大丽轮枝菌微菌核形成能力的遗传

带有硝酸盐利用缺陷型遗传标记的大丽轮技菌Verticilliumdahliae黑色菌核型和白色菌丝型菌株在25℃下配对培养,形成野生型融合菌落带,对融合带的分生孢子后代进行遗传分析的结果表明,融合带中的异核体表现不稳定,分布不均匀。微菌核遗传因子可随亲本细胞质在异核体中的运动和交换而发生迁移。     

大丽轮枝菌微菌核形成能力的遗传

带有硝酸盐利用缺陷型遗传标记的大丽轮枝菌Ｖｅｒｔｉｃｉｌｌｉｕｍ ｄａｈｌｉａｅ黑色菌核型和白色菌丝型菌株在２５℃下配对培养,形成野生型融合菌落带,对融合带的分生孢子后代进行遗传分析的结果表明,融合带中的异核体表现不稳定,分布不均匀。微菌核遗传因子可随亲本细胞质在异核体中的运动和交换而发生迁移。     

大丽轮枝菌异核体及其后代的形态与致病力变异

以硝酸盐利用缺陷型突变（ｎｉｔ突变）和抗杀菌剂突变两种遗传标记,对大丽轮枝菌（Ｖｅｒｔｉｃｉｌｌｉｕｍｄａｈｌｉａｅ）异核体后代的形态和致病力进行研究,结果表明,菌核型菌株与菌丝型菌株经菌丝融合形成异核体后,菌丝型菌株能恢复形成微菌核,其后代单孢菌落形成微菌核的数量明显低于菌核型亲本,且遗传性状不稳定;随着转代次数的增多,微菌核形成能力的丧失较菌核型亲本菌株快,异核体后代对棉苗的致病力变化较大,一     

大丽轮枝菌异核体及其后代的形态与致病力变异

以硝酸盐利用缺陷型突变（nit突变）和抗杀菌剂突变两种遗传标记,对大丽轮枝菌（Verticilliumdahliae）异核体后代的形态和致病力进行研究,结果表明,菌核型菌株与菌丝型菌株经菌丝融合形成异核体后,菌丝型菌株能恢复形成微菌核,其后代单孢菌落形成微菌核的数量明显低于菌核型亲本,且遗传性状不稳定;随着转代次数的增多,微菌核形成能力的丧失较菌核型亲本菌株快。异核体后代对棉苗的致病力变化较大,一般均低于致病力强的亲本菌株,或介于两个亲本致病力之间,或与亲本致病力相近。     

大丽轮枝菌异核体及其后代的形态与致病力变异*

以硝酸盐利用缺陷型突变（nit突变）和抗杀菌剂突变两种遗传标记,对大丽轮枝菌（Verticilliumdahliae）异核体后代的形态和致病力进行研究,结果表明,菌核型菌株与菌丝型菌株经菌丝融合形成异核体后,菌丝型菌株能恢复形成微菌核,其后代单孢菌落形成微菌核的数量明显低于菌核型亲本,且遗传性状不稳定;随着转代次数的增多,微菌核形成能力的丧失较菌核型亲本菌株快。异核体后代对棉苗的致病力变化较大,一般均低于致病力强的亲本菌株,或介于两个亲本致病力之间,或与亲本致病力相近。     

真菌异核体不亲和性机理研究进展

异核体不亲和性是普遍存在于真菌中的一种生物学现象,由特定的不亲和性位点所控制。从真菌异核体不亲和性分子机理的研究进展以及不亲和融合细胞死亡过程和相关蛋白的关系展开论述。对真菌异核体不亲和性机理的深入研究,将有助于揭示生物中非己识别系统的演化规律,并为解决原生质体融合子的不稳定性探索出有效的方法和途径。     

葱鳞葡萄孢菌的异核现象

本文报导了葱鳞葡萄孢菌(Botrytis squamosa)的异核现象。采用荧光染色法证实了该菌的分生孢子和菌丝细胞均是多核的。以该菌的自然形态和生长特征为遗传标记,在193个单孢子培养物中获得了三种菌型:菌核型(S)、菌丝型(M)、界于菌核和菌丝之间的菌核菌丝型(SM)。SM型在形态上与原始菌株相同。S型和M型在连续的传代过程中其特征保持稳定,因而被认为是同核型。SM型在连续传代过程中是不稳定的,且一再分离出S型和M型,因此被认为是异核型。用传代稳定的S型和M型重新合成的异核体在特征上相同于原始菌株和SM型。用这个合成的异核体再次进行单孢子分离,三种菌型重新被获得。三种菌型在致病力方面没有表现出显著的差异。     

羊肚菌菌核的交配型

交配型是调控真菌子实体发育和生活史的重要因子.作为羊肚菌生活史中一个有争议的阶段,菌核与交配型的关系研究甚少.以六妹羊肚菌Morchella sextelata和梯棱羊肚菌M.importuna为研究对象,以栽培菌包内的菌核和实验室单孢交配产生的菌核为研究材料,以Morchella sp.Mes-20单孢交配产生的菌核为参考,进行交配型分析.结果 发现:1)菌核的交配型呈明显的偏分离现象,六妹羊肚菌和梯棱羊肚菌中交配型为MAT1-1的菌核远多于交配型为MAT1-2的菌核,而Morchella sp.Mes-20中MAT1-2菌核较多于MAT1-1的菌核;2)部分菌核含有2种交配型,但无法确定菌核是由异核体可育菌丝还是由2个交配型的单倍体不育菌丝缠绕组织化形成;3)部分菌核既不具有MAT1-1,也没有MAT1-2,出现交配型全部缺失现象.交配型缺失在羊肚菌菌种退化检测中的应用值得探寻,而羊肚菌菌核的偏分离和交配型缺失的机制及其与菌种选育和子实体产量之间的关系也有待深入研究.     

生物钟基因突变型prd—4对野生型prd—4^＋呈不完全显性

运用异核体对生物钟基因突变型ｐｒｄ－４和野生型ｐｒｄ－４＋的显隐性关系进行了研究。经多次回交除去１０多个异核体不相容位点的等位基因差异。在回交前分别导入肌醇缺陷型和泛酸缺陷型突变。回交后代分别含有上述两遗传突变,并分别含有ｐｒｄ－４和ｐｒｄ－４＋位点。由于遗传互补,从回交后代构建的异核体能在基本培养基上生长。异核体的ｐｒｄ－４／ｐｒｄ－４＋核比例即为肌醇缺陷型／泛酸缺陷型核比例。分别在２５℃和２９℃确定异核体近似日周期。结果表明ｐｒｄ－４对ｐｒ－４＋呈不完全显性。     

球孢白僵菌种群野生菌株异核现象的分子验证*

球孢白僵菌28S rRNA基因D11域由于一内含子的存在与否,引起这一区域的扩增产物有两种片段类型(约520bp及120bp),将此作为分子标记分析了球孢白僵菌的117个菌株,发现17%菌株的扩增产物为双片段类型,即为异核体。异核体在不同种群中的分布频率因地理生态条件而异。单孢分离分析表明,异核体的分离频率约为35%。RAPD检测结果说明异核体在准性循环中发生了不同程度的遗传交换及遗传重组。球孢白僵菌异核体在继代培养中极不稳定。     

Biosurfactants are involved in the biological control of Verticillium microsclerotia by Pseudomonas spp

AIMS: To examine the effect of previously described bacterial antagonists on the viability of Verticillium microsclerotia in vitro and to elucidate the possible modes of action of bacterial strains in the suppression of Verticillium microsclerotia viability. METHODS AND RESULTS: A microplate assay was developed to test the suppressive effect of well-defined Pseudomonas spp. on the viability of Verticillium microsclerotia in vitro. Experiments using phenazine- and biosurfactant-deficient mutants indicated that biosurfactants and phenazine-1-carboxylic acid play a role in the suppression of microsclerotia viability by Pseudomonas spp. In addition, microsclerotia colonization tests revealed that Pseudomonas spp. are able to colonize the surface of the microsclerotia, but not the inner matrix. Growth response curves showed that the population levels of Pseudomonas spp. increased when they were in the vicinity of Verticillium microsclerotia, indicating that Pseudomonas spp. may utilize nutrients from the microsclerotia for their growth. CONCLUSIONS: Pseudomonas spp. seem to be good candidates for Verticilllium microsclerotia biocontrol. Biosurfactant production is one of the main mechanisms involved in their mode of action. SIGNIFICANCE AND IMPACT OF THE STUDY: This line of work may contribute to a better understanding of biological control agents and their working mechanisms.     

Microsclerotial germination of Verticillium dahliae as affected by rape rhizosphere

Abstract The germination of nylon net-trapped microsclerotia of Verticillium dahliae pathogenic to rape ( Brassica napus ) was assessed in different systems by fluorescence microscopy using fluorescein diacetate. The influence of the culture's age and the size of the microsclerotia on germination percentages was assessed in water, mineral salts solution and mineral salts solution plus sucrose for 3 V. dahliae isolates. Large microsclerotia germinated better than smaller ones. The microsclerotia of 2 isolates showed decreased germination percentages with culture age over a 4–11-week period. Microsclerotial germination percentages were always higher in mineral salts solution plus sucrose than in mineral salts solution alone or water. In a sand culture system with the intact rape plant, microsclerotial germination percentages were high close to the root and decreased in a steep gradient to background levels within 5 mm from the root.     

Formation and ultrastructure of microsclerotia of Pyrenochaeta lycopersici

The formation of microsclerotia of Pyrenochaeta lycopersici Schneider & Gerlach was observed in culture, in isolated tomato root segments and in roots of intact tomato plants. In culture microsclerotia formed in the aerial mycelium, and submerged in the agar; in tomato roots they formed in single cells of the outer cortex. The microsclerotium of P. lycopersici is structurally unspecialized and has cell walls of similar thickness throughout. Microsclerotia are thought to have powers of long survival in soil. In isolation the microsclerotia of P. lycopersici maintained viability over 2 years with consistently high germinability. Transverse hyphae were never seen in sections of microsclerotia, suggesting that germ-tubes arise from the outermost layers of cells.     

Morphological and pathogenic variability in Macrophomina phaseolina isolates of pigeonpea (Cajanus cajan L.) and their relatedness using principle component analysis

Sixty-one isolates of Macrophomina phaseolina were characterised on the basis of different morphological characters to study the variability in the population. PCA analysis extracted three main components, microsclerotia, texture and colour, from the population that described the variability in the population most appropriately. Colour of the isolates and the presence of the microsclerotia have a significant effect on the area under disease progress curve. Isolates with the production of microsclerotia (M⁺) were more aggressive as compared to isolates with no production of microsclerotia (M^?). The study has brought out pathogenic variation in M. phaseolina, thus better understanding of host pathogen relationship to identify physiologic races in the breeding programme.     

Effect of Temperature on the Formation of Microsclerotia of Verticillium dahliae

Microsclerotium formation by six isolates of Verticillium dahliae was studied at different temperatures both in vitro and in Arabidopsis thaliana . In vitro mycelial growth was optimal at 25°C, but microsclerotium formation was greatest at 20°C (two isolates) or 15–20°C (one isolate). Seedlings of A. thaliana were root-dipped in a conidial suspension, planted, and either placed at 5, 10, 15, or 25°C, or left at 20°C until the onset of senescence, after which some of the plants were placed at 5, 10, 15, or 25°C. The amount of microsclerotia per unit of shoot weight was assessed in relation to isolate and temperature. The optimal temperature for production of microsclerotia was 15–25°C. Two isolates each produced about 10 times more microsclerotia than each of the other four isolates. For these isolates, high R ²_adj.-values of 0.77 and 0.66 were obtained, with temperature and its square as highly significant (P   < 0.001) independent variables. R ²_adj.-values for the other isolates varied between 0.28 and 0.39. Moving plants to different temperatures at the onset of senescence led to microsclerotial densities that were intermediate between densities on plants that had grown at constantly 20°C and plants grown at other temperatures. This suggests that vascular colonization rate and rate of microsclerotium formation are similarly affected by temperature. The senescence rate of plants appeared unimportant except for plants grown at 25°C, which showed the highest amounts of microsclerotia per unit of plant weight in the most rapidly senescing plants.     

Liquid culturing of microsclerotia of Mycoleptodiscus terrestris,a potential biological control agent for the management of hydrilla

Mycoleptodiscus terrestris has potential as an inundative biological control agent for the management of hydrilla, one of the world’s worst aquatic weeds. Essential to producing a marketable bioherbicidal product was the development of liquid culture procedures that would yield propagules that maintained biocontrol efficacy. Since M. terrestris did not produce conidia in liquid culture, various nutritional conditions were evaluated as a means to produce high concentrations of stable fungal propagules such as microsclerotia. Evaluations of propagule formation and biomass yield were carried out in liquid culture media containing a basal salts solution amended with corn steep liquor powder or cottonseed meal combined with 4% or 6% glucose. Hyphal aggregation was observed by day 2, and by day 8 abundant melanized microsclerotia were present in the broth cultures. When applied as a liquid inoculum to hydrilla at rates of 0.1 and 0.2 ml/l, the microsclerotial matrix was capable of significantly reducing hydrilla shoot biomass by as much as 99%. Air-dried microsclerotia were capable of hyphal germination in 24 h and sporogenic germination in 72 h. These capabilities have significance for the use of microsclerotia of M. terrestris as the preferred inoculum for biocontrol purposes. Hyphae germinating from microsclerotia on hydrilla plant surfaces can establish initial infection sites followed several days later by secondary infections resulting from the development and release of spores from the surface of the microsclerotia. The capability of microsclerotia of M. terrestris to remain stable as a dry preparation and to germinate both hyphally and sporogenically upon rehydration enhances the potential of this fungus for use as a nonchemical, biological control agent for hydrilla.     

VdToxD1和VdToxD3调控大丽轮枝菌微菌核的萌发

大丽轮枝菌Verticillium dahliae是一种土传性植物病原真菌,可侵染660多种植物,引发黄萎病。微菌核是大丽轮枝菌在侵染后期形成的特殊休眠结构,可在土壤中存活14年,是黄萎病的主要初侵染来源。本研究在前期大丽轮枝菌萌发与休眠微菌核转录组分析的基础上,选择微菌核休眠状态高表达的VdToxD1和VdToxD3基因进行功能研究。结果发现,VdToxD1和VdToxD3敲除突变体菌株的微菌核萌发率分别为87.3%和86.0%,显著高于野生型菌株的64.0%;负调控微菌核萌发的转录因子VdCrz1与VdToxD1上游的-937 bp到-349 bp中的特定基序结合,与VdToxD3上游的-752 bp到-496 bp中的特定基序结合,结合位点均位于2个基因的启动子区。     

大丽轮枝菌 VdCPMO基因克隆与功能分析

大丽轮枝菌 Verticillium dahliae是一种典型的土传病原真菌,可侵染400多种植物引致黄萎病,造成巨大的经济损失。微菌核是大丽轮枝菌的特殊休眠结构,能够在土壤中存活14年之久,是病害的主要初侵染来源。本研究在前期微菌核萌发表达谱的基础上,选择上调倍数最高的环戊酮1,2-单加氧酶基因（ VDAG_03943）,进行克隆与功能分析。结果表明,该基因全长1 929bp,cDNA序列全长1 668bp,编码555个氨基酸组成的蛋白,命名为 VdCPMO。与野生型菌株JY相比,敲除突变体菌株的菌落生长减慢,微菌核萌发率显著降低,芽管平均长度明显较短,而互补突变体菌株与野生型菌株JY间无显著性差异。致病力测定结果显示,敲除突变体菌株的微菌核丧失了对棉花的致病力。     

贵州栽培笃斯越橘DSE真菌、AM真菌与ERM真菌空间分布研究

为探明贵州省栽培笃斯越橘根系深色有隔内生真菌（dark septate endophytes,DSE）、丛枝菌根（arbuscular mycorrhiza,AM）真菌与欧石南菌根（ericoid mycorrhiza,ERM）真菌的定殖及地理分布情况,揭示共生真菌在栽培笃斯越橘生长中的地位,本研究在贵州省笃斯越橘主栽区麻江县、凤岗县和高坡乡分别选取主栽品种圆蓝、粉蓝、奥尼尔和莱格西的根样及根围土样,观测不同地区不同品种根样DSE真菌、AM真菌和ERM真菌的定殖结构和定殖率,并测定土样土壤理化性质,分析不同真菌与土壤因子相关性。结果表明,3个地区的4个笃斯越橘品种均有DSE真菌、AM真菌和ERM真菌定殖,栽培笃斯越橘能与3类真菌形成共生关系,平均定殖率分别为61.11%、25.55%和22.50%。DSE真菌定殖率：高坡（62.50%）>麻江（61.66%）>凤岗（59.16%）;AM真菌定殖率：凤岗（34.14%）>麻江（25.83%）>高坡（16.66%）;ERM真菌定殖率：高坡（35.00%）>凤岗（20.00%）>麻江（12.5%）。相关性分析表明,DSE真菌中的菌丝与微菌核的定殖率呈负相关,AM真菌的总定殖率及定殖强度与微菌核的定殖率呈极显著正相关,与DSE真菌菌丝的定殖率呈负相关。ERM真菌总定殖率与DSE真菌菌丝的定殖率及AM真菌定殖强度呈极显著正相关,与DSE真菌总定殖率呈显著正相关。土壤有效磷与DSE真菌和ERM真菌总定殖率呈显著正相关,与AM真菌定殖率呈显著负相关。土壤铵态氮与DSE真菌中微菌核结构定殖率及AM真菌定殖率呈极显著正相关,与ERM真菌定殖率呈极显著负相关。土壤pH值与DSE定殖强度呈显著负相关,与ERM定殖强度呈极显著正相关。本研究分析比较贵州省3个笃斯越橘种植基地不同品种栽培笃斯越橘DSE真菌、AM真菌和ERM真菌的定殖及其与土壤理化性质之间的相关性,为贵州省栽培笃斯越橘的管理和发展提供技术基础和理论依据。