First Report of Fusarium Wilt of Iholena Banana (Musa spp.) Caused by Fusarium oxysporum f. sp. cubense Tropical Race 4 in China.

Fusarium wilt, caused by Fusarium oxysporum f. sp. cubense (Foc), has been considered as the most devastating disease affecting bananas (Musa spp.) worldwide. A highly virulent strain of Foc, known as tropical race 4 (TR4), has been detected in the southeast Asia in the 1990s, and has since spread to western Asia, Australia, the Middle East, southern Africa, and South America (Viljoen et al. 2020). Foc TR4 can cause severe yield losses in Cavendish (AAA), Gros Michel (AAA), Silk (AAB), Pisang Awak (ABB) and Bluggoe (ABB) bananas (Ploetz et al. 2006). However, cooking bananas such as plantain (AAB) and Matooke (AAA) bananas, appear to be resistant (Zuo et al. 2017). Iholena bananas (AAB), a subgroup of varieties related to plantains (also known as Pacific plantains), is an important staple food in the Pacific Islands where it was domesticated. It is also popular in Peru, probably due to its nutritional value (Kepler et al. 2011) and is wildly cultivated in other South American countries (Dita et al. 2013). In December 2019, typical symptoms of banana Fusarium wilt were observed on Iholena accession 'Pacific Plantain' (ITC0210) in experimental fields located in Dongguan, Guangdong Province of China. The symptoms included leaf yellowing and pseudostem splitting. The vascular tissue inside the pseudostems was dark red to brown, and the inner rhizomes yellowish-brown. Vascular tissues from three diseased plants were sampled aseptically and placed on potato dextrose agar (PDA) containing 0.05 g/liter kanamycin. Fungal colonies typical of F. oxysporum developed rapidly, with purple-tinged white aerial mycelia and an abundance of microconidia borne in false heads on short microconidia (Nelson et al. 1983). Chlamydospores were produced singly or in pairs in hyphae and macroconidia. Molecular identification was performed using Foc race 4-specific primers (Lin et al. 2009), Foc TR4-specific primers (Dita et al. 2010), Foc race 1 and Foc STR4-specific primers (Ndayihanzamaso et al. 2020). Amplicons of expected sizes were obtained for Foc TR4 and race 4, but not for Foc race 1 and STR4. Sequencing of the ITS and 18S rDNA from the three Iholena isolates and BLAST result showed a 100% similarity to the Foc TR4 reference sequences in GenBank (Foc II5, PRJNA73539 and PRJNA56513) to prove that the isolates were Foc TR4. Pathogenicity of the three isolates from Iholena bananas was determined by infecting 4-month-old Cavendish cv. 'Grand Nain' bananas and three Iholena accessions, 'Pacific Plantain' 'Tigua' and 'Uzakan', under greenhouse conditions by root immersion in a Foc conidial suspension and soil drenching at 106 conidia/ml (Dita, 2010). Control plants were treated with sterile distilled water. Three replications of five plantlets were used for each accession. After 35 days, the inoculated plantlets developed typical Fusarium wilt symptoms such as yellowing of the older leaves and discoloration of the inner rhizome. The control plants did not develop symptoms. To complete Koch's postulates, the fungus was re-isolated from inoculated plants and identified as Foc TR4 by PCR (Dita et al, 2010). The susceptibility of 'Tigua' and 'Uzakan' was also confirmed in Foc TR4-infested field trials, with both accessions developing severe Fusarium wilt symptoms. The susceptibility of Iholena bananas to Foc TR4 is of significant concern to all countries where this subgroup is cultivated for major food source, including Peru and other South American countries.

Transcriptome and metabolome profiling provide insights into molecular mechanism of pseudostem elongation in banana.

BACKGROUND: Banana plant height is an important trait for horticultural practices and semi-dwarf cultivars show better resistance to damages by wind and rain. However, the molecular mechanisms controlling the pseudostem height remain poorly understood. Herein, we studied the molecular changes in the pseudostem of a semi-dwarf banana mutant Aifen No. 1 (Musa spp. Pisang Awak sub-group ABB) as compared to its wild-type dwarf cultivar using a combined transcriptome and metabolome approach. RESULTS: A total of 127 differentially expressed genes and 48 differentially accumulated metabolites were detected between the mutant and its wild type. Metabolites belonging to amino acid and its derivatives, flavonoids, lignans, coumarins, organic acids, and phenolic acids were up-regulated in the mutant. The transcriptome analysis showed the differential regulation of genes related to the gibberellin pathway, auxin transport, cell elongation, and cell wall modification. Based on the regulation of gibberellin and associated pathway-related genes, we discussed the involvement of gibberellins in pseudostem elongation in the mutant banana. Genes and metabolites associated with cell wall were explored and their involvement in cell extension is discussed. CONCLUSIONS: The results suggest that gibberellins and associated pathways are possibly developing the observed semi-dwarf pseudostem phenotype together with cell elongation and cell wall modification. The findings increase the understanding of the mechanisms underlying banana stem height and provide new clues for further dissection of specific gene functions.

MaMAPK3-MaICE1-MaPOD P7 pathway, a positive regulator of cold tolerance in banana.

BACKGROUND: Banana is a tropical fruit with a high economic impact worldwide. Cold stress greatly affects the development and production of banana. RESULTS: In the present study, we investigated the functions of MaMAPK3 and MaICE1 involved in cold tolerance of banana. The effect of RNAi of MaMAPK3 on Dajiao (Musa spp. 'Dajiao'; ABB Group) cold tolerance was evaluated. The leaves of the MaMAPK3 RNAi transgenic plants showed wilting and severe necrotic symptoms, while the wide-type (WT) plants remained normal after cold exposure. RNAi of MaMAPK3 significantly changed the expressions of the cold-responsive genes, and the oxidoreductase activity was significantly changed in WT plants, while no changes in transgenic plants were observed. MaICE1 interacted with MaMAPK3, and the expression level of MaICE1 was significantly decreased in MaMAPK3 RNAi transgenic plants. Over-expression of MaICE1 in Cavendish banana (Musa spp. AAA group) indicated that the cold resistance of transgenic plants was superior to that of the WT plants. The POD P7 gene was significantly up-regulated in MaICE1-overexpressing transgenic plants compared with WT plants, and the POD P7 was proved to interact with MaICE1. CONCLUSIONS: Taken together, our work provided new and solid evidence that MaMAPK3-MaICE1-MaPOD P7 pathway positively improved the cold tolerance in monocotyledon banana, shedding light on molecular breeding for the cold-tolerant banana or other agricultural species.

Reclaiming Inactive Lithium with a Triiodide/Iodide Redox Couple for Practical Lithium Metal Batteries.

High-energy-density lithium (Li) metal batteries suffer from a short lifespan owing to apparently ceaseless inactive Li accumulation, which is accompanied by the consumption of electrolyte and active Li reservoir, seriously deteriorating the cyclability of batteries. Herein, a triiodide/iodide (I3 - /I- ) redox couple initiated by stannic iodide (SnI4 ) is demonstrated to reclaim inactive Li. The reduction of I3 - converts inactive Li into soluble LiI, which then diffuses to the cathode side. The oxidation of LiI by the delithiated cathode transforms cathode into the lithiation state and regenerates I3 - , reclaiming Li ion from inactive Li. The regenerated I3 - engages the further redox reactions. Furthermore, the formation of Sn mitigates the corrosion of I3 - on active Li reservoir sacrificially. In working Li | LiNi0.5 Co0.2 Mn0.3 O2 batteries, the accumulated inactive Li is significantly reclaimed by the reversible I3 - /I- redox couple, improving the lifespan of batteries by twice. This work initiates a creative solution to reclaim inactive Li for prolonging the lifespan of practical Li metal batteries.

Overexpression of MaTPD1A impairs fruit and pollen development by modulating some regulators in Musa itinerans.

BACKGROUND: Pollen formation and development is important for crop fertility and is a key factor for hybrid development. Previous reports have indicated that Arabidopsis thaliana TAPETUM DETERMINANT1 (AtTPD1) and its rice (Oryza sativa) homolog, OsTPD1-like (OsTDL1A), are required for cell specialization and greatly affect pollen formation and development. Little is known about the role of the TPD1 homolog in banana pollen development. RESULTS: Here, we report the identification and characterization of TPD1 homologs in diploid banana (Musa itinerans) and examine their role in pollen development by overexpressing the closest homolog, MaTPD1A. MaTPD1A exhibits high expression in stamen and localizes in the plasma membrane. MaTPD1A-overexpressing plants produce no pollen grains and smaller and seedless fruit compared to wild-type plants. Transcriptome analysis showed that in plant hormone, starch and sucrose metabolism, and linolenic acid metabolism-related pathways were affected by overexpression of MaTPD1A, and the expression of several key regulators, such as PTC1 and MYB80, which are known to affect anther development, is affected in MaTPD1A-overexpressing lines. CONCLUSIONS: Our results indicate that MaTPD1A plays an important role in pollen formation and fruit development in diploid banana, possibly by affecting the expression of some key regulators of pollen development.

Fusaric acid instigates the invasion of banana by Fusarium oxysporum f. sp. cubense TR4.

Fusaric acid (FSA) is a phytotoxin produced by several Fusarium species and has been associated with plant disease development, although its role is still not well understood. Mutation of key genes in the FSA biosynthetic gene (FUB) cluster in Fusarium oxysporum f. sp. cubense tropical race 4 (Foc TR4) reduced the FSA production, and resulted in decreased disease symptoms and reduced fungal biomass in the host banana plants. When pretreated with FSA, both banana leaves and pseudostems exhibited increased sensitivity to Foc TR4 invasion. Banana embryogenic cell suspensions (ECSs) treated with FSA exhibited a lower rate of O2 uptake, loss of mitochondrial membrane potential, increased reactive oxygen species (ROS) accumulation, and greater nuclear condensation and cell death. Consistently, transcriptomic analysis of FSA-treated ECSs showed that FSA may induce plant cell death through regulating the expression of genes involved in mitochondrial functions. The results herein demonstrated that the FSA from Foc TR4 functions as a positive virulence factor and acts at the early stage of the disease development before the appearance of the fungal hyphae in the infected tissues.

Genome-Wide Identification and Characterization of the NAC Transcription Factor Family in Musa Acuminata and Expression Analysis during Fruit Ripening.

Banana (Musa acuminata, AAA group) is a representative climacteric fruit with essential nutrients and pleasant flavors. Control of its ripening determines both the fruit quality and the shelf life. NAC (NAM, ATAF, CUC2) proteins, as one of the largest superfamilies of transcription factors, play crucial roles in various functions, especially developmental processes. Thus, it is important to conduct a comprehensive identification and characterization of the NAC transcription factor family at the genomic level in M. acuminata. In this article, a total of 181 banana NAC genes were identified. Phylogenetic analysis indicated that NAC genes in M. acuminata, Arabidopsis, and rice were clustered into 18 groups (S1-S18), and MCScanX analysis disclosed that the evolution of MaNAC genes was promoted by segmental duplication events. Expression patterns of NAC genes during banana fruit ripening induced by ethylene were investigated using RNA-Seq data, and 10 MaNAC genes were identified as related to fruit ripening. A subcellular localization assay of selected MaNACs revealed that they were all localized to the nucleus. These results lay a good foundation for the investigation of NAC genes in banana toward the biological functions and evolution.

Genome-Wide Identification of Long Non-coding RNA in Trifoliate Orange (Poncirus trifoliata (L.) Raf) Leaves in Response to Boron Deficiency.

Long non-coding RNAs (lncRNAs) play important roles in plant growth and stress responses. As a dominant abiotic stress factor in soil, boron (B) deficiency stress has impacted the growth and development of citrus in the red soil region of southern China. In the present work, we performed a genome-wide identification and characterization of lncRNAs in response to B deficiency stress in the leaves of trifoliate orange (Poncirus trifoliata), an important rootstock of citrus. A total of 2101 unique lncRNAs and 24,534 mRNAs were predicted. Quantitative real-time polymerase chain reaction (qRT-PCR) experiments were performed for a total of 16 random mRNAs and lncRNAs to validate their existence and expression patterns. Expression profiling of the leaves of trifoliate orange under B deficiency stress identified 729 up-regulated and 721 down-regulated lncRNAs, and 8419 up-regulated and 8395 down-regulated mRNAs. Further analysis showed that a total of 84 differentially expressed lncRNAs (DELs) were up-regulated and 31 were down-regulated, where the number of up-regulated DELs was 2.71-fold that of down-regulated. A similar trend was also observed in differentially expressed mRNAs (DEMs, 4.21-fold). Functional annotation of these DEMs was performed using Gene Ontology (GO) and the Kyoto Encyclopedia of Genes and Genomes (KEGG) pathway analyses, and the results demonstrated an enrichment of the categories of the biosynthesis of secondary metabolites (including phenylpropanoid biosynthesis/lignin biosynthesis), plant hormone signal transduction and the calcium signaling pathway. LncRNA target gene enrichment identified several target genes that were involved in plant hormones, and the expression of lncRNAs and their target genes was significantly influenced. Therefore, our results suggest that lncRNAs can regulate the metabolism and signal transduction of plant hormones, which play an important role in the responses of citrus plants to B deficiency stress. Co-expression network analysis indicated that 468 significantly differentially expressed genes may be potential targets of 90 lncRNAs, and a total of 838 matched lncRNA-mRNA pairs were identified. In summary, our data provides a rich resource of candidate lncRNAs and mRNAs, as well as their related pathways, thereby improving our understanding of the role of lncRNAs in response to B deficiency stress, and in symptom formation caused by B deficiency in the leaves of trifoliate orange.

Proteomic analysis of conidia germination in Fusarium oxysporum f. sp. cubense tropical race 4 reveals new targets in ergosterol biosynthesis pathway for controlling Fusarium wilt of banana.

Conidial germination is a crucial step of the soilborne fungus Fusarium oxysporum f. sp. cubense tropical race 4 (Foc TR4), a most important lethal disease of banana. In this study, a total of 3659 proteins were identified by isobaric tags for relative and absolute quantitation (iTRAQ)-based comparative proteomic approach, of which 1009 were differentially expressed during conidial germination of the fungus at 0, 3, 7, and 11 h. Functional classification and bioinformatics analysis revealed that the majority of the differentially expressed proteins are involved in six metabolic pathways. Particularly, all differential proteins involved in the ergosterol biosynthesis pathway were significantly upregulated, indicating the importance of the ergosterol biosynthesis pathway to the conidial germination of Foc TR4. Quantitative RT-PCR, western blotting, and in vitro growth inhibition assay by several categories of fungicides on the Foc TR4 were used to validate the proteomics results. Four enzymes, C-24 sterol methyltransferase (ERG6), cytochrome P450 lanosterol C-14α-demethylase (EGR11), hydroxymethylglutaryl-CoA synthase (ERG13), and C-4 sterol methyl oxidase (ERG25), in the ergosterol biosynthesis pathway were identified and verified, and they hold great promise as new targets for effective inhibition of Foc TR4 early growth in controlling Fusarium wilt of banana. To the best of our knowledge, this report represents the first comprehensive study on proteomics profiling of conidia germination in Foc TR4. It provides new insights into a better understanding of the developmental processes of Foc TR4 spores. More importantly, by host plant-induced gene silencing (HIGS) technology, the new targets reported in this work allow us to develop novel transgenic banana leading to high protection from Fusarium wilt and to explore more effective antifungal drugs against either individual or multiple target proteins of Foc TR4.

Quantitative proteomic analysis reveals that antioxidation mechanisms contribute to cold tolerance in plantain (Musa paradisiaca L.; ABB Group) seedlings.

Banana and its close relative, plantain are globally important crops and there is considerable interest in optimizing their cultivation. Plantain has superior cold tolerance compared with banana and a thorough understanding of the molecular mechanisms and responses of plantain to cold stress has great potential value for developing cold tolerant banana cultivars. In this study, we used iTRAQ-based comparative proteomic analysis to investigate the temporal responses of plantain to cold stress. Plantain seedlings were exposed for 0, 6, and 24 h of cold stress at 8 °C and subsequently allowed to recover for 24 h at 28 °C. A total of 3477 plantain proteins were identified, of which 809 showed differential expression from the three treatments. The majority of differentially expressed proteins were predicted to be involved in oxidation-reduction, including oxylipin biosynthesis, whereas others were associated with photosynthesis, photorespiration, and several primary metabolic processes, such as carbohydrate metabolic process and fatty acid beta-oxidation. Western blot analysis and enzyme activity assays were performed on seven differentially expressed, cold-response candidate plantain proteins to validate the proteomics data. Similar analyses of the seven candidate proteins were performed in cold-sensitive banana to examine possible functional conservation, and to compare the results to equivalent responses between the two species. Consistent results were achieved by Western blot and enzyme activity assays, demonstrating that the quantitative proteomics data collected in this study are reliable. Our results suggest that an increase of antioxidant capacity through adapted ROS scavenging capability, reduced production of ROS, and decreased lipid peroxidation contribute to molecular mechanisms for the increased cold tolerance in plantain. To the best of our knowledge, this is the first report of a global investigation on molecular responses of plantain to cold stress by proteomic analysis.

Analysis of therapeutic effect and safety of target-dose metoprolol in the treatment of patients with diabetes mellitus with chronic heart failure.

OBJECTIVE: To explore the therapeutic effect and safety of target-dose metoprolol in treating chronic heart failure (CHF) patients complicated with diabetes mellitus (DM). Method s : One hundred and fifty-four elderly patients were randomly divided into an observation group and a control group (n=77), which were treated with target-dose metoprolol and conventional therapy, and routinely treated respectively. The New York Heart Association (NYHA) classification, left ventricular end-systolic diameter (LVESD), left ventricular end-diastolic diameter (LVEDD), left ventricular ejection fraction (LVEF), 6-min walking distance and medication safety of the two groups were compared. RESULTS: Compared with the results before treatment, the NYHA classification, LVESD, LVEDD, LVEF and 6-minutes walking distance of both groups were significantly improved (P<0.05), with significantly better results in the observation group than those in the control group after treatment (P<0.05). In the 6 months of follow-up, the incidence of cardiac events in the observation group (3.90%) was significantly lower than that of the control group (14.29%) (P<0.05). The levels of average fasting blood sugar and glycosylated hemoglobin in the groups showed no significant differences (P>0.05). CONCLUSION: Treating CHF patients complicated with DM with target-dose metoprolol can obviously boost the cardiac function and exercise tolerance, leading to satisfactory clinical therapeutic effect, high security and moderate tolerance.

Genome-wide analysis of the trihelix gene family reveals that MaGT21 modulates fruit ripening by regulating the expression of MaACO1 in Musa acuminata.

The trihelix transcription factor (GT) gene family members play vital roles in plant growth and development, responses to abiotic or biotic stress, and fruit ripening. However, its role in banana fruit ripening remains unclear. Here, 59 MaGT gene members were identified in banana and clustered into five subfamilies, namely GT1, GT2, GTγ, SIP1, and SH4. This classification is completely supported by their gene structures and conserved motif analysis. Transcriptome data analysis indicated that MaGT14, MaGT21, and MaGT27 demonstrated significant differential expression during fruit ripening. Quantitative real-time PCR analysis revealed that these three genes were highly induced by ethylene treatment, responded to cold and heat stress, and had a high expression abundance in ripe fruit. Subcellular localization demonstrated that MaGT21 and MaGT27 functioned as nuclear proteins, while MaGT14 functioned as a nuclear and cell membrane protein. Further investigation indicated MaGT21 could positively stimulate the transcription of the key ethylene biosynthesis gene MaACO1 by directly targeting the GT motif in the promoter. MaGT21 transient overexpression in banana fruit upregulated MaACO1 and accelerated fruit ripening. Our findings provide comprehensive and valuable information for further functional studies of MaGT genes in banana, help to understand the roles of MaGTs during banana fruit ripening.

High-quality genome assemblies for two Australimusa bananas (Musa spp.) and insights into regulatory mechanisms of superior fiber properties.

Bananas (Musa spp.) are monocotyledonous plants with high genetic diversity in the Musaceae family that are cultivated mainly in tropical and subtropical countries. The fruits are a popular food, and the plants themselves have diverse uses. Four genetic groups (genomes) are thought to have contributed to current banana cultivars: Musa acuminata (A genome), Musa balbisiana (B genome), Musa schizocarpa (S genome), and species of the Australimusa section (T genome). However, the T genome has not been effectively explored. Here, we present the high-quality TT genomes of two representative accessions, Abaca (Musa textilis), with high-quality natural fiber, and Utafun (Musa troglodytarum, Fe'i group), with abundant ß-carotene. Both the Abaca and Utafun assemblies comprise 10 pseudochromosomes, and their total genome sizes are 613 Mb and 619 Mb, respectively. Comparative genome analysis revealed that the larger size of the T genome is likely attributable to rapid expansion and slow removal of transposons. Compared with those of Musa AA or BB accessions or sisal (Agava sisalana), Abaca fibers exhibit superior mechanical properties, mainly because of their thicker cell walls with a higher content of cellulose, lignin, and hemicellulose. Expression of MusaCesA cellulose synthesis genes peaks earlier in Abaca than in AA or BB accessions during plant development, potentially leading to earlier cellulose accumulation during secondary cell wall formation. The Abaca-specific expressed gene MusaMYB26, which is directly regulated by MusaMYB61, may be an important regulator that promotes precocious expression of secondary cell wall MusaCesAs. Furthermore, MusaWRKY2 and MusaNAC68, which appear to be involved in regulating expression of MusaLAC and MusaCAD, may at least partially explain the high accumulation of lignin in Abaca. This work contributes to a better understanding of banana domestication and the diverse genetic resources in the Musaceae family, thus providing resources for Musa genetic improvement.

Two haplotype-resolved genome assemblies for AAB allotriploid bananas provide insights into banana subgenome asymmetric evolution and Fusarium wilt control.

Bananas (Musa spp.) are one of the world's most important fruit crops and play a vital role in food security for many developing countries. Most banana cultivars are triploids derived from inter- and intraspecific hybridizations between the wild diploid ancestor species Musa acuminate (AA) and M. balbisiana (BB). We report two haplotype-resolved genome assemblies of the representative AAB-cultivated types, Plantain and Silk, and precisely characterize ancestral contributions by examining ancestry mosaics across the genome. Widespread asymmetric evolution is observed in their subgenomes, which can be linked to frequent homologous exchange events. We reveal the genetic makeup of triploid banana cultivars and verify that subgenome B is a rich source of disease resistance genes. Only 58.5% and 59.4% of Plantain and Silk genes, respectively, are present in all three haplotypes, with >50% of genes being differentially expressed alleles in different subgenomes. We observed that the number of upregulated genes in Plantain is significantly higher than that in Silk at one-week post-inoculation with Fusarium wilt tropical race 4 (Foc TR4), which confirms that Plantain can initiate defense responses faster than Silk. Additionally, we compared genomic and transcriptomic differences among the genes related to carotenoid synthesis and starch metabolism between Plantain and Silk. Our study provides resources for better understanding the genomic architecture of cultivated bananas and has important implications for Musa genetics and breeding.

Metabolic profiling reveals genotype-associated alterations in carotenoid content during banana postharvest ripening.

Banana fruits have attracted considerable attention for health-promoting effects attributed to ubiquitous functional metabolites. However, genotype-dependent accumulation patterns of carotenoids in banana remain largely unclear. Here, we performed a systematic metabolomic investigation of 18 banana cultivars of the AAA, AAB, or ABB genome groups. Our results indicate that the levels of soluble sugars increase during postharvest ripening regardless of genotype, whereas amino acids (AAs) and tricarboxylic acid (TCA) cycle-derived organic acids display genotype-dependent patterns. The levels of AAs derived from the glycolytic pathway increased, whereas those derived from the TCA cycle significantly decreased during ripening. The carotenoid composition in banana pulp was genotype-specific, and the contents of α-carotene were the highest in AAA-genome bananas. Moreover, high α-carotene and ß-carotene contents in banana were correlated with elevated levels of TCA cycle-derived AAs and decreased levels of glycolysis-derived AAs. Taken together, these findings provide a comprehensive understanding of genotype-associated carotenoid accumulation, thereby facilitating the breeding of future high carotenoid banana cultivars.

Genome-wide analysis of the polyphenol oxidase gene family reveals that MaPPO1 and MaPPO6 are the main contributors to fruit browning in Musa acuminate.

Introduction: Polyphenol oxidases (PPOs), which are widely present in plants, play an important role in the growth, development, and stress responses. They can catalyze the oxidization of polyphenols and result in the browning of damaged or cut fruit, which seriously affects fruit quality and compromises the sale of fruit. In banana (Musa acuminata, AAA group), 10 PPO genes were determined based on the availability of a high-quality genome sequence, but the role of PPO genes in fruit browning remains unclear. Methods: In this study, we analyzed the physicochemical properties, gene structure, conserved structural domains, and evolutionary relationship of the PPO gene family of banana. The expression patterns were analyzed based on omics data and verified by qRT-PCR analysis. Transient expression assay in tobacco leaves was used to identify the subcellular localization of selected MaPPOs, and we analyzed the polyphenol oxidase activity using recombinant MaPPOs and transient expression assay. Results and discussion: We found that more than two-thirds of the MaPPO genes had one intron, and all contained three conserved structural domains of PPO, except MaPPO4. Phylogenetic tree analysis revealed that MaPPO genes were categorized into five groups. MaPPOs did not cluster with Rosaceae and Solanaceae, indicating distant affinities, and MaPPO6/7/8/9/10 clustered into an individual group. Transcriptome, proteome, and expression analyses showed that MaPPO1 exhibits preferential expression in fruit tissue and is highly expressed at respiratory climacteric during fruit ripening. Other examined MaPPO genes were detectable in at least five different tissues. In mature green fruit tissue, MaPPO1 and MaPPO6 were the most abundant. Furthermore, MaPPO1 and MaPPO7 localized in chloroplasts, and MaPPO6 was a chloroplast- and Endoplasmic Reticulum (ER)-localized protein, whereas MaPPO10 only localized in the ER. In addition, the enzyme activity in vivo and in vitro of the selected MaPPO protein showed that MaPPO1 had the highest PPO activity, followed by MaPPO6. These results imply that MaPPO1 and MaPPO6 are the main contributors to banana fruit browning and lay the foundation for the development of banana varieties with low fruit browning.