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Fusarium wilt is currently spreading in banana growing regions around the world leading to substantial losses. The disease is caused by the fungus Fusarium oxysporum f. sp. cubense (Foc), which is further classified into distinct races according to the banana varieties that they infect. Cavendish banana is resistant to Foc race 1, to which the popular Gros Michel subgroup succumbed last century. Cavendish effectively saved the banana industry, and became the most cultivated commercial subgroup worldwide. However, Foc tropical race 4 (TR4) subsequently emerged in Southeast Asia, causing significant yield losses due to its high level of aggressiveness to cultivars of Cavendish, and other commonly grown cultivars. Preventing further spread is crucially important in the absence of effective control methods or resistant market-acceptable banana cultivars. Implementation of quarantine and containment measures depends on early detection of the pathogen through reliable diagnostics. In this study, we tested the hypothesis that secreted in xylem (SIX) genes, which currently comprise the only known family of effectors in F. oxysporum , contain polymorphisms to allow the design of molecular diagnostic assays that distinguish races and relevant VCGs of Foc. We present specific and reproducible diagnostic assays based on conventional PCR targeting SIX genes, using as templates DNA extracted from pure Foc cultures. Sets of primers specifically amplify regions of: SIX6 in Foc race 1, SIX1 gene in TR4, SIX8 in subtropical race 4, SIX9 / SIX10 in Foc VCG 0121, and SIX13 in Foc VCG 0122. These assays include simplex and duplex PCRs, with additional restriction digestion steps applied to amplification products of genes SIX1 and SIX13. Assay validations were conducted to a high international standard including the use of 250 Fusarium spp. isolates representing 16 distinct Fusarium species, 59 isolates of F. oxysporum , and 21 different vegetative compatibility groups (VCGs). Tested parameters included inter and intraspecific analytical specificity, sensitivity, robustness, repeatability, and reproducibility. The resulting suite of assays is able to reliably and accurately detect R1, STR4, and TR4 as well as two VCGs (0121 and 0122) causing Fusarium wilt in bananas. [ABSTRACT FROM AUTHOR]

Occasional strategic tillage (ST) has been suggested as a possible solution to manage herbicide-resistant weeds and control crop diseases in Australia's northern grain-growing regions. We evaluated the impact of ST on microbial communities as indicators of soil quality for two distinct tillage systems that have been applied to a Vertosol for 43 years (no-tillage – NT or conventional tillage – CT) and two stubble management practices (retention – SR or burning – SB). Soil samples were collected 15 weeks after ST and analysed for total enzymatic activity (fluorescein diacetate assay), metabolic diversity (Ecoplates Biolog ® ) and bacterial community structure (terminal restriction fragment length polymorphism). There was no significant effect of ST on the measured biological attributes. However, total enzymatic activity for treatments under CTSR and CTSR-ST were significantly higher compared with NTSR-ST (+0.8 fluorescein μg ml −1 g −1 soil h −1 , P < 0.01). Differences may be attributed to a significant increase in bulk density for CTSR treatment ( P < 0.05) and an increment in bulk density on CTSR-ST plots. The lack of changes may be attributed to a high resistance and/or resilience of soil microbial communities after 15 weeks of tillage. More studies on the long-term effect of ST are required to assess the impact on soil biological properties. [ABSTRACT FROM AUTHOR]

Soil microbial communities play an important role in plant health and soil quality. Researchers have developed a wide range of methods for studying the structure, diversity, and activity of microbes to better understand soil biology and plant-microbe interactions. Functional microbiological analyses of the rhizosphere have given new insights into the role of microbial communities in plant nutrition and plant protection against diseases. In this review, we present the most commonly used traditional as well as new culture-independent molecular methods to assess the diversity and function of soil microbial communities. Furthermore, we discuss advantages and disadvantages of these techniques and provide a perspective on emerging technologies for soil microbial community profiling. [ABSTRACT FROM AUTHOR]

• Bacterial community composition differed between NT and CT. • Fungal diversity and bacterial alpha diversity did not differ between NT and CT. • Microbial biomass and activity did not differ between NT and CT. • No effects of ST were detected for any of the measured parameters. • Microbial diversity and substrate utilisation potential differed between depths. Strategic tillage (ST) has been proposed to help control weeds that germinate during fallow-periods in continuous no till (NT) farming systems, but its effects on soil biological properties are unclear. ST is a type of reduced tillage (RT) that places particular emphasis during the decision-making process on agro-climatic conditions, and the frequency, timing and implements used to till. Here, we assessed the impacts of one-time ST following 44 years of continuous NT or conventional tillage (CT) on soil microbial diversity, biomass, enzymatic activity and substrate utilisation potential at two depths. The composition of bacterial and archaeal communities differed significantly between contrasting land management (CT vs NT); albeit albeit no differences were observed after ST. Meanwhile, fungal community composition was not affected by contrasting land management or ST. Biological indicators for catabolic diversity, microbial biomass and enzymatic activity showed no significant changes after ST operation. Our results suggest that, in the short-term (13 months), ST exerts only minor impacts on a range of biological properties, as well as, bacterial, archaeal and fungal diversity. Hence, ST has the potential to be used as a strategy to control weeds and diseases without compromising soil health. [ABSTRACT FROM AUTHOR]

Moko disease in banana is a bacterial wilt caused by strains within Ralstonia solanacearum sensu stricto . The disease is endemic to Central and South America but has spread to the Philippines and peninsular Malaysia. Detecting new incursions early in Moko-free banana production regions is of utmost importance for containment and eradication, as Moko management significantly increases costs of banana production. Molecular studies have supported the classification of R. solanacearum sensu stricto into phylotypes IIA, IIB and IIC, each comprising of various sequevars based on nucleotide divergence of a partial sequence within the endoglucanase gene. Moko disease in banana is caused by strains classified as sequevars 6, 24, 41, and 53 within phylotype IIA, and sequevars 3, 4, and 25 within phylotype IIB. To ensure accurate diagnostic assays are available to detect all Moko sequevars, we systematically validated previously published assays for Moko diagnostics. To be able to identify all sequevars, including the latest described sequevars, namely IIB-25, IIA-41, and IIA-53, we developed and validated two novel assays using genome-wide association studies on over 100 genomes of R. solanacearum sensu stricto . Validations using 196 bacterial isolates confirmed that a previous multiplex PCR based assay targeting sequevars IIB-3, IIB-4, IIA-6 and IIA-24 and our two novel assays targeting sequevars IIB-25, IIA-41 and IIA-53 were specific, reproducible, and accurate for Moko diagnostics.

Blood disease in bananas caused by Ralstonia syzygii subsp. celebesensis is a bacterial wilt causing significant crop losses in Indonesia and Malaysia. Disease symptoms include wilting of the plant and red-brown vascular staining, internal rot, and discoloration of green banana fruit. There is no known varietal resistance to this disease in the Musa genus, although variation in susceptibility has been observed, with the popular Indonesian cooking banana variety Kepok being highly susceptible. This study established the current geographic distribution of Blood disease in Indonesia and confirmed the pathogenicity of isolates by Koch's postulates. The long-distance distribution of the disease followed an arbitrary pattern indicative of human-assisted movement of infected banana materials. In contrast, local or short-distance spread radiated from a single infection source, indicative of dispersal by insects and possibly contaminated tools, water, or soil. The rapid expansion of its geographical range makes Blood disease an emerging threat to banana production in Southeast Asia and beyond.