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Abstract Since publication of a draft genome of the doubled-haploid ‘Pahang’ banana (Musa acuminata, DH-Pahang), a new era for banana biology research has begun. With the release of genomic data from some important Musa species and subspecies and the continuous development of molecular biology techniques, significant progress has been made. Here, we summarize the achievements and advances in the banana molecular biology and breeding over the past decade covering origin and domestication, fruit biology, stress biology, and breeding aspects, and highlight their challenges and future perspectives. This review is intended to provide researchers with the latest information on the complex genetic background and evolutionary relationship of bananas, the biology of fruit ripening, and multi-omics-based stress biology research. We especially focus on recent advances in the molecular breeding of bananas, offering an informative research direction and providing valuable technical references for future research in the field.

Banana, a monocotyledonous plant in the genus Musa of the family Musaceae, is one of the most important tropical fruit crops. Globally, banana is also the fourth most important staple food, followed by rice, wheat, and maize. Cultivated bananas, comprising dessert and cooking types, are mostly triploids with complicated genetic background. Researches on origins and domestication of cultivated bananas are essential for accelerating the process of banana breeding. In recent years, great progress has been made in molecular phylogenetics of banana germplasm resources, providing valuable genetic information to facilitate further uses in the design of breeding strategies. This review updated the sectional taxonomy of the wild species in the genus Musa closely related to cultivated bananas, and summarized the common cultivated groups in the world and their characteristics, encompassing 18 cultivated types in 7 genomic groups. Moreover, we concluded the recent updates on the origins and domestication of the most important cultivated groups including AAA, AAB, ABB and AB, the evolutionary biology of wild M. acuminata subspecies and the existence of undefined ancestral genepools which both contributed to the diversity of the cultivated bananas. Besides, we proposed the prospect for future researches on the cultivated bananas. This review, to the best knowledge of the authors, is systematically summarize the recent advances in the origins and domestication of the cultivated bananas, with a view to seeking elite germplasm and novel gene resources for providing references for the researches on banana breeding.

Abstract Low temperature is a major environmental factor that limits the growth, yield, and geographical distribution of Cavendish and Dajiao bananas (Musa spp.). Dajiao bananas exhibit a significantly higher cold tolerance than the Cavendish cultivar. However, the underlying mechanisms involved in cold tolerance regulation in Dajiao plants have not been determined. In this study, we investigated the mechanisms underlying the differences in cold tolerance at the cellular level between the cold-sensitive Cavendish and cold-tolerant Dajiao banana types through comparative metabolomics and transcriptomics analyses in calli exposed to cold treatment under dark conditions. A higher accumulation of lipids was observed in Dajiao calli cells compared to Cavendish cells under cold stress. After cold treatment, 4,626 and 5,516 differentially expressed genes (DEGs) were identified in Cavendish and Dajiao banana cells, respectively. By integrating the transcriptomic and metabolomic datasets, we discovered that the linoleic acid and α-linoleic acid metabolism and the Abscisic acid (ABA)-independent Mitogen-activated protein kinase (MAPK) cascade-Inducer of CBF expression 1 (ICE1) signal transduction pathway (including Mitogen-activated protein kinase kinase kinase 1 [MEKK1], MAPK5, ICE1, and Cold-regulated 47 [COR47]) played crucial roles in the cold tolerance of Dajiao bananas. Our study provides new insights into cold response regulation and novel cold tolerance mechanisms, providing valuable leads and targets for the genetic improvement of cold tolerance in bananas.

Banana fruits are rich in starch, and unripe banana flour is considered a beneficial ingredient in the food industry because it has high levels of resistant starch, which significantly aids in promoting gut health and regulating blood sugar and lipid levels. However, the associations between banana cultivars with various genotypes cultivated globally and their resistant starch properties remain unclear. Herein, we investigated resistant starches from 16 banana cultivars covering three genome groups (ABB, AAB, and AAA) in order to reveal the differences and similarities among these cultivars. The results showed that there was a genotype-specific pattern in banana resistant starch (BRS) degradation. The AAA genome BRS exhibited a high degree of resistant starch degradation. The genotypes of the banana cultivars also impacted the granular morphology of the resistant starch. The ABB and AAB genome BRS were more conducive to forming resistant starch. The BRS samples from the three genome groups displayed either B-type or C-type structures. Even within the same genome group, the BRS samples exhibited differences in thermal and pasting properties. These findings reveal the impact of genotypes on BRS content and characteristics, providing a basis for future breeding and resistant starch utilization.

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This study investigated the ameliorative effects of plantain flour on the organs of diabetic rats, and explored the mechanism from the perspective of kidney transcription. Diabetes was induced in rats by streptozotocin (STZ) and a high-sugar and high-fat diet. The alleviation effects and molecular mechanism of diabetes by plantain flour at different doses (1 g/kg·Bw, 2 g/kg·Bw, and 4 g /kg·Bw) were evaluated based on the analysis of biochemical indicators, histological observations, homeostasis model, and kidney transcriptome. The findings demonstrated that the intervention of plantain flour can reduce the level of alanine aminotransferase (ALT), aspartate aminotransferase (AST), and urea (UREA). The pathological study revealed that plantain flour can alleviate the pathological symptoms of fat accumulation in liver and kidney tissue. According to the homeostasis model, plantain flour was able to reduce insulin resistance in diabetic rats. Transcriptome analysis showed that plantain flour improved dyslipidemia by up-regulating Acsl1, down-regulating Fabp4 and Plin1, and reduced kidney injury by up-regulating Slco1a6, Hsd17b2, Cyp2c24, and Cyp2c11 genes. These results suggested that plantain flour could improve the organ damage caused by diabetes and has the potential to be developed into healthy food.

Banana is a kind of high-potassium and low-sodium fruit with extremely high nutritional value, which is widely cultivated in tropical and subtropical regions. Banana is an important fruit and food crop, and it is the fruit with the largest production, trade volume and trade value among fresh fruits in the world. China plays an important role in banana production in the world and is the second largest banana producer in the world. According to the statistics of the Food and Agriculture Organization of the United Nations, in 2020, the banana production was 11.8726 million tons, accounting for about 9.01% of the world. Banana germplasm resources are abundant, and there are thousands of banana cultivars in the world. The types and contents of nutrients of banana in different varieties, regions, development and post ripening stages in bananas vary greatly. The main nutritional components in bananas such as mineral elements, resistant starch and dietary fiber were introduced, and phenolic compounds, carotenoids, fatty acids, phytosterols, amines, antioxidants and other active ingredients were summarized. The functional properties of bananas were reviewed, including preventing cardiovascular diseases, relaxing bowel, reducing weight, lowering blood sugar, anti-depression and so on. Its main processed products like banana drink and banana powder were summarized. The intensive processing of banana can improve the resource utilization rate and the product added value, and promote the sustainable development of banana industry and world food security.

IntroductionPolyphenol 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.MethodsIn 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 discussionWe 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.

IntroductionReconstructive breeding based on autotetraploids to generate triploid varieties is a promising breeding strategy in banana (Musa spp.). Therefore understanding the molecular mechanisms underlying the phenotypic differences between the original diploid and its autopolyploid derivatives is of significant importance in such breeding programs of banana.MethodsIn this study, a number of non-chimeric autotetraploid plants, confirmed by flow cytometry and chromosome counting were obtained using colchicine treatment of ‘Pisang Berlin' (AA Group), a diploid banana cultivar highly resistant to Fusarium wilt Tropical Race 4 (Foc TR4) and widely cultivated in Asia.Results and discussionThe autotetraploids showed significant increase in plant height, pseudostem diameter, root length, leaf thickness, leaf area, and leaf chlorophyll content. Transcriptomic analysis indicated that differentially expressed genes were mainly enriched in plant hormone signal transduction, mitogen-activated protein kinase (MAPK) signaling pathway, and carbon fixation in photosynthetic organelles. The genes related to the metabolism, transport or signaling of auxin, abscisic acid (ABA), cytokinin (CTK) and gibberellin (GA), as well as the genes encoding essential enzymes in photosynthetic CO2 fixation were differentially expressed in leaves of autotetraploids and most of them were up-regulated. Metabolomic analysis revealed that the differentially accumulated metabolites were mainly involved in plant hormone signal transduction, porphyrin and chlorophyll metabolism, indole alkaloid biosynthesis, and carbon fixation in photosynthetic organelles. The results therefore, demonstrate that the hormones IAA, ABA, and photosynthetic regulation may play a vital role in the observed enhancement in the autotetraploids. These could be used as molecular and biochemical markers to facilitate the generation of triploid progenies as suitable new varieties for cultivation.

IntroductionDevelopment of functional food ingredients with anti-obesity is a growing interest in the global food industry. Plantain (Musa spp. AAB), a special type of cooking/starchy banana, is widely growing in African and Latin American countries. The flour made from unripe plantain pulp, which is considered as a natural source of indigestible carbohydrates such as resistant starch (RS), could be used in the formulation of diverse functional foods due to its anti-obesity properties. However, the mechanisms underlying the anti-obesity properties of plantain flour are not explored.MethodsIn this study, we investigated the changes in serum hormone levels, liver transcriptome profiles, and the modulation of gut microbiota in high-fat-fed Sprague-Dawley (SD) rats. The male SD rats were divided into six groups, viz. two control groups [non-obese (NC) or obese (OC)] which were not given the supplementation, one positive control (PC) group which received orlistat supplementation (60 mg/kg body weight/day), and three groups of obese rats which were supplemented with unripe plantain flour (UPF) at a dosage (body weight/day) of 1.25 g/kg (low-dose, LD), 2.50 g/kg (intermediate-dose, MD) or 5.0 g/kg (high-dose, HD).Results and discussionIt was found that UPF supplementation could lower the insulin levels of the obese rats. Moreover, UPF supplementation had a positive impact on gut microbiota, decreasing the relative abundances of Blautia, Parasutterella and Fusicatenibacter which were closely related to obesity, and increasing the relative abundances of probiotics (Allobaculum, Romboutsia, Staphylococcus, and Bacteroides). The spearman correlation analysis revealed that UPF supplementation reduced the relative abundance of Parasutterella and possibly decreased the blood sugar levels, leading to a decrease in the relative abundances of Blautia and Fusicatenibacter and a subsequent decrease in insulin levels. Furthermore, transcriptomic analysis of the liver tissues displayed that the peroxisome proliferator activated receptor-1α (PPAR) and AMP-activated protein kinase (AMPK) signaling pathway genes (Pparaa, Cpt1a, Prkaa1, Prkab1, Prkaa2, and Ppargc1a) were upregulated in those groups supplemented with UPF. These results indicated that UPF could mediate the glucolipid metabolism in the obese rats. Taken together, our findings suggested that the anti-obesity properties of UPF could be achieved by decreasing the insulin levels, positive-regulating of the gut microbiota composition as well as altering gene expression related to glucolipid metabolism.

Abstract 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.

Abstract 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.

Abstract 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.

An efficient biolistic transformation system of banana combined with a liquid medium selection system was developed during this study. An embryogenic cell suspension (ECS) of Musa acuminata cv. Baxi (AAA) was bombarded with a particle delivery system. After 7 days of restoring culture in liquid M2 medium, embryogenic cells were transferred to a liquid selection M2 medium supplemented with 10 μg/mL hygromycin for resistance screening. The untransformed cell clusters were inhibited or killed, and a small number of transformants proliferated in the liquid selection medium. After the 0th, first, second, and third generation of antibiotic screening, there were 0, 65, 212, and 320, respectively, vitality-resistant buds obtained from a 0.5-mL packed cell volume (PCV) of embryogenic cell suspension. The β-glucuronidase (GUS) staining, polymerase chain reaction (PCR) analysis, and Southern blot hybridization results all demonstrated a 100% positive rate of regenerated resistant seedlings. Interestingly, the number of buds obtained through third-generation screening was almost equal to that obtained from the original ECS in M2 medium without antibiotics. These results suggested that the liquid medium selection system facilitated the proliferation of a positive transgenic ECS, which significantly improved the regeneration rate of transformants. This protocol is suitable for the genetic transformation of all banana genotypes and is highly advantageous to varieties with low callusing potential.

The CRISPR/Cas9-mediated genome editing system has been used extensively to engineer targeted mutations in a wide variety of species. Its application in banana, however, has been hindered because of the species’ triploid nature and low genome editing efficiency. This has delayed the development of a DNA-free genome editing approach. In this study, we reported that the endogenous U6 promoter and banana codon-optimized Cas9 apparently increased mutation frequency in banana, and we generated a method to validate the mutation efficiency of the CRISPR/Cas9-mediated genome editing system based on transient expression in protoplasts. The activity of the MaU6c promoter was approximately four times higher than that of the OsU6a promoter in banana protoplasts. The application of this promoter and banana codon-optimized Cas9 in CRISPR/Cas9 cassette resulted in a fourfold increase in mutation efficiency compared with the previous CRISPR/Cas9 cassette for banana. Our results indicated that the optimized CRISPR/Cas9 system was effective for mutating targeted genes in banana and thus will improve the applications for basic functional genomics. These findings are relevant to future germplasm improvement and provide a foundation for developing DNA-free genome editing technology in banana.

Banana cultivars with the AAB genome group comprise diverse subgroups, such as Plantain, Silk, Iholena, and Pisang Raja, among others, which play an important role in food security in many developing countries. Some of these cultivars are susceptible to Fusarium oxysporum f. sp. cubense tropical race 4 (Foc TR4), the most destructive pathogen threatening banana production worldwide, and some of them are still largely unknown. We evaluated the resistance of 37 banana genotypes, including Plantain, Silk, Iholena, Maia Maoli/Popoulu, Pisang Raja, Pome, and Mysore, to Foc TR4 under both greenhouse and field conditions. Genotypes from the Silk and Iholena subgroups were highly susceptible to Foc TR4. Pome and Mysore showed resistance and intermediate resistance, respectively. However, Pisang Raja ranged from susceptible to intermediate resistance. One cultivar from the Maia Maoli/Popoulu subgroup was highly susceptible, while the other displayed significant resistance. Most Plantain cultivars exhibited high resistance to Foc TR4, except two French types of cultivar, ‘Uganda Plantain’ and ‘Njombe N°2’, which were susceptible. The susceptibility to Foc TR4 of some of the AAB genotypes evaluated, especially Plantain and other cooking bananas, indicates that growers dependent on these varieties need to be included as part of the prevention and integrated Foc TR4 management strategies, as these genotypes play a crucial role in food security and livelihoods.

Anthocyanins spatiotemporally accumulate in certain tissues of particular species in the banana plant, and MYB transcription factors (TFs) serve as their primary regulators. However, the precise regulatory mechanism in banana remains to be determined. Here, we report the identification and characterization of MaMYB4, an R2R3-MYB repressor TF, characterized by the presence of EAR (ethylene-responsive element binding factor–associated amphiphilic repression) and TLLLFR motifs. MaMYB4 expression was induced by the accumulation of anthocyanins. Transgenic banana plants overexpressing MaMYB4 displayed a significant reduction in anthocyanin compared to wild type. Consistent with the above results, metabolome results showed that there was a decrease in all three identified cyanidins and one delphinidin, the main anthocyanins that determine the color of banana leaves, whereas both transcriptome and reverse transcription–quantitative polymerase chain reaction analysis showed that many key anthocyanin synthesis structural genes and TF regulators were downregulated in MaMYB4 overexpressors. Furthermore, dual-luciferase assays showed that MaMYB4 was able to bind to the CHS, ANS, DFR, and bHLH promoters, leading to inhibition of their expression. Yeast two-hybrid analysis verified that MaMYB4 did not interact with bHLH, which ruled out the possibility that MaMYB4 could be incorporated into the MYB-bHLH-WD40 complex. Our results indicated that MaMYB4 acts as a repressor of anthocyanin biosynthesis in banana, likely due to a two-level repression mechanism that consists of reduced expression of anthocyanin synthesis structural genes and the parallel downregulation of bHLH to interfere with the proper assembly of the MYB-bHLH-WD40 activation complex. To the best of our knowledge, this is the first MYB TF that regulates anthocyanin synthesis that was identified by genetic methods in bananas, which will be helpful for manipulating anthocyanin coloration in banana programs in the future.

A leaf blight disease with an incidence level of about 50% was found on Robusta banana in Guangdong province of China in September 2020. The early symptom appeared as pale gray to black brown, irregular, small, necrotic lesions mainly on the top 3–5 leaves. Severely infected leaves were withered and necrotic. Two representative fungus strains, strain L1 and strain L2, were isolated from affected banana leaves, and morphological and molecular identification analysis confirmed that the two fungi were both Alternaria jacinthicola. Many Alternaria species have been reported to cause wilting, decay, leaf blight and leaf spots on plants and lead to serious economic losses in their production, including A. alternata, causing leaf blight and leaf sport diseases on banana. The Koch’s postulates of A. jacinthicola causing the leaf blight disease was further fulfilled, which confirmed that it is the causal agent of this disease. To our knowledge, this is the first report of A. jacinthicola causing leaf blight on banana in China.

Fungal glucosylceramide plays important role in cell division, hyphal formation and growth, spore germination and the modulation of virulence and has recently been considered as target for small molecule inhibitors. In this study, we characterized CgGCS, a protein encoding a glucosylceramide synthase (GCS) in Colletotrichum gloeosporioides. Disruption of CgGCS resulted in a severe reduction of mycelial growth and defects in conidiogenesis. Sphingolipid profile analysis revealed large decreases in glucosylceramide production in the mutant strains. Pathogenicity assays indicated that the ability of the ΔCgGCS mutants to invade both tomato and mango hosts was almost lost. In addition, the expression levels of many genes, especially those related to metabolism, were shown to be affected by the mutation of CgGCS via transcriptome analysis. Overall, our results demonstrate that C. gloeosporioides glucosylceramide is an important regulatory factor in fungal growth, conidiation, and pathogenesis in hosts.

In our living environment, we frequently encounter natural disasters such as typhoons, floods, earthquakes, drought, petrochemical plant safety accident, stripping accident and so on. Due to global climate change, natural disasters have become even more devastating. Thus, it is important to look for ways to mitigate disasters. This study employs the concept of resilience to examine the ability of parks to recover from natural disasters. According to previous disaster history, the damage caused by natural disasters in the parks is mainly water and wind. Therefore, this study first conducted a literature review to find out relevant resilience criteria that are advantageous for combating water and wind disasters, and to recognize the degree of resilience achieved by each criterion. Then, based on the elements of the resilience criteria, the study establishes a preliminary resilience grading system. Furthermore, three different types of parks are evaluated under the grading system. This study provides a theoretical basis for resilience grading and suggestions for improving parks in the future.

Banana is an important tropical fruit with high economic value. One of the main cultivars (‘Cavendish’) is susceptible to low temperatures, while another closely related specie (‘Dajiao’) has considerably higher cold tolerance. We previously reported that some membrane proteins appear to be involved in the cold tolerance of Dajiao bananas via an antioxidation mechanism. To investigate the early cold stress response of Dajiao, here we applied comparative membrane proteomics analysis for both cold-sensitive Cavendish and cold-tolerant Dajiao bananas subjected to cold stress at 10°C for 0, 3, and 6 h. A total of 2,333 and 1,834 proteins were identified in Cavendish and Dajiao, respectively. Subsequent bioinformatics analyses showed that 692 Cavendish proteins and 524 Dajiao proteins were predicted to be membrane proteins, of which 82 and 137 differentially abundant membrane proteins (DAMPs) were found in Cavendish and Dajiao, respectively. Interestingly, the number of DAMPs with increased abundance following 3 h of cold treatment in Dajiao (80) was seven times more than that in Cavendish (11). Gene ontology molecular function analysis of DAMPs for Cavendish and Dajiao indicated that they belong to eight categories including hydrolase activity, binding, transporter activity, antioxidant activity, etc., but the number in Dajiao is twice that in Cavendish. Strikingly, we found peroxidases (PODs) and aquaporins among the protein groups whose abundance was significantly increased after 3 h of cold treatment in Dajiao. Some of the PODs and aquaporins were verified by reverse-transcription PCR, multiple reaction monitoring, and green fluorescent protein-based subcellular localization analysis, demonstrating that the global membrane proteomics data are reliable. By combining the physiological and biochemical data, we found that membrane-bound Peroxidase 52 and Peroxidase P7, and aquaporins (MaPIP1;1, MaPIP1;2, MaPIP2;4, MaPIP2;6, MaTIP1;3) are mainly involved in decreased lipid peroxidation and maintaining leaf cell water potential, which appear to be the key cellular adaptations contributing to the cold tolerance of Dajiao. This membrane proteomics study provides new insights into cold stress tolerance mechanisms of banana, toward potential applications for ultimate genetic improvement of cold tolerance in banana.

The psychology and physiology of miners would be damaged if they continue to stay in uncomfortable environments for long time. When waiting for rescue in emergency refuge facilities, numerous factors can affect comfortable feelings of miners. In this paper, four crucial factors, including oxygen concentration, carbon dioxide concentration, temperature and relative humidity, were selected as the study objects. Adopting the grading method, a series of human live experiments were performed to obtain vote data under both a single and multiple environmental variables. The functional relation between human comfort degree and environmental variables were got by statistically analyzing these vote data. A mathematical model was proposed to predict the human comfort degree of miners in refuge confined space. The allowable ranges of oxygen level, carbon dioxide level, temperature and relative humidity along with their controlling principles were discussed in the end.

The development of organized, informative, robust, user-friendly, and freely accessible molecular markers is imperative to the Musa marker assisted breeding program. Although several hundred SSR markers have already been developed, the number of informative, robust, and freely accessible Musa markers remains inadequate for some breeding applications. In view of this issue, we surveyed SSRs in four different data sets, developed large-scale non-redundant highly informative therapeutic SSR markers, and classified them according to their attributes, as well as analyzed their cross-taxon transferability and utility for the genetic study of Musa and its relatives. A high SSR frequency (177 per Mbp) was found in the Musa genome. AT-rich dinucleotide repeats are predominant, and trinucleotide repeats are the most abundant in transcribed regions. A significant number of Musa SSRs are associated with pre-miRNAs, and 83% of these SSRs are promising candidates for the development of therapeutic SSR markers. Overall, 74% of the SSR markers were polymorphic, and 94% were transferable to at least one Musa spp. Two hundred forty-three markers generated a total of 1047 alleles, with 2-8 alleles each and an average of 4.38 alleles per locus. The PIC values ranged from 0.31 to 0.89 and averaged 0.71. We report the largest set of non-redundant, polymorphic, new SSR markers to be developed in Musa. These additional markers could be a valuable resource for marker-assisted breeding, genetic diversity and genomic studies of Musa and related species.

BACKGROUND: Fusarium wilt, caused by the fungal pathogen Fusarium oxysporum f. sp. cubense (Foc), is one of the most destructive diseases of banana. Toxins produced by Foc have been proposed to play an important role during the pathogenic process. The objectives of this study were to investigate the contamination of banana with toxins produced by Foc, and to elucidate their role in pathogenesis. METHODOLOGY/PRINCIPAL FINDINGS: Twenty isolates of Foc representing races 1 and 4 were isolated from diseased bananas in five Chinese provinces. Two toxins were consistently associated with Foc, fusaric acid (FA) and beauvericin (BEA). Cytotoxicity of the two toxins on banana protoplast was determined using the Alamar Blue assay. The virulence of 20 Foc isolates was further tested by inoculating tissue culture banana plantlets, and the contents of toxins determined in banana roots, pseudostems and leaves. Virulence of Foc isolates correlated well with toxin deposition in the host plant. To determine the natural occurrence of the two toxins in banana plants with Fusarium wilt symptoms, samples were collected before harvest from the pseudostems, fruit and leaves from 10 Pisang Awak 'Guangfen #1' and 10 Cavendish 'Brazilian' plants. Fusaric acid and BEA were detected in all the tissues, including the fruits. CONCLUSIONS/SIGNFICANCE: The current study provides the first investigation of toxins produced by Foc in banana. The toxins produced by Foc, and their levels of contamination of banana fruits, however, were too low to be of concern to human and animal health. Rather, these toxins appear to contribute to the pathogenicity of the fungus during infection of banana plants.

There is cycling leisure in various landscapes where different management challenges emerge needing unique, context-specific strategies. Managers must be critical of these landscape nuances to produce suitable planning directives corresponding to the case of these certain environments. In this study, the Recreation Opportunity Spectrum (ROS) was adapted to the bike activity to create a Bicycle Opportunity Spectrum (BOS) framework. These cycling landscapes were processed into a regional-scale tool based on the compatibilities between landscape types and cyclist needs to produce satisfactory experiences. The development of the tool was based on diverse cycling experiences elaborated during focus group discussions (FGDs). This resulted in a framework of four landscape classifications (Urban, Rural, Natural with Road, and Natural with Trail) and criteria of seven (Bike Route, Path Remoteness, Infrastructure Development, Viewshed, Path Interactivity, Support Facilities and Services, and Pathway Management). The framework was tested by evaluating the cycling landscapes of Taichung City, Taiwan. The results, validated by the FGD participants verified the fitness of the BOS towards evaluating bikeways, creating a tool for bikeway management applicable to various landscape classes. [ABSTRACT FROM AUTHOR]

AIM: To evaluate the value of the ratio of axial length to corneal radius of curvature(AL/CR)in the diagnosis of myopia in children and adolescents.METHODS: The refraction and ocular biometric parameters of 2 182 cases of children and adolescents(4 364 eyes)who initially visited the optometry clinic of the Second People's Hospital of Beihai from January 2022 to December 2023 were collected and analyzed.RESULTS: The receiver operating characteristic curve(ROC curve)showed that AL/CR had a higher area under the curve(AUC=0.925, 95%CI: 0.917-0.933)in diagnosing myopia. When AL/CR was 3.053, the Youden index was the largest. As the threshold of AL/CR decreased, the sensitivity of myopia diagnosis increased, while the specificity decreased. Compared with screening myopia, the sensitivity of AL/CR=3.053 in diagnosing myopia was low, but the specificity was high, especially in the cases of hyperopia and pre-myopia.CONCLUSION: The accuracy of AL/CR in diagnosing myopia is superior to that of axial length and average corneal curvature, with higher specificity. As the threshold of AL/CR decreases, the sensitivity of diagnosing myopia increases, but the specificity decreases. In cases of hyperopia and pre-myopia, the specificity of AL/CR in diagnosing myopia is higher than that of screening myopia.