Crop Rotation and Minimal Tillage Selectively Affect Maize Growth Promotion under Late Wilt Disease Stress.

In recent years, worldwide scientific efforts towards controlling maize late wilt disease (LWD) have focused on eco-friendly approaches that minimize the environmental impact and health risks. This disease is considered to be the most severe threat to maize fields in Israel and Egypt, and a major growth restraint in India, Spain, and Portugal. Today's most commonly used method for LWD control involving resistant maize genotypes is under constant risk from aggressive pathogen lines. Thus, this study's objectives were to evaluate the effect of crop rotation and avoiding tillage on restraining the disease. Such an agrotechnical approach will support the continuity of soil mycorrhiza networks, which antagonize the disease's causal agent, Magnaporthiopsis maydis. The method gained positive results in previous studies, but many knowledge gaps still need to be addressed. To this end, a dual-season study was conducted using the LWD hyper-susceptible maize hybrid, Megaton cv. The trials were performed in a greenhouse and in the field over full dual-growth seasons (wheat or clover as the winter crop followed by maize as the summer crop). In the greenhouse under LWD stress, the results clearly demonstrate the beneficial effect of maize crop rotation with clover and wheat on plant weight (1.4-fold), height (1.1-1.2-fold) and cob yield (1.8-2.4-fold), especially in the no-till soil. The clover-maize growth sequence excels in reducing disease impact (1.7-fold) and pathogen spread in the host tissues (3-fold). Even though the wheat-maize crop cycle was less effective, it still had better results than the commercial mycorrhizal preparation treatment and the uncultivated non-infected soil. The results were slightly different in the field. The clover-maize rotation also achieved the best growth promotion and disease restraint results (2.6-fold increase in healthy plants), but the maize rotation with wheat showed only minor efficiency. Interestingly, pre-cultivating the soil with clover had better results in no-till soil in both experiments. In contrast, the same procedure with wheat had a better impact when tillage was applied. It may be concluded that crop rotation and soil cultivation can be essential in reducing LWD, but other factors may affect this approach's benefits in commercial field growth.

Cultivars Resistance Assay for Maize Late Wilt Disease.

Magnaporthiopsis maydis late wilt disease (LWD) in corn is considered to be the most severe in Israel and Egypt and poses a significant threat in other countries. Research efforts extending over a period of five decades led to the development of chemical, biological, agrotechnical, physical (solar disinfection) and other means for controlling late wilt disease. Today, some applications can reduce damage even in severe cases. However, cultivating disease-resistant maize varieties is the primary means for reducing the disease's impact. The current work uses a rapid (six days) laboratory seedling pathogenicity test and a full-season open encloser semi-field conditioned pots assay (101 days) to classify maize varieties according to their LWD resistance. To better evaluate differences between the cultivars, a real-time based molecular assay was applied to track the pathogen's presence in the plants' tissues, and visible light aerial imaging was used in parallel. The findings show that in cases of extreme sensitivity or tolerance (for example, in the highly susceptible Megaton cultivar (cv.) or the resistant Hatai cv.), a similarity in the results exists between the different methods. Thus, a reliable estimate of the varieties' sensitivity can be obtained in a seed assay without the need for a test carried out throughout an entire growing season. At the same time, in most situations of partial or reduced LWD sensitivity/resistance, there is no match between the various tests, and only the entire growing season can provide the most reliable results. Tracking the amount of M. maydis DNA in the plants' bodies is a precise, sensitive scientific tool of great importance for studying the development of the disease and the factors affecting it. Yet, no complete overlap exists between the fungal DNA amount and symptom severity. Such a correlation exists in high sensitivity or resistance cases but not in intermediate situations. Still, the valuation of the pathogen's establishment in asymptomatic corn hybrids can indicate the degree of LWD immunity and the chance of susceptibility development.

Crop Cycle and Tillage Role in the Outbreak of Late Wilt Disease of Maize Caused by Magnaporthiopsis maydis.

The destructive maize late wilt disease (LWD) has heavy economic implications in highly infected areas such as Israel, Egypt, and Spain. The disease outbreaks occur near the harvest, leading to total yield loss in severe cases. Crop rotation has long been known as an effective means to reduce plant diseases. Indeed, agricultural soil conservation practices that can promote beneficial soil and root fungi have become increasingly important. Such methods may have a bioprotective effect against Magnaporthiopsis maydis, the LWD causal agent. In this two-year study, we tested the role of crop rotation of maize with either wheat or clover and the influence of minimum tillage in restricting LWD. In the first experiment, wheat and clover were grown in pots with LWD infected soil in a greenhouse over a full winter growth period. These cultivations were harvested in the spring, and each pot's group was split into two subgroups that underwent different land processing practices. The pots were sown with LWD-sensitive maize cultivar and tested over a whole growth period against control soils without crop rotation or soil with commercial mycorrhizal preparation. The maize crop rotation with wheat without tillage achieved prominent higher growth indices than the control and the clover crop cycle. Statistically significant improvement was measured in the non-tillage wheat soil pots in sprout height 22 days after sowing, in the healthy plants at the season's end (day 77), and in shoot and cob wet weight (compared to the control). This growth promotion was accompanied by a 5.8-fold decrease in pathogen DNA in the plant stems. The tillage in the wheat-maize growth sequence resulted in similar results with improved shoot wet-weight throughout the season. In contrast, when maize was grown after clover, the tillage reduced this parameter. The addition of commercial mycorrhizal preparation to the soil resulted in higher growth measures than the control but was less efficient than the wheat crop cycle. These results were supported by a subsequent similar experiment that relied on soil taken from commercial wheat or clover fields. Here too, the wheat-maize growth cycle (without permanent effect for the tillage) achieved the best results and improved the plants' growth parameters and immunity against LWD and lowered pathogen levels. In conclusion, the results of this study suggest that wheat and perhaps other crops yet to be inspected, together with the adjusted tillage system, may provide plants with better defense against the LWD pathogen.

Trichoderma asperellum Secreted 6-Pentyl-α-Pyrone to Control Magnaporthiopsis maydis, the Maize Late Wilt Disease Agent.

Late wilt disease (LWD) is a destructive vascular disease of maize (Zea mays L.) caused by the fungus Magnaporthiopsis maydis. Restricting the disease, which is a significant threat to commercial production in Israel, Egypt, Spain, India, and other countries, is an urgent need. In the past three years, we scanned nine Trichoderma spp. isolates as biological control candidates against M. maydis. Three of these isolates showed promising results. In vitro assays, seedlings pathogenicity trials, and field experiments all support the bio-control potential of these isolates (or their secretions). Here, a dedicated effort led to the isolation and identification of an active ingredient in the growth medium of Trichoderma asperellum (P1) with antifungal activity against M. maydis. This Trichoderma species is an endophyte isolated from LWD-susceptible maize seeds. From the chloroform extract of this fungal medium, we isolated a powerful (approx. 400 mg/L) active ingredient capable of fully inhibiting M. maydis growth. Additional purification using liquid chromatography-mass spectrometry (LC-MS) and gas chromatography-mass spectrometry (GC-MS) separation steps enabled identifying the active ingredient as 6-Pentyl-α-pyrone. This compound is a potential fungicide with high efficiency against the LWD causal agent.

Trichoderma longibrachiatum and Trichoderma asperellum Confer Growth Promotion and Protection against Late Wilt Disease in the Field.

Late wilt disease (LWD) of maize, caused by Magnaporthiopsis maydis, is considered a major threat to commercial fields in Israel, Egypt, Spain, and India. Today's control methods include chemical and agronomical intervention but rely almost solely on resistant maize cultivars. In recent years, LWD research focused on eco-friendly biological approaches to restrain the pathogen. The current study conducted during two growing seasons explores the potential of three Trichoderma species as bioprotective treatments against LWD. These species excelled in preliminary assays performed previously under controlled conditions and were applied here in the field by directly adding them to each seed with the sowing. In the first field experiment, Trichoderma longibrachiatum successfully rescued the plants' growth indices (weight and height) compared to T. asperelloides and the non-treated control. However, it had no positive effect on yield and disease progression. In the subsequent season, this Trichoderma species was tested against T. asperellum, an endophyte isolated from susceptible maize cultivar. This experiment was conducted during a rainy autumn season, which probably led to a weak disease burst. Under these conditions, the plants in all treatment groups were vivid and had similar growth progression and yields. Nevertheless, a close symptoms inspection revealed that the T. longibrachiatum treatment resulted in a two-fold reduction in the lower stem symptoms and a 1.4-fold reduction in the cob symptoms at the end of the seasons. T. asperellum achieved 1.6- and 1.3-fold improvement in these parameters, respectively. Quantitative Real-time PCR tracking of the pathogen in the host plants' first internode supported the symptoms' evaluation, with 3.1- and 4.9-fold lower M. maydis DNA levels in the two Trichoderma treatments. In order to induce LWD under the autumn's less favorable conditions, some of the plots in each treatment were inoculated additionally, 20 days after sowing, by stabbing the lower stem section near the ground with a wooden toothpick dipped in M. maydis mycelia. This infection method overrides the Trichoderma roots protection and almost abolishes the biocontrol treatments' protective achievements. This study suggests a biological Trichoderma-based protective layer that may have significant value in mild cases of LWD.