| Abstract: | The global yield of bananas—one of the most important food crops—is severely hampered by parasites, such as nematodes, which cause yield losses up to 75%. Plant–nematode interactions of two banana cultivars differing in susceptibility to Radopholus similis were investigated by combining the conventional and spatially resolved analytical techniques 1H NMR spectroscopy, matrix-free UV-laser desorption/ionization mass spectrometric imaging, and Raman microspectroscopy. This innovative combination of analytical techniques was applied to isolate, identify, and locate the banana-specific type of phytoalexins, phenylphenalenones, in the R. similis-caused lesions of the plants. The striking antinematode activity of the phenylphenalenone anigorufone, its ingestion by the nematode, and its subsequent localization in lipid droplets within the nematode is reported. The importance of varying local concentrations of these specialized metabolites in infected plant tissues, their involvement in the plant’s defense system, and derived strategies for improving banana resistance are highlighted.Bananas and plantains (Musa spp.) are among the world’s most important food and cash crops, with a global production of about 138 million tons in 2010. These crops are part of a well-balanced human diet and are a major food staple for more than 400 million people in the tropics (1, 2). About 82% of the world’s banana production is consumed locally, particularly in India, China, and many African countries (Table S1) (1, 2). Export of bananas to the northern hemisphere represents an important source of employment in countries such as Costa Rica, Ecuador, Colombia, and the Philippines (Table S2) (1, 2). Banana yields are severely hampered by fungi, insects, and plant-parasitic nematodes. The burrowing nematode, Radopholus similis (Cobb, 1893) Thorne, 1949, is the key nematode pathogen, causing yield losses up to 75% (3). R. similis is found in all major banana-producing regions of the world; its best-known hosts are bananas, black pepper, Citrus spp. (4), and coffee (5). R. similis causes extensive root lesions that can lead to toppling of banana plants (6).Plant-parasitic nematodes have been effectively managed through the use of nematicides. However, their high toxicity has adverse effects on humans and their toxic residues are known to accumulate through nontarget organisms in the food chain (7). After the withdrawal of many effective nematicides, such as methyl bromide, from the market (8), organophosphate and carbamate nematicides are still intensively applied to banana and therefore continue to threaten the health of agricultural workers and the environment (9). Although several biological control approaches, including the application of both single and multiple control organisms—such as Fusarium oxysporum, Paecilomyces lilacinus, Trichoderma atroviride isolates, and Bacillus firmus—have proved promising under greenhouse conditions, the control they confer to banana plants most probably does not protect plants for more than one cycle in the field, and most of these organisms have yet to be tested under field conditions (10).The in-depth investigation of the plant–nematode interactions at the cellular and molecular level could lead to the development of more rational and efficient control strategies (11). The production of toxic, herbivore-deterrent or -repellent secondary metabolites, which is typical for many plant defense systems, is particularly interesting in this context. Musa cultivars resistant to R. similis have been identified, especially the cultivar Yangambi km5 (Ykm5) (12). Histochemical and ultrastructural investigations of lesions caused by R. similis in Ykm5 revealed the accumulation of phenolic compounds in response to infection (13). Unfortunately, many of these studies were based solely on histochemical staining methods and did not identify the chemical structures of nematicidal secondary metabolites (7, 14, 15). Initial phytochemical analyses of R. similis-infected roots of the Musa cultivar Pisang sipulu identified the phenylphenalenone anigorufone (1) as a phytoalexin produced in response to nematode damage and confirmed earlier suggestions of the significant role of phytoalexins in the plant defense system (16). Phenylphenalenones are a group of special phenylpropanoid-derived natural products (17), which are known as Musaceae phytoalexins (18). The activity of phenylalanine ammonia lyase (EC 4.3.15), the entry-point enzyme of the phenylpropanoid pathway, is correlated to the biosynthesis of specific phenylpropanoids involved in defense and was substantially induced in nematode infected roots of Ykm5 (19). Phenylphenalenone-related compounds show biological activity against bacteria, fungi, algae, and diatoms (18, 20–22). The formation of these compounds has been elicited in banana leaves by Mycosphaerella fijiensis (Black Sigatoka leaf streak disease), in the fruit peels by Colletotrichum musae (anthracnose disease), and in roots and rhizomes by F. oxysporum f. sp. cubense (Panama disease) and R. similis (16, 18, 21, 23). |
