 μg·mL^－1时,显著降低黄萎病菌分生孢子的大小;当其质量浓度大于5.0 μg·mL^－1时,显著抑制棉花黄萎病菌、枯萎病菌和红腐病菌在PDA培养基上生长及黄萎病菌分生孢子的大小,促进黄萎病菌产孢和粗毒素分泌。     

影响向日葵菌核菌（Sclerotinia sclerotiorum）生长和毒素产生的条件研究

为探讨向日葵菌核菌生长和毒素产生的营养和环境条件。笔者研究了在不同培养液、培养时间、培养温度、pH值、碳源、氮源条件下菌核菌生长和毒素的产生情况。结果表明：向日葵菌核菌菌丝生长和产生毒素所需的营养和环境条件不同。有利于菌丝生长的最佳培养液有PD、马铃薯麦芽糖培养液和Richard,最适生长温度为20℃,最适pH为4.0~8.0,最佳碳源为淀粉,最佳氮源为NH4Cl。含有琥珀酸钠的培养液B最有利于菌核菌产生草酸毒素,草酸毒素积累的最适温度为25℃,最适pH为6.0~7.0,最佳碳源为乳糖,最佳氮源有天门冬酰胺和谷氨酸。菌核菌在最佳培养条件下培养10d时菌丝干重和草酸含量都达到最大值。     

五倍子粗提液对棉花黄萎病的防治效果

以五倍子为对象,研究其粗提液对大丽轮枝菌的抑菌效果及对棉花黄萎病的田间防治效果,为研制用于棉花黄萎病防治的新型植物源制剂提供技术支持。结果表明,当五倍子粗提液体积分数达到0.6%时,其对分离自不同棉区的大丽轮枝菌强致病力菌株Vd080、Vd147、Vd021的菌落生长抑制率和产孢量抑制率均达到100%。田间叶面喷施五倍子粗提液对棉花黄萎病的防治效果最高可达到58.8%,40、80倍液处理能显著提高棉花纤维的断裂伸长率,且前者能显著提高籽棉产量。推荐生产上在棉花黄萎病发生前或发生初期叶面喷施40～80倍的五倍子粗提液进行防治。     

一种快速检测向日葵地块土壤中黄萎病菌微菌核的新方法

由大丽轮枝孢菌侵染引起的向日葵黄萎病是一种重要的土传病害,微菌核是该病害主要的初侵染来源。目前,土壤中大丽轮枝孢菌微菌核的定量检测方法多操作步骤复杂繁琐,如利用PCR方法进行检测,对仪器设备和操作人员的素质都有较高的要求,而常规的土壤梯度稀释湿筛法的实验周期长且检测效率低,因此,建立一种快速定量检测土壤中大丽轮枝孢菌微菌核的方法,对于向日葵黄萎病的预报预测和防控非常重要。为了能够快速的定量检测土壤中微菌核的数量,以期探明不同耕作方式地块中土壤微菌核数量的差异,本实验建立了一套操作相对简单,实验周期较短的微菌核快速分离和定量检测的方法,即采样器—干筛法。该方法将微生物采样器和选择性培养基相结合,基于微生物采样器的撞击法原理,使土壤微生物粒子加速撞击到选择性培养基的培养皿表面,经培养后可见单菌落形成。利用该方法对内蒙古巴彦淖尔市不同的向日葵黄萎病发病地块中采集到的土壤样本中微菌核进行了定量检测,结果表明：两年向日葵连作地（样地1）土壤中微菌核的数量最多,平均每克土样中含有微菌核32.80个;与非寄主作物玉米轮作地块（样地2）土样中微菌核的数量最少,平均每克土样中含有微菌核11.80个,与寄主作物打籽葫芦轮作地块（样地3）微菌核数量介于二者之间。利用该方法能够明显区分不同地块土壤中微菌核的数量。通过和荧光定量PCR检测的结果进行相关性分析发现,该方法能够准确检测土壤中大丽轮枝孢菌微菌核的数量。     

向日葵黄萎病菌营养亲和性及致病力分化

为进一步研究向日葵黄萎病病原菌的变异规律及不同菌株致病力差异,本研究以采自于黑龙江、内蒙古东部和西部、河北、陕西、宁夏、新疆等向日葵黄萎病发生严重的地块,并经分离纯化后得到的120株菌株为材料,开展了菌株间营养亲和性、致病力分化研究。结果表明：利用硝酸盐营养缺陷型突变体( nit )技术可将120株菌株分为3个亲和群（VCGs）,3个亲和群分别有89株、13株、7株,此外,还有11株未能鉴定出亲和群。通过对3个鉴别寄主的致病力测定,120株菌株可分为3个致病类型,强致病力（I）占7.3%,中等致病力（II）占52.7%,弱致病力（III）占40%,菌株致病力存在明显的差异。研究同时表明菌株间营养亲和性与地理位置没有明显的相关性,但与致病性类型具有一定的相关性。       

The Low Potential of Teff (<Emphasis Type="Italic">Eragrostis tef</Emphasis>) as an Inoculum Source for <Emphasis Type="Italic">Verticillium dahliae</Emphasis>

Teff (Eragrostis tef) is a fine stemmed annual grass and gluten free small grain that is of interest as a forage, cover, or a rotation crop. Little is known about the susceptibility of teff to many diseases. Teff could be grown in rotation with potato in the northwestern United States provided teff cultivation is economical and does not increase soil populations for pathogens affecting rotation crops such as Verticillium dahliae. Verticillium dahliae infects a wide range of dicotyledonous plants, making it one of the most important fungal pathogens of crop plants in North America, including potato. The objective of this study was to quantify the susceptibility of teff to eight V. dahliae isolates and compare the susceptibility of teff to eggplant. Teff was confirmed as a host for V. dahliae, as indicated by the presence of microsclerotia in teff stems and roots after artificial inoculation in two years of greenhouse studies. The number of microsclerotia produced in teff did not differ between mint and potato pathotypes of V. dahliae. No V. dahliae isolate produced significantly greater numbers of microsclerotia than any of the seven other isolates tested in a two-year study. Microsclerotia production of V. dahliae in teff was consistently less than in susceptible eggplant cv. Night shadow in both greenhouse experiments (P?<?0.02). It is unlikely that teff infected by V. dahliae will proliferate microsclerotia of mint or potato-aggressive pathotypes, especially when compared to susceptible eggplant cultivars.     

10种无机盐对橡胶树多主棒孢病菌的抑菌作用及对粗毒素的钝化

以橡胶树棒孢霉落叶病病原菌及其产生的粗毒素为供试材料,研究了10种无机盐对多主棒孢菌株菌丝生长、分生孢子萌发的抑制作用和对其粗毒素的钝化作用。结果表明:CuSO4.5H2O、FeCl3和FeSO4.6H2O在2～5mg/mL浓度范围内对多主棒孢病菌菌丝生长及其孢子萌发都有很强抑制作用,菌丝生长抑制率为100%,孢子萌发率为0。KMnO4在0.5～5 mg/mL浓度范围内能够完全抑制孢子的萌发,而1～5 mg/mL浓度范围内却促进菌丝的生长。KMnO4和FeCl3在浓度为0.5 mg/mL时对该毒素均有很强的钝化作用,萎蔫指数分别为2.04%和3.97%。     

Production profile and comparison analysis of main toxin components of Fusarium oxysporum f. sp. sesami isolates with different pathogenicity levels

Fusarium wilt is a common fungal disease in sesame caused by Fusarium oxysporum f. sp. sesami (FOS). To determine the toxin production profiles of the FOS isolates with different pathogenicity levels under various culture conditions, we assessed the content variation of fusaric acid (FA) and 9, 10-dehydrofusaric acid (9, 10-DFA) produced by the four representative FOS isolates. Results indicated that the concentration of FA reached to a maximum of 2848.66 ​μg/mL in Czapek medium, while 9, 10-DFA was mainly produced in Richard and Low-carbon Richard medium. The concentration of 9, 10-DFA on Richard culture medium varied from 0 ​μg/mL to 716.89 ​μg/mL. Of the five culture media used in this study, Czapek culture medium was the most conductive to produce FA. FA production was significantly affected by culture medium, culture time, and their interactions. Results suggest that there is no correlation between toxin production and pathogenicity level of FOS isolates. These findings provide key information for the mechanism analysis of FOS- sesame interaction and pathogen control.     

Recovery and Biological Properties of Nitrate Non-utilizing Mutants of Rice Blast, Magnaporthe grisea

Eleven nitrate non-utilizing (nit) mutants were recovered from six isolates of Magnaporthe grisea cultured on MM media -amended with 60 g/L potassium chlorate, with a frequency of 1.42 %. Some biological properties, such as growth rate, growth biomass, cultural characters, conidial production, sexual reproduction ability, and pathogenicity were compared between nit mutants and their parent isolates. Results showed that all the nit mutants were resistant to chlorate. Some important biological properties such as the growth rate on YPSA, conidial production ability on TPSA, pathogenicity, had no significant differences between nit mutants and their parent isolates. Mating type didn't change, but perithecia production ability of fertile isolates changed significantly as compared with that of their parent isolates. Therefore, the nit can be used as a genetic marker to study the genetics such as pathogenicity, fungicide resistance in Magnaporthe grisea.     

稻瘟病菌nit突变体的诱导及其生物学性状

将6个稻瘟病菌（Magnaporthe grisea）菌株分别在含60 g/L氯酸钾的MM培养基上培养40 d后,共得到11个硝酸盐利用缺陷突变体（Nitrate nonutilizing mutants,简称nit突变体）,频率为1.42 %。比较了各nit突变体与亲本之间在无性和有性阶段的主要生物学性状,结果表明,所有的nit突变体均对氯酸钾产生了抗性。在YPSA平板上的生长速率、在TPSA平板上的产孢量和致病能力等重要的生物学性状都与亲本没有显著差异,交配型没有发生改变,但可交配菌株产子囊壳的能力与亲本有显著差异。因此,可以用nit作为遗传标记研究稻瘟病菌有关性状的遗传学。     

Formation of microsclerotia ofVerticillium dahliae Kleb. on various plant parts of two potato cultivars

Summary Potato plants of cvs Element and Mirka were artificially infected withV. dahliae in two greenhouse experiments. Leaf blade, petiole, aerial stem, subterranean stem, stolon and root mass were separately harvested both when the canopy was still green and at maturity. After 4 weeks incubation, the plant tissue was air-dried and the numbers of microsclerotia per mg tissue and per plant were determined. The highest numbers of microsclerotia were observed in the haulm when harvest took place at maturity. Cultivar Element yielded significantly more microsclerotia in the haulm than cv. Mirka, whilst there were no cultivar differences in the microsclerotial production on subterranean parts. The petiole and the aerial stem contributed most to the total microsclerotial production, whereas roots were much more important for formation of microsclerotia than stolons.     

QuantifyingVerticillium dahliae in soils collected from potato fields using a competitive PCR assay

A quantitative PCR assay based on the competitive PCR technique was compared to the classical soil dilution (SD) method for its ability to estimateV. dahliae propagules directly in soils collected from fields under potato production. A strong correlation (r = 0.97) was observed betweenV. dahliae propagules estimated using the quantitative PCR assay and those using the SD method. Coamplification ofV. dahliae DNA with competitor DNA provided accurate quantification in the range of 10² to 10⁷ spores and 1 to 100 microsclerotia/g of soil. The number ofV. dahliae propagules detected in PEI soils ranged from 4.9 to 15.6 and 0.06 to 0.5 microsclerotia/g of soil for PCR assay and SD method, respectively. The strong correlation between PCR assay and SD method and the non significant differences between replications of PCR estimates ofV. dahliae propagules in soils (P< 0.05) show that the PCR assay is reliable and reproducible, and comparable to the SD method. This method is fast, does not depend on the subjectiveness of the traditional plating method, and offers an improvement in speed and precision over currently used methods. In addition, it can be extended to estimateV. dahliae propagules in other pathosystems and finds immediate and practical use in epidemiological studies to determine the effects of various crop management strategies on the dynamics and level of fungal propagules in the soil in order to establish threshold levels for assessing disease risks and develop disease prediction systems.     

香蕉枯萎病菌粗毒素的毒性及其模型

以香蕉组织培养苗为受试植物,蕉苗受毒素作用后的病情指数为指标,测定香蕉枯萎病菌(Fusarium oxysporum f.sp.cubense)粗毒素对蕉苗的毒性。结果表明,毒素处理72,96,120和144h引致香蕉苗受害程度(以病情指数表示)达90的剂量(TD90)的质量浓度分别为2057.20,1245.49,549.54,380.19μgmL,蕉苗的受害程度存在着随处理时间延长和处理剂量的增加而加重的趋势。根据毒素对蕉苗的毒性测定结果,建立了蕉苗受毒素作用的“时间-剂量-受害程度”模型。根据模型的分析结果认为,粗毒素引致蕉苗受害的有效时间为96~120h,有效剂量质量浓度为260.0~130.0μgmL。