Expression of ethylene biosynthetic and signaling genes in relation to ripening of banana fruit after cold storage

Banana fruit are highly sensitive to chilling injury (CI), while the effect of different degrees of CI on the subsequent fruit ripening is largely unknown. In the present work, ripening characteristic of banana fruit after storage at 7 °C for 3 days or for 8 days, and expression levels of eight genes associated with ethylene biosynthetic and signaling, including MaACS1, MaACO1, MaERS1, MaERS3, and MaEIL1–4, were investigated. The results showed that banana fruit stored at 7 °C for 8 days exhibited more severe chilling symptoms than those at 7 °C for 3 days. Compared with banana fruit stored at 7 °C for 8 days, which showed abnormal ripening, more decrease in fruit firmness, while higher increase in ethylene production and hue angle were observed in banana fruit stored at 7 °C for 3 days, which could ripening normally. Moreover, gene expression profiles during ripening revealed that ethylene biosynthetic and signaling genes were differentially expressed in peel and pulp of banana fruit after storage at 7 °C for 3 days and 7 °C for 8 days. In the peel of fruit storage at 7 °C for 3 days, expression levels of MaACS1, MaACO1, MaEIL1, and MaEIL2 increased remarkably while MaERS3, MaEIL1, and MaEIL4 were enhanced in the fruit after storage at 7 °C for 8 days. In the pulp, with the exception of MaACO1 and MaERS3, expression levels of other genes did not exhibit a significant difference, between the banana fruit storage at 7 °C for 3 days and 7 °C for 8 days. Taken together, our results suggest that differential expression of ethylene biosynthetic and signaling genes such as MaERS3, MaACO1, and MaEIL2, may be related to ripening behavior of banana fruit with different degrees of CI after cold storage.     

Ozone-induced inhibition of kiwifruit ripening is amplified by 1-methylcyclopropene and reversed by exogenous ethylene

Background

Understanding the mechanisms involved in climacteric fruit ripening is key to improve fruit harvest quality and postharvest performance. Kiwifruit (Actinidia deliciosa cv. ‘Hayward’) ripening involves a series of metabolic changes regulated by ethylene. Although 1-methylcyclopropene (1-MCP, inhibitor of ethylene action) or ozone (O₃) exposure suppresses ethylene-related kiwifruit ripening, how these molecules interact during ripening is unknown.

Results

Harvested ‘Hayward’ kiwifruits were treated with 1-MCP and exposed to ethylene-free cold storage (0?°C, RH 95%) with ambient atmosphere (control) or atmosphere enriched with O₃ (0.3?μL?L^??1) for up to 6?months. Their subsequent ripening performance at 20?°C (90% RH) was characterized. Treatment with either 1-MCP or O₃ inhibited endogenous ethylene biosynthesis and delayed fruit ripening at 20?°C. 1-MCP and O₃ in combination severely inhibited kiwifruit ripening, significantly extending fruit storage potential. To characterize ethylene sensitivity of kiwifruit following 1-MCP and O₃ treatments, fruit were exposed to exogenous ethylene (100?μL?L^??1, 24?h) upon transfer to 20?°C following 4 and 6?months of cold storage. Exogenous ethylene treatment restored ethylene biosynthesis in fruit previously exposed in an O₃-enriched atmosphere. Comparative proteomics analysis showed separate kiwifruit ripening responses, unraveled common 1-MCP- and O₃-dependent metabolic pathways and identified specific proteins associated with these different ripening behaviors. Protein components that were differentially expressed following exogenous ethylene exposure after 1-MCP or O₃ treatment were identified and their protein-protein interaction networks were determined. The expression of several kiwifruit ripening related genes, such as 1-aminocyclopropane-1-carboxylic acid oxidase (ACO1), ethylene receptor (ETR1), lipoxygenase (LOX1), geranylgeranyl diphosphate synthase (GGP1), and expansin (EXP2), was strongly affected by O₃, 1-MCP, their combination, and exogenously applied ethylene.

Conclusions

Our findings suggest that the combination of 1-MCP and O₃ functions as a robust repressive modulator of kiwifruit ripening and provide new insight into the metabolic events underlying ethylene-induced and ethylene-independent ripening outcomes.

     

Brassinosteroid signaling modulates submergence-induced hyponastic growth in <Emphasis Type="Italic">Arabidopsis thaliana</Emphasis>

The role of brassinosteroids (BRs) in hyponastic growth induced by submergence was investigated in Arabidopsis thaliana. Under flooding conditions, exogenously applied BRs increased hyponastic growth of rosette leaves. This hyponastic growth was reduced in a BR insensitive mutant (bri1-5), while it was increased in a BR dominant mutant (bes1-D). Further, expression of hypoxia marker genes, HRE1 and HRE2, was elevated in submerged bes1-D. These results indicate that BRs exert a positive action on hyponastic growth of submerged Arabidopsis leaves. Expression of ethylene biosynthetic genes, such as ACS6, ACS8 and ACO1, which are up-regulated by submergence, was also activated by application of BRs and in bes1-D. The enhanced hyponastic growth in submerged bes1-D was significantly reduced by application of cobalt ion, suggesting that BRs control hyponastic growth via ethylene, which seems to be synthesized by ACO6 and ACO8 followed by ACO1 in submerged leaves. A double mutant, bes1-Dxaco1-1, showed hyponastic growth activity similar to that seen in aco1-1, demonstrating that the BR signaling for regulation of hyponastic growth seems to be an upstream event in ethylene-induced hyponastic growth under submergence in Arabidopsis.     

Interaction of karrikinolide and ethylene in controlling germination of dormant <Emphasis Type="Italic">Avena fatua</Emphasis> L. caryopses

Caryopses of Avena fatua L. are dormant after harvest and germinate poorly at 20 °C. Dormancy was released by after-ripening the dry caryopses in the dark at 25 °C for 3 months. Karrikinolide (butenolide, 3-methyl-2H-furo[2,3-c]pyran-2-one, KAR₁), in contrast to exogenous ethylene and the precursor of ethylene biosynthesis 1-aminocyclopropane-1-carboxylic acid (ACC), completely overcame dormancy. The effect of KAR₁ was not affected by aminoethoxyvinylglycine (AVG), α-aminoisobutyric acid (AIB) and CoCl₂, inhibitors of ACC synthase and oxidase, respectively. 2,5-Norbornadiene (NBD), a reversible inhibitor of ethylene binding to its receptor, counteracted the stimulatory effect of KAR₁. Ethylene, ethephon and ACC counteracted and AVG reinforced inhibition caused by norbornadiene. Inhibition due to norbornadiene, applied during the first 3 days of imbibition in the presence of KAR₁, disappeared after transfer to air or ethylene. The obtained results confirm that KAR₁ breaks dormancy and indicate that ethylene alone plays no role in releasing dormancy of Avena fatua caryopses. KAR₁ probably did not relieve dormancy via the stimulation of ethylene biosynthesis. Some level of endogenous ethylene is probably required for ethylene action, which might be required for releasing dormancy by KAR₁ or for subsequent germination of caryopses after removing dormancy.     

Temperature-induced changes in treatment efficiency and microbial structure of aerobic granules treating landfill leachate

This paper investigates the effect of temperature on nitrogen and carbon removal by aerobic granules from landfill leachate with a high ammonium concentration and low concentration of biodegradable organics. The study was conducted in three stages; firstly the operating temperature of the batch reactor with aerobic granules was maintained at 29 °C, then at 25 °C, and finally at 20 °C. It was found that a gradual decrease in operational temperature allowed the nitrogen-converting community in the granules to acclimate, ensuring efficient nitrification even at ambient temperature (20 °C). Ammonium was fully removed from leachate regardless of the temperature, but higher operational temperatures resulted in higher ammonium removal rates [up to 44.2 mg/(L h) at 29 °C]. Lowering the operational temperature from 29 to 20 °C decreased nitrite accumulation in the GSBR cycle. The highest efficiency of total nitrogen removal was achieved at 25 °C (36.8 ± 10.9 %). The COD removal efficiency did not exceed 50 %. Granules constituted 77, 80 and 83 % of the biomass at 29, 25 and 20 °C, respectively. Ammonium was oxidized by both aerobic and anaerobic ammonium-oxidizing bacteria. Accumulibacter sp., Thauera sp., cultured Tetrasphaera PAO and Azoarcus–Thauera cluster occurred in granules independent of the temperature. Lower temperatures favored the occurrence of denitrifiers of Zooglea lineage (not Z. resiniphila), bacteria related to Comamonadaceae, Curvibacter sp., Azoarcus cluster, Rhodobacter sp., Roseobacter sp. and Acidovorax spp. At lower temperatures, the increased abundance of denitrifiers compensated for the lowered enzymatic activity of the biomass and ensured that nitrogen removal at 20 °C was similar to that at 25 °C and significantly higher than removal at 29 °C.     

A role of ETR1 in regulating leaf petiole elongation mediated by elevated temperature in <Emphasis Type="Italic">Arabidopsis</Emphasis>

Temperature fluctuation profoundly affects the plant growth and development. In this study, we show that ethylene receptor ETR1 is involved in regulating leaf petiole elongation mediated by higher temperatures (at 32 °C in this study). ETR1 loss-of-function mutant etr1-7 cannot elongate the leaf petiole at 32 °C as much as wild-type seedlings (WT). Overexpression of ETR1 in etr1-7 not only fully rescued the deficient in petiole elongation under higher temperature conditions but also caused longer petiole length under normal temperature conditions (22 °C). Plants with different mutant ETR1 alleles including etr1-7 etr1-1, and etr1-9 but not etr1-3 impair the petiole elongation mediated by elevated temperature. RNA-Seq analysis showed that hundreds of genes induced by elevated temperature in WT were not differentially expressed in etr1-7. Gene ontology enrichment analysis reveals that the molecular functions of these genes primarily relate to photosynthesis and protein degradation. Furthermore, genes involved in regulating organ elongation (such as BRI1-EMS-SUPPRESSOR 1, BES1), are significantly up-regulated in WT rather than in etr1-7 after the treatment of higher temperature. The results from this study suggest ETR1 is involved in regulating Arabidopsis response to elevated ambient temperature in both molecular and morphological levels.     

The eurythermal adaptivity and temperature tolerance of a newly isolated psychrotolerant Arctic <Emphasis Type="Italic">Chlorella</Emphasis> sp.

A new strain of Chlorella sp. (Chlorella-Arc), isolated from Arctic glacier melt water, was found to have high specific growth rates (μ) between 3 and 27 °C, with a maximum specific growth rate of 0.85 day^?1 at 15 °C, indicating that this strain was a eurythermal strain with a broad temperature tolerance range. To understand its acclimation strategies to low and high temperatures, the physiological and biochemical responses of the Chlorella-Arc to temperature were studied and compared with those of a temperate Chlorella pyrenoidosa strain (Chlorella-Temp). As indicated by declining F _v/F _m, photoinhibition occurred in Chlorella-Arc at low temperature. However, Chlorella-Arc reduced the size of the light-harvesting complex (LHC) to alleviate photoinhibition, as indicated by an increasing Chl a/b ratio with decreasing temperatures. Interestingly, Chlorella-Arc tended to secrete soluble sugar into the culture medium with increasing temperature, while its intracellular soluble sugar content did not vary with temperature changes, indicating that the algal cells might suffer from osmotic stress at high temperature, which could be adjusted by excretion of soluble sugar. Chlorella-Arc accumulated protein and lipids under lower temperatures (<15 °C), and its metabolism switched to synthesis of soluble sugar as temperatures rose. This reflects a flexible ability of Chlorella-Arc to regulate carbon and energy distribution when exposed to wide temperature shifts. More saturated fatty acids (SFA) in Chlorella-Arc than Chlorella-Temp also might serve as the energy source for growth in the cold and contribute to its cold tolerance.     

Properties of Probiotics <Emphasis Type="Italic">Kocuria</Emphasis> SM1 and <Emphasis Type="Italic">Rhodococcus</Emphasis> SM2 Isolated from Fish Guts

This study characterized probiotics Kocuria SM1 and Rhodococcus SM2, which were recovered from the intestinal microbiota of rainbow trout (Oncorhynchus mykiss, Walbaum). The cultures were Gram-positive, non-motile, catalase-positive and oxidase-negative cocci or rods. Cell multiplication of SM1 and SM2 was observed at 4–37 °C (45 °C for SM1), in 0–20% (w/v) NaCl and at pH 2–11. The viability was not affected when exposed to pepsin at pH 2.0 and 3.0, and pancreatin at pH 8.0. Neither isolates were chrome azurol S-positive for siderophore production. Of the 19 common enzymes analysed using the API-ZYM system, only 8 were evident in the culture of SM1 compared to 11 enzymes for SM2. The secondary metabolites of both probiotics were inhibitory to Acinetobacter baumannii, Vibrio anguillarum and V. ordalii; SM2 inhibited Escherichia coli, Klebsiella pneumoniae, Pseudomonas aeruginosa and Staphylococcus aureus. SM2 was resistant to penicillin and sulphatriad, out of six antimicrobial agents; SM1 was resistant to sulphatriad. These results suggest that Kocuria SM1 and Rhodococcus SM2 are able to grow over a wide range of temperature, salinity and pH, including in conditions that mimic the gastrointestinal environment of fish and produce extracellular enzymes that may have a role in the host digestive processes. Importantly, Rhodococcus SM2 displays a high degree of bacteriocinogenic potential against multi-drug-resistant human pathogens that have never been documented among the gut microbiota of fish.     

Effect of Temperature on Development and Reproduction of <Emphasis Type="Italic">Epilachna dodecastigma</Emphasis> (Wied.) (Coleoptera: Coccinellidae)

The effect of five constant temperatures of 21, 24, 27, 30 and 33 °C on adult life span, reproduction, oviposition behavior and larval developmental time of a bitter gourd inhabited coleopteran insect Epilachna dodecastigma (Wied.) (Coccinellidae) was determined in laboratory conditions under 70 ± 5 % relative humidity and a photoperiod of 12 L : 12 D. Larval developmental time of E. dodecastigma decreased as temperature increased from 21 to 33 °C. Life table data revealed that overall mortality was lowest at 27 °C and highest at 21 °C. Females lived longer than males at all temperatures; but longevity decreased with increase in temperature. Pre-oviposition period decreased significantly with increase in temperature up to 27 °C and thereafter increased at a slower rate; whereas oviposition period decreased significantly with increase in temperature. Fecundity and egg viability increased significantly with an increase in temperature up to 27 °C and thereafter decreased at a slower rate. The intrinsic rate of increase (r _m ) was 0.1703, 0.1984, 0.2235, 0.2227 and 0.2181 day^?1 at 21, 24, 27, 30 and 33 °C, respectively. The net reproductive rate and finite rate of increase was highest at 27 °C (R _o  = 112.05; λ = 1.4233) and lowest at 21 °C (R _o  = 51.23; λ = 1.2581).     

Jasmonic acid regulates the ascorbate–glutathione cycle in <Emphasis Type="Italic">Malus baccata</Emphasis> Borkh. roots under low root-zone temperature

Jasmonic acid (JA), which is an important phytohormone, plays a key role in plant growth, development and stress responses. Here, Malus baccata Borkh. seedlings were used to study the mechanism by which JA alleviates the oxidative damage induced by low root-zone temperature (5 °C) through regulating the ascorbate–glutathione (AsA–GSH) cycle. The roots of M. baccata Borkh. were subjected to three treatments [5 °C, 5 °C + JA, and 5 °C + ibuprofen (IBU)] for 0, 12, 24, and 48 h. The results showed that treatment with low root-zone temperature could modulate the non-enzymatic and enzymatic components of the AsA–GSH cycle, significantly inducing the accumulation of MDA and H₂O₂. Additionally, the endogenous JA content changed dramatically, and the expression levels of the related genes [lipoxygenase (LOX), allene oxide synthase (AOS), and allene oxide cyclase (AOC)] showed different trends. In plants pretreated with JA, the endogenous JA content increased at 24 h, and the gene expression levels of LOX, AOS, and AOC were upregulated. We also found a marked increase in the activities of antioxidant enzymes [ascorbate peroxidase (APX), monodehydroascorbate reductase (MDHAR), and glutathione reductase (GR)], a decrease in oxidized glutathione (GSSG) and an increased GSH/GSSG ratio, which resulted in lower MDA and H₂O₂ contents. Thus, the oxidative stress was alleviated. Plants pretreated with IBU experienced an opposite effect on the function of the AsA–GSH cycle and the gene expression in the JA synthesis route relative to those subjected to exogenous JA treatment, indicating that endogenous JA can alleviate oxidative damage by regulating the function of the AsA–GSH cycle under low root-zone temperature.     

Low temperature and <Emphasis Type="Italic">Daphnia</Emphasis>-associated infochemicals promote colony formation of <Emphasis Type="Italic">Scenedesmus obliquus</Emphasis> and its harvesting

Objective

To explore the combined effects of temperature and Daphnia-associated infochemicals on colony formation of Scenedesmus obliquus to faciliate harvesting the algal biomass.

Results

A three-parameter modified Gaussian model fitted the changes of the number of cells per particle in S. obliquus induced by Daphnia culture filtrate well under any temperature. Decreases in temperature enhanced the induced–colony formation of Scenedesmus. The maximum colony size at 15–25 °C was significantly larger than those at 30–35 °C. An additional 1 or 2 days at low temperature was needed to reach the maximum colony size, which indicates the best harvest time for algal biomass.

Conclusion

Induced-colony formation of Scenedesmus by Daphnia culture filtrate at 15–25 °C is recommended to settle algal cells. This condition facilitates harvesting the biomass.

     

Potential Outdoor Cultivation of Green Microalgae Based on Response to Changing Temperatures and by Combining with Air Temperature Occurrence

In this study, our working hypothesis was to examine whether temperature alters biomass and metabolite production by microalgae according to strain. We also addressed whether it is possible to choose a strain suitable for growing in each season of a given region. A factorial experiment revealed a significant interaction between chlorophylls a and b (Chl a and Chl b), carotenoid/Chl (a?+?b) ratio, biomass and total lipid productivity of six green microalgae (four Chlorella spp., Chlorella sorokiniana and Neochloris oleoabundans) after 15 days at four temperatures. At 39/35 °C, two Chlorella sp. strains (IPR7115 and IPR7117) showed higher total carotenoids/Chl (a?+?b) (0.578 and 0.830), respectively. N. oleoabundans had the highest Chl a (8210 μg L^?1) and Chl b (1909 μg L^?1) at 19/15 °C and highest maximum dry biomass (2900 mg L^?1), specific growth rate (0.538 day^?1) and total lipids (1003 mg L^?1) at 15/8 °C. We applied a method to infer the growth of these six green microalgae in outdoor ponds, as based on their response to changing temperatures and by combining with historical data on day/night air temperature occurrence for a given region. We conclude that the use of regionalized maps based on air temperature is a good strategy for predicting microalgal cultivation in outdoor ponds based on their features and tolerance to changing temperature.     

Diversity in growth patterns among strains of the lethal fungal pathogen <Emphasis Type="Italic">Batrachochytrium dendrobatidis</Emphasis> across extended thermal optima

The thermal sensitivities of organisms regulate a wide range of ecological interactions, including host–parasite dynamics. The effect of temperature on disease ecology can be remarkably complex in disease systems where the hosts are ectothermic and where thermal conditions constrain pathogen reproductive rates. Amphibian chytridiomycosis, caused by the pathogen Batrachochytrium dendrobatidis (Bd), is a lethal fungal disease that is influenced by temperature. However, recent temperature studies have produced contradictory findings, suggesting that our current understanding of thermal effects on Bd may be incomplete. We investigated how temperature affects three different Bd strains to evaluate diversity in thermal responses. We quantified growth across the entire thermal range of Bd, and beyond the known thermal limits (T _max and T _min). Our results show that all Bd strains remained viable and grew following 24 h freeze (?12 °C) and heat shock (28 °C) treatments. Additionally, we found that two Bd strains had higher logistic growth rates (r) and carrying capacities (K) at the upper and lower extremities of the temperature range, and especially in low temperature conditions (2–3 °C). In contrast, a third strain exhibited relatively lower growth rates and carrying capacities at these same thermal extremes. Overall, our results suggest that there is considerable variation among Bd strains in thermal tolerance, and they establish a new thermal sensitivity profile for Bd. More generally, our findings point toward important questions concerning the mechanisms that dictate fungal thermal tolerances and temperature-dependent pathogenesis in other fungal disease systems.     

Temperature-dependent gastric evacuation rate of the Japanese delicate loach <Emphasis Type="Italic">Niwaella delicata</Emphasis> (Cobitidae)

The gastric evacuation rate (GER) of the delicate loach Niwaella delicata was estimated experimentally under four temperature conditions (10, 15, 20, and 25 °C). The GER was relatively high under the ≥20 °C condition and was nearly zero under the ≤15 °C condition. The relationship between GER and water temperature was estimated following the method of Elliot (1972) as follows: GER = exp (0.436 × temperature ? 10.59). This physical characteristic might affect the seasonal activity of this species.