Transcriptome and Gene Co-Expression Network Analysis Identifying Differentially Expressed Genes and Signal Pathways Involved in the Height Development of Banana (Musa spp.).

Plant height is an important and valuable agronomic trait associated with yield and resistance to abiotic and biotic stresses. Dwarfism has positive effects on plant development and field management, especially for tall monocotyledon banana (Musa spp.). However, several key genes and their regulation mechanism of controlling plant height during banana development are unclear. In the present study, the popular cultivar 'Brazilian banana' ('BX') and its dwarf mutant ('RK') were selected to identify plant height-related genes by comparing the phenotypic and transcriptomic data. Banana seedlings with 3-4 leaves were planted in the greenhouse and field. We found that the third and fourth weeks are the key period of plant height development of the selected cultivars. A total of 4563 and 10507 differentially expressed genes (DEGs) were identified in the third and fourth weeks, respectively. Twenty modules were produced by the weighted gene co-expression network analysis (WGCNA). Eight modules were positively correlated with the plant height, and twelve other modules were negatively correlated. Combining with the analysis of DEGs and WGCNA, 13 genes in the signaling pathway of gibberellic acid (GA) and 7 genes in the signaling pathway of indole acetic acid (IAA) were identified. Hub genes related to plant height development were obtained in light of the significantly different expression levels (|log2FC| ≥ 1) at the critical stages. Moreover, GA3 treatment significantly induced the transcription expressions of the selected candidate genes, suggesting that GA signaling could play a key role in plant height development of banana. It provides an important gene resource for the regulation mechanism of banana plant development and assisted breeding of ideal plant architecture.

First report of Neopestalotiopsis clavispora causing leaf spot disease on banana (Musa acuminate L.) in China.

Banana (Musa acuminate L.) is an important tropical fruit in China. In October 2020, a new leaf spot disease was observed on banana plants at an orchard of Zhenkang county (23°45'23.46″ N, 98°48'46.52″ E), Lincang city, Yunnan province, China. The disease incidence was about 1%. The leaf spots occurred sporadically and the percentage of the leaf area covered by lesions was less than 5%. Symptoms on the leaves were initially small, irregular, reddish-brown spots that gradually expanded to fusiform-shaped lesions with a yellow halo and eventually become necrotic, dry, and cracked. To isolate the pathogen, thirty symptomatic leaves (15 mm2) from five plants were surface disinfected in 70% ethanol (10 s) and 0.8% NaClO (2 min), rinsed in sterile water three times, and transferred to potato dextrose agar (PDA) at 28°C for 5 days. Twenty-five colonies formed on the PDA plates were white with cottony aerial mycelium, round with a light orange underside. Abundant black globular acervuli semi-immersed on PDA were observed after a week. Conidia were straight or slightly curved, clavate to spindle, five cells, four septa with dimensions of 17.49 to 34.51 µm × 4.24 to 7.28 µm (avg. 23.83 × 5.62 µm; n=50). The apical and basal cells were hyaline, whereas the three median cells were dark brown. Conidia had a single basal appendage with lengths of 2.95 to 17.7 µm (avg. 7.18 µm; n=50) and two to three apical appendages with lengths of 10.7 to 53.84 µm (avg. 17.36 µm; n=50). These morphological characteristics are consistent with those of Neopestalotiopsis spp. (Maharachchikumbura et al. 2014). To confirm species, single-spore cultures of two representative isolates CATAS-102001 and CATAS-102002 were selected for further identification. The internal transcribed spacer (ITS) region, translation elongation factor 1-α (TEF1-α) and ß-tubulin (TUB2) genes of the two isolates were amplified with primers ITS1/ITS4 (White et al. 1990), EF1-728/EF2 (O'Donnell et al. 1998; Carbone and Kohn, 1999) and T1/Bt2b (Glass and Donaldson, 1995; O'Donnell and Cigelnik, 1997), respectively, and sequenced. The sequences were deposited in GenBank (ITS: OM281005 and OM281006; TEF1-α: OM328820 and OM328821; TUB2: OM328818 and OM328819). A maximum likelihood phylogenetic tree was constructed using the MEGA 7.0 (Kumar et al. 2016) based on the concatenated sequences ITS region, EF1-α and TUB2 gene, and the cluster analysis placed the representative isolates CATAS-102001 and CATAS-102002 within a clade comprising Neopestalotiopsis clavispora. The pathogenicity of two isolates was conducted on six 7-leaf-old banana seedlings. Two leaves from each potted plants were stab inoculated by puncturing into 1-mm using a sterilized needle, and stabbing three points at both sides of leaf midrib, and then placing 10 µl conidial suspension (1×106 conidia/ml) on one side of wounded points and the other side of wounded points were inoculated with sterile water as control. Inoculated plants were kept inside a plastic bag for 72 h and maintained in the greenhouse (12 h/12 h light/dark, 28°C, 90% relative humidity). The experiments were repeated twice. Irregular necrotic lesions on inoculated leaves appeared 7 days after inoculation, whereas controls were asymptomatic. The fungus was recovered from inoculated leaves, and its taxonomy was confirmed morphologically and molecularly, fulfilling Koch's postulates. Neopestalotiopsis clavispora has been reported to cause leaf spot on Mangifera indica (Shu et al. 2020), Macadamia integrifolia (Santos et al. 2019) and Ligustrum lucidum (Chen et al. 2020). To our knowledge, this is the first report of N. clavispora on banana in China. The identification of N. clavispora as the causal agent of the observed leaf spot disease on banana is critical to the prevention and control of this disease in the future.

The mediating effect of pain on the association between multimorbidity and disability and impaired physical performance among community-dwelling older adults in southern China.

AIM: To investigate the association between multimorbidity and disability and impaired physical performance, and to further evaluate the mediating effect of physical pain in this association. METHODS: 1321 community-dwelling older adults, who were over 60 years old in southern China, were regarded as participants in this cross-sectional study. Subjects completed a multi-instrument questionnaire including essential characteristics and physical function assessments. Physical function was assessed by activities of daily living (ADL), instrumental activities of daily living (IADL), index of mobility scale (NAGI), index of basic physical activities scale (RB), and short physical performance battery (SPPB). Multimorbidity was defined as the simultaneous presence of two or more chronic conditions. Multivariable regression and mediation analyses were conducted and gender differences were explored. RESULTS: The prevalence of multimorbidity was 44.6% in our study. In gender stratification analysis, multimorbidity was significantly associated with ADL disability (OR = 2.16), IADL disability (OR = 1.97), NAGI disability (OR = 2.84), RB disability (OR = 2.65) and lower SPPB score (ß = - 0.83) in women. The rate of pain increased with the number of chronic diseases and the multimorbidity patients with higher pain prevalence. Moreover, the presence of pain was also significantly associated with disability and impaired physical performance. Mediation analysis illustrated that pain was accounted for 16.5% to 22.1% of the adverse effects of multimorbidity on disability and impaired physical performance in women. CONCLUSIONS: Multimorbidity was significantly associated with disability and impaired physical performance, and pain might be a mediating factor for adverse effects of multimorbidity on disability and impaired physical performance in women.

Association of handgrip strength with hypertension among middle-aged and elderly people in Southern China: A cross-sectional study.

Backgroud and Purpose: Hypertension has been regarded as one of the most common chronic diseases reported in different studies, and handgrip strength is a good indicatorof anindividual's overall health. However, few studies have concentrated on investigating the relationship between hypertension and handgrip strength, especially for the middle-aged and elderly population in the community. Therefore, the purpose of this study was to explore the association of handgrip strength with the risk of hypertension.Methods: A cross-sectional study was conducted using a multi-instrument questionnaire. A total of 1152 participants aged 45 and older were included in this study. Handgrip strength, social-demographiccharacteristics, behavioral lifestyle and health-related variables were collected. Binary logistic regression was employed to analyse the relationship.Results: Handgrip strength was positively related to the risk of hypertension. Binary logistic regression models revealed that the increase of handgrip strength was significantly associated with the reduction of hypertension risk in female after adjusting forsocial-demographic characteristics, behavioral lifestyle and health-related variables (OR [95%CI] =0.265 [0.089-0.787]). In addition, after stratifying by age groups, the significant association was still existing in 60-74 years and ≥75 years of female groups, respectively(OR [95%CI] =0.158 [0.032-0.779]; (OR [95%CI] =0.009 [0.000-0.409]). No significant associations were observed in male after adjusting variables.Conclusion: stronger handgrip strength was association with the lower risk ofhypertension for the elderly female population.Abbreviations: BMI: body mass index; DBP: diastolic blood pressure; HC: hip circumference; SBP, systolic blood pressure; WC: waist circumference; WHC: hip-waist relation.

Interaction of lipid accumulation product and family history of hypertension on hypertension risk: a cross-sectional study in the Southern Chinese population.

OBJECTIVES: This study aimed at investigating the applicability of a novel index based on waist circumference (WC) and triglyceride (TG) which was named lipid accumulation product (LAP) in the Southern Chinese population, and compared the predictive effects of LAP and other obesity indicators on hypertension risk. Moreover, this study investigated the interactive effects of LAP and family history of hypertension. METHODS: A total number of 2079 of community-dwelling adults in Southern China were enrolled in this cross-sectional study. The participants underwent questionnaire surveys, anthropometric tests and laboratory examinations. Themultinomial logistic regression model and receiver operating characteristic curves, including LAP, body mass index (BMI), waist-to-hip ratio (WHR), WC and TG, were used to assess the association between hypertension risk and obesity indexes. The interaction effects were evaluated by relative excess risk of interaction (RERI), attributable proportion due to interaction (AP) and synergy index (SI). RESULTS: Higher LAP levels have a relatively higher risk of having hypertension in both sexes (males: adjusted OR=2.79 per SD increase, 95% CI 1.43 to 5.44, p<0.001; females: adjusted OR=3.15, 95% CI 1.56 to 6.39, p<0.001). LAP (area under the curve=0.721; 95% CI 0.680 to 0.761) is a better indicator in identifying hypertension risk than BMI, WHR and TG in females, but WC performed better in males. A significant interaction between LAP and family history of hypertension was observed in males (RERI=1.652, 95% CI 0.267 to 3.037; AP=0.516, 95% CI 0.238 to 0.794; SI=3.998, 95% CI 0.897 to 17.820), but there is no statistically significant difference in females. CONCLUSIONS: LAP significantly associates with hypertension risk in the Southern Chinese population. It has better performance than BMI, WHR and TG on predicting hypertension risk of the Southern Chinese female population. Moreover, LAP and family history of hypertension might synergistically increase the risk of hypertension.

The association between multimorbidity and health-related quality of life: a cross-sectional survey among community middle-aged and elderly residents in southern China.

BACKGROUND: Multimorbidity is common among the middle-aged and elderly residents. And it is associated to the reduction of health-related quality of life (HRQoL), including physical and psychological dimensions. However, there are few studies that have paid attention to the HRQoL of residents with multimorbidity in China. Therefore, this study aims to investigate the relationships between different multimorbidity patterns and HRQoL among middle-aged and elderly adults in China. METHODS: Based on a cross-sectional survey, the information regarding 18,137 adults, who were at least 45 years of age, was collected through interviews. Self-perceived HRQoL was assessed with the EQ-5D-3 L instrument, and the EQ-5D-3 L index score was calculated using the Chinese EQ-5D-3 L value set. The Tobit regression was used to explore the impacts of multimorbidity groups on HRQoL. RESULTS: Of 18,137 respondents, more than a fifth (3773,20.8%) of people had multimorbidity. Mean (SD) of EQ-5D index and VAS values were 0.95(0.14) and 76.02(13.66), respectively. Significant correlations were found between a lower HRQoL and increasing numbers of chronic conditions (P < 0.001). Most of chronic diseases co-occurred frequently, and the association between hypertension and diabetes mellitus was the strongest (adjusted OR = 3.82). The most prevalent disease is hypertension (5052,27.9%), and the most prevalent chronic diseases pair is hypertension and diabetes mellitus (841,4.6%). Among those chronic diseases with high prevalence, the effects on HRQoL ranged from chronic pain to hypertension (adjust b = - 0.036 to - 0.008). In the common multimorbidity patterns, co-occurrence of chronic pain and bone disease (adjust b = - 0.039) had the greatest impact on HRQoL. CONCLUSIONS: The HRQoL of middle-aged and elderly adults declines by multimorbidity. More attention should be paid to the HRQoL of residents with multimorbidity in China. The effect of different multimorbidity patterns on HRQoL is not simply added by two diseases, but changes by the different combination. Identifying different multimorbidity patterns of residents can provide more targeted measures to improve the HRQoL.

Essential Oil Extracted from Cymbopogon citronella Leaves by Supercritical Carbon Dioxide: Antioxidant and Antimicrobial Activities.

To improve essential oil quality, especially to reserve the thermal instability of compounds, supercritical CO2 extraction (SFE) was applied to recover essential oil from Cymbopogon citronella leaves. A response surface methodology was applied to optimize the extraction process. The highest essential oil yield was predicted at extraction time 120 min, extraction pressure 25 MPa, extraction temperature 35°C, and CO2 flow 18 L/h for the SFE processing. Under these experimental conditions, the mean essential oil yield is 4.40%. In addition, the chemical compositions of SFE were compared with those obtained by hydrodistillation extraction (HD). There were 41 compounds obtained of SFE, while 35 compounds of HD. Alcohols and aldehydes were the main compositions in the essential oils. Furthermore, the antioxidant activities and antimicrobial of essential oils obtained by HD and the evaluated condition of SFE were compared. Results showed that the antioxidant activities of SFE oil are better than those of HD. Minimum inhibitory concentrations (MICs) were determined by the microdilution method. Essential oil obtained from SFE and HD exhibited a significant antimicrobial activity against all tested microorganisms. It is confirmed that the SFE method can be an alternative processing method to extract essential oils from Cymbopogon citronella leaves.

Associations between objectively assessed physical fitness levels and sleep quality in community-dwelling elderly people in South China.

PURPOSE: The aim of this study was to explore associations between objectively assessed physical fitness levels and sleep quality in community-dwelling elderly people in South China. METHODS: One thousand one hundred thirty-six (504 males and 632 females) community-dwelling adults aged ≥ 50 years old in Dongguan City, South China, were included in the cross-sectional study. All the participants were asked to complete all prepared multi-instrument questionnaire, including the Pittsburgh Sleep Quality Index (Chinese version), for the assessment of the sleep quality and information regarding socio-demographic characteristics, lifestyle, and physical health data. Physical fitness was measured by grip strength, one-leg standing test (OLST) with eyes open, back scratch test, and the forced vital capacity (FVC). RESULTS: The percentage of poor sleep quality among elderly people (≥ 50 years old) was up to 18.2%. Lower FVC was associated with the poorer sleep quality (adjusted OR = 0.74 per SD increase; P = 0.009), and participants with lower performance in back scratch test were more likely to suffer poor sleep quality (adjusted OR = 1.17 per SD increase; P = 0.035). The independent contribution of physical fitness tests results on the risk of poor sleep quality was 22.1%. CONCLUSIONS: Our results indicated that sleep quality was strongly associated with physical fitness among community-dwelling elderly people; the lower of the physical fitness predicted poorer sleep quality.

The characterization of 1-(4-bromophenyl)-5-phenyl-1H-1,2,3-triazole on acute toxicity, antimicrobial activities, photophysical property, and binding to two globular proteins.

1-(4-Bromophenyl)-5-phenyl-1H-1,2,3-triazole (BPT) was a newly synthesized compound. The acute toxicities of BPT to mice by intragastric administration have been determined and the result indicates that the intragastric administration of BPT did not produce any significant toxic effect on Kunming strain mice. It is also evaluated for the antimicrobial activity of BPT against three kinds of plant mycoplasma, Fusarium Wilt (race 4), Colletotrichum gloeosporioides Penz. and Xanthomonas oryzae by different method in vitro. The compound exhibited distinct inhibitory activities against Fusarium Wilt (race 4) and Colletotrichum gloeosporioides Penz. by mycelium growth rate test and the values of EC50 were 29.34 and 12.53µg/mL respectively. And BPT had also the most potent inhibitory activities against Xanthomonas oryzae when compared with that of control drugs by the agar well diffusion method. In addition, the structural and photophysical properties of BPT including ionization energy, electron affinities, and theoretical spectrum was studied by quantum-chemical methods. Then the interaction of BPT with two kinds of globular proteins, human immunoglobulin (HIg) and bovine hemoglobin (BHg) was investigated by using UV-vis absorption spectra, synchronous fluorescence, 3D fluorescence spectra, and fluorescence titration in combination with molecular modeling. UV-vis absorption, 3D and synchronous fluorescence measurements show that BPT has influence on the microenvironment surrounding HIg or BHg in aqueous solution and the fluorescence experiments show that BPT quenches the fluorescence intensity of HIg or BHg through a static mechanism. The binding parameters including the binding constants, the number of binding site and average binding distance between BPT and HIg or BHg at different temperatures were calculated. The thermodynamic parameters suggest that the hydrophobic interaction is the predominant intermolecular forces in stabilizing the BPT-HIg or BPT-BHg complex. Molecular docking was performed to reveal that the BPT moiety binds to the hydrophobic cavity of HIg or BHg and they are in good agreement with the spectroscopic measurements.

[Genetic diversity analysis of Fusarium oxysporum f. sp. cubense populations from China using Inter-Simple Sequence Repeats-PCR (ISSR-PCR) technique].

OBJECTIVE: We used Inter-Simple Sequence Repeats (ISSR) markers to reveal the genetic diversity of 95 Fusarium oxysporum f. sp. cubense ( FOC ) isolates from banana in China, for the rational control of the disease. METHODS: Eight primers were chosen for analyzing FOC isolates to study their genetic diversity by ISSR-PCR. All isolates were clustered using Unweighted Pair-Group Method with Arithmetic means (UPGMA) analysis by NTSYSpc v2.10e software. RESULTS: A total of 52 sites were generated, among them 92.3% were polymorphic. Genetic distance was 0.57 to 1.00 based on the Nei's standard. Isolates were grouped into six distinct clusters (A, B, C, D, E and F) based on ISSR analysis using a genetic distance threshold of 0.68, the proportion of 51.06%, 39.58%, 5.20%, 2.08%, 1.04%, and 1.04%, respectively. CONCLUSION: There were high levels of genetic variation among the FOC isolates, and the ISSR clustering groups had obvious correlation with hosts and races of the pathogen.

Rapid and quantitative detection of Fusarium oxysporum f. sp. cubense race 4 in soil by real-time fluorescence loop-mediated isothermal amplification.

AIMS: In this study, a real-time fluorescence loop-mediated isothermal amplification (RealAmp) was developed and evaluated for the rapid and quantitative detection of Fusarium oxysporum f. sp. cubense race 4 (R4) in soil. METHODS AND RESULTS: The LAMP primer set was designed based on previously verified RAPD marker sequences, and the RealAmp assay could specifically detect and distinguish R4 isolates from other related species. The detection sensitivity of the RealAmp assay was approx. 3·82 × 10(3) copies of plasmid DNA or 10(3) of spores per gram in artificially infested soil, indicating that the method is highly tolerant to inhibitor substances in soil compared to real-time PCR. Combining previously published TR4-specific detection methods with the newly established R4-specific RealAmp assay, an indirect approach to detect and differentiate ST4 isolates was achieved by comparing the detection results of R4 and TR4 simultaneously. The existence of ST4 isolates in China was subsequently confirmed through the developed approach. CONCLUSION: The developed RealAmp assay has been confirmed to be a simple, rapid and effective method to detect R4 in soil, which facilitates to further identify and distinguish ST4 isolates through the comparative analysis of detection results between TR4 and R4 simultaneously. SIGNIFICANCE AND IMPACT OF THE STUDY: The technique is an alternative quantitative detection method, which will be used for a routine detection service for the soil-borne pathogen in China.

Development of a real-time fluorescence loop-mediated isothermal amplification assay for rapid and quantitative detection of Fusarium oxysporum f. sp. cubense tropical race 4 in soil.

Fusarium oxysporum f. sp. cubense (Foc), the causal agent of Fusarium wilt (Panama disease), is one of the most devastating diseases of banana (Musa spp.). The Foc tropical race 4 (TR4) is currently known as a major concern in global banana production. No effective resistance is known in Musa to Foc, and no effective measures for controlling Foc once banana plants have been infected in place. Early and accurate detection of Foc TR4 is essential to protect banana industry and guide banana planting. A real-time fluorescence loop-mediated isothermal amplification assay (RealAmp) was developed for the rapid and quantitative detection of Foc TR4 in soil. The detection limit of the RealAmp assay was approximately 0.4 pg/µl plasmid DNA when mixed with extracted soil DNA or 10(3) spores/g of artificial infested soil, and no cross-reaction with other relative pathogens were observed. The RealAmp assay for quantifying genomic DNA of TR4 was confirmed by testing both artificially and naturally infested samples. Quantification of the soil-borne pathogen DNA of Foc TR4 in naturally infested samples was no significant difference compared to classic real-time PCR (P>0.05). Additionally, RealAmp assay was visual with an improved closed-tube visual detection system by adding SYBR Green I fluorescent dye to the inside of the lid prior to amplification, which avoided the inhibitory effects of the stain on DNA amplification and makes the assay more convenient in the field and could thus become a simple, rapid and effective technique that has potential as an alternative tool for the detection and monitoring of Foc TR4 in field, which would be a routine DNA-based testing service for the soil-borne pathogen in South China.

Molecular and pathogenic variation identified among isolates of Corynespora cassiicola.

Pathogenicity tests and inter simple sequence repeat (ISSR) molecular fingerprinting markers were utilized to analyze 24 Corynespora cassiicola isolates obtained from a lot of Hevea clones grown in most rubber nurseries and a few plantations in China. The C. cassiicola isolates were collected from Hainan and Yunnan provinces, China, from 2006 to 2008. The assay of 24 C. cassiicola isolates on detached leaves of four different Hevea rubber clones (genotypes PR 107, Dafeng 95, RRIM 600, and Reyan 7-33-97) indicated that 23 of the isolates were susceptible to RRIM 600, and were therefore considered race 1 except for CC-023. ISSR analysis grouped 24 C. cassiicola isolates into four clusters (A, B, C, and D). Unweighted pair-group method with arithmetic averaging (UPGMA) analysis based on Nei and Li's coefficient (calculated from the binary matrix data of 103 DNA fragments generated from 16 ISSR primers) indicated that cluster A included 19 isolates from Hainan and Yunnan (this cluster was further divided into two sub clusters (I, II), sub cluster II contained isolate CC-023); clusters B and C comprised of 1 isolates from Hainan, respectively; while cluster D encompassed 3 isolates from Hainan and Yunnan. Pathogenicity tests and ISSR analysis showed that there was no correlation between race structure, the geographical origin of the pathogen and their ISSR clusters because 23 of the isolates belonging to four distinct clusters were considered race 1 except for isolate CC-023. However, most of the isolates with different pathogenicity levels shared the same clades, and furthermore, the ISSR clusters and cology color had an exact correlation. These results should facilitate the development of rubber clones with enhanced resistance against all genetic clusters of C. cassiicola.