Combining the oomycete Pythium oligandrum with two other antagonistic fungi: Root relationships and tomato grey mold biocontrol

To reduce Pythium oligandrum biocontrol variability and improve its efficacy, experiments were performed by combining the oomycete with two other antagonistic fungi, Fusarium oxysporum strain Fo47 and Trichoderma harzianum. In Petri dishes, Fo47 or T. harzianum hyphae destroyed P. oligandrum cells by antibiosis and mycoparasitism processes; in the rhizosphere of tomato plants (Lycopersicon esculentum), the same antagonistic features were observed. However, in the rhizosphere, hyphae are frequently separated by a certain distance; this allows the coexistence and the persistence of the three microorganisms on the root systems. When introduced in the rhizosphere, Fo47 and P. oligandrum were able to penetrate the root tissues with Fo47 limited to the epidermal and upper layers of cortical cells while P. oligandrum colonized deeper tissue at a faster rate. The two antagonists were killed in few days within roots following elicited plant-defense reactions. T. harzianum was not able to penetrate root tissues. Root colonization with either P. oligandrum alone or in combination with Fo47 and/or T. harzianum resulted in systemic plant resistance which provided plant protection against Botrytis cinerea infection of leaves. The level of control and the expression of pathogenesis-related proteins (PR-proteins) in leaves were similar whatever the antagonistic microbial treatment applied to roots.     

Interactions of Neotyphodium gansuense, Achnatherum inebrians, and plant-pathogenic fungi

Interactions of Neotyphodium gansuense, Achnatherum inebrians, and nine fungal pathogens were studied by tests of inhibition of four fungal pathogens by Neotyphodium endophytes in vitro and by inoculation of nine fungal pathogens on detached leaves of endophyte-infected (E+) and endophyte-free (E−) plants. Compared with the controls, most isolates of N. gansuense significantly inhibited the growth in vitro of, in decreasing order of inhibition, Bipolaris sorokiniana, Curvularia lunata, Fusarium acuminatum, and Alternaria alternata. Inhibition zones appeared between pathogens and some isolates of N. gansuense. Some isolates of N. gansuense significantly inhibited sporulation of B. sorokiniana, A. alternata, and C. lunata. However, there was no significant inhibition of F. acuminatum and a few isolates significantly increased sporulation. The leaf inoculation trial indicated that almost all fungal pathogens were able to cause lesions on detached leaves regardless of endophyte status. Both the number and size of disease lesions on E+ A. inebrians leaves caused by A. alternata, F. chlamydosporum, F. oxysporum, and F. solani were reduced compared with those on E− leaves. Only lesion numbers (not size) of Ascochyta leptospora leaf spots were significantly reduced on E+ leaves compared with E− leaves. Conversely, only the length of Ascochyta leptospora leaf spots were significantly smaller on E+ leaves than on E− leaves; numbers of lesions were not significantly affected. C. lunata was strongly pathogenic to both E+ and E− leaves and numerous lesions developed and merged into patches, the leaf surface was covered and the leaf rotted away.     

农杆菌介导的携带egfp基因载体转化寡雄腐霉

【目的】寡雄腐霉(Pythium oligandrum Drechsler)是一种对动、植物和环境无害,兼具杀菌和增产效果的生防真菌。通过研究建立农杆菌介导的寡雄腐霉遗传转化体系。【方法】选用EHA105、AGL-1、LBA4404三种农杆菌菌株对寡雄腐霉进行遗传转化研究,通过对影响遗传转化效果的条件参数试验优化,确立适宜寡雄腐霉遗传转化的农杆菌菌株及转化条件,建立农杆菌介导的寡雄腐霉遗传转化体系。【结果】经研究发现,所选3种农杆菌菌株中EHA105菌株对寡雄腐霉的遗传转化效果最好,其次是AGL-1菌株,LBA4404菌株转化效果不好。EHA105菌株经IM(含300μmol/L AS)诱导培养至OD_(600)=0.6时,与浓度为10~6–10~7个/m L的寡雄腐霉孢子悬浮液以1–10:1的比例混合,在25–26°C以液体振荡的方式避光共培养72 h(pH 5.0,含300μmol/L AS),寡雄腐霉菌体液体振荡恢复培养24 h,涂布抗性选择平板筛选寡雄腐霉转化子,即可得到寡雄腐霉基因工程菌株,其转化率可达到130个转化子/106个孢子。【结论】本研究首次构建了农杆菌介导的寡雄腐霉遗传转化体系,研究结果可为寡雄腐霉的生防机制及分子育种研究提供技术支撑。     

寡雄腐霉发酵液的动物安全性及其对黄瓜的防病促生效应

【目的】生物农药安全无害,环境友好。为了评价寡雄腐霉发酵液(Pythium oligandrum broth,POB)的动物安全性和防病促生效应,研制高效、无害的生物农药。【方法】试验利用自主分离的寡雄腐霉生防菌株(P.oligandrum CQ2010)制备POB,通过动物试验、拮抗试验、盆栽和生产试验,研究了POB的动物毒性,对黄瓜蔓枯病的防治作用,以及对黄瓜生长、产量和品质的影响。【结果】用大剂量的POB灌胃给药对小鼠体重增长无显著影响,其外观和行为均无异常,组织器官也未见病理改变。POB对甜瓜球腔菌的抑制率为51.95%,药效介于1∶800的百菌清溶液和1∶200的甲基托布津溶液之间。在黄瓜幼苗接种甜瓜球腔菌前后喷施POB,叶片丙二醛含量下降,过氧化物酶和超氧化物歧化酶活性增强,发病率和病情指数均显著降低,相对防治效果达到54.8%–64.1%,故POB能减轻病原菌对细胞膜的伤害,激发防御性生理反应,增强黄瓜植株的抗病能力。此外,POB处理提高叶绿素含量,增强根系活力,增加植株氮、磷、钾吸收量,促进黄瓜植株生长,生物量和果实产量分别提高81.10%和11.58%。POB还使黄瓜果实Vc和可溶性糖提高,硝酸盐含量降低。【结论】寡雄腐霉发酵液对动物安全无毒,能有效防治黄瓜蔓枯病,促进黄瓜生长,提高产量品质。     

Interactions between two rust fungi and their host plant Anemone nemorosa

In a field study we found that two rust fungi. Tranzchelia anemones and Ochropsora ariae. on the shared host Anemone nemorosa varied in abundance among populations. Tranzchelia anemones was favoured in high light habitats and was particularly common in mixed deciduous forests as well as productive grasslands, whereas the abundance and occurrence of O. ariae was dependent on the presence of the alternate host, Sorbus aucuparia When found in the same population, the two rusts rarely attacked the same ramet. Both T. anemones and O. ariae had a negative effect on plant performance by reducing flower production, and rhizome length. Diseased shoots were also elongated.     

Interactions between mammals and ectomycorrhizal fungi

Many ectomycorrhizal (ECM) fungi produce fruit-bodies below ground and rely on animals, especially mammals, for dispersal of spores. Mammals may therefore play an important role in the maintenance of mycorrhizal symbiosis and biodiversity of ECM fungi in many forest ecosystems. Given the pivotal role played by mycorrhizal fungi In the nutrition of their plant hosts and, possibly, in the determination of plant community structure, the ecological significance of mycophagous mammals may extend to the productivity and diversity of plant communities. Mycologists and mammalogists have been aware of the interaction between their study organisms for many years, but recent research has produced new insights Into the evolution of mammal-vectored spore dispersal among ECM fungi, the ecological importance of mycophagy to small mammals, and the effectiveness of mammals as spore-dispersal agents.     

Virulence genes and the evolution of host specificity in plant-pathogenic fungi

In the fungal kingdom, the ability to cause disease in plants appears to have arisen multiple times during evolution. In many cases, the ability to infect particular plant species depends on specific genes that distinguish virulent fungi from their sometimes closely related nonvirulent relatives. These genes encode host-determining "virulence factors," including small, secreted proteins and enzymes involved in the synthesis of toxins. These virulence factors typically are involved in evolutionary arms races between plants and pathogens. We briefly summarize current knowledge of these virulence factors from several fungal species in terms of function, phylogenetic distribution, sequence variation, and genomic location. Second, we address some issues that are relevant to the evolution of virulence in fungi toward plants; in particular, horizontal gene transfer and the genomic organization of virulence genes.     

Interactions between vesicular-arbuscular mycorrhizal fungi and asparagus

Summary The vesicular-arbuscular mycorrhizal (VAM) fungus,Glomus versiforme increased significantly the growth ofAsparagus officinalis under controlled conditions using Turface as the growth medium. The growth responses, including increases in root fresh weight, numbers of shoots, shoot dry weight, and shoot height follow a pattern similar to other mycorrhizal systems. Indigenous VAM fungi appeared to have negative effects on average shoot fresh and dry weight, number of shoots per pot and average shoot height on one year oldA. officinalis seedlings obtained from the field and grown under controlled conditions. These results may be due either to the high levels of soluble phosphate present in the soil or the ineffectiveness of the particular indigenous fungi as mycorrhizal fungi in asparagus. Indigenous mycorrhizal fungi overwinter in asparagus root crown as vesicles and as external and internal hyphae. Soil obtained from the same fields as the one year old crowns was a good source of mycorrhizal inoculum for sterile seedlings.     

Histopathological studies of sclerotia of phytopathogenic fungi parasitized by a GFP transformed Trichoderma virens antagonistic strain

The gfp gene from the jellyfish Aequorea victoria, coding for the Green Fluorescent Protein (GFP), was used as a reporter gene to transform a Trichoderma virens strain I10, characterized as having a promising biocontrol activity against a large number of phytopathogenic fungi. On the basis of molecular and biological results, a stable GFP transformant was selected for further experiments. In order to evaluate the effects of GFP transformation on mycoparasitic ability of T. virens I10, sclerotia of Sclerotium rolfsii, Sclerotinia sclerotiorum and S. minor were inoculated with the T. virens strain I10 GFP transformant or the wild type strain. Statistical analysis of percentages of decayed sclerotia showed that the transformation of the antagonistic isolate with the GFP reporter gene did not modify mycoparasitic activity against sclerotia. Sclerotium colonization was followed by fluorescent microscopy revealing intracellular growth of the antagonist in the cortex (S. rolfsii) and inter-cellular growth in the medulla (S. rolfsii, and S. sclerotiorum). The uniformly distributed mycelium of T. virens just beneath the rind of sclerotia of both S. rolfsii and S. sclerotiorum suggests that the sclerotia became infected at numerous randomly distributed locations without any preferential point of entry.     

Interactions between killer yeasts and pathogenic fungi

Abstract A total of 17 presumptive killer yeast strains were tested in vitro for growth inhibitory and killing activity against a range of fungal pathogens of agronomic, environmental and clinical significance. Several yeasts were identified which displayed significant activity against important pathogenic fungi. For example, isolates of the opportunistic human pathogen, Candida albicans , were generally very sensitive to Williopsis mrakii killer yeast activity, whilst killer strains of Saccharomyces cerevisiae and Pichia anomala markedly inhibited the growth of certain wood decay basidiomycetes and plant pathogenic fungi. Results indicate that such yeasts, together with their killer toxins, may have potential as novel antimycotic biocontrol agents.     

Mechanics of Solid Tissue Invasion by the Mammalian Pathogen Pythium insidiosum

The relative significance of mechanical penetration versus the action of substrate-degrading enzymes during solid tissue invasion has not been established for any fungal disease. Pythium insidiosum is an oomycete fungus (or stramenopile) that causes a rare, but potentially lethal infection in humans and other mammalian hosts. Experiments with miniature strain gauges showed that single hyphal apices of this pathogen exert forces of up to 6.9 μN, corresponding to maximum pressures of 0.3 μN μm⁻² or MPa. Samples of cutaneous and subcutaneous tissue from fresh human cadavers displayed a mean strength (resistance to needle puncture) of 24 μN μm⁻², and a mean pressure of 30 μN μm⁻² was necessary to penetrate skin strips from slaughtered horses. These experiments demonstrate that P. insidiosum does not exert sufficient pressure to penetrate undamaged skin by mechanics alone, but must effect a decisive reduction in tissue strength by proteinase secretion.     

不同叶型维西堇菜叶片结构及光合特征比较

为探讨叶片斑纹的光合适应意义,该文选择有斑和无斑两种叶型的维西堇菜(Viola monbeigii)作为研究材料,采用石蜡切片制片和显微镜观察叶片结构及GFS 3000便携式光合测量仪测量光合参数并进行比较。结果表明：(1)两种叶型维西堇菜气孔均为不等型气孔,有斑叶片上、下表皮气孔密度、栅栏组织(PT)厚度、栅栏组织与海绵组织的比值(PT/ST)均显著低于无斑叶片;而上表皮气孔较大,表现出更适应弱光环境的结构特征。(2)两种叶型维西堇菜暗呼吸速率(R_d)、初始荧光(F_o)、最大荧光产量(F_m)、光化学猝灭系数(q_P)、非光化学猝灭系数(q_N)、PSⅡ最大光化学量子产量(F_v/F_m)、实际光化学反应量子效率(Yield)均无显著差异,有斑植株叶片叶绿素含量、最大净光合速率(P_max)显著低于无斑植株;有斑叶片表观光合电子传递速率(ETR)在光合有效辐射(PAR)处于400～2 000μmol·m^-2     

Intra-isolate heterogeneity of the ITS region of rDNA in Pythium helicoides

Heterogeneity of the rDNA ITS region in Pythium helicoides and the phylogenetic relationship between P. helicoides and closely related species were investigated. In PCR-RFLP analysis of the rDNA ITS region of six P. helicoides isolates investigated, including the type culture, intraspecific variation was found at the HhaI site. The total length of fragments was longer than before cutting, indicating sequence heterogeneity within isolates. Digestion of the cloned rDNA ITS region derived from seven isolates with HhaI revealed polymorphisms among and within single zoospore isolates, and variability of the region was also present among the clones derived from the same isolate. To test whether the rDNA ITS region of closely related species and other regions in the genome of P. helicoides are also variable, the rDNA ITS region of P. ultimum and the cytochrome oxydase II (cox II) gene encoded in mitochondria were sequenced. P. ultimum had little variation in the rDNA ITS region. The cox II gene sequences of both species revealed only a low intraspecific variability and no intra-isolate variation. In the phylogenic tree based on the rDNA ITS sequences, all clones of P. helicoides formed one large clade that was distinct from the clades comprising morphologically similar species, such as P. oedochilum and P. ostracodes, and was closely related to P. chamaehyphon rather than the other species.