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1.
新疆棉花黄萎病菌病原种群监测研究   总被引:2,自引:0,他引:2  
采用致病性测定、营养亲和群测定及PCR特异性扩增3种方法,对新疆棉区采集的35个棉花黄萎病菌代表性菌系进行致病型鉴定。结果表明,新疆棉花黄萎病菌存在强、中、弱3种不同的致病类型,其中以中等致病类型居多。利用营养亲和群方法将新疆棉花黄萎病菌分为2个亲和群,其中1个亲和群中的所有菌系均与标准落叶型菌系相亲和,另1个亲和群中的所有菌系均与标准非落叶型菌系相亲和。利用棉花黄萎病菌落叶型特异引物进行PCR特异性扩增,有3个菌系扩增出550bp的落叶型黄萎病菌特异性片段。用营养亲和群及PCR特异性扩增均进一步证实目前新疆存在落叶型黄萎病菌菌系。  相似文献   

2.
新疆棉花黄萎病发生调查及病原菌系统进化分析   总被引:1,自引:0,他引:1  
[目的]调查新疆主要植棉地区棉花黄萎病发生情况,对新疆棉花黄萎病菌系系统进化及落叶型菌系分布进行分析和检测.[方法]抽样调查棉花黄萎病发病情况,利用Verticillium dahliae特异性引物D1/D2进行落叶型检测;利用真菌通用引物ITS1和ITS4对新疆菌系进行扩增,测序后进行BLASTN分析.[结果]棉花黄萎病在新疆主要植棉地区普遍发生,棉花黄萎病发生中度及以上病田占比达48.1;,严重发生病田占比为24.1;;BLASTN分析证实采集的棉花黄萎病菌系均为大丽轮枝菌;供试棉花黄萎病菌系检测出22株落叶型菌系,占供试菌系的37.2;.在分离到棉花黄萎病菌系的15个采集点中有9个点检测到了落叶型菌系,占比为60.0;.[结论]新疆部分棉区棉花黄萎病发生比较严重,强致病力的棉花黄萎病落叶型菌系在新疆呈现逐年增加的趋势,且地理分布较广.  相似文献   

3.
石河子地区棉花黄萎病菌培养特性和致病力分化研究   总被引:1,自引:0,他引:1  
[目的]明确当前石河子地区棉花黄萎病菌致病力分化的情况,从病原角度入手搞清病害明显加重的原因.[方法]采用鉴别寄主法和分子生物学的方祛,对采自石河子地区的29个黄萎病菌系进行培养特性观察、致病力测定和特异性引物PCR检测,分析不同致病类型菌系的致病力及接种后落叶情况.[结果]各供试菌系的生长速度存在一定差别,其菌落形态主要为菌核型和菌丝型,分别占75.9;和17.2;.鉴别寄主法测定表明,供试菌系存在明显的致病性分化,强、中、弱菌系分别占51.7;、31.0;和17.3;.致病型测定显示,供试菌系中落叶型菌系和非落叶型菌系分别占37.9;和58.6;,1个菌系未检测出致病类型.落叶型菌系的致病性明显强于非落叶型菌系,其平均病情指数分别为39.5和27.6,二者接种后的落叶程度虽有明显差异,但都可以产生落叶症状,其落叶症状的轻重不仅与菌系的致病类型有关,同时与品种的抗病性也密切相关.[结论]石河子地区棉花黄萎病菌的菌落形态主要以菌核型为主.落叶型菌系的致病力明显高于非落叶型菌系,强致病力落叶型菌系的明显增多是当前病害明显加重的原因之一.  相似文献   

4.
[目的]建立一种能从棉花植株中快速检测黄萎病菌致病类型的方法,为黄萎病菌致病类型的大面积普查打下基础.[方法]利用Dneasy Plant Mini Kit试剂盒对落叶型标准菌株592和非落叶型标准菌株511及病圃中显症前后不同时期棉花植株DNA进行提取,然后用落叶型引物DB19/DB22、DB19/espdef01和非落叶型引物NDf/ NDr、InND2f/InND2r对所提DNA进行巢式PCR扩增.[结果]使用落叶型引物只在592菌株和落叶型病圃的病株中检测到目标条带,使用非落叶型引物只在511菌株和非落叶型病圃的病株中检测到目标条带.[结论]该试剂盒可以有效地提取棉花植株中的DNA,供试的两套引物对检测植株中棉花黄萎病菌致病类型具有高度特异性,该法可直接检测棉花植株中黄萎病菌的致病类型.  相似文献   

5.
【目的】 研究新疆北疆棉区棉花黄萎病菌的致病类型、培养特性及致病力分化。【方法】 从北疆主要植棉区采集棉花黄萎病病株,利用大丽轮枝菌特异引物VDS-F/VDS-R、落叶型菌系特异性引物D1/D2、非落叶型菌系特异性引物ND1/ND2对所有分离的菌株进行检测;根据落叶型和非落叶型菌株在单个条田中所占的比例对田间地块进行分析;选取18个代表菌株对其在PDA上的培养特性及对棉花的致病力分析。【结果】 从北疆棉田病株上分离到644个大丽轮枝菌菌株,67.5%的菌株为落叶型菌系,27.8%为非落叶型菌系,4.7%菌株不能归类,供试菌株的致病类型存在分化,其中以落叶型菌系为主要致病类型。对田间地块进行分析,单一落叶型菌株发生的地块最多,其次是落叶型和非落叶型菌株混合发生地块,田间以落叶型菌株为优势菌株的地块占70.1%,落叶型菌系为田间地块的主要致病类型。18个代表性菌株形成菌核型、菌丝型和中间型3种菌落类型,菌核型为主要的培养类型,并进一步分化形成3种不同的类型。对棉花感病品种的致病力进行测定,落叶型菌株引起的病指普遍高于非落叶型菌株引起的病指,非落叶型菌株中也有病指很高的菌株,北疆棉花黄萎病菌的致病力存在明显分化。【结论】 北疆棉田采集的棉花黄萎病菌,其致病类型、培养特性及致病力均存在明显分化。  相似文献   

6.
山东部分地区棉花黄萎病菌的致病类型及毒性分析   总被引:1,自引:0,他引:1  
从德州、东营和泰安等棉花黄萎病田采集病株样本,经分离培养和鉴定,共获得Verticillium dahliae Kleb.分离物202株,分别用落叶型和非落叶型菌系特异引物INTD2f/INTD2r和INTND2f/INTND2r对各分离物进行PCR扩增,并随机选择8个菌株分析其产毒素及致萎能力。结果表明,202株分离物中,落叶型菌系201株,非落叶型菌系1株;落叶菌株HDF4-3-1产毒素能力最强,毒素浓度约50.82μg/m L,非落叶菌系HLX5-1-1产毒素能力较弱,毒素浓度约13.85μg/m L;落叶菌系HDF5-3-1的致萎指数最高,为72.5%,非落叶菌系HLX5-1-1的致萎指数最低,为42.5%。表明,落叶型菌系产毒素和致萎能力总体较高。  相似文献   

7.
新疆棉花黄萎病菌致病性分化监测研究   总被引:21,自引:9,他引:12  
对新疆棉区采集的35个棉花黄萎病菌代表性菌系进行了生物学特性研究、鉴别寄主法测定及营养亲和群研究.依据鉴别寄主法测定结果,新疆棉花黄萎病菌可分为强、中、弱三种致病型,其中以中等致病类型居多.营养亲和群测定将新疆棉花黄萎病菌分为2个亲合群,其中一个亲合群中的所有菌系均与标准落叶型菌系相亲和,另一亲和群中的所有菌系与标准非落叶型菌系相亲和,从而证实在新疆存在落叶型黄萎菌系.  相似文献   

8.
商丘地区棉花黄萎菌的分离与鉴定   总被引:1,自引:0,他引:1  
从河南商丘各县区采集棉花黄萎病病株,对其进行黄萎病菌的分离和鉴定。采用连续稀释法和单孢分离法分离得到16个单菌系,并以这些单菌系作为研究对象,利用真菌通用引物ITS1和ITS4对所分离的单菌系内转录间隔区(ITS)进行PCR扩增,得到543 bp大小的片段,经过克隆测序和在GenBank中进行BLASTn分析,与安阳棉花黄萎菌的ITS序列的同源性均达到98%以上,证明了这些片段来自大丽轮枝菌(Verticillium dahliae)的ITS区。同时将这些序列在NCB I的GenBank进行了登记,获得5个登记号:FJ715500、FJ715501、FJ715502、FJ715503、FJ715504。  相似文献   

9.
测定了源于新疆棉区的22个棉花黄萎病菌单孢菌株和3个落叶型棉黄萎病菌菌株T9、VD8和V151对4个棉花品种上的致病力.采用随机引物聚合酶链反应(RAPD)技术分析了这些菌株的遗传多样性.结果表明:供试的25个棉花黄萎病菌之间的致病力存在显著差异(P<0.05).在各棉花品种上,22个新疆菌株中都有与T9、VD8或V151的致病力差异不显著的菌株,说明新疆存在较强致病力的棉花黄萎病菌菌系.RAPD分析表明,供试的8个随机引物中6个可以从25个菌株的基因组DNA中稳定地扩增出多态性DNA片段.对扩增片段统计结果表明,供试菌株间遗传相似系数变化幅度为0.57~1.00.聚类分析表明,在阈值0.625处可将25个菌株分为4个RAPD群(命名为RG1、RG2、RG3和RG4),RG1包括2个菌株,即B、BL-19菌株; RG2 包括5个菌株,即BL-17、BL-13、N-5、H、N-16;RG3为最大一个亚群,包括T9、SL等16个菌株;RG4包括2个菌株,即VD8和V151.综合分析表明新疆棉花黄萎病菌的致病力强弱和菌株的采集地等遗传背景存在一定的差异,这可能与各主产棉区的频繁引种有着直接的联系.  相似文献   

10.
[目的]研究棉花黄萎病原菌的入侵机理.[方法]以带有潮霉素抗性筛选基因的PUCCATPH载体以及pCAMBIA1304载体作为骨架,构建trpc启动子驱动的GFP-GUS基因的新疆棉花黄萎病菌表达载体1304-P-ORF,导入农杆菌GV3101和AGL-1中.通过农杆菌介导(ATMT)法分别转入新疆棉花黄萎病原菌VD-1和VD-278中,对表达效果进行检测.[结果]筛选的潮霉素B浓度为30 μg/mL,农杆菌AGL-1对黄萎病菌的转化效率比GV3101高40;.T0代经含有潮霉素B的PDA培养基筛选,获得了T1代转基因大丽轮枝菌VD-12株,VD-2 784株.通过GUS组织染色,在转基因VD-1和VD-278的菌丝和孢子中均可观察到GUS基因的表达.对转基因黄萎菌进行PCR分子检测,均可检测到GFP和潮霉索B基因.GUS-GFP和潮霉素B基因已成功转入新疆棉花黄萎菌VD-1和VD-278中.[结论]建立了新疆棉花黄萎菌的遗传转化体系,为进一步利用GFP-GUS基因研究新疆棉花黄萎菌对棉花侵染的过程奠定基础.  相似文献   

11.
抗·感棉花品种体内棉花黄萎菌的动态分布研究   总被引:3,自引:0,他引:3  
[目的]检测接种后不同时间抗、感棉花品种体内棉花黄萎菌的分布情况。[方法]应用双抗体夹心酶联免疫吸附测定(DAS-ELISA)方法监测棉花黄萎菌在较抗棉花黄萎病的海岛棉品种海7124和较感棉花黄萎病的陆地棉品种鄂荆1号体内的传导情况。[结果]棉花黄萎菌可以有效侵入抗、感棉花品种的根部,但病原菌很少再向抗病品种海7124的上部茎和叶部传导,而感病品种鄂荆1号向上传导的菌量很大。[结论]初步确定海7124的抗病特性表现为不抗侵入但可有效抑制病原菌在体内的传导,感病品种鄂荆1号既不抗侵入也不抗传导。  相似文献   

12.
棉花黄萎病病叶光谱特征与病情严重度的估测   总被引:6,自引:0,他引:6  
 【目的】阐明棉花黄萎病病叶光谱特征并对其病情严重度进行估测,为今后通过航空、航天遥感大面积监测棉花黄萎病提供依据。【方法】试验采用不同品种棉花黄萎病的病叶材料,在不同发病时期的病谱田和大田同步测定其光谱和发病严重度,定性和定量地分析病叶光谱反射特征、微分光谱特征差异。【结果】棉花不同品种、不同发病时期的黄萎病的病叶光谱均随发病严重度的增加而表现出有规律的变化,可见光(400~700 nm)到近红外区(700~1 300 nm)波段,光谱反射率随病情加重呈现上升趋势,可见光520~680 nm波段范围内尤为明显。当病叶严重度达到b2(25%)时,可作为病害识别的临界,对其进行早期诊断。对光谱一阶微分特征研究表明,在红边范围内(680~780 nm)处理间变幅最大,分析后发现红边斜率减小,红边位置发生了“蓝移”,表现出了病害特有的特征。试验证明:434~724和909~1 600 nm为棉花黄萎病病叶光谱敏感波段,建立的多个遥感估测模型均达到极显著水平。【结论】棉花黄萎病病叶光谱特征明显,建立的相应病害反演模型中利用一阶微分光谱723 nm建立的模型精度最高,可用来定量反演棉花单叶黄萎病的发生情况。  相似文献   

13.
[目的]筛选、鉴定植物枯萎病菌拮抗细菌,为植物枯萎病生防制剂的研究奠定基础。[方法]采用对峙培养法筛选对植物枯萎病菌具有良好抑制作用的生防细菌,通过Biolog细菌自动鉴定系统和16SrDNA序列分析法鉴定其分类地位,并研究该菌株对棉花和小鼠的安全性。[结果]从棉花根际土壤中分离出的12株细菌中,有5株对多隔镰刀菌、香石竹尖孢镰刀菌、棉花枯萎病菌具有拮抗活性;其中,KL-1菌株对3株目标病原菌的抑菌活性分别达69.09%、80.78%、78.89%。Biolog细菌自动鉴定系统和16SrDNA序列分析法鉴定结果表明,KL-1菌株为解淀粉芽孢杆菌;生物安全性初步测定结果表明,KL-1菌株对棉花和小鼠安全无毒。[结论]解淀粉芽孢杆菌KL-1菌株具有对多种植物枯萎病菌的抑菌活性,且对棉花和小鼠安全无毒,是一株具有研究和开发潜能的生防菌株。  相似文献   

14.
[目的]探索实现常规棉高产的适宜施钾水平。[方法]以转基因常规棉邯棉802为试验材料,设氯化钾施肥量0(不施肥,CK)、120、180、240、300 kg/hm~2共5个处理,在最佳施肥方式(基施和花铃期追施各占50%)和最佳种植密度(52 500株/hm~2)条件下,研究不同施钾量对棉花黄萎病发病株率和产量的影响。[结果]邯棉802对钾肥需求十分敏感,施用钾肥能有效降低棉花黄萎病的发生程度,显著促进棉花总铃数和单铃重的增加,明显提高棉花产量,且效果均随施钾量的增加而逐渐提高。其中,施钾量≤240 kg/hm~2时,不同施钾量处理的效果差异均达到了显著水平;施钾量240 kg/hm~2时,继续加大施肥水平,增效作用不再明显。[结论]该研究条件下,邯棉802的氯化钾经济施肥量为240 kg/hm~2,该处理下,棉花黄萎病发病株率为15.0%,较CK降低64.29%;产量为4 630.4 kg/hm~2,较CK增产35.00%。  相似文献   

15.
分析、测定内生细菌73 a、拮抗细菌JB52和化学药剂黄腐酸绿源宝单独使用、混合使用对棉苗生长、棉花产量的影响以及对棉花黄萎病的防治效果。结果表明:73 a和JB52都具有促进棉苗生长和提高棉花产量的作用,特别是73 a的作用最强,苗期株高可增长29.9%,鲜重增加45.2%,田间试验结果籽棉增产11.5%;73 a菌液灌根对棉花黄萎病的防治效果最好,达到50%;黄腐酸绿源宝对棉苗生长和棉花产量没有促进作用,但对防治棉花黄萎病有一定的效能;内生细菌73 a与拮抗细菌JB52或黄腐酸绿源宝混用都不如单独使用效果好。  相似文献   

16.
用大丽轮枝菌(Verticillium dahlida)落叶和非落叶致病型菌株特异性引物扩增,鉴定了浙江省绍兴市茄子黄萎病菌菌株的致病型,并研究了其生物学特性。特异性引物的聚合酶链式反应扩增结果表明,来自浙江省绍兴市的5个代表性分离菌株(VD-sy1、VD-sy2、VD-sy3、VD-sy4和VD-sy5)均属于落叶型菌株。生物学特性研究结果表明,菌株VD-sy1菌丝生长的最适条件为温度25 ℃,24 h黑暗,有较宽的酸碱度范围(pH 4.0~9.0);在以蔗糖为碳源和以酵母膏为氮源的培养基上菌丝生长较快,线性生长率分别为4.25和4.06 mm·d-1;在以半乳糖为碳源和以氯化铵为氮源的培养基上生长较慢,线性生长率分别为2.24和2.42 mm·d-1。  相似文献   

17.
【Objective】The objective of this study is to research the effects of broccoli residues (BR) on cotton verticillium wilt and soil bacterial community structure at different growth stages, and to provide new ways and ideas for green ecological control of cotton verticillium wilt and reduction of chemical pesticides.【Method】In the field plot experiment, the soil planted with cotton was treated with broccoli residues and no broccoli residues (CK). The incidence dynamics of cotton verticillium wilt in different treatments were monitored at different growth stages. The number of DNA copies of Verticillium dahliae was determined by real-time quantitative PCR, and the structure of soil bacterial communities was measured by high-throughput sequencing (Illumina MiSeq). The effects of broccoli residues returning to soil on the quantity of pathogen and the community of soil bacteria were analyzed. Principal component analysis (PCA) was used to explore the characteristics and rules of structural changes of soil bacterial community at the level of phylum and genus in different growth stages.【Result】The incidence and disease index of cotton verticillium wilt showed a downward trend after the return of broccoli residues to soil, respectively. Moreover, the peak period of verticillium wilt was delayed, and 70.77% control efficacy was achieved at the peak period. The development curve of disease index and time course of disease in the whole growth period was established, and the average control efficacy was 57.21%. Compared to the blank control, the number of DNA copies of V. dahliae was decreased by 10.96%, 11.11%, 25.95% and 11.25% at the 4 stages of cotton growing and non-growing season, respectively, after the return of broccoli residues to soil. Illumina MiSeq analysis showed that broccoli residues treatment significantly increased soil bacterial diversity. The richness index Chao1 was increased significantly at all growth stages, and the ACE index was significantly increased at the pre-sowing stage, flowering and boll stage and boll-opening stage. Compared to the blank control, the relative abundance of Actinobacteria was increased significantly at different growth stages after the return of broccoli residues to soil. Meanwhile, the results showed that there were differences in the composition of dominant microorganisms at different growth stages after the return of broccoli residues to soil. For example, Firmicutes increased to the dominant microflora at the pre-sowing stage and seedling stage, and Cyanobacteria became the dominant microflora at the bud stage and flowering and boll stage. Principal component analysis showed that broccoli residues to soil changed the structure of bacterial community at seedling stage, bud stage, flowering and boll stage and boll-opening stage. Further analysis showed that the relative abundance of Streptomyces and Bacillus was significantly increased after the return of broccoli residues to soil.【Conclusion】Broccoli residues returning to soil can effectively inhibit the occurrence of cotton verticillium wilt, reduce the number of DNA copies of V. dahliae in soil, change the structure of soil bacterial community, and increase the relative abundance of beneficial microorganisms, which is an effective green measure to control cotton verticillium wilt.  相似文献   

18.
Hyper spectrum remote sensing with fine spectrum information is an efficient method to estimate the verticillium wilt of cotton. The research was conducted in Xinjiang, the largest cotton plant region of China, by using the data which were collected both by canopy spectrum infected with verticillium wilt and severity level (SL) in the year 2005-2006. The quantitative correlation was analyzed between SL and canopy of reflectance spectrum or derivative spectrum reflectance. The results indicated that spectrum characteristics of cotton canopy infected with verticillium wilt changed regularly with the increase of SL in different periods and varieties, Spectrum reflectance increased in the visible light region (620-700 nm) with the increase of the SL, which inverted in near-infrared region and was extremely significant in the region of (780-1 300 nm). When SL attained b2 (DI = 25), cotton canopy infected with verticillium wilt was used as a watershed and diagnosed index in the beginning stages of the disease. The results also indicated that there were marked different characteristics of the first derivative spectrum in these SL, it changed significantly in the red edge ranges (680-760 nm) with different SL, i.e., red edge swing decreased, and red edge position equally moved to the blue. In this study 1 001-1 110 nm and 1 205- 1 320 nm were selected out as sensitive bands for SL of canopy. Inversion models established for estimating cotton canopy infected with verticillium wilt reached the most significant level. Finally, the different spectrum characteristics of cotton canopy infected with verticillium wilt were marked, some inversion models were established, which could estimate SL of canopy infected with verticillium wilt. The best recognized model was the first derivative spectra at (FD 731 nm- FD 1317 nm), and it might be used to forecast the position of cotton canopy infected with verticillium wilt quantitatively.  相似文献   

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