Pisang Awak Research Articles

Peduncle Rot of Banana Caused by Fusarium petroliphilum and F. pernambucanum in Guangxi, China.

Banana is one of the main fruit crops worldwide. In October 2020, peduncles with rot were observed on bananas (Musa sp. ABB, Pisang Awak subgroup) at a about 1600 square meter commercial banana plantation in Dayu Town (23.17° N, 109.80° E), Guigang, Guangxi, China. The incidence of the disease was about 40%. The interior of the peduncle initially appeared reddish-brown and gradually turned black, and the peduncle eventually rotted. Two whole diseased bunches diseased samples were collected from banana plantations. Small pieces of tissues from the peduncle at the junction of disease and health were surface-disinfected in 75% ethanol for 10 s, 2% NaClO for 1 min, and rinsed three times in sterile water, then placed on potato dextrose agar (PDA) for incubation at 25°C. Forty-nine fungal isolates with similar morphology were recovered from diseased tissues, with 82% isolation frequency. Six isolates (GG3-1, GG3-2, GG3-3, GG4-1, GG4-2 and GG4-3) were selected for further study. Genomic DNAs of these isolates were extracted from 7-day-old mycelia. The internal transcribed spacer (ITS), translation elongation factor (TEF1), calmodulin (CAM), and partial RNA polymerase second largest subunit (RPB2) of six representative isolates were amplified and sequenced (O'Donnell et al. 2000, 2010; White et al. 1990). Sequences were deposited in GenBank. (accessions PP087392-PP087397 for ITS; PP102792-PP102797 for TEF1; PP102798-PP102803 for CAM; PP102804-P102809 for RPB2). A phylogenetic tree based on concatenated sequences of all markers using the Maximum Likelihood algorithm (Xia et al. 2019; Schroers et al. 2016). Based on phylogenetic analyses, GG3-1, -2 and -3 were identified as F. petroliphilum with 100% bootstrap support, and GG4-1, -2 and -3 were tightly clustered with F. pernambucanum with 95% bootstrap support. The two representative isolates GG3-2 and GG4-2 were selected for morphology and pathogenicity observation. Colonies of GG3-2 were light yellow and flat mycelium. They produced falciform macroconidia of 46.1 ± 5.3 × 2.6 ± 0.4 μm with 3 to 5 septates, and hyaline, ovoid microconidia of 7.6 ± 0.9 × 4.0 ± 0.6 μm with 0 septate (Brown et al. 2022). Mycelia were whitish to yellowish aerial mycelium for GG4-2. Their macroconidia were falcate of 31.6 ± 3.0 × 4.3 ± 0.3 μm with curved apical cells, foot-shaped basal cells, and 3 to 5 septates. The microconidia were fusoid of 8.9 ± 1.0 × 2.7 ± 0.3 μm with 0 to 1 septate (Santos et al. 2019). For pathogenicity tests, the ends of the banana peduncles were cut off. Needle punctures were made on the ethanol-treated peduncle pieces, followed by inoculation with 20 μL of conidial suspension (106 spores/ml) of each of the two isolates with three replications each. Sterilized water was used as a control. Peduncle pieces were placed in a humid box and incubated at 28ºC. After 7 days, reddish-brown to black lesions were observed on all inoculated peduncle pieces, while no symptoms were observed on the control pieces. The fungus was isolated from the inoculated peduncle pieces and found to match the morphological characteristics and marker sequences of the original isolates, confirming Koch's postulates. To our knowledge, this is the first report of peduncle rot on banana caused by F. petroliphilum and F. pernambucanum in China. This study will provide valuable information on causal pathogens of this disease which can contribute to improving prevention and disease management strategies for growers.

Genetic diversity of the banana Fusarium wilt pathogen in Cuba and across Latin America and the Caribbean.

Fusarium wilt of bananas (FWB) is a severe plant disease that leads to substantial losses in banana production worldwide. It remains a major concern for Cuban banana cultivation. The disease is caused by members of the soil-borne Fusarium oxysporum species complex. However, the genetic diversity among Fusarium species infecting bananas in Cuba has remained largely unexplored. In our comprehensive survey, we examined symptomatic banana plants across all production zones in the country, collecting 170 Fusarium isolates. Leveraging genotyping-by-sequencing and whole-genome comparisons, we investigated the genetic diversity within these isolates and compared it with a global Fusarium panel. Notably, typical FWB symptoms were observed in Bluggoe cooking bananas and Pisang Awak subgroups across 14 provinces. Our phylogenetic analysis revealed that F. purpurascens, F. phialophorum, and F. tardichlamydosporum are responsible for FWB in Cuba, with F. tardichlamydosporum dominating the population. Furthermore, we identified between five and seven distinct genetic clusters, with F. tardichlamydosporum isolates forming at least two subgroups. This finding underscores the high genetic diversity of Fusarium spp. contributing to FWB in the Americas. Our study sheds light on the population genetic structure and diversity of the FWB pathogen in Cuba and the broader Latin American and Caribbean regions.

Effects of the Ripening Time of Porcelain Banana (Musa spp. Abb cv. Pisang awak) on the total Polyphenol Contents and Bioactivities

To investigate the influence of the banana ripening stage on the change in the content of bioactive components and their bioactivities, ripening stages of porcelain banana peels were screened for the total phenolic content (TPC) and antioxidant activities using three methods, including DPPH scavenging assay, ferric reducing/antioxidant power (FRAP) assay, and inhibition of polyphenol oxidase activity (PPO assay). The Pearson correlation analysis of antioxidant parameters of banana peel extracts for each ripening stages with TPC data showed that the TPC values in the sample extracts were strongly negatively correlated with the IC50 values of the DPPH assay (r = -0.905). They positively correlated with the Fe3+ reduction capacity of Fe2+ (r = 0.723) and the ability to inhibit PPO enzyme at a concentration extract of 100 µg/mL (r = 0.786). During the ripening of bananas, the TPC at stage 1 (M1, light green, 228 mg GAE/100 g DW) was highest, and decreased to a minimum at stage 4 (M4, yellow color, accounts for 50% to 85% of the total banana peel surface area), and increase in stage 6 (M6, yellow color and appearance of black points, 199.9 mg GAE/100 g DW). The banana peel extract in stage 6 showed intense activities, having the highest extraction yield (15.18%) and was analyzed chemical constituents by HPLC-MS, which showed that it contains valuable group compounds such as phenolic acids, flavonols, flavan-3-ols, catecholamines, lignans, and antibacterial compounds. This shows the potential of using banana peels to prepare polyphenol extracts containing high antioxidant activities, enhancing the use value of bananas.

1-methylcyclopropene combined with ethylene absorbent delays the ripening of ‘Fenjiao’ banana (Musa ABB Pisang Awak)

The 'Fenjiao' banana (Musa ABB Pisang Awak) has a high commercial and nutritional value and is harvested when it is highly plump at between 85 and 90 % commercial maturity. The fruit are less angular at this stage. However, these fruit soften rapidly after harvest. 1-Methylcyclopropene (1-MCP) significantly delays ripening, but it can cause ripening disorders when used improperly. Ethylene absorbent (EA) is also widely used to increase the postharvest life of fruit, but it has less of an effect on 'Fenjiao' banana. This study was conducted to optimize the concentrations of 1-MCP combined with EA to increase the shelf life of these bananas. The results showed that a low concentration of 1-MCP (7 g L−1) combined with EA (2 g kg−1) significantly prolonged the shelf life of the 'Fenjiao' banana and maintained its high quality. The combination of 1-MCP and EA repressed the activities of 1-aminocyclopropane-1-carboxylic acid (ACC) oxidase, ACC synthase, pectin methylesterase, polygalacturonase and pectate lyase in the peel and pulp of 'Fenjiao' banana, as well as the corresponding levels of expression of MbACS1, MbACO1, MbPG1, MbPL1, except for MbPME1, which are induced by 1-MCP. This study provides a useful technique for preserving 'Fenjiao' fruit with a study on its possible physiological mechanism, which could be applied in the postharvest production of 'Fenjiao' banana.

Lipid dynamics of 'Fenjiao' bananas (Musa ABB Pisang Awak) during post-ripening based on lipidomics analysis

Changes in cellular lipid homeostasis during ripening of ‘Fenjiao’ bananas (Musa ABB Pisang Awak) significantly affected its storage tolerance, and an in-depth understanding of the changes in cellular lipids during fruit ripening could provide more information and basic data for post-harvest quality regulation of 'Fenjiao' bananas. In this study, a comprehensive metabolomics approach was employed to annotate and characterize a total of 184 lipids, which encompassed glycerides, glycero-phospholipids, sphingolipids, and sterols. In the pulp of 'Fenjiao' bananas at three different stages of ripening (color-transferring, yellow-ripening, and over-ripening), of which glycerides and glycerophospho-lipids were the most predominant lipid constituents in the pulp of ‘Fenjiao’ bananas, with the percentages of the species accounting for 60.33% and 30.43%, respectively. During the post-ripening process of ‘Fenjiao’ bananas, the percentage of glycerides in the pulp exhibited a decreasing trend followed by an increasing trend. In contrast, the trend of glycerophospholipids was opposite to that of glycerides, showing an increasing trend followed by a decreasing trend. The tria-cylglycerol (TAG) content will increase significantly during the senescence stage of the 'Fenjiao' bananas, whereas the levels of diglycerides (DAG), Phosphatidylinositol (PI), Pho-sphatidylcholine (PC), and Phosphatidylethanolamine (PE) will gradually decrease. Through analysis of the differential lipid metabolites, it was discovered that these lipids were primarily enriched in the glycerolipid metabolism, glycerophospholipid metabolism, biosynthesis of unsaturated fatty acids, saturated fatty acid biosynthesis, and phosphatidylinositol signaling pathway. This study is the first to describe and analyze the lipid dynamics during the post-ripening process of 'Fenjiao' bananas, providing valuable insights into its cellular lipid metabolism during this stage.

First Report of Banana mild mosaic virus Infecting Banana in China.

Banana (Musa spp.) is an important fruit in tropical and subtropical regions and an essential food crop in some developing countries. China has a long history of banana cultivation and ranks second in global banana production, with a planting area exceeding 11 million hectares (FAOSTAT, 2023). Banana mild mosaic virus (BanMMV) is a flexuous filamentous virus infecting bananas and a banmivirus in the Betaflexiviridae family. Its infection often results in symptomless plants of Musa spp., and the virus has a worldwide distribution, which can explain its high prevalence (Kumar et al., 2015). BanMMV infection often causes transitory symptoms, such as mild chlorotic streaks and mosaics, on young leaves (Thomas, 2015). The mixed infection of BanMMV with other banana-infecting viruses such as banana streak viruses (BSV) and cucumber mosaic virus (CMV), can exacerbate the mosaic symptoms of BanMMV (Fidan et al., 2019). In October 2021, we collected twenty-six leaf samples of suspected viral disease of bananas from four cities (Huizhou, Qingyuan, Zhanjiang, and Yangjiang) in Guangdong province, two cities (Hekou and Jinghong) in Yunnan province, two cities (Yulin and Wuming) in Guangxi Zhuang autonomous region. After fully mixing these infected samples, we divided them into two pools and sent them to Shanghai Biotechnology Corporation (China) for metatranscriptome sequencing. Each sample contained about 5 g of leaves in total. Zymo-Seq RiboFree Total RNA Library Prep Kit (Zymo Research, USA) was used for ribosomal RNA depletion and library preparations. Illumina sequencing (Illumina NovaSeq 6000) was carried out by Shanghai Biotechnology Corporation (China). Paired-end (150 bp) sequencing of the RNA library was performed on an Illumina HiSeq 2000/2500 platform. Clean reads were assembled by a metagenomic de novo assembly using the CLC Genomics Workbench (version: 6.0.4). Then the non-redundant protein database in the National Center for Biotechnology Information (NCBI) was used for BLASTx annotation. A total of 79,528 contigs were generated from the clean reads (68,878,162) through de novo assembly. A contig of 7265 nucleotides (nts) showed the highest nucleotide sequence identity (90.08%) to the genome of BanMMV isolate EM4-2 (GenBank accession no. OL826745.1). We designed specific primers according to the BanMMV CP gene (Table S1), tested the twenty-six leaf samples collected from the above-mentioned eight cities, and found that only one sample of Fenjiao (Musa ABB Pisang Awak) in Guangzhou city was infected with this virus. The symptoms of banana leaves containing BanMMV were slight chlorosis and yellowing of leaf edges (Fig. S1). We failed to detect other banana viruses, such as BSV, CMV, and banana bunchy top virus (BBTV) in the BanMMV-infected banana leaves. RNA from the infected leaves was extracted, and the assembled contig was confirmed by overlapping PCR amplification across the whole sequence (Table S1). All ambiguous regions were amplified by PCR and RACE, and the products were subjected to Sanger sequencing. The complete genome of the virus candidate was 7310 nts in length, excluding the poly (A) tail. The sequence was deposited in GenBank under accession number ON227268 (isolate BanMMV-GZ from Guangzhou). A schematic representation of the genome organization of BanMMV-GZ is shown in Fig. S2. Its genome has five open reading frames (ORF) encoding RNA-dependent RNA polymerase (RdRp), three triple gene block proteins necessary for cell-to-cell movement (TGBp1 to TGBp3) and a coat protein (CP), similar to other BanMMV isolates (Kondo et al., 2021). Phylogenetic analyses of the complete nt sequence of the full genome and RdRp gene using the neighbor-joining (NJ) method also clearly placed the BanMMV-GZ firmly within all isolates of BanMMV (Fig. S3). To our knowledge, this is the first report of BanMMV infecting bananas in China, extending the geographical range of this viral disease around the world. Accordingly, larger-scale BanMMV investigations must be conducted to determine the distribution and prevalence of BanMMV in China.

Distribution and diversity of Fusarium oxysporum f.sp. cubense TR4 causing banana wilt in Sub-tropics of India and comparative analysis of TR4 specific molecular detection methods

Fusarium wilt caused by Fusarium oxysporum f.sp. cubense (Foc) is regarded as the most recurring and deadliest threat to banana cultivation worldwide. Among three physiological races of the pathogen capable of infecting Musa spp., Tropical race 4 (TR4) is considered the most virulent race of Foc without any known source for resistance. In India, race 1 was commonly observed to affect the cultivars of Pisang Awak, Rasthali (Silk), and Malbhog (Silk) while the Grand Naine (G9) the most popular and extensively grown cultivar of Cavendish banana in subtropics of Uttar Pradesh (U.P.) and Bihar was free from the disease till 2018. But in 2018, the incidence of wilt was first time observed in Sohawal, Ayodhya (U.P.) later in Katihar (Bihar) of India. The Association of TR4 was later confirmed with diseased samples. In the current study, the distribution and incidence of TR4 was assessed in the subtropics covering the states of U.P. and Bihar. Forty-one isolates pathogenic to the banana plant were collected from 6 states, which varied in terms of aggressiveness, and morphological characteristics like colony color, spore size, and shape. The isolates from U.P. and Bihar were characterized as TR4 using two primer sets targeting the SIX1a (secreted in the xylem) gene however isolates from Gujarat, Assam, Andhra Pradesh, and Tamil Nadu (TN) were identified as Race 1 using two primer sets targeting SIX6b and a hypothetical protein (XM_018394505.1). Genetic variability was assessed using 8 ISSR markers and divided all 41 isolates into two major groups, with a set of isolates from the U.P. and Bihar in group A and isolates from other states in group B. In the comparative analysis of three TR4-specific primers, only 1 out of 3 primer sets was confirmed to be more accurate in the characterization of the TR4. Analysis of molecular variance revealed that genetic variations among the regions and within the region accounted for 37.0 per cent and 63.0 per cent. The current study signified that the spread of the TR4 in U.P. and Bihar suggests the need for efforts for management with emphasis on biosecurity practices and the use of antagonistic biological agents to prevent further spread.

English

Ninety backyard gardens in the south-eastern Ethiopian highlands with enset (Ensete ventricosum (Welw.) Cheesman), banana and various annual crops were assessed for the presence of enset root mealybugs (Paraputo ensete (Williams & Matile-Ferrero)). This study presents the first observation of enset root mealybugs on banana. This pest has until now been exclusively recorded on enset in Ethiopia. In the Dilla Zuria district of the Gedeo zone, southern Ethiopia, infested banana mats of the ‘Pisang Awak’ (ABB genome) landrace were observed adjacent to infested enset plants in three small-holder backyard gardens. As roots of banana mats and enset plants were overlapping and intertwined, and large numbers of mealybugs were observed on enset roots, possibly representing an overpopulation, the observed mealybugs on banana might have represented a “chance infestation". The smaller size of mealybugs on banana roots might indicate a non-optimal host status of this crop. Experimental choice and no-choice pot trials however provided another indication of the possible host potential of ‘Pisang Awak’ and of an additional banana cultivar ‘Matooke’ (AAA-East African Highland). The enset root mealybug was able to fully develop, produce viable offspring and survive on both banana cultivars. Not all investigated banana cultivars presented this host status, and the susceptibility of most Musa cultivars remains low. Key words: Choice trials, corm, crawlers, enset suckers, field survey, Musa cultivars, no-choice trials, roots, screenhouse pot trials.

A study of starch resources with high-amylose content from five Chinese mutant banana species.

Investigation on staple crop starch of new species has been becoming the research focus of scholars at present. Based on this, the physicochemical properties and microstructural characteristics of starches isolated from Chinese mutant Musa acuminata Colla acuminata and double balbisiana (MA), Musa double acuminata cv. Pisang Mas (MAM), Musa acuminata cv. Pisang Awak (MAA), and Musa Basjoo Siebold (MBS), and Musa double acuminata and balbisiana-Prata (MAP) were investigated. Results exhibited that all starches exhibited high content of amylose (34.04-42.59%). According to the particle size, they were divided into medium (MA, MAM) (14.54-17.71 μm) and large (MAA, MBS, MAP) (23.01-23.82 μm) group. The medium group with A-type crystallization showed higher peak viscosity (PV), final viscosity, gel fracturability and gel hardness. For large group with B-type crystallization, the compact particle morphology, higher degree of crystallinity, short range order, gelatinization enthalpy, pasting temperature, lower porosity, water absorption capacity (WAC) and oil absorption capacity were found. In addition, the medium group with higher PV and gel hardness could be used as food thickening or gelling agents. The large group with higher Rc, short-range order, lower porosity and WAC could be potential to become raw material for resistant starch. All results showed the amylose content, had significant effect on the microstructure and physicochemical properties of starch samples. Outcomes in this investigation might provide a basis of theoretical application for industrial food production.

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 theITS 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.

First Report of Stem-end Rot Caused by Lasiodiplodia brasiliensis on Banana in USA.

US banana producers are looking for the organic banana market in the southeastern US including Florida and the coastal region of Georgia (Schupska, 2008). In December of 2020, a 6-hand bunched banana (cv. Pisang Awak, belongs to tetraploid AABB genome) with nearly 50% infection (with 15-20% disease severity, <1% of the total harvest) was received from the UGA Banana Research Plot, Tifton, GA with typical stem end rot symptoms of softened and water-soaked flesh. To identify the pathogen, the infected tissues were separated with a sterilized blade, surface disinfested with 10% bleach solution for 1 min, and subsequently washed in three changes of sterile distilled water. The sterilized tissues were aseptically placed on potato dextrose agar (PDA) medium and incubated at 25°C in the dark for 5-10 days. Two isolates of the pathogen with similar colony morphology were obtained and initially identified morphologically using a Botryosphaeriaceae taxonomic key (Phillips et al., 2013). The first growth phase for the isolates documented on PDA, gave rise to white colonies, followed by a dense, black mycelium. The mycelium was fast-spreading, immersed, branched, and septate. The shiny black pycnidia were viewed on the PDA surface after 8-10 days of incubation. Initially, the morphological features of the isolates were identified as Lasiodiplodia spp. (Phillips et al., 2013). To identify to species level, genomic DNA was extracted from two isolates (SW1 & SW2) and amplified by PCR for sequencing using ITS1/ITS4 (White et al., 1990), EF1- 688F/ EF1- 1251R (Alves et al., 2008), Bt2a/Bt2b (Glass & Donaldson, 1995) and rpb2-LasF/ rpb2-LasR (Cruywagen et al., 2017). The ITS (MZ293097 and MZ293114), EF1(OL657173 and OL657174) and rpb2 (OL704860 and OL704861) sequences showed 100% identity and Bt (OL657175 and OL657176) sequences showed 99.5% and 99.7% identity to the corresponding sequences of Lasiodiplodia brasiliensis type strain CMW35884 in GenBank (ITS: KU887094, EF1: KU886972, Bt: KU887466 and rpb2: KU696345). To further affirm the identity, a concatenated phylogenetic analysis was executed with ITS, EF1, Bt, and rpb2 sequences of both isolates and 31 reference strains using Geneious Prime 2019.2.3 Tamura-Nei Neighbor-joining method with 1,000 bootstrap replications, and the outcome was consistent with the conclusion above. To fulfill Koch's postulates, a pathogenicity test was performed with bunched bananas. Two whole bunched bananas surface sterilized with 10% bleach solutions and subsequent washing with sterilized water were cut into 3 bananas per brunch. The inoculum was prepared with 105 spores/ml. The conidial suspension was inoculated on the on-cut surface of the banana crown (300 µl per crown) using a micropipette. Sterile distilled water was applied as a control. The fruit was then packed and sealed in plastic bags and incubated at 25°C. Stem end rot symptoms were first appeared at 5 dpi and increased 7 days later. Two weeks post-inoculation, typical blackened and softened rot tissues were observed, and control fruits remained asymptomatic. To the best of our knowledge, this is the first report of Lasiodiplodia brasiliensis causing stem-end rot of bananas in the USA. This report would be valuable to the banana growers in the southeastern US by taking suitable control measures to confront this fungal disease.

First Report of Crown Rot of Banana Caused by Fusarium proliferatum in Georgia, U.S.A.

Crown rot is one of the most damaging disease of banana fruit characterized by rot and necrosis of crown tissues. In severe cases, the disease can spread to the pedicel and banana pulp. Crown rot can be infected by several common fungi, including Lasiodiplodia theobromae, Musicillium theobromae, Colletotrichum musae, and a complex of Fusarium spp. and lead to softening and blackening of tissues (Lassois et al., 2010; Kamel et al., 2016; Triest et al., 2016; Snowdon, 1990). In November 2020, typical crown rot of banana fruits (cv. Pisang Awak, belonging to the tetraploid AABB genome) were observed from UGA Banana Research 12 Plots, Tifton, GA, with incidence rates of 15%. Initial symptoms appeared in the infected crown of green banana fruits. As the infection progressed, the crown tissues became blackened and softened, followed by an internal development of infection affecting the peduncle and the fruit, triggered early ripening of bananas. At last, the development of necrosis on the pedicels and fruits appeared and caused the fingers to fall off. To identify the pathogen, tissue pieces (~0.25 cm2) from the infected crown and pedicles were surface-sterilized in a 10% bleach solution for 1 min, followed by 30 s in 70% EtOH. The disinfected tissues were rinsed in sterile water 3 times and cultured on potato dextrose agar (PDA) amended with 50 µg/ml streptomycin at 25°C in the dark for 5-10 days. Isolates of the pathogen were purified using the single-spore isolation method (Leslie and Summerell 2006). Colonies on PDA produced fluffy aerial mycelium and developed an intense purple pigment when viewed from the underside. A range of colony pigmentation and growth rates were observed among the isolates. The microconidia were ovoid, hyaline, or ellipse in shape. The morphological features of the isolates were identified as Fusarium proliferatum (Leslie and Summerell, 2006). To further identify the isolates, genomic DNA was extracted from a representative isolate. And the internal transcribed spacer (ITS) region, the partial elongation factor (TEF1-α) gene and the β-tubulin gene (TUB2)were amplified and sequenced using the primers ITS1/ITS4 (Yin et al. 2012), EF-1 /EF-2 (O'Donnell et al. 1998) and B-tub1 /B-tub2 (O'Donnell and Cigelnik, 1997), respectively. The amplicons were sequenced and deposited in NCBI (accessions no. MZ292989, MZ293071 for ITS: MZ346602, MZ346603 for TEF1-α and MZ346600 and MZ346601 for B-tub). The ITS, TEF1-α, and B-tub sequences of the isolates showed 100% sequence similarity with Fusarium proliferatum isolates (accessions no. MT560212, LS42312, and LT575130, respectively) using BLASTn in Genbank. For pathogenicity testing, three whole bunched bananas sterilized with 10% bleach solutions and washed by sterilized water, were cut into 5 bananas per brunch. The cut surface of the banana crown was inoculated with conidial suspension (1.0 × 107 cfu/ml) of the pathogen with pipette tips. Equal number of bananas were treated with sterilized water in the same volume as a control. All bananas were sealed in a plastic bag and incubated at 25°C. After 7 days post inoculation, all inoculated bananas showed initial crown rot symptoms while no symptoms were observed on the control bananas. The fungus was re-isolated from the symptomatic tissues of infected bananas and confirmed to be genetically identical to F. proliferatum of the original inoculated strains according to morphological characteristics and molecular identification, fulfilling Koch's postulates. To the best of our knowledge, this is the first report of F. proliferatum causing crown rot on bananas in Georgia, USA.

Endophytic Bacillus subtilis TR21 Improves Banana Plant Resistance to Fusarium oxysporum f. sp. cubense and Promotes Root Growth by Upregulating the Jasmonate and Brassinosteroid Biosynthesis Pathways.

The banana (Musa spp.) industry experiences dramatic annual losses from Fusarium wilt of banana disease, which is caused by the fungus Fusarium oxysporum f. sp. cubense (FOC). Pisang Awak banana 'Fenza No. 1' (Musa spp. cultivar Fenza No. 1), a major banana cultivar with high resistance to F. oxysporum f. sp. cubense race 4, is considered to be ideal for growth in problematic areas. However, 'Fenza No. 1' is still affected by F. oxysporum f. sp. cubense race 1 in the field. TR21 is an endophytic Bacillus subtilis strain isolated from orchids (Dendrobium sp.). Axillary spraying of banana plants with TR21 controls Fusarium wilt of banana, decreasing the growth period and increasing yields in the field. In this study, we established that TR21 increases root growth in different monocotyledonous plant species. By axillary inoculation, TR21 induced a similar transcriptomic change as that induced by F. oxysporum f. sp. cubense race 1 but also upregulated the biosynthetic pathways for the phytohormones brassinosteroid and jasmonic acid in 'Fenza No. 1' root tissues, indicating that TR21 increases Fusarium wilt of banana resistance, shortens growth period, and increases yield of banana by inducing specific transcriptional reprogramming and modulating phytohormone levels. These findings will contribute to the identification of candidate genes related to plant resistance against fungi in a nonmodel system and facilitate further study and exploitation of endophytic biocontrol agents.

Geographical Distribution and Genetic Diversity of the Banana Fusarium Wilt Fungus in Laos and Vietnam.

Fusarium wilt, caused by the fungus Fusarium oxysporum f. sp. cubense (Foc), poses a major threat to global banana production. The tropical race 4 (TR4) variant of Foc is a highly virulent form with a large host range, and severely affects Cavendish bananas. Foc TR4 was recently observed within the Greater Mekong Subregion, after Chinese private companies expanded Cavendish production to the region. In this study, extensive surveys conducted across Laos and Vietnam show that Foc TR4 is still mainly constricted to the northern regions of these countries and is limited to Cavendish cultivation settings. In Laos, Foc TR4 is associated with large-scale Cavendish plantations owned by or involved with Chinese companies through which infected planting material could have been imported. In Vietnam, mostly small-holder Cavendish farmers and backyard gardens were affected by Foc TR4. In Vietnam, no direct link is found with Chinese growers, and it is expected the pathogen mainly spreads through local and regional movement of infected planting materials. Foc TR4 was not recorded on banana cultivars other than Cavendish. The extensively cultivated ‘Pisang Awak’ cultivar was solely infected by VCGs belonging to Foc race 1 and 2, with a high occurrence of VCG 0123 across Laos, and of VCG 0124/5 in Vietnam. Substantial diversity of Foc VCGs was recorded (VCGs 0123, 0124/5, 01218 and 01221) from northern to southern regions in both countries, suggesting that Fusarium wilt is well established in the region. Interviews with farmers indicated that the local knowledge of Fusarium wilt epidemiology and options for disease management was limited. Clear communication efforts on disease epidemiology and management with emphasis on biosecurity practices need to be improved in order to prevent further spread of Foc TR4 to mixed variety smallholder settings.

Physico-chemical Properties of Mechanically Extracted Banana Juice at Different Ripening Stages

This paper presents the findings of a study on physico-chemical properties of low viscosity banana juice from five banana cultivars: Pisang Awak (PSA) an ABB genome, Yangambi km5 (YKM5) an AAA genome and cultivars from AAA-EA genome, namely Mbilabile, Ndeshi and Mlonga. Low viscosity banana juice was extracted using improved mechanical method at five ripening stages. The physico-chemical properties significantly varied according to cultivars and ripening stages (p &lt; 0.05). Yields, total soluble solids (TSS), titratable acidity (TA) and pH were in the range of 19.50–74.03%, 7.33–27.64 ºBrix, 0.37–0.76% malic acid equivalent and 4.09–4.70, respectively. Yields decreased in the order of Mbilabile &gt; Ndeshi &gt; Mlonga &gt; YKM5 &gt; PSA whereas the total soluble solids increased as follows Mlonga &lt; Mbilabile &lt; Ndeshi &lt; YKM5 &lt; PSA. Furthermore, ashes and mineral contents decreased with ripening stages whereas the low viscosity banana juice was rich in potassium and magnesium. Finally, the study recommends stage 5 of the ripeness as ideal for producing low viscosity banana juice.&#x0D; Keywords: Mechanical extraction, Banana cultivar, Ripening stage, Tannin, Banana juice

Effects of Ripeness and Blending Speed on the Extraction Yield and Physical Chemical Properties of Low Viscosity Banana Juice

Low viscosity banana juice can be extracted from banana by blending and pressing the resulted semi-solid pulp to separate the juice. This juice extraction technology is relatively new and further studies to understand key parameters for juice release is vital for scaling up production and commercialisation. This study, investigated the influence of blending speed and ripeness stage on banana juice yield using two banana cultivars; the East African highland banana (Mbile) and the exotic banana (Pisang awak). Types of sugars and organic acids in the juice were analysed using an HPLC. Changes with ripening of fruit firmness, juice pH and soluble solids were also examined. The juice yield increased significantly (p &lt; 0.05) with increasing blending speed and ripeness stage for all cultivars. The Mbile cultivar from Kagera had the highest juice yield of 82.35%, followed by Mbile cultivar from Kilimanjaro with 75.30% and Pisang awak cultivar with juice yield of 62.50%. The highest juice yield for all cultivars was achieved at 3500 rpm blending speed and ripeness stage 5. Fructose and glucose sugars increased significantly (p &lt; 0.05) with ripeness, while sucrose showed insignificant changes. Malic and citric acids were observed to increase significantly (p &lt; 0.05) during ripening in all banana cultivars.&#x0D; Keywords: East African highland banana, Banana juice extraction, Blending speed, Low viscosity banana juice.

Physicochemical Characteristics, Microstructure and Health Promoting Properties of Green Banana Flour.

This study aimed to investigate the proximate composition, mineral content, functional properties, molecular structure, in vitro starch digestibility, total phenolic content (TPC), total flavonoid content (TFC) and antioxidant activity (DPPH, FRAP) of green banana flour (GBF) cultivars grown in South Africa. With proximate composition, Finger Rose and Pisang Awak had the highest protein (4.33 g/100 g) and fat (0.85 g/100 g) content, respectively. The highest ash content (3.50 g/100 g) occurred with both Grand Naine and FHIA-01 cultivars. Potassium and copper were the most abundant and least minerals, respectively. Pisang Awak cultivar had the highest water absorption capacity (67.11%), while Du Roi had the highest swelling power (0.83 g/g) at 90 °C. Scanning electron microscopy (SEM) images revealed that starch granules from all GBF cultivars were irregular in shape and they had dense surfaces with debris. All the GBF cultivars had similar diffraction patterns with prominent peaks from 15°–24° diffraction angles. The resistant starch (RS) and amylose content of the FHIA-01 cultivar indicates that the GBF has the potential to lower risks of type 2 diabetes and obesity. The highest TPC, TFC and antioxidant activity occurred with the Grande Naine cultivar. Based on their functional characteristics, the Grand Naine and FHIA-01 GBF cultivars could potentially be used as raw materials for bakery products as well as for the fortification of snacks.

Corrigendum: A Novel Banana Mutant “RF 1” (Musa spp. ABB, Pisang Awak Subgroup) for Improved Agronomic Traits and Enhanced Cold Tolerance and Disease Resistance

[This corrects the article DOI: 10.3389/fpls.2021.730718.].

Application of Pisang Awak Bunch-Derived Heterogenous Base Catalyst in Transesterification of Palm Oil into Biodiesel

Biodiesel is an alternative fuel for diesel machine comprosied of alkyl monoesters deriving from vegetable oils or animal fats. Cooking oil is an oil originated from vegetable or animal fat which has been priorly purified, where it appears in liquid form at room temperature and is usually used to fry food ingredients. Heterogenous catalyst is a catalyst present in different phase with the reagent in a reaction it catalyzes. Kalium content in banana in a banana bunch is sufficiently high reaching 94.4%. The aim of this study was to utilize banana bunch which has been priorly ashed using furnace at 700°C for 4 hours, thereafter, applied as a heterogenous catalyst in a the preparation process of biodiesel from cooking oil. Processing variables investigated in this research included the influences of the number of catalyst (3, 4, 5, 6, and 7%) and molar rasio of oil and methanol (1:5, 1:6, 1:7, 1:8, and 1:9) against the properties of produced biodiesel, namely density, viscosity, and water content which later compred with Indonesian standard (SNI). From the study, it was obtained maximum yield of 90.97% with methanol:oil rasio of 1:7 at processing temperature of 60°C with reaction time of 90 minutes and catalyst as much as 3 % w/w. The characteristics of the cooking oil-based biodiesel obtained from the a reaction with oil: methanol rasio of 1:6 and catalyst as much as 3% w/w were density 850 kg/m3 and viscosity 621 mm2/s. This research showed that the obtained biodiesel characteristics had been sufficient according to the SNI, and the use of calcinated banana bunch was very potential in the production of biodiesel acting as solid catalyst person.

Identification of MaWRKY40 and MaDLO1 as Effective Marker Genes for Tracking the Salicylic Acid-Mediated Immune Response in Bananas.

Bananas are among the world's most important cash and staple crops but are threatened by various devastating pathogens. The phytohormone salicylic acid (SA) plays a key role in the regulation of plant immune response. Tracking the expression of SA-responsive marker genes under pathogen infection is important in pathogenesis elucidation. However, the common SA-responsive marker genes are not consistently induced in different banana cultivars or different organs. Here, we conducted transcriptome analysis for SA response of a banana cultivar, 'Pei-Chiao' (Cavendish, AAA genome), and identified three genes, MaWRKY40, MaWRKY70, and Downy Mildew Resistant 6 (DMR6)-Like Oxygenase 1 (MaDLO1) that are robustly induced upon SA treatment in both the leaves and roots. Consistent induction of these three genes by SA treatment was also detected in both the leaves and roots of bananas belonging to different genome types such as 'Tai-Chiao No. 7' (Cavendish, AAA genome), 'Pisang Awak' (ABB genome), and 'Lady Finger' (AA genome). Furthermore, the biotrophic pathogen cucumber mosaic virus elicited the expression of MaWRKY40 and MaDLO1 in infected leaves of susceptible cultivars. The hemibiotrophic fungal pathogen Fusarium oxysporum f. sp. cubense tropical race 4 (TR4) also consistently induced the expression of MaWRKY40 and MaDLO1 in the infected roots of the F. oxysporum f. sp. cubense TR4-resistant cultivar. These results indicate that MaWRKY40 and MaDLO1 can be used as reliable SA-responsive marker genes for the study of plant immunity in banana. Revealing SA-responsive marker genes provides a stepping stone for further studies in banana resistance to pathogens.