All Foc isolates were collected from the ABB Pisang Awak cultivar Kluai Namwa [45]. 5. It was originally isolated from Pisang Keling and Pisang Ambon in Malaysia, respectively [33]. This large diversity could be attributed to the polyphyletic nature of Foc [47], its possible history of sexual reproduction [59], parasexuality [60, 61] and horizontal gene transfer [62]. To simplify the identification process, the isolates were first divided into DNA lineages using PCR-RFLP analysis. No location information was available for these isolates. The banana Fusarium wilt fungus Foc is particularly diverse within the F. oxysporum species complex when compared to other formae speciales. VCG diversity in Cambodia, Vietnam and Bangladesh was reported for the first time. A total of 615 Fusarium isolates were obtained from Asia, which are all maintained in 15% glycerol at -80°C in the culture collection of the Department of Plant Pathology at the University of Stellenbosch in South Africa (S1 Table). Whilst most of Asia’s bananas are consumed locally, the region also produces bananas for export. Clade A affects mainly M. acuminata hybrids, whereas Clade B affects mainly M. acuminata x M. balbisiana hybrids. PCR cycling conditions consisted of 35 cycles at 94°C for 45 sec, 50°C (IGS) or 59°C (MtR) for 45 sec, and 72°C for 90 sec. PCR cycle conditions for the Foc TR4 primer set consisted of 30 cycles at 95°C for 1 min, 60°C for 1 min and 72°C for 3 min. VCG 0128 was isolated in the northwest and VCG 01217 in the north-central region (Fig 7). Writing – review & editing, Affiliation (1998) [28] found 13 putative VCGs or genotypes when investigating a worldwide population of Foc. VCGs 01217 and 01218 were found on the Silk banana Pisang Rastali (AAB) and the Bluggoe cultivar Pisang Abu Keling (ABB) in northern Peninsular Malaysia, VCG 0123 on Pisang Awak and Pisang Rastali in the northeast and northwest of Peninsular Malaysia, and VCG 0128 in the Kelantan area (Table 1; Fig 4), for which no cultivar information was available. Production on the continent is affected by a number of biotic constraints, of which banana Fusarium wilt is one of the most devastating. VCG 01213/16, for instance, is well-established in areas where Cavendish bananas are grown in monoculture, such as the Philippines, mainland China and Taiwan. originated in Asia following interspecific hybridization between two species, M. acuminata Colla and M. balbisiana Colla. Is the Subject Area "Bananas" applicable to this article? Each PCR was preceded by an initial denaturation step of 94°C for 5 min and concluded with a final extension step of 72°C for 5 min. When Foc VCG 01213/16 began to cause large-scale epidemics to the expanding Cavendish industry in China [17] and the Philippines [18], a renewed interest and concerns of the threat of Foc to the Asian banana industry was generated. Indonesian Agency for Agriculture Research and Development, Jakarta, Indonesia, Affiliation Fusarium oxysporum formae specialis cubense (Foc) is a soil-borne fungus that causes Fusarium wilt, which is considered to be the most destructive disease of bananas. Conceptualization, For cultural and morphological identification, each isolate was grown on PDA and carnation leaf agar (CLA), respectively. Given the association of these fungi with plant roots, a form that is able to grow beyond the cortex and into the xylem could exploit this ability and hopefully gain an advantage over fungi that are restricted to the cortex. Despite the significant Cavendish industry, no Foc race 4 VCGs, such as VCG 0120/15 and VCG 01213/16, were detected. Guangdong Academy of Agricultural Sciences, Institution of Fruit Tree Research, Guangzhou, Guangdong Province, China, Roles Individual isolates of . These incompatible isolates should be tested for pathogenicity on banana, vegetative compatibility, and relatedness to previously described genotypes of the pathogen. They are pervasive plant endophytes that can colonize plant roots[13][14] and may even protect plants or form the basis of disease suppression. It was also affected by an isolate not compatible to known VCGs (S1 Table). causes devastating diseases of more than 100 plant species, including . Each PCR was preceded by an initial denaturation step at 95°C for 5 min and concluded by a final extension step at 72°C for 10 min. VCG diversity in Foc has been determined for strains collected in Malaysia, Indonesia, Thailand, India, China and the Philippines; all countries where a large diversity of bananas exists [28, 40, 41, 42, 43, 44, 45,46]. A mutation at a single vic locus, thus, could place closely related individuals in different VCGs [28, 31]. Writing – review & editing. e0181630. Nit mutants were successfully generated for all the Asian Foc isolates. [25], The fungus has the ability to dissolve gold, then excrete it onto the surface of the fungus, encrusting itself with gold. Given the association of these fungi with plant roots, a form that is able to grow beyond the cortex and into the xylem could exploit this ability and hopefully gain an advantage over fungi that are restricted to the cortex. Several Foc VCGs were not found in Asia. Validation, Briefly, the 1500-bp intergenic spacer (IGS) region was first amplified using the primer set PNFo and PN22 [54] with an Eppendorf Mastercycler Gradient PCR machine (Eppendorf Scientific, Hamburg, Germany). The extracted DNA was quantified using a Nanodrop spectrophotometer (NanoDrop, Wilmington, USA) and stored at -20°C until further use. These associations may be indicators of the environmental contribution to the outcome of these interactions. F. oxysporum has received considerable attention from plant pathologists for more than a century owing to its broad host range and the economic losses it causes. The population was dominated by the VCG 0124/5 complex, which was obtained from all affected banana cultivars in the country; including the Silk type banana Bangla Kola, the AAB bananas Sabri and Plantain, and the AB Ney Poovan banana Champa (Table 2). The dominant VCG found in India was the VCG 0124/5 complex (Fig 1), and isolates from this VCG were collected from ABB bananas including Monthan and the Pisang Awak cultivar Karpooravalli, AB bananas such as Ney Poovan, and the Silk cultivars Rasthali, Malbhog and Mortaman (AAB) (Table 2). Whereas vegetative compatibility provides a clear measure of phenotypic relatedness, the technique does not measure the genetic distance between phenotypes [28, 29]. 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