Search Results (9)

Pine wilt disease (PWD), which poses a significant risk to pine plantations across the globe, is caused by the pathogenic agent Bursaphelenchus xylophilus, also referred to as the pine wood nematode (PWN). A droplet digital PCR (ddPCR) assay was developed for the quick identification of the PWN in order to improve detection sensitivity. The research findings indicate that the ddPCR assay demonstrated significantly higher analysis sensitivity and detection sensitivity in comparison to traditional quantitative PCR (qPCR). However, it had a more limited dynamic range. High specificity was shown by both the ddPCR and qPCR techniques in the diagnosis of the PWN. Assessments of reproducibility revealed that ddPCR had lower coefficients of variation at every template concentration. Inhibition tests showed that ddPCR was less susceptible to inhibitors. There was a strong linear association between standard template measurements obtained using ddPCR and qPCR (Pearson correlation = 0.9317; p < 0.001). Likewise, there was strong agreement (Pearson correlation = 0.9348; p < 0.001) between ddPCR and qPCR measurements in the evaluation of pine wood samples. Additionally, wood samples from symptomatic (100% versus 86.67%) and asymptomatic (31.43% versus 2.9%) pine trees were diagnosed with greater detection rates using ddPCR. This study’s conclusions highlight the advantages of the ddPCR assay over qPCR for the quantitative detection of the PWN. This method has a lot of potential for ecological research on PWD and use in quarantines. Full article

Leaf wilting is one of the most intuitive morphological manifestations of plants under drought stress, and it is useful in drought resistance screening. However, existing quantitative leaf-wilting measurement methods lack simplicity and high-throughput capacity under field conditions, and there is a gap in the systematic drought resistance assessments. The present study was conducted in 2020, 2021, and 2022 using 100 inbred maize lines. The maize lines were subjected to three different water stress treatments: normal irrigation, moderate drought, and severe drought. The findings led to the design of a simplified image acquisition and processing platform for measuring the visible green leaf area. A new measurement index and quantitative formula for wilting have been proposed, which effectively reflect leaf wilting and facilitate a systematic analysis of the relationship between yield and drought resistance. The results showed that the daily variation pattern of the visible green leaf area followed a trend of wilting first and then recovery, with maximum wilting occurring at noon (14:00–16:00 local time). The period of maximum wilting throughout the entire growth stage was the flowering stage. The maize plants with leaf wilt exceeding 1/2 (wilt ratio > 0.5) during the flowering stage were all low-yielding or low-tolerance inbred lines. In conclusion, this study emphasizes that the flowering stage is crucial for monitoring leaf wilting and introduces a quick high-throughput method to quantify leaf wilting. Our findings not only provide details about key indicators for identifying drought and heat resistance but also facilitate research on sustainable screening methods in maize, which will expedite the selection and accelerate the breeding of new varieties. Full article

In 2013, an outbreak of Xylella fastidiosa (Xf) was identified for the first time in Europe, in the extreme south of Italy (Apulia, Salento territory). The locally identified subspecies pauca turned out to be lethal for olive trees, starting an unprecedented phytosanitary emergency for one of the most iconic cultivations of the Mediterranean area. Xf pauca (Xfp) is responsible for a severe disease, the olive quick decline syndrome (OQDS), spreading epidemically and with dramatic impact on the agriculture, the landscape, the tourism and the cultural heritage of this region. The bacterium, transmitted by insects that feed on xylem sap, causes rapid wilting in olive trees due to biofilm formation, which obstructs the plant xylematic vessels. The aim of this review is to perform a thorough analysis that offers a general overview of the published work, from 2013 to December 2023, related to the Xfp outbreak in Apulia. This latter hereto has killed millions of olive trees and left a ghostly landscape with more than 8000 square kilometers of infected territory, that is 40% of the region. The majority of the research efforts made to date to combat Xfp in olive plants are listed in the present review, starting with the early attempts to identify the bacterium, the investigations to pinpoint and possibly control the vector, the assessment of specific diagnostic techniques and the pioneered therapeutic approaches. Interestingly, according to the general set criteria for the preliminary examination of the accessible scientific literature related to the Xfp outbreak on Apulian olive trees, fewer than 300 papers can be found over the last decade. Most of them essentially emphasize the importance of developing diagnostic tools that can identify the disease early, even when infected plants are still asymptomatic, in order to reduce the risk of infection for the surrounding plants. On the other hand, in the published work, the diagnostic focus (57%) overwhelmingly encompasses all other possible investigation goals such as vectors, impacts and possible treatments. Notably, between 2013 and 2023, only 6.3% of the literature reports addressing the topic of Xfp in Apulia were concerned with the application of specific treatments against the bacterium. Among them, those reporting field trials on infected plants, including simple pruning indications, were further limited (6%). Full article

For about a decade, olive groves in Apulia (Southern Italy) have been progressively destroyed by Olive Quick Decline Syndrome (OQDS), a disease caused by the bacterium Xylella fastidiosa subsp. pauca (Xfp). Recently, we described an additional wilting syndrome affecting olive trees in that area. The botryosphaeriaceous fungus Neofusicoccum mediterraneum was found associated with the diseased trees, and its high virulence toward olive trees was demonstrated. Given the common features with Branch and Twig Dieback (BTD) of olive tree, occurring in Spain and California, we suggested that the observed syndrome was BTD. During our first survey, we also found a botryosphaeriaceous species other than N. mediterraneum. In the present article, we report the morphological and molecular characterization of this fungal species which we identified as Neofusicoccum stellenboschiana. In the study, we also included for comparison additional N. stellenboschiana isolates obtained from olive trees in Latium and Tuscany region (Central Italy). The occurrence of N. stellenboschiana in olive trees is reported here for the first time in the northern hemisphere. The pathogenicity and virulence were tested in nine inoculation trials, where the Apulian N. stellenboschiana isolate was compared with the isolate from Latium and with the Apulian isolate of N. mediterraneum. Both isolates of N. stellenboschiana proved pathogenic to olive trees. They caused evident bark canker and wood discolouration when inoculated at the base of the stem of two/three-year-old trees and on one-year-old twigs. However, virulence of N. stellenboschiana was significantly lower, though still remarkable, compared with N. mediterraneum in term of necrosis progression in the bark and the wood and capacity of wilting the twigs. Virulence of N. stellenboschiana and N. mediterraneum did not substantially change when inoculations were performed in spring/summer and in autumn, suggesting that these fungal species have the potential to infect and damage olive trees in all seasons. The high thermotolerance of N. stellenboschiana was also revealed with in vitro growth and survival tests. The high virulence of these Botryosphaeriaceae species highlights their contribution in BTD aetiology and the necessity to investigate right away their diffusion and, possibly, the role of additional factors other than Xfp in the general decline of olive groves in Apulia. Hence the importance of assessing the degree of overlap of BTD/Botryosphariaceae with OQDS/Xfp is discussed. Full article

Peanut foliar diseases and disorders can be difficult to rapidly diagnose with little experience because some abiotic and biotic symptoms present similar symptoms. Developing algorithms for automated identification of peanut foliar diseases and disorders could potentially provide a quick, affordable, and easy method for diagnosing peanut symptoms. To examine this, images of peanut leaves were captured from various angles, distances, and lighting conditions using various cameras. Color space data from all images was subsequently extracted and subjected to logistic regression. Separate algorithms were developed for each symptom to include healthy, hopperburn, late leaf spot, Provost injury, tomato spotted wilt, paraquat injury, or surfactant injury. The majority of these symptoms are not included within currently available disease identification mobile apps. All of the algorithms developed for peanut foliar diagnostics were ≥ 86% accurate. These diagnostic algorithms have the potential to be a valuable tool for growers if made available via a web-accessible platform, which is the next step of this work. Full article

Olive trees are infected and damaged by Botryosphaeriaceae fungi in various countries. The botryosphaeriaceous fungus Neofusicoccum mediterraneum is highly aggressive and is a major concern for olive groves in Spain and California (USA), where it causes ‘branch and twig dieback’ characterized by wood discoloration, bark canker, and canopy blight. During surveys of olive groves in Apulia (southern Italy), we noticed that—in some areas—trees were heavily affected by severe branch and twig dieback. In addition, chlorosis and the appearance of red-bronze patches on the leaf preceded the wilting of the foliage, with necrotic leaves persisting on the twigs. Given the severity of the manifestation in zones also subject to olive quick decline syndrome (OQDS) caused by Xylella fastidiosa subsp. pauca, we investigated the etiology and provide indications for differentiating the symptoms from OQDS. Isolation from diseased wood samples revealed a mycete, which was morphologically and molecularly identified as N. mediterraneum. The pathogenicity tests clearly showed that this fungus is able to cause the natural symptoms. Therefore, also considering the low number of tested samples, N. mediterraneum is a potential causal agent of the observed disease. Specifically, inoculation of the twigs caused complete wilting in two to three weeks, while inoculation at the base of the stem caused severe girdling wedge-shaped cankers. The growth rate of the fungus in in vitro tests was progressively higher from 10 to 30 °C, failing to grow at higher temperatures, but keeping its viability even after prolonged exposure at 50 °C. The capacity of the isolate to produce catenulate chlamydospores, which is novel for the species, highlights the possibility of a new morphological strain within N. mediterraneum. Further investigations are ongoing to verify whether additional fungal species are involved in this symptomatology. Full article

Phytophthora capsici is one of the most destructive pathogens causing quick wilt (foot rot) disease in black pepper (Piper nigrum L.) to which no effective resistance has been defined. To better understand the P. nigrum–P. capsici pathosystem, we employed metabolomic approaches based on flow-infusion electrospray–high-resolution mass spectrometry. Changes in the leaf metabolome were assessed in infected and systemic tissues at 24 and 48 hpi. Principal Component Analysis of the derived data indicated that the infected leaves showed a rapid metabolic response by 24 hpi whereas the systemic leaves took 48 hpi to respond to the infection. The major sources of variations between infected leaf and systemic leaf were identified, and enrichment pathway analysis indicated, major shifts in amino acid, tricarboxylic acid cycle, nucleotide and vitamin B6 metabolism upon infection. Moreover, the individual metabolites involved in defensive phytohormone signalling were identified. RT-qPCR analysis of key salicylate and jasmonate biosynthetic genes indicated a transient reduction of expression at 24 hpi but this increased subsequently. Exogenous application of jasmonate and salicylate reduced P. capsici disease symptoms, but this effect was suppressed with the co-application of abscisic acid. The results are consistent with abscisic acid reprogramming, salicylate and jasmonate defences in infected leaves to facilitate the formation of disease. The augmentation of salicylate and jasmonate defences could represent an approach through which quick wilt disease could be controlled in black pepper. Full article

Black pepper (Piper nigrum L.) is one of the most important crops and global demand continues to increase, giving it a high export value. However, black pepper cultivation has been seriously affected by a number of pathogenic diseases. Among them, “quick wilt” caused by Phytophthora sp., “slow decline” caused by Fusarium sp., and root-knot nematode Meloidogyne sp. have a serious negative effect on black pepper growth and productivity. There have been different chemical and biological methods applied to control these diseases, but their effectiveness has been limited. The aim of this research was to evaluate different combinations of rhizosphere bacteria and endophytic bacteria isolated from black pepper farms in the Central Highland of Vietnam for their ability to suppress pathogens and promote black pepper growth and yield. Formula 6, containing the strains Bacillus velezensis KN12, Bacillus amyloliquefaciens DL1, Bacillus velezensis DS29, Bacillus subtilis BH15, Bacillus subtilis V1.21 and Bacillus cereus CS30 exhibited the largest effect against Phytophthora and Fusarium in the soil and in the roots of black pepper. These bio-products also increased chlorophyll a and b contents, which led to a 1.5-fold increase of the photosynthetic intensity than the control formula and a 4.5% increase in the peppercorn yield (3.45 vs. 3.30 tons per hectare for the control). Our results suggest that the application of rhizosphere and endophytic bacteria is a promising method for disease control and growth-promotion of black pepper. Full article

Fusarium wilt caused by Fusarium oxysporum f.sp. cubense (Foc) is one of the most destructive diseases for banana. For their risk assessment and hazard characterization, it is vital to quickly determine the virulence of Foc isolates. However, this usually takes weeks or months using banana plant assays, which demands a better approach to speed up the process with reliable results. Foc produces various mycotoxins, such as fusaric acid (FSA), beauvericin (BEA), and enniatins (ENs) to facilitate their infection. In this study, we developed a linear regression model to predict Foc virulence using the production levels of the three mycotoxins. We collected data of 40 Foc isolates from 20 vegetative compatibility groups (VCGs), including their mycotoxin profiles (LC-MS) and their plant disease index (PDI) values on Pisang Awak plantlets in greenhouse. A linear regression model was trained from the collected data using FSA, BEA and ENs as predictor variables and PDI values as the response variable. Linearity test statistics showed this model meets all linearity assumptions. We used all data to predict PDI with high fitness of the model (coefficient of determination (R² = 0.906) and adjust coefficient (R²_adj = 0.898)) indicating a strong predictive power of the model. In summary, we developed a linear regression model useful for the prediction of Foc virulence on banana plants from the quantification of mycotoxins in Foc strains, which will facilitate quick determination of virulence in newly isolated Foc emerging Fusarium wilt of banana epidemics threatening banana plantations worldwide. Full article