Further Characterization of Host Preference of Acidovorax citrulli Based on Growth Competition between Group I and Group II Strains

Acidovorax citrulli, the causal agent of bacterial fruit blotch, can be divided into two groups. Group I is mainly isolated from melon, and group II is mainly isolated from watermelon. In this study, group I and II strains were used to assess competition in vivo and in vitro by evaluating inhibition activity assays and by measuring population growth dynamics. Our results indicated that there is no mutual inhibition of growth between the two groups of strains in King’s B medium. The bacterial populations of A. citrulli strains were counted within 7 days after inoculation in melon and watermelon cotyledons and plotted against time to generate population growth curves. Area under the population growth curves was calculated. When the two groups of strains were inoculated separately into melon and watermelon cotyledons, the results of analysis of variance show that there was no significant difference. In this study, for the first time in an experimental setting, we inoculated two A. citrulli groups into melon and watermelon cotyledons at the same time and determined the population dynamics of each strain. The results showed that there was no significant difference between two group strains in melon cotyledons. However, in watermelon cotyledons, the area under population growth curves of group II strain were significantly higher than those of group I strain. Three-way analysis of variance results showed that there was interaction between host and grouping on the influence of strain population level (p < 0.0001). These data provide additional information on the host preference of different groups of A. citrulli

Essential Acidovorax citrulli Virulence Gene hrpE Activates Host Immune Response against Pathogen

Bacterial fruit blotch (BFB) caused by Acidovorax citrulli (Ac) is a devastating watermelon disease that severely impacts the global watermelon industry. Like other Gram-negative bacteria, the type three secretion system (T3SS) is the main pathogenicity factor of A. citrulli. The T3SS apparatus gene hrpE codes for the Hrp pilus and serves as a conduit to secret effector proteins into host cells. In this study, we found that the deletion of hrpE in A. citrulli results in the loss of pathogenicity on hosts and the hypersensitive response on non-hosts. In addition, the A. citrulli hrpE mutant showed a reduction in in vitro growth, in planta colonization, swimming and twitching motility, and displayed increases in biofilm formation ability compared to the wild type. However, when HrpE was transiently expressed in hosts, the defense responses, including reactive oxygen species bursts, callose deposition, and expression of defense-related genes, were activated. Thus, the A. Citrulli growth in HrpE-pretreated hosts was suppressed. These results indicated that HrpE is essential for A. citrulli virulence but can also be used by hosts to help resist A. citrulli. Our findings provide a better understanding of the T3SS pathogenesis in A. citrulli, thus providing a molecular basis for biopesticide development, and facilitating the effective control of BFB

BSCL2/Seipin deficiency in hearts causes cardiac energy deficit and dysfunction via inducing excessive lipid catabolism

Abstract Background Heart failure (HF) is one of the leading causes of death worldwide and is associated with cardiac metabolic perturbations. Human Type 2 Berardinelli‐Seip Congenital Lipodystrophy (BSCL2) disease is caused by mutations in the BSCL2 gene. Global lipodystrophic Bscl2−/− mice exhibit hypertrophic cardiomyopathy with reduced cardiac steatosis. Whether BSCL2 plays a direct role in regulating cardiac substrate metabolism and/or contractile function remains unknown. Methods We generated mice with cardiomyocyte‐specific deletion of Bscl2 (Bscl2cKO) and studied their cardiac substrate utilisation, bioenergetics, lipidomics and contractile function under baseline or after either a treatment regimen using fatty acid oxidation (FAO) inhibitor trimetazidine (TMZ) or a prevention regimen with high‐fat diet (HFD) feeding. Mice with partial ATGL deletion and cardiac‐specific deletion of Bscl2 were also generated followed by cardiac phenotyping. Results Different from hypertrophic cardiomyopathy in Bscl2−/− mice, mice with cardiac‐specific deletion of Bscl2 developed systolic dysfunction with dilation. Myocardial BSCL2 deletion led to elevated ATGL expression and FAO along with reduced cardiac lipid contents. Cardiac dysfunction in Bscl2cKO mice was independent of mitochondrial dysfunction and oxidative stress, but associated with decreased metabolic reserve and ATP levels. Importantly, cardiac dysfunction in Bscl2cKO mice could be partially reversed by FAO inhibitor TMZ, or prevented by genetic abolishment of one ATGL allele or HFD feeding. Lipidomic analysis further identified markedly reduced glycerolipids, glycerophospholipids, NEFA and acylcarnitines in Bscl2cKO hearts, which were partially normalised by TMZ or HFD. Conclusions We identified a new form of cardiac dysfunction with excessive lipid utilisation which ultimately causes cardiac substrate depletion and bioenergetics failure. Our findings also uncover a crucial role of BSCL2 in controlling cardiac lipid catabolism and contractile function and provide novel insights into metabolically treating energy‐starved HF using FAO inhibitor or HFD

The effects of vermicompost and shell powder addition on Cd bioavailability, enzyme activity and bacterial community in Cd-contaminated soil: A field study

Cadmium (Cd) contamination has become serious in soil and in situ stabilization technology has been widely used for heavy metal remediation. A field study was conducted to determine the effect of amendments with the doses of 3 kg/m2, including single vermicompost (A1), a 95% vermicompost mixed with 5% shell powder composite (A2) and a 95% vermicompost mixed with 5% modified shell powder composite (A3), on the Cd bioavailability, enzyme activity and bacterial community in soil, and the experiment was conducted with lettuce (Lactuca sativa L.) grown in a Cd-contaminated farmland soil. The results showed that the application of amendments increased the pH, cation exchange capacity (CEC), organic matter (OM), available nutrients, catalase (S-CAT), invertase (S-SC) and urease (S-UE) activities in soil, while significantly reduced the Cd bioavailability with the lowest Cd bioavailability being observed in the soil with A3 application. The soil bacterial richness and diversity increased after amendments application, and the bacterial community was characterized by an increase in metal-tolerant bacteria but a decrease in Proteobacteria, Acidobacteria and Gemmatimonadetes. In addition, the application of amendments significantly improved the growth of lettuce (Lactuca sativa L.) and inhibited Cd accumulation in its edible parts, especially, the Cd content in lettuce (Lactuca sativa L.) grown in soil with A3 application was below the limit of the National Food Safety Standard of China (maximum level ≤ 0.2 mg/kg). Thus, composite amendment obtained from vermicompost mixed with modified shell powder can be used as potential remediation material in Cd-contaminated soil. Capsule: Composite amendment obtained from vermicompost and modified shell powder had good effects on remediation of Cd-contaminated soil

Further Characterization of Host Preference of <i>Acidovorax citrulli</i> Based on Growth Competition between Group I and Group II Strains

Acidovorax citrulli, the causal agent of bacterial fruit blotch, can be divided into two groups. Group I is mainly isolated from melon, and group II is mainly isolated from watermelon. In this study, group I and II strains were used to assess competition in vivo and in vitro by evaluating inhibition activity assays and by measuring population growth dynamics. Our results indicated that there is no mutual inhibition of growth between the two groups of strains in King’s B medium. The bacterial populations of A. citrulli strains were counted within 7 days after inoculation in melon and watermelon cotyledons and plotted against time to generate population growth curves. Area under the population growth curves was calculated. When the two groups of strains were inoculated separately into melon and watermelon cotyledons, the results of analysis of variance show that there was no significant difference. In this study, for the first time in an experimental setting, we inoculated two A. citrulli groups into melon and watermelon cotyledons at the same time and determined the population dynamics of each strain. The results showed that there was no significant difference between two group strains in melon cotyledons. However, in watermelon cotyledons, the area under population growth curves of group II strain were significantly higher than those of group I strain. Three-way analysis of variance results showed that there was interaction between host and grouping on the influence of strain population level (p A. citrulli

Hcp of the Type VI Secretion System (T6SS) in <i>Acidovorax citrulli</i> Group II Strain Aac5 Has a Dual Role as a Core Structural Protein and an Effector Protein in Colonization, Growth Ability, Competition, Biofilm Formation, and Ferric Iron Absorption

A type VI secretion system (T6SS) gene cluster has been reported in Acidovorax citrulli. Research on the activation conditions, functions, and the interactions between key elements in A. citrulli T6SS is lacking. Hcp (Hemolysin co-regulated protein) is both a structural protein and a secretion protein of T6SS, which makes it a special element. The aims of this study were to determine the role of Hcp and its activated conditions to reveal the functions of T6SS. In virulence and colonization assays of hcp deletion mutant strain Δhcp, tssm (type VI secretion system membrane subunit) deletion mutant strain Δtssm and double mutant ΔhcpΔtssm, population growth was affected but not virulence after injection of cotyledons and seed-to-seedling transmission on watermelon. The population growth of Δhcp and Δtssm were lower than A. citrulli wild type strain Aac5 of A. citrulli group II at early stage but higher at a later stage. Deletion of hcp also affected growth ability in different culture media, and the decline stage of Δhcp was delayed in KB medium. Biofilm formation ability of Δhcp, Δtssm and ΔhcpΔtssm was lower than Aac5 with competition by prey bacteria but higher in KB and M9-Fe3+ medium. Deletion of hcp reduced the competition and survival ability of Aac5. Based on the results of Western blotting and qRT-PCR analyses, Hcp is activated by cell density, competition, ferric irons, and the host plant. The expression levels of genes related to bacterial secretion systems, protein export, and several other pathways, were significantly changed in the Δhcp mutant compared to Aac5 when T6SS was activated at high cell density. Based on transcriptome data, we found that a few candidate effectors need further identification. The phenotypes, activated conditions and transcriptome data all supported the conclusion that although there is only one T6SS gene cluster present in the A. citrulli group II strain Aac5, it related to multiple biological processes, including colonization, growth ability, competition and biofilm formation

Long-Term Field Biochar Application for Rice Production: Effects on Soil Nutrient Supply, Carbon Sequestration, Crop Yield and Grain Minerals

Research is still under-represented for the long-term impacts of field biochar application on soil fertility, rice production and mineral nutrition and soil C sequestration. To investigate these effects, a field trial was established in the fall of 2010 with low (2.4 t ha&minus;1), intermediate (6 t ha&minus;1) and high (12 t ha&minus;1) biochar doses. The biochar effect on soil nutrients is more pronounced in the surface soil and is dose dependent. In the surface soil, biochar application increased the availability of soil N, P and K. High dose decreased soil total Fe by 11.6% and Mn by 20.7%, leading to a slight but insignificant decrease in grain Fe and Mn concentration. The intermediate and high doses significantly increased the soil available Zn by 96.2% and 227.9% but did not affect the grain Zn concentration. Grain Cu concentration was significantly decreased by the intermediate (&minus;19.6%) and high (&minus;14.3%) dose of biochar. Biochar had a slight but insignificant boost on rice yield. Soil total C and N increased linearly with biochar application for the surface soil only where biochar was applied. In conclusion, with great soil C sequestration capacity, the long-term field return of biochar is beneficial for improving the soil macronutrient and Zn fertility, but caution is needed, as higher doses of biochar could potentially lead to a decrease in soil Fe and Mn and a reduction in grain Cu concentration

Long-Term Field Biochar Application for Rice Production: Effects on Soil Nutrient Supply, Carbon Sequestration, Crop Yield and Grain Minerals

Research is still under-represented for the long-term impacts of field biochar application on soil fertility, rice production and mineral nutrition and soil C sequestration. To investigate these effects, a field trial was established in the fall of 2010 with low (2.4 t ha−1), intermediate (6 t ha−1) and high (12 t ha−1) biochar doses. The biochar effect on soil nutrients is more pronounced in the surface soil and is dose dependent. In the surface soil, biochar application increased the availability of soil N, P and K. High dose decreased soil total Fe by 11.6% and Mn by 20.7%, leading to a slight but insignificant decrease in grain Fe and Mn concentration. The intermediate and high doses significantly increased the soil available Zn by 96.2% and 227.9% but did not affect the grain Zn concentration. Grain Cu concentration was significantly decreased by the intermediate (−19.6%) and high (−14.3%) dose of biochar. Biochar had a slight but insignificant boost on rice yield. Soil total C and N increased linearly with biochar application for the surface soil only where biochar was applied. In conclusion, with great soil C sequestration capacity, the long-term field return of biochar is beneficial for improving the soil macronutrient and Zn fertility, but caution is needed, as higher doses of biochar could potentially lead to a decrease in soil Fe and Mn and a reduction in grain Cu concentration

<i>Acidovorax citrulli</i> Effector AopV Suppresses Plant Immunity and Interacts with Aromatic Dehydratase ADT6 in Watermelon

Bacterial fruit blotch (BFB) is a disease of cucurbit plants caused by Acidovorax citrulli. Although A. citrulli has great destructive potential, the molecular mechanisms of pathogenicity of A. citrulli are not clear, particularly with regard to its type III secreted effectors. In this study, we characterized the type III secreted effector protein, AopV, from A. citrulli strain Aac5. We show that AopV significantly inhibits reactive oxygen species and the expression of PTI marker genes, and helps the growth of Pseudomonas syringae D36E in Nicotiana benthamiana. In addition, we found that the aromatic dehydratase ADT6 from watermelon was a target of AopV. AopV interacts with ADT6 in vivo and in vitro. Subcellular localization indicated ADT6 and AopV were co-located at the cell membrane. Together, our results reveal that AopV suppresses plant immunity and targets ADT6 in the cell membrane. These findings provide an new characterization of the molecular interaction of A. citrulli effector protein AopV with host cells