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Charcoal rot disease (CRD), caused by the phytopathogenic fungus Macrophomina phaseolina, is a significant threat to cotton production in Israel and worldwide. The pathogen secretes toxins and degrading enzymes that disrupt the water and nutrient uptake, leading to death at the late stages of growth. While many control strategies were tested over the years to reduce CRD impact, reaching that goal remains a significant challenge. The current study aimed to establish, improve, and deepen our understanding of a new approach combining biological agents and chemical pesticides. Such intervention relies on reducing fungicides while providing stability and a head start to eco-friendly bio-protective Trichoderma species. The research design included sprouts in a growth room and commercial field plants receiving the same treatments. Under a controlled environment, comparing the bio-based coating treatments with their corresponding chemical coating partners resulted in similar outcomes in most measures. At 52 days, these practices gained up to 38% and 45% higher root and shoot weight and up to 78% decreased pathogen root infection (tracked by Real-Time PCR), compared to non-infected control plants. Yet, in the shoot weight assessment (day 29 post-sowing), the treatment with only biological seed coating outperformed (p < 0.05) all other biological-based treatments and all Azoxystrobin-based irrigation treatments. In contrast, adverse effects are observed in the chemical seed coating group, particularly in above ground plant parts, which are attributable to the addition of Azoxystrobin irrigation. In the field, the biological treatments had the same impact as the chemical intervention, increasing the cotton plants' yield (up to 17%), improving the health (up to 27%) and reducing M. phaseolina DNA in the roots (up to 37%). When considering all treatments within each approach, a significant benefit to plant health was observed with the bio-chemo integrated management compared to using only chemical interventions. Specific integrated treatments have shown potential in reducing CRD symptoms, such as applying bio-coating and sprinkling Azoxystrobin during sowing. Aerial remote sensing based on high-resolution visible-channel (RGB), green–red vegetation index (GRVI), and thermal imaging supported the above findings and proved its value for studying CRD control management. This research validates the combined biological and chemical intervention potential to shield cotton crops from CRD. [ABSTRACT FROM AUTHOR]

The fungus Macrophomina phaseolina causes charcoal rot disease (CRD) in cotton, whose symptoms develop in the late stages of growth and result in wilting and death. Despite significant research efforts to reduce disease incidences, effective control strategies against M. phaseolina are an ongoing scientific effort. Today's CRD control tends toward green options to reduce the chemicals' environmental footprint and health risks. Here, different Trichoderma species were examined separately and in combination with Azoxystrobin (AS) in semi-field open-enclosure pots and a commercial field throughout a full season. In the pot experiment, the T. asperellum (P1) excelled and led to improvement in growth (13%-14%, day 69) and crops (the number of capsules by 36% and their weight by 78%, day 173). The chemical treatment alone at a low dose had no significant impact. Still, adding AS improved the effect of T. longibrachiatum (T7507) and impaired P1 efficiency. Real-time PCR monitoring of the pathogen DNA in the plants' roots at the harvest (day 176), revealed the efficiency of the combined treatments: T. longibrachiatum (T7407 and T7507) + AS. In a commercial field, seed dressing with a mixture of Trichoderma species (mix of P1, T7407, and Trichoderma sp. O.Y. 7107 isolate) and irrigation of their secreted metabolites during seeding resulted in the highest yields compared with the control. Applying only AS irrigation at a low dose (2,000 cc/ha), with the sowing, was the second best in promoting crops. The molecular M. phaseolina detection showed that the AS at a high dose (4,000 cc/ha) and the biological mix treatments were the most effective. Reducing the AS chemical treatment dosages by half impaired its effectiveness. Irrigation timing, also studied here, is proven vital. Early water opening during the late spring suppresses the disease outburst and damages. The results demonstrated the benefits of CRD bioshielding and encouraged to explore the potential of a combined bio-chemo pest control approach. Such interphase can be environmentally friendly (reducing chemical substances), stabilize the biological treatment in changing environmental conditions, achieve high efficiency even in severe CRD cases, and reduce the development of fungicide resistance. [ABSTRACT FROM AUTHOR]

Simple Summary: Maize late wilt disease (LWD) is considered the most severe corn disease in Israel and Egypt and a significant threat in other countries. The utilization of disease-resistant maize cultivars is currently our best LWD control method. Here we evaluated the predictive ability of a rapid test to examine the susceptibility/tolerance of corn varieties to the disease. The fastest assay is based on the sensitivity of corn seeds to metabolites secreted by the pathogen. However, it is only reliable in detecting highly resistant or extremely susceptible maize varieties. To identify LWD immunity of mildly tolerant hybrid plants, a full-season pot assay in the open air, under field conditions, is needed. New detection methods, including aerial visible dehydration symptom monitoring and real-time PCR-based tracking of M. maydis DNA inside the plants' tissues, were evaluated here and proved to provide a great advantage. No complete overlap exists between the fungal DNA amount and the severity of symptoms. 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 is a powerful means to predict each maize variety's LWD immunity. 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. [ABSTRACT FROM AUTHOR]

6-pentyl-α-pyrone (6-PP) is a powerful Magnaporthiopsis maydis antifungal compound, recently discovered when the potent growth medium of Trichoderma asperellum was analyzed. Despite its high potential in plate assay, it was not inspected for plant treatment prevention. Late wilt disease, caused by the fungus M. maydis, threatens commercial maize production in high-risk areas. Thus, the search for control options against the pathogen is one of the top priorities in Israel, Egypt, and other countries. Disease-resistant maize genotypes can reduce the damages. Yet, aggressive variants of the fungus can overcome host resistance. The current study aimed at inspecting T. asperellum and its secreted metabolite, pure 6-PP, against the pathogen in plants over a full growth period. First, adding T. asperellum directly to seeds with sowing provides significant protection to sprouts (up to 42 days) in a growth room, with more than two-fold growth promotion and reduced pathogen root infection (detected by real-time PCR). The same procedure applied in a commercial field was less beneficial in rescuing the plants' growth and yield. Still, it reduced the cobs' symptoms by 11% and resulted in nine-fold lower levels of the pathogen's DNA in the stem tissue. Second, the T. asperellum purified 6-PP compound (30 µg/seed) was used in seed coating and tested against the T. asperellum secretory metabolites' crude (diluted to 50%). At the season's end, these treatments improved plant biomass by 90–120% and cob weight by 60%. Moreover, the treatments significantly (p < 0.05) reduced the symptoms (up to 20%) and pathogen infection (94–98%). The current study's results reveal the potential of 6-PP as a new fungicide against M. maydis. Such a treatment may protect maize plants from other soil diseases. [ABSTRACT FROM AUTHOR]

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. [ABSTRACT FROM AUTHOR]

Late wilt (LWD) is a vascular wilt disease that outbursts late in maize development, usually during or after flowering. The disease causal agent, the soil and seed-borne fungi, Magnaporthiopsis maydis, causes significant economic losses in Egypt, Israel, Spain, Portugal, and India. Since its discovery in the early 1960s in Egypt, the knowledge base of the disease was significantly expanded. This includes basic information on the pathogen and its mode of action, disease symptoms and damages, methods to study and monitor the pathogen, and above all, control strategies to restrain M. maydis and reduce its impact on commercial maize production. Three approaches stand out from the various control methods inspected. First, the traditional use of chemical pesticides was investigated extensively. This approach gained attention when, in 2018–2020, a feasible and economical treatment based on Azoxystrobin (alone or in combination with other fungicides) was proven to be effective even in severe cases of LWD. Second, the growing trend of replacing chemical treatments with eco-friendly biological and other green protocols has become increasingly important in recent years and has already made significant achievements. Last but not least, today’s leading strategy to cope with LWD is to rely on resistant maize genotypes. The past two decades’ introduction of molecular-based diagnostic methods to track and identify the pathogen marked significant progress in this global effort. Still, worldwide research efforts are progressing relatively slowly since the disease is considered exotic and unfamiliar in most parts of the world. The current review summarizes the accumulated knowledge on LWD, its causal agent, and the disease implications. An additional important aspect that will be addressed is a future perspective on risks and knowledge gaps. [ABSTRACT FROM AUTHOR]

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. [ABSTRACT FROM AUTHOR]

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-timePCR tracking of the pathogen in the host plants’ first internode supported the symptoms’ evaluation,with 3.1- and 4.9-fold lower M. may dis DNA levels in the two Trichoderma treatments. In order toinduce 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. may dismycelia. This infection method overrides the Trichodermaroots protection and almost abolishes the biocontrol treatments’ protective achievements.This study suggests a biologicalTrichoderma-based protective layer that may have significant value in mild cases of LWD. [ABSTRACT FROM AUTHOR]

Late wilt, a disease severely affecting maize fields throughout Israel, is characterized by the relatively rapid wilting of maize plants from the tasseling stage to maturity. The disease is caused by the fungus Magnaporthiopsis maydis, a soil and seed-borne pathogen. The pathogen is controlled traditionally through the use of maize cultivars having reduced sensitivity to the disease. Nevertheless, such cultivars may lose their immunity after several years of intensive growth due to the presence of high virulent isolates of M. maydis. Alternative effective and economical chemical treatment to the disease was recently established but is dependent on the use of a dripline assigned for two adjacent rows and exposes the risk of fungicide resistance. In the current work, eight marine and soil isolates of Trichoderma spp., known for high mycoparasitic potential, were tested as biocontrol agents against M. maydis. An in vitro confront plate assay revealed strong antagonistic activity against the pathogen of two T. longibrachiatum isolates and of T. asperelloides. These species produce soluble metabolites that can inhibit or kill the maize pathogen in submerged and solid media culture growth assays. In greenhouse experiments accompanied by real-time PCR tracking of the pathogen, the Trichoderma species or their metabolites managed to improve the seedlings’ wet biomass and reduced the pathogen DNA in the maize roots. A follow-up experiment carried out through a whole growth session, under field conditions, provided important support to the Trichoderma species’ beneficial impact. The direct addition of T. long ibrachiatum and even more T. asperelloides to the seeds, with the sowing, resulted in a yield improvement, a significant increase in the growth parameters and crops, to the degree of noninfected plants. These bioprotective treatments also restricted the pathogen DNA in the host tissues (up to 98%) and prevented the disease symptoms. The results encourage more in-depth research to uncover such biological agents’ potential and the methods to implement them in commercial fields. If adequately developed into final products and combined with other control methods, the biological control could play an important role in maize crop protection against Late wilt. [ABSTRACT FROM AUTHOR]

Late wilt is a destructive disease of corn: outbreaks occur at the advanced growth stage and lead to severe dehydration of susceptible hybrids. The disease’s causal agent is the fungus Magnaporthiopsis maydis, whose spread relies on infested soils, seeds, and several alternative hosts. The current study aimed at advancing our understanding of the nature of this plant disease and revealing new ways to monitor and control it. Two field experiments were conducted in a heavily infested area in northern Israel seeded with highly sensitive corn hybrid. The first experiment aimed at inspecting the Azoxystrobin (AS) fungicide applied by spraying during and after the land tillage. Unexpectedly, the disease symptoms in this field were minor and yields were high. Nevertheless, up to 100% presence of the pathogen within the plant’s tissues was measured using the quantitative real-time PCR method. The highest AS concentration tested was the most effective treatment, and resulted in a 6% increase in cob yield and a 4% increase in A-class yield. In the second experiment conducted in the following summer of the same year in a nearby field, the disease outbreak was dramatically higher, with about 350 times higher levels of the pathogen DNA in the untreated plots’ plants. In this field, fungicide mixtures were applied using a dripline assigned for two coupling rows. The most successful treatment was AS and the Difenoconazole mixture, in which the number of infected plants decreased by 79%, and a 116% increase in crop yield was observed, along with a 41% increase in crop quality. Evaluation of the effectiveness of the treatments on the plants’ health using a remote, thermal infra-red sensitive camera supported the results and proved to be an essential research tool. [ABSTRACT FROM AUTHOR]

The soil fungus Macrophomina phaseolina, the charcoal rot disease agent, poses a major threat to cotton fields. In Israel, highly infected areas are also inhabited by the maize pathogen Magnaporthiopsis maydis. This study reveals the relationships between the two pathogens and their impact on cotton sprouts. Infecting the soil 14 days before sowing (DBS) with each pathogen or with M. phaseolina before M. maydis caused a strong inhibition (up to 50–65%) of the sprouts' development and survival, accompanied by each pathogen's high DNA levels in the plants. However, combined or sequence infection with M. maydis first led to two distinct scenarios. This pathogen acted as a beneficial protective endophyte in one experiment, leading to significantly high emergence and growth indices of the plants and a ca. 10-fold reduction in M. phaseolina DNA in the sprouts' roots. In contrast, M. maydis showed strong virulence potential (with 43–69% growth and survival suppression) in the other experiment, proving its true nature as an opportunist. Interestingly, soil inoculation with M. phaseolina first, 14 DBS (but not at sowing), shielded the plants from M. maydis' devastating impact. The results suggest that the two pathogens restrict each other, and this equilibrium may lead to a moderate disease burst. [ABSTRACT FROM AUTHOR]

Control of maize late wilt disease (LWD) has been at the forefront of research efforts since the discovery of the disease in the 1960s. The disease has become a major economic restraint in highly affected areas such as Egypt and Israel, and is of constant concern in other counties. LWD causes dehydration and collapsing at a late stage of maize cultivation, starting from the male flowering phase. The disease causal agent, Magnaporthiopsis maydis, is a seed- and soil-borne phytoparasitic fungus, penetrating the roots at sprouting, colonizing the vascular system without external symptoms, and spreading upwards in the xylem, eventually blocking the water supply to the plant's upperparts. Nowadays, the disease's control relies mostly on identifying and developing resistant maize cultivars. Still, host resistance can be limited because M. maydis undergoes pathogenic variations, and virulent strains can eventually overcome the host immunity. This alarming status is driving researchers to continue to seek other control methods. The current review will summarize the various strategies tested over the years to minimize the disease damage. These options include agricultural (crop rotation, cover crop, no-till, flooding the land before sowing, and balanced soil fertility), physical (solar heating), allelochemical, biological, and chemical interventions. Some of these methods have shown promising success, while others have contributed to our understanding of the disease development and the environmental and host-related factors that have shaped its outcome. The most updated global knowledge about LWD control will be presented, and knowledge gaps and future aims will be discussed. [ABSTRACT FROM AUTHOR]

Maize late wilt disease caused by the fungus Magnaporthiopsis maydis significantly damages crops in Israel and in other countries. Resistant maize cultivars are the preferred method for disease restraining. However, the pathogen populations of Spain and Egypt have varying aggressiveness, and virulent strains can overcome host resistance. In 2001 and from 2016 to -2019, 17 M. maydis strains were isolated from infected maize fields in Israel. The isolates' effects on seed germination, plant development, and disease symptoms severity were evaluated. The isolates from Israel display a diverse degree of aggressiveness that is not linked to their geographic distribution. The virulent strains are found in mixed populations, whereas less virulent M. maydis isolates exist. Aggressive strains harmed the development of plants and ears and caused severe wilting and death. In contrast, plants inoculated with less virulent strains exhibited only mild dehydration signs, and crop yield was similar to that of the non-infected control. Interestingly, different host cultivars can evoke specific virulence of M. maydis strains. Moreover, some pathogen strains significantly repress plant development, while the impact of other strains was evidenced by wilting symptoms. The current research further increases our understanding of the pathogen and our ability to control it. [Display omitted] • Israel is considered one of the world's most affected by maize late wilt disease. • This is the first work to describe Israel's pathogen dispersal and aggressiveness. • Virulent M. maydis strains scattered throughout the country in mixed populations. • Some strains mainly affect plant growth, while others disease symptoms. • Different host cultivars evoke specific M. maydi s strains' virulence. [ABSTRACT FROM AUTHOR]

The maize pathogen Magnaporthiopsis maydis causes severe damage to commercial fields in the late growth stages. This late wilt disease has spread since its discovery (the 1980s) and is now common in most corn-growing areas in Israel. In some fields and sensitive plant species, the disease can affect 100% of the plants. The M. maydis pathogen has a hidden endophytic lifecycle (developed inside the plants with no visible symptoms) in resistant corn plants and secondary hosts, such as green foxtail and cotton. As such, it may also be opportunist and attack the host in exceptional cases when conditions encourage it. This work aims to study the pathogen's interactions with maize endophytes (which may play a part in the plant's resistance factors). For this purpose, 11 fungal and bacterial endophytes were isolated from six sweet and fodder corn cultivars with varying susceptibility to late wilt disease. Of these, five endophytes (four species of fungi and one species of bacteria) were selected based on their ability to repress the pathogen in a plate confrontation test. The selected isolates were applied in seed inoculation and tested in pots in a growth room with the Prelude maize cultivar (a late wilt-sensitive maize hybrid) infected with the M. maydis pathogen. This assay was accompanied by real-time qPCR that enables tracking the pathogen DNA inside the host roots. After 42 days, two of the endophytes, the Trichoderma asperellum, and Chaetomium subaffine fungi, significantly (p < 0.05) improved the infected plants' growth indices. The fungal species T. asperellum, Chaetomium cochliodes, Penicillium citrinum, and the bacteria Bacillus subtilis treatments were able to reduce the M. maydis DNA in the host plant's roots. Studying the maize endophytes' role in restricting the invasion and devastating impact of M. maydis is an essential initial step towards developing new measures to control the disease. Such an environmentally friendly control interface will be based on strengthening the plants' microbiome. [ABSTRACT FROM AUTHOR]

Fungal pathogens are a significant threat to crops worldwide. The soil fungus, Magnaporthiopsis maydis, severely affects sensitive maize hybrids by causing the rapid wilting of plants at the maturity stage. Similarly, the soil fungus, Macrophomina phaseolina, develops in a variety of host plants, which leads to rot and plant mortality. The presence of both pathogens together in diseased cotton plants in Israel suggests possible interactions between them. Here, these relationships were tested in a series of experiments accompanied by real-time PCR tracking in maize and cotton. Despite the fact that neither of the pathogens was superior in a growth plate confrontation assay, their co-inoculum had a significant influence under field conditions. In maize sprouts and fully matured plants, infection by both pathogens (compared to inoculation with each of them alone) led to lesser amounts of M. maydis DNA but to increased amounts of M. phaseolina DNA levels. These results were obtained under a restricted water regime, while optimal water irrigation led to less pronounced differences. In water-stressed cotton sprouts, infection with both pathogens led to an increase in DNA amounts of each of the pathogens. Whereas the M. maydis DNA levels in the double infection remain high at the end of the season, a reduction in the amount of M. phaseolina DNA was observed. The double infection caused an increase in growth parameters in maize and cotton and decreased levels of dehydration in maize plants accompanied by an increase in yield production. Dehydration symptoms were minor in cotton under an optimal water supply. However, under a restricted water regime, the double infection abolished the harmful effect of M. phaseolina on the plants' development and yield. These findings are the first report of interactions between these two pathogens in maize and cotton, and they encourage expanding the study to additional plant hosts and examining the potential involvement of other pathogens. [ABSTRACT FROM AUTHOR]

Late wilt, a destructive vascular disease of maize caused by the fungus Magnaporthiopsis maydis, is characterized by relatively fast wilting of maize plants closely before the physiological maturity stage. Previously, traditional microbiology-based methods have been used to isolate the pathogen and to characterize its traits. More recently, several molecular methods have been developed, enabling accurate and sensitive examination of the pathogen spread within the host. Here, we review the methods developed in the past 10 years in Israel, which include new or modified microbial and molecular techniques to identify, monitor, and study M. maydis in controlled environments and in the field. The assays inspected are exemplified with new findings and include microbial isolation methods, microscopic and PCR or qPCR identification, spore germination evaluation, root pathogenicity assay, M. maydis hyphae or filtrate effects on grain germination and sprout development, and a field assay. These diagnostic protocols enable rapid and reliable detection and identification of the pathogen in plants and seeds and studying the pathogenesis of M. maydis in susceptible and relatively resistant maize cultivars in a contaminated field. Moreover, these techniques are important for studying the population structure, and for future development of new strategies to restrict the disease's outburst and spread. [ABSTRACT FROM AUTHOR]