丛枝菌根真菌对不同番茄品种抗根结线虫病的影响

【背景】根结线虫病害严重制约我国设施蔬菜的生产。丛枝菌根真菌(Arbuscular Mycorrhizal Fungi,AMF)作为土壤中最重要的有益真菌之一,可以促进植物生长,提高植物抗病性,减轻土传真菌和线虫病害的发生。在蔬菜保护地栽培中,AMF对于植物线虫病防治作用的研究受到了广泛关注。【目的】针对番茄生产中危害最严重的南方根结线虫(Meliodogyne incognita)病害问题,研究AMF和番茄品种不同组合的抗线虫效应,以期为菌根真菌作为生物防治剂和生物菌肥应用于实际生产提供技术基础。【方法】在灭菌土壤中,人工接种根结线虫,比较不同菌种Rhizophagus intraradices(Ri)、Acaulosporamellea(Am)及菌种组合Rhizophagusintraradices+Acaulosporamellea(Ri+Am)在不同番茄品种(感病品种蒙特卡罗和抗线虫品种仙客1号)上对南方根结线虫侵染和繁殖的影响,研究AMF对根结线虫的拮抗效应。另外,采用南方根结线虫连作发病的土壤,在感病品种蒙特卡洛上接种AMF混合菌种Ri+Am,番茄苗移栽入连作土壤中,测定各生长指标和调查根结和卵块数量,评价接种AMF处理对根结线虫病的防治效果。【结果】在灭菌土壤中,普通番茄品种蒙特卡罗的菌根效应显著优于抗线虫番茄品种仙客1号,表现为前者单位根重的根结和卵块的数量均比对照显著降低,而后者仅降低了卵块数量;蒙特卡罗上接种Ri+Am混合菌种的效果优于接种单一菌种Am和Ri;而仙客1号上接种Ri的效果更好。接种线虫也显著影响了AMF的侵染,而且对抗性品种仙客1号的影响更为明显。但除了接种Am的处理,大多数处理收获时菌根侵染率仍维持较高的水平(70%以上)。在连作土壤中,感病品种蒙特卡罗接种混合菌种Ri+Am具有较好的抗/耐线虫效应,主要表现为促进植株生长,显著降低根结和卵块数量,但菌根侵染率较灭菌土壤低,约为40%。【结论】综合以上结果,表明菌根化苗能够在一定程度上减轻根结线虫病的危害。土壤灭菌条件下,在感病和抗线虫番茄品种上接种AMF能够减轻线虫的侵染和繁殖,而且在感病品种上的防治效果更加显著。在连作土壤中,在番茄感病品种上接种AMF也表现较好的抗线虫效果。     

In vitro studies of antagonistic fungi against the root-knot nematode,Meloidogyne incognita

The present study was carried out in vitro to determine the efficacy of indigenous fungi isolated from egg masses of root-knot nematode, Meloidogyne incognita on egg parasitism, egg hatching, mobility and mortality against root-knot nematode, M. incognita. The tested fungi were Acremonium strictum, Aspergillus terreus, A. nidulans, A. niger, Chetomium aubense, Chladosporium oxysporum, Fusarium chlamydosporium, F. dimarum, F. oxysporum, F. solani, Paecilomyces lilacinus, Pochonia chlamydosporia, Trichoderma viride and T. harzianum. All tested fungi showed varied effects against the nematodes. Culture filtrates of A. strictum was very effective against the nematode in regards to egg parasitism (53%), egg hatching inhibition (86%) and mortality (68%) compared to controls. A. strictum was found to have an advantage over P. lilacinus, P. chlamydosporia, T. viride and T. harzianum in that it caused greater mortality of the second stage juveniles (J₂). A. terreus did not show egg parasitism but was found to be highly toxic against second stage juveniles (J₂) causing high mortality (around 68%). Thus, A. strictum and A. terreus showed good biocontrol potential against root-knot nematode, M. incognita under in vitro conditions.     

Interactions between the root-knot nematode Meloidogyne incognita and Glomus mosseae in banana

The effects of the interaction between the arbuscular mycorrhizal fungus Glomus mosseae and the root-knot nematode Meloidogyne incognita on growth and nutrition of micropropagated ;Grand Naine banana (Musa AAA) cultivar was studied under greenhouse conditions. Inoculation with two G. mosseae isolates significantly increased growth of plants in relation to non-mycorrhizal plants. Response to mycorrhizae was as effective as with an optimum P fertilization in promoting plant development for most growth parameters. Meloidogyne incognita had no apparent effect on the percentage of root colonization in mycorrhizal plants. In contrast, G. mosseae suppressed root galling and nematode buildup in the roots. The percentage of mycorrhizal colonization was high (over 80%) in low P fertilized plants, but optimum P rates for bananas (four times higher than low P) significantly reduced mycorrhizal colonization. Most elements were within sufficiency levels for banana with exception of N which was low for all treatments. Mycorrhizal plants fertilized with a low P rate showed higher N, P, K, Ca, and Mg contents as compared to non-mycorrhizal plants low in P with or without the nematode. Inoculation with G. mosseae favours growth of banana plants by enhancing plant nutrition and by suppressing nematode reproduction and galling during the early stages of plant development.     

Suppression of the root-knot nematode [Meloidogyne incognita (Kofoid & White) Chitwood] on tomato by dual inoculation with arbuscular mycorrhizal fungi and plant growth-promoting rhizobacteria

Arbuscular mycorrhizal (AM) fungi and plant growth-promoting rhizobacteria (PGPR) have potential for the biocontrol of soil-borne diseases. The objectives of this study were to quantify the interactions between AM fungi [Glomus versiforme (Karsten) Berch and Glomus mosseae (Nicol. & Gerd.) Gerdemann & Trappe] and PGPR [Bacillus polymyxa (Prazmowski) Mace and Bacillus sp.] during colonization of roots and rhizosphere of tomato (Lycopersicon esculentum Mill) plants (cultivar Jinguan), and to determine their combined effects on the root-knot nematode, Meloidogyne incognita, and on tomato growth. Three greenhouse experiments were conducted. PGPR increased colonization of roots by AM fungi, and AM fungi increased numbers of PGPR in the rhizosphere. Dual inoculations of AM fungi plus PGPR provided greater control of M. incognita and greater promotion of plant growth than single inoculations, and the best combination was G. mosseae plus Bacillus sp. The results indicate that specific AM fungi and PGPR can stimulate each other and that specific combinations of AM fungi and PGPR can interact to suppress M. incognita and disease development.     

Biocontrol of root-knot nematode Meloidogyne incognita by the isolates of Bacillus on tomato

Fifteen isolates of Bacillus, isolated from the root-knot nematode suppressive soils, were used for the biocontrol of Meloidogyne incognita on tomato. Bacillus isolates B1, B4, B5 and B11 caused greater inhibitory effect on hatching of M. incognita than caused by other isolates. In addition, these isolates (B1, B4, B5 and B11) caused greater colonisation of tomato roots and also caused greater increase in the growth of tomato seedling than caused by other isolates. All the isolates of Bacillus were able to increase growth of tomato and caused reduction in galling and nematode multiplication in green house tests. Isolates B1, B4, B5 and B11 caused a greater increase in growth of tomato and higher reduction in galling and nematode multiplication than other isolates in a green house test. These isolates were also tested for hydrogen cyanide (HCN) and indole acetic acid productions. Only one isolate (B13) produced HCN out of 15 tested. On the other hand, isolates B5, B11, B4 and B1 showed greater production of IAA than the other 11 isolates tested. This study suggests that Bacillus isolates B5, B11, B4 and B1 may be used for the biocontrol of M. incognita on tomato.     

丛枝菌根真菌和植物寄生线虫

本文综述了土壤微生物中丛枝菌根真菌和植物寄生线虫的互作关系及其互作机理,并阐述了丛枝菌根真菌在防治植物线虫病害方面的应用前景和实际操作中应注意的技术环节。     

Biocontrol of root-knot nematode Meloidogyne incognita by the isolates of Pseudomonas on tomato

Biocontrol of root-knot nematode Meloidogyne incognita was studied on tomato using 15 isolates of fluorescent Pseudomonads isolated from pathogen suppressive soils. Pseudomonas aeruginosa (isolates Pa8, Pa9 and Pa3) caused greater inhibitory effect on hatching of M. incognita than other isolates. In addition, isolates Pa8, Pa9 and Pa3 caused greater colonisation of tomato roots and also caused a greater increase in the growth of tomato seedlings. These isolates also caused a greater increase in growth of tomato and higher reduction in galling and nematode multiplication in a green house test than is caused by other isolates. Isolates Pf1, Pf5, Pf6 and Pa13 were unable to increase growth of tomato and caused less reduction in galling and nematode multiplication compared to other isolates. Only 10 isolates produced siderophores on chromo-azurol sulfonate (CAS) agar medium and isolate Pa12 showed greater production of siderophore followed by Pa11, Pa9, Pf10, Pa3 and Pf5. Similarly, isolates Pa14, Pa12, Pf10, Pa9, Pa8, Pa7 and Pa6 produced greater amount of HCN than the other isolates tested. Isolates Pa8 and Pa9 showed greater production of IAA than the other 13 isolates tested. This study suggests that P. aeruginosa isolates Pa8 and Pa9 may be used for the biocontrol of M. incognita on tomato.     

Histopathology of roots of Glycine max (L.) Merrill induced by root-knot nematode (Meloidogyne incognita)

In Glycine max, the second-stage juveniles of Meloidogyne incognita entered the roots through the apical meristem or elongation zone. The juveniles induced giant cells in the zone of vascular strands. Near the head of the nematode and adjacent to the giant cells, the vascular strands exhibited abnormalities in their shapes and structures; both xylem and phloem were found to be affected. The giant cells had dense and granular cytoplasm, and large nuclei with large nucleoli. Some parenchyma cells exhibited hypertrophy, while others exhibited hyperplasia. The distinctive feature of the study is reporting the occurrence of abnormal xylem, abnormal phloem and abnormal parenchyma.     

Ralstonia solanacearum colonization of tomato roots infected by Meloidogyne incognita

Bacterial wilt, caused by Ralstonia solanacearum, is one of the most serious diseases of tomato (Solanum lycopersicum). Concomitant infection of R. solanacearum and root‐knot nematode Meloidogyne incognita increases the severity of bacterial wilt in tomato, but the role of this nematode in disease complexes involving bacterial pathogens is not completely elucidated. Although root wounding by root‐knot nematode infection seems to play an important role, it might not entirely explain the increased susceptibility of plants to R. solanacearum. In the present study, green fluorescent protein (GFP)‐labelled R. solanacearum distribution was observed in the root systems of the tomato cultivar Momotaro preinoculated with root‐knot nematode or mock‐inoculated with tap water. Fluorescence microscopy revealed that GFP‐labelled R. solanacearum mainly colonized root‐knot nematode galls, and little or no green fluorescence was observed in nematode‐uninfected roots. These results suggest that the gall induced by the nematode is a suitable location for the growth of R. solanacearum. Thus, it is crucial to control both R. solanacearum and root‐knot nematode in tomato production fields to reduce bacterial wilt disease incidence and effects.     

Identification of arbuscular mycorrhizal fungi in soils and roots of plants colonizing zinc wastes in southern Poland

 Analysis of the community of arbuscular mycorrhizal (AM) fungi in roots of Fragaria vesca growing in a heavy metal contaminated site was carried out on a Zn waste site near Chrzanow (southern Poland). The waste substratum was characterized by high contents of Pb, Zn, Cd, Cu and As, and by low levels of N, P and organic matter. Spores of Glomales were isolated by wet sieving and DNA was isolated from individual spores. Nested polymerase chain reaction (PCR) with taxon-specific primers was used to identify the species Glomus mosseae, Glomus intraradices and Glomus claroideum. Spores of other fungi were morphologically characterized and new taxon-discriminating molecular probes were developed for two of them (Glomus sp. HM-CL4 and HM-CL5) based on variations in the large ribosomal subunit (25S rDNA). High sequence similarities were found between Glomus sp. HM-CL4 and Glomus gerdemanii, and between Glomus sp. HM-CL5 and Glomus occultum. The designed primers were used to characterize the population of AM fungi colonizing the roots of F. vesca collected from the Zn waste site. The analysis, carried out on roots stained with trypan blue, showed that the most effective colonizer was closely related to G. gerdemannii. G. claroideum and the G. occultum-like fungus were slightly less common whilst frequencies of G. intraradices and G. mosseae in roots were much lower. The analysis of mycorrhiza stained with rhodizoniate to localize heavy metal accumulation showed that the stain does not influence the PCR reaction. Seventy percent of the root samples containing positively stained fungal hyphae were found to be colonized by G. mosseae. The data obtained demonstrate the usefulness of nested PCR for studies carried out in polluted areas. It will enable selection of AM fungi which are able to colonize plant roots under heavy metal stress conditions, as well as the identification of fungi showing high in situ accumulation of potentially toxic elements. Accepted: 7 July 2000     

Integrated management of root-knot nematode, Meloidogyne incognita infestation in tomato and grapevine

An integrated approach with the obligate bacterial parasite, Pasteuria penetrans and nematicides was assessed for the management of the root-knot nematode, Meloidogyne incognita infestation in tomato and grapevine. Seedlings of tomato cv. Co3 were transplanted into pots filled with sterilized soil and inoculated with nematodes (5000 juveniles/pot). The root powder of P. penetrans at 10 mg/pot was applied alone and in combination with carbofuran at 6 mg/pot. Application of P. penetrans along with carbofuran recorded lowest nematode infestation (107 nematodes/200 g soil) compared to control (325 nematodes/200 g soil). The rate of parasitization was 83.1% in the carbofuran and P. penetrans combination treatment as against 61.0% in the P. penetrans treatment only. The plant growth was also higher in the combination treatment compared to all other treatments. A field trial was carried out to assess the efficacy of P. penetrans and nematicides viz., carbofuran and phorate in the management of root-knot nematode, M. incognita infestation of grapevine cv. Muscat Hamburg. A nematode and P. penetrans infested grapevine field was selected and treatments either with carbofuran or phorate at 1 g a.i/vine was given. The observations were recorded at monthly interval. The results showed that the soil nematode population was reduced in nematicide treated plots. Suppression of nematodes was higher under phorate (117 nematodes/200 g soil) than under carbofuran (126.7 nematodes/200 g soil) treatment. The number of juveniles parasitized was also influenced by nematicides and spore load carried/juvenile with phorate being superior and the increase being 17.0 and 29.0% respectively over the control. The results of these experiment confirmed the compatibility of P. penetrans with nematicides and its biological control potential against the root-knot nematode.     

Temporal dynamics of arbuscular mycorrhizal fungi colonizing roots of representative shrub species in a semi-arid Mediterranean ecosystem

Arbuscular mycorrhizal (AM) symbiosis plays an important role in improving plant fitness and soil quality, particularly in fragile and stressed environments, as those in certain areas of Mediterranean ecosystems. AM fungal communities are usually affected by dynamic factors such as the plant community structure and composition, which in turn are imposed by seasonality. For this reason, a one-year-round time-course trial was performed by sampling the root system of two representative shrubland species (Rosmarinus officinalis and Thymus zygis) within a typical Mediterranean ecosystem from the Southeast of Spain. The 18S rDNA gene, of the AM fungal community in roots, was subjected to PCR-SSCP, sequencing, and phylogenetic analysis. Forty-three different AM fungal sequence types were found which clustered in 16 phylotypes: 14 belonged to the Glomeraceae and two to the Diversisporaceae. Surprisingly, only two of these phylotypes were related with sequences of morphologically defined species: Glomus intraradices and Glomus constrictum. Significant differences were detected for the relative abundance of some phylotypes while no effects were found for the calculated diversity indices. These results may help to design efficient mycorrhizal-based revegetation programs for this type of ecosystems.     

Control of root-knot nematode,Meloidogyne incognita and reniform nematode,Rotylenchulus reniformis on pigeonpea and linseed

Abstract

Pigeonpea (Cajanus cajan) and linseed (Linum usitatissimum) are susceptible to Meloidogyne incognita and Rotylenchulus reniformis nematodes. Reduction in different growth parameters (length and weight of plant, number of pods), bulk density of pigeonpea stem, oil content of linseed, chlorophyll content of leaf and water absorption of roots caused by M. incognita and R. reniformis were statistically significant. Similar effects were also observed in plants raised from seeds soaked in different concentrations of water soluble fractions (WSF) of rice polish and pyridoxine solutions, however, the reductions were of a comparatively lesser extent. Higher concentrations of the solutions were more effective when compared to lower ones and pyridoxine was more beneficial than WSF for improving plant growth and reducing disease incidence.