Studies on arbuscular mycorrhiza (AM) in the Alentejo (Portugal) using pea mutants resistant to AM fungi as a control tool for field conditions

The utilization and management of arbuscular mycorrhiza (AM) symbiosis may improve production and sustainability of the cropping system. For this purpose, native AM fungi (AMF) were sought and tested for their efficiency to increase plant growth by enhanced P uptake and by alleviation of drought stress. Pot experiments with safflower (Carthamus tinctorius) and pea (Pisum sativum) in five soils (mostly sandy loamy Luvisols) and field experiments with peas were carried out during three years at four different sites. Host plants were grown in heated soils inoculated with AMF or the respective heat sterilized inoculum. In the case of peas, mutants resistant to AMF colonization were used as non-mycorrhizal controls. The mycorrhizal impact on yields and its components, transpiration, and P and N uptake was studied in several experiments, partly under varying P and N levels and water supply. Screening of native AMF by most probable number bioassays was not very meaningful. Soil monoliths were placed in the open to simulate field conditions. Inoculation with a native AMF mix improved grain yield, shoot and leaf growth variables as compared to control. Exposed to drought, higher soil water depletion of mycorrhizal plants resulted in a haying-off effect. The growth response to this inoculum could not be significantly reproduced in a subsequent open air pot experiment at two levels of irrigation and P fertilization, however, safflower grew better at higher P and water supply by multiples. The water use efficiency concerning biomass was improved by the AMF inoculum in the two experiments. Transpiration rates were not significantly affected by AM but as a tendency were higher in non-mycorrhizal safflower. A fundamental methodological problem in mycorrhiza field research is providing an appropriate (negative) control for the experimental factor arbuscular mycorrhiza. Soil sterilization or fungicide treatment have undesirable side effects in field and greenhouse settings. Furthermore, artificial rooting, temperature and light conditions in pot experiments may interfere with the interpretation of mycorrhiza effects. Therefore, the myc- pea mutant P2 was tested as a non-mycorrhizal control in a bioassay to evaluate AMF under field conditions in comparison to the symbiotic isogenetic wild type of var. FRISSON as a new integrative approach. However, mutant P2 is also of nod- phenotype and therefore unable to fix N2. A 3-factorial experiment was carried out in a climate chamber at high NPK fertilization to examine the two isolines under non-symbiotic and symbiotic conditions. P2 achieved the same (or higher) biomass as wild type both under good and poor water supply. However, inoculation with the AMF Glomus manihot did not improve plant growth. Differences of grain and straw yields in field trials were large (up to 80 per cent) between those isogenetic pea lines mainly due to higher P uptake under P and water limited conditions. The lacking N2 fixation in mutants was compensated for by high mineral N supply as indicated by the high N status of the pea mutant plants. This finding was corroborated by the results of a major field experiment at three sites with two levels of N fertilization. The higher N rate did not affect grain or straw yields of the non-fixing mutants. Very efficient AMF were detected in a Ferric Luvisol on pasture land as revealed by yield levels of the evaluation crop and by functional vital staining of highly colonized roots. Generally, levels of grain yield were low, at between 40 and 980 kg ha-1. An additional pot trial was carried out to elucidate the strong mycorrhizal effect in the Ferric Luvisol. A triplication of the plant equivalent field P fertilization was necessary to compensate for the mycorrhizal benefit which was with five times higher grain yield very similar to that found in the field experiment. However, the yield differences between the two isolines were not always plausible as the evaluation variable because they were also found in (small) field test trials with apparently sufficient P and N supply and in a soil of almost no AMF potential. This similarly occurred for pea lines of var. SPARKLE and its non-fixing mycorrhizal (E135) and non-symbiotic (R25) isomutants, which were tested in order to exclude experimentally undesirable benefits by N2 fixation. In contrast to var. FRISSON, SPARKLE was not a suitable variety for Mediterranean field conditions. This raises suspicion putative genetic defects other than symbiotic ones may be effective under field conditions, which would conflict with the concept of an appropriate control. It was concluded that AMF resistant plants may help to overcome fundamental problems of present research on arbuscular mycorrhiza, but may create new ones.

Pathogens occuring in the winter pea-maize-winter wheat rotation, their host specificity and the potential of compost in suppressing foot and root disease of peas

The overall aim of the work presented was to evaluate soil health management with a specific focus on soil borne diseases of peas. For that purpose field experiments were carried out from 2009 until 2013 to assess crop performance and pathogen occurrence in the rotation winter pea-maize-winter wheat and if the application of composts can improve system performance. The winter peas were left untreated or inoculated with Phoma medicaginis, in the presence or absence of yard waste compost at rate of 5 t dry matter ha-1. A second application of compost was made to the winter wheat. Fusarium ssp. were isolated and identified from the roots of all three crops and the Ascochyta complex pathogens on peas. Bioassays were conducted under controlled conditions to assess susceptibility of two peas to Fusarium avenaceum, F. solani, P. medicaginis and Didymella pinodes and of nine plant species to F. avenaceum. Also, effects of compost applications and temperature on pea diseases were assessed. Application of composts overall stabilized crop performance but it did not lead to significant yield increases nor did it affect pathogen composition and occurrence. Phoma medicaginis was dominating the pathogen complex on peas. F. graminearum, F. culmorum, F. proliferatum, Microdochium nivale, F. crookwellense, F. sambucinum, F. oxysporum, F. avenaceum and F. equiseti were frequently isolated species from maize and winter wheat with no obvious influence of the pre-crop on the Fusarium species composition. The spring pea Santana was considerably more susceptible to the pathogens tested than the winter pea EFB33 in both sterile sand and non-sterilized field soil. F. avenaceum was the most aggressive pathogen, followed by P. medicaginis, D. pinodes, and F. solani. Aggressiveness of all pathogens was greatly reduced in non-sterile field soil. F. avenaceum caused severe symptoms on roots of all nine plant species tested. Especially susceptible were Trifolium repens, T. subterraneum, Brassica juncea and Sinapis alba in addition to peas. Reduction of growing temperatures from 19/16°C day/night to 16/12°C and 13/10°C did not affect the efficacy of compost. It reduced plant growth and slightly increased disease on EFB33 whereas the highest disease severity on Santana was observed at the highest temperature, 19/16°C. Application of 20% v/v of compost reduced disease on peas due to all four pathogens depending on pea variety, pathogen and growing media used. Suppression was also achieved with lower application rate of 3.5% v/v. Tests with γ sterilized compost suggest that the suppression of disease caused by Fusarium spp. is biological in origin, whereas chemical and physical properties of compost are playing an additional role in the suppression of disease caused by D. pinodes and P. medicaginis.