江苏棉田中黄萎病菌微菌核的检测

为获取江苏省棉田中黄萎病菌微菌核数据以供预测预报和防治参考,通过传统的选择性培养基计数法,利用棉选一号选择性培养基和水筛法分离江苏南京和大丰棉田0～40 cm土壤中的微菌核。结果显示江苏省大丰市的5块棉田中都存在黄萎病菌的微菌核,集中分布在0～20 cm的土层中,占0～40 cm土层微菌核总数的79.9%;其中0～10 cm土层中的每克土壤微菌核数量平均为8.2 个,10～20 cm土层中的每克土壤微菌核数量平均为6.8个。     

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。     

棉田复种芥菜对土壤中大丽轮枝菌和黄萎病的影响

为明确棉田复种芥菜（Brassica juncea）对黄萎病及土壤中大丽轮枝菌（Verticillium dahliae Kleb.）的影响，通过定量检测土壤中V. dahliae微菌核数量，监测棉花黄萎病发生程度，研究棉田复种芥菜对土壤中V. dahliae微菌核数量季节性动态的影响和棉花黄萎病的防治效果。试验设棉花单作和3月复种芥菜2个处理，每个处理7个重复，随着棉花的生长在不同时期取试验田中的土壤样品用于土壤中V. dahliae微菌核密度检测，同时调查黄萎病发生程度。试验结果表明，在整个试验期间，复种芥菜的棉田土壤中V. dahliae微菌核密度低于单种棉花的，两者在8月份存在显著性差异；棉田复种芥菜对棉花黄萎病有明显的防治效果，6月和8月的防治效果分别达到42.4%和19.6%。综上，初步认为棉田复种芥菜作为绿肥可以有效减少土壤中V. dahliae，并且减轻黄萎病的发生。这些研究结果可为利用合理的农业措施防治棉花黄萎病提供依据。     

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.）。     

品种抗性与接种棉株中大丽轮枝菌的分布及与茎切面上单孢生长的关系

不同棉花品种,茎基针刺接种大丽轮枝菌中等毒力小种(SS—4)或强毒力小种(T—1)3日内,经室内培养,自棉茎端部维管束和叶柄组织分离到该病原菌.在所有供试的感耐,抗病品种温室培养的幼苗中均观察到大丽轮枝菌在棉茎中的这种传导。7—9日后叶部显现症状。自抗病品种SBSl 上分离到病原菌的频率低于自耐、感病陆地棉品种分离到的频率。在茎段上接种萌发单孢,抗病品种棉茎段比之感、耐病品种茎段能更强烈抑制大丽轮枝菌菌丝的生长。在抗病品种茎干切段上,即使病菌能够生长,产生的微菌核也少,很晚。而感病品种切段上,菌丝生长旺盛,产生的微菌核也多。接种后不久,抗、感病品种棉茎端部和叶柄组织中大丽轮枝菌的存在,及抗、感病品种段上菌丝生长和微菌核形成的差异表明,这种抗性是一种被激活的寄主化学反应。这些结果与下一篇关于类倍半萜烯植保素的积累的论文中所报道的资料相一致。这种植保素可能是棉花抗黄萎病的关键因子。     

向日葵黄萎病菌不同培养型产毒能力和致病力分化

为了明确向日葵黄萎病菌间毒素产生能力以及致病力存在着分化现象,本文选取5种不同培养型的向日葵黄萎病菌对其菌落生长速度、粗毒素含量、毒素致萎活性及致病力进行测定。结果表明,不同培养型的黄萎病菌分生孢子的形态没有显著差异,但是菌株生长速度、粗毒素产生量及其致萎能力存在一定的差异。粗毒素产生能力高的菌株,其致萎能力也强,如菌株V33(产生大量微菌核)、V21(不产生微菌核)、V27(产生一定量的微菌核及气生菌丝)的粗毒素分泌量分别为0.36、0.34、0.23mg/mL,其72h的致萎指数为58.33、52.08、37.50。采用纸钵撕底蘸根法对供试菌株的致病力进行测定,结果表明,不同培养型黄萎病菌的致病力存在一定的分化现象。其中致病力最强的菌株是V33,其病情指数为53.97;其次是菌株V21和V39(产生大量微菌核)其病情指数分别为45.91和43.35;菌株V27的致病力最弱,其病情指数为37.44。相关性分析的结果表明,菌株致病力的强弱与其分泌的粗毒素含量呈显著的正相关关系（R2=0.8169,P<0.05）,但与其菌落生长速度以及微菌核的数量没有相关性。向日葵黄萎病菌毒素的分泌量是决定其致病力分化的关键因子。     

根据土壤黄萎病病原体水平确定黄萎发病率和棉花产量损失之间的关系

根据土壤黄萎病病原体水平确定黄萎发病率和棉花产量损失之间的关系播种期大丽花轮枝孢病原体密度（每克土繁殖体个数），用于两个品种连续３年的试验结果绘制而成的发病率（致萎品种叶部症状百分数）曲线非常接近反指数曲线Ｙ＝５７．０９（１－ｅ－０．３５１：ｘ）。当...     

不同茶园土壤微生物数量状况调查初报

调查了全省14个茶场17块茶园土壤微生物的数量状况,结果表明：茶园土壤微生物总数为每克于土1000万个以上,多的达到5800多万个,远较一般农田和旱土的土壤微生物数量大。三大类群微生物中以细菌数量最多,约占总数的90％左右,其次是放线菌,最少的是真菌。凡土层深厚、土质疏松、物理性能较好、有效养分较丰富的茶园,土壤中微生物总量也较高。     

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。     

品种抗性与接种棉株中大丽轮枝菌的分布及与茎切面上单孢生长的关系(续上期)

结果1.接种棉株中大丽轮枝菌的分布。接种大丽轮枝菌中等菌力非落叶型小种SS-4的抗病品种SBSI 和感病品种岱15的棉株,经过连续培养,证明该小种在其茎干组织中的分布无差异(图2A)。两品种取自茎基的切段均于接种当天产生病菌,中部切段接种后24小时产生病菌,端部切段接种后48小时产生病菌。接种后第3天,两品种各重复茎段切面上均出现了大丽轮枝菌,但观察到茎段切面上的菌丝生长及产孢活动有些差异。接种后4—5天进行抽样调查表明,感病品种岱15培养至第5或6天时产生     

Detection methods forVerticillium species in naturally infested and inoculated soils

The detection and identification of threeVerticillium species in field soils with a polymerase chain reaction (PCR)-based assay was compared to the traditional plating assay method. The two methods were both able to detect the commonVerticillium species in soils although the PCR method detectedV. tricorpus in three soil samples that the traditional method did not. In addition, the PCR assay was rapid, efficient, and required only 1 to 2 days for positive identification whereas the traditional methods required 4 to 8 weeks. The traditional method provided a quantitative measure of pathogen propagules in the soil with population levels ranging from 0 to 21, 625 colony-forming units per gram of soil. However, it was not able to differentiate between the weakly pathogenicV. albo-atrum strain 2 and the more aggressiveV. albo-atrum strain 1, but these two were distinguished with the PCR assay. Results from this study demonstrate that when symptoms of verticillium wilt are observed in potato plants in the field, the major verticillium wilt pathogens present in field soils can be rapidly and reliably detected by the PCR assay.     

Population dynamics of Verticillium species in cauliflower fields: Influence of crop rotation,debris removal and ryegrass incorporation

Verticillium wilt, caused by the soil-borne fungus Verticillium longisporum, leads to economic losses in cauliflower production in Belgium. Development of sustainable control measures of the disease is necessary. Previous studies in our laboratory have shown that incorporation of ryegrass can reduce the viability of V. longisporum microsclerotia in soil. However, field experiments are lacking in Belgian conditions. To gain knowledge about the population dynamics of V. longisporum and its relationship with Verticillium wilt, experiments were conducted from 2006 to 2010 in two cauliflower fields (Oppuurs and Ardooie) with history of Verticillium wilt. Three main crop systems (fallow, cauliflower and cauliflower followed by removal of debris) and two cover crop systems (no cover crop and ryegrass) were tested. The results of this study showed that (i) crop rotation intensity of cauliflower cultivation does not affect the inoculum density of microsclerotia forming Verticillium species in soil, (ii) amendment of lignin-rich ryegrass may reduce the soil inoculum, and (iii) seasonal fluctuations of inoculum densities occur but any soil inoculum variation above a level of one microsclerotium per gram of soil does not affect disease levels. Furthermore, we found that Verticillium wilt of cauliflower in Ardooie was reduced in soil containing Verticillium tricorpus-like organisms. The Verticillium isolate Vt305 obtained from the cauliflower field in Ardooie was morphologically similar to V. tricorpus, but its rDNA ITS region showed 100% identity with the non-pathogenic species Verticillium isaacii, recently described in literature.     

The Effect of Alfalfa Residue Incorporation on Soil Bacterial Communities and the Quantity of <Emphasis Type="Italic">Verticillium dahliae</Emphasis> Microsclerotia in Potato Fields in the Columbia Basin of Washington State,USA

Verticillium wilt, caused by the soil-borne fungus Verticillium dahliae, is one of the most important diseases of potato in North America. Soil incorporation of alfalfa residues prior to planting potato could be a nonchemical Verticillium wilt management tactic by reducing the number of viable microsclerotia in field soil. Verticillium dahliae microsclerotia were quantified in field soils where organic material from alfalfa was incorporated, and numbers of microsclerotia were compared to fields where alfalfa residue was not incorporated. In addition, bacterial metagenomics was utilized to characterize soils where organic material from alfalfa was or was not incorporated to determine if alfalfa residue incorporation facilitates the formation of soils that suppress or kill V. dahliae microsclerotia. The number of V. dahliae microsclerotia in soil was greater (P = 0.0003) in fields where crop residue was incorporated than fields without incorporation when chloropicrin was used as a fumigant. Conversely, the number of V. dahliae microsclerotia observed in potato plants did not differ (P = 0.4020) between fields where residues were or were not incorporated if chloropicrin was used. Alfalfa residue incorporation did not significantly alter the soil bacterial metagenome compared to fields not subject to residue incorporation in both years of study. Despite these conclusions, the method can be employed to analyze the effect of grower practices with the intent of linking a field practice to increasing soil bacterial diversity and decreasing Verticillium wilt severity on potato.     

Induction of soil suppressiveness to verticillium wilt of potato by successive croppings

Nine soils collected in New Jersey and Maryland were infested with microsclerotia ofVerticillium dahliae and planted with potato cv. Superior for five successive crops in the greenhouse to induce suppressiveness to Verticillium wilt. Potatoes were harvested 9–12 wk after planting and disease incidence was determined by recovery ofV. dahliae from stem segments on selective medium. Six of the nine soils had high disease incidence during the first cropping and remained conducive to Verticillium wilt throughout the experiment. Disease incidence increased in soils 3 and 9 over time, while disease incidence decreased nonlinearly in soil 1. Viability of microsclerotia buried in soil 9 for 4 wks was reduced compared to the viability of microsclerotia buried in soils 1, 3, and 6. Addition of 50 or 100 g hydrated lime/10 kg soil to soil 9 eliminated its suppressiveness to Verticillium wilt. Repeated cropping of the same soil can induce suppressiveness to Verticillium wilt. Soils exhibiting induced suppressiveness may be a source of new antagonists againstV. dahliae.     

Potato production and incidence ofVerticillium dahliae following rotation to nonhost crops and soil fumigation in the state of Washington

For three years, potato yields, tuber quality, and incidence ofVerticillium dahliae were measured in field plots previously cropped for one year to the following nonhosts: sudan grass, green peas followed by sudan grass (same year), spring wheat, spring wheat followed by sudan grass (same year), sweet corn and field corn. One year rotation to nonhosts did not reduce the population ofV. dahliae propagules in the soil and in only one of the three years wereV. dahliae propagule numbers in potato stems significantly reduced in plots following nonhost crops, compared with plots following potatoes. Verticillium wilt symptoms in potatoes were not reduced by one year rotation to any of these crops and only in one year in three was yield significantly increased. In two of three years, percent U.S. No. 1 tubers was increased following one-year rotation with green peas plus sudan grass, and in one of the two years, specific gravity was increased by rotation. Rotational cropping to nonhosts for two years between potatoes significantly reduced preplant soil populations ofV. dahliae propagules in the soil and in potato stems the following fall. However, soil populations in these two year rotational plots the spring following potato were not reduced compared to plots previously cropped to potatoes two consecutive years. Cropping to nonhosts for two years had no consistent effect on incidence of Verticillium wilt in subsequent potato crops. Two years’ rotation to nonhosts increased plant height and yield compared to continuous cropping to potato but not percent U.S. No. 1 tubers. The various nonhost crops all had about the same non-significant effect on yield. In two out of three years’ trials, fumigation significantly reduced both the incidence of Verticillium wilt and number ofV. dahliae propagules in stems in plots compared to plots non-fumigated. In only one trial, fumigation significantly increased tuber yields and percent U.S. No. 1 tubers.     

棉花黄萎病研究进展浅述

本文通过时棉花黄萎病传播史、危害、致病机理及遗传特点等研究现状描速。突出抗黄萎病育种在防治黄萎病的关键作用，同时指出拂花抗黄萎病育种研究的限制因素．探讨了针对黄萎病的特定育种方案，并对我国抗黄萎病育种前景作了展望。     

Effects of Soil Flooding on the Survival of Two Potato Pathogens, Sclerotinia sclerotiorum and Verticillium dahliae

Results of studies on survival of sclerotia of Sclerotinia and microsclerotia of Verticillium, demonstrated that soil flooding in western Washington is a possible alternative field rotation practice for S. sclerotiorum (white mold), but not for V. dahliae (Verticillium wilt). Cone-tainer experiments in the greenhouse showed that flooding at 16.5 °C caused S. sclerotiorium sclerotia to lose viability between 12 and 24 weeks while a growth chamber experiment revealed that flooding for 18 weeks at 11 °C or 20 °C was sufficient. V. dahliae microsclerotia appeared resistant to flooding under greenhouse and field settings; recovery ranged within 5 to 10 % of the initial soil population after 6 and 12 months. Potatoes planted into field microplots either flooded or fallowed the previous summer had similar Verticillium wilt ratings and potato yield. Lack of control of V. dahliae by flooding may be due partly to relatively low soil temperatures in a cool, marine climate.     

Disease Tolerance in Helianthus petiolaris : A Genetic Resource for Sunflower Breeding

Abstract

Argentina is a major sunflower producer in the world, with crop acreage of 2?2.7 million ha in the last four years. Sunflower crop yield is often influenced by sanitary constraints, mainly fungal pathogens. Helianthus petiolaris is a wild species native to North America established in central Argentina displays a high tolerance to a number of fungal diseases and insects. Controlled crosses of this species with sunflower demonstrated that H. petiolaris constitutes a valuable genetic variability source for sunflower breeding to improve tolerance to rust (Puccinia helianthi), white rust (Albugo tragopogonis), verticillium wilt (Verticillium dahliae), powdery mildew (Erisiphe sp.) and the sunflower moth (Rachiplusia nu). This places H. petiolaris in an outstanding position as a genetic resource since different important traits could be transferred to the crop through interspecific hybridization.     

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。       

近十年来参加江西省棉花区试品种的抗病性分析

通过对江西省2010~2019年207份材料抗枯萎病性分析及对2016~2019年87份材料的抗黄萎病性分析,鉴定结果表明,参试品种的抗枯萎病性水平较高,近10年来高抗以及抗病品种占总参试品种的37.20%,且抗枯萎病性总体呈上升趋势;参试品种的抗黄萎病性水平较低,近4年来抗病品种占参试品种的17.24%,缺乏高抗品种,但近两年来对黄萎病抗性水平总体呈增强趋势。共筛选出5个对枯萎病和黄萎病兼抗的品种。