DIA-based quantitative proteomic analysis on porcine meat quality at different chilling rates

The objective of this study was to evaluate the effects of chilling rate on porcine meat quality from the perspective of proteome using data independent acquisition(DIA)-based quantitative proteomic strategy. M. longissimus thoracis et lumborum(n = 9) was assigned randomly to the control group(3.72 ℃/h), very fast chilling-Ⅰ group(VFC-Ⅰ, 9.31℃/h) and VFC-Ⅱ group(14.43 ℃/h). The DIA was used to analyze the difference in proteins under different chilling rates. Results showed that tenderness was improved significantly in meat at the chilling rate of 14.43 ℃/h. Seventy-nine differential abundant proteins(fold change > 1.5, P < 0.05), including 46 up-regulated and 33 down-regulated proteins, were identified and mainly involved in carbon metabolism, pyruvate metabolism and proteasome pathways. These pathways indicated that VFC delayed cell metabolism and glycolysis by down-regulating the expression of metabolic enzymes. The tenderness was improved by up-regulating the expression of proteasome and m-calpain.

Effects of Cyclic Electron Flow Inhibitor(Antimycin A)on Photosystem Photoinhibition of Sweet Pepper Leaves upon Exposure to Chilling Stress Under Low Irradiance

In chloroplast, there were two pathways involved in the cyclic electron flow around photosystem 1 （PS 1）. One was the NADH dehydrogenase （NDH）-dependent flow and the other was the ferredoxin quinone reductase-dependent flow. It was proposed that the NDH-dependent cyclic electron flow around PSI was related to the xanthophyll cycle-dependent non-photochemical quenching （NPQ） at chilling temperature under low irradiance （CL）. The function of the chloroplastic cyclic electron flow around PS 1 was examined by comparing sweet pepper （Capsicum annuum L.） control with its antimycin A （AA）-fed leaves upon exposure to CL stress. During CL stress, the maximum photochemical efficiency of PS2 （Fv/Fm） decreased markedly in both controls and AA-fed leaves, and P700＋ was also lower in AA-fed leaves than in controls. These results implied that cyclic electron flow around PS 1 functioned to protect the photosynthetic apparatus from CL stress. Under such stress, NPQ and PS2-driven electron transport rate were different between AA-fed leaves and controls. The lower NPQ in AA-fed leaves might be related to an inefficient proton gradient across thylakoid membranes （ApH） because of inhibiting cyclic electron flow around PS 1 under CL stress.

In-situ observation of crystal detachment behavior in NH_4Cl-H_2O system from chilling metal surface with vibration

The crystal detachment behaviors from a chilling solid surface with vibration were investigated using a transparent NH4Cl-70%H2O alloy.The nucleation experiments of NH4Cl-70%H2O alloy were performed on a chilling surface generator with various vibration frequency from 20 to 1 000 Hz and vibration acceleration from 10 to 100 m/s2.The results indicate that the crystal detach rate increases and the grains are refined with increasing vibration acceleration under a certain vibration frequency.It is interesting to note that,when the frequency is 50 Hz,acceleration is 100 m/s2,the crystal detach rate increases sharply and the grain refinement effect is strengthened.A special grain refinement phenomenon can be observed,when the vibration acceleration and frequency are up to a critical value.A power-based definition was introduced in order to describe the relationship between the vibration parameters and imposed power on the system.It is found that a power criterion exists for the grain refinement in the current system.

Effects of Short-Term Chilling Stress on the Photosystems and Chloroplast Ultrastructure in Sweet Pepper

By measuring chlorophyll fluorescence parameters, composition of fatty acids, active oxygen species and activities ofsome antioxidant enzymes, effects of chilling stress (4C) in the low light (100 mmol m-2 s-1) on chilling-sensitive plants werestudied. After 6 h chilling stress (4C) in the low light, the maximal photochemical efficiency of PSII (Fv/Fm) of sweetpepper leaves decreased by 35.6%, and the oxidizable P700 decreased by 60%. However, chilling stress in the dark had noeffect on both of them. Unsaturation of fatty acids in thylakoid membrane was accelerated, which might be helpful tostabilize photosynthetic apparatus. Distortion and swelling of grana caused by chilling in the dark probably decreasedactivities of antioxidant enzymes, which resulted in the accumulation of active oxygen species. On the contrary,photooxidation might be related to the disintegration and unstacking of grana. Chilling stress induced photoinhibition ofPSII and PSI, and active oxygen species might be one of the factors causing the decrease of the oxidizable P700. PSIseemed to be more sensitive to chilling stress in the low light than PSII.

Exogenous melatonin improves the chilling tolerance and preharvest fruit shelf life in eggplant by affecting ROS-and senescence-related processes

Low temperature is the most common abiotic stress factors during the eggplant cultivation in solar greenhouses.Melatonin plays important roles in plant resistance to low temperature.However,the role of melatonin in regulating chilling tolerance and extending the preharvest shelf life of eggplant fruits is still unknown.In this study,we investigated the effects of exogenous melatonin on eggplant plants and fruits in response to low temperature.Under simulated low-temperature conditions,exogenous melatonin significantly relieved the chilling symptoms of seedlings by reducing reactive oxygen species (ROS) and malondialdehyde (MDA) levels and relative leakage rates.These reductions were caused by higher superoxide dismutase (SOD) and catalase (CAT) activities and increased endogenous polyamine and melatonin levels compared with those in untreated seedlings.Notably,the expression levels of SOD,CAT1/2,and polyamine synthesis genes (ADC and ODC) were also increased by 100μmol·L~(-1)melatonin,as well as those of genes involved in melatonin synthesis (TDC,T5H,SNAT,ASMT,and COMT) and cold regulation (COR1,CBFa/b,and ZAT2/6/12).To further investigate the effects of melatonin on eggplant leaves and fruits under natural low temperature conditions,100μmol·L~(-1)melatonin was sprayed on the functional leaves at three days before commodity maturation.Melatonin significantly alleviated chilling injury in the leaves and pericarp and extended the preharvest shelf life of the fruit by increasing the expression of COR1,CBF,ZAT2/6/12,and API5 and decreasing the expression of senescence-related genes (NCED1/2 and SAG12).Therefore,100μmol·L~(-1)melatonin improved chilling tolerance and fruit shelf life by upregulating ZAT2/6/12 to affect ROS-and senescence-related processes,which provides a reference for alleviating cold stress and extending the preharvest fruit shelf life in eggplant.

Genome-wide profiling of histone H3 lysine 27 trimethylation and its modification in response to chilling stress in grapevine leaves

Histone H3 lysine 27 trimethylation(H3K27me3) is a histone modification associated with transcriptional repression. However, insights into the genome-wide pattern of H3K27me3 in grapevines are limited. Here, anti-H3K27 chromatin immunoprecipitation(ChIP), high-throughput sequencing, and transcriptome analysis were performed using leaves of Vitis amurensis. The leaves were treated at 4°C for 2 h and 24 h and used to investigate changes in H3K27me3 under chilling treatment. The results show that H3K27me3 is well-distributed both in gene regions(-50%) and in the intergenic region(-50%) in the grapevine genome(Vitis vinifera ‘Pinot Noir PN40024'). H3K27me3 was found to be localized in8 368 annotated gene regions in all detected samples(leaves at normal temperature and under chilling treatments) and mainly enriched in gene bodies with the adjacent promoter and downstream areas. The short-term chilling treatments(4°C for 2 h) induced 2 793 gains and 305losses in H3K27me3 modification. Subsequently, 97.3% of the alterations were restored to original levels after 24 h treatment. The ChIP-qPCR for five differential peaks showed similar results to the data for ChIP-seq, indicating that the chilling-induced H3K27me3 modification is reliable.Integrative analysis of transcriptome and ChIP-seq results showed that the expression of H3K27me3 target genes was significantly lower than those of non-target genes, indicating transcriptional repression of H3K27me3 in grapevine leaves. Furthermore, histone methylation alterations were detected in 82 genes and were related to either repression or activation of their expression during chilling stress. The findings provide the genome-wide H3K27me3 patterns in grapevines and shed light on uncovering its regulation in chilling stress responses.

Melatonin treatment alleviates chilling injury in mango fruit 'Keitt' by modulating proline metabolism under chilling stress

Mangoes often suffer from low temperature-induced chilling injury (CI) during postharvest cold storage.Therefore,advanced techniques are crucial and in high demand to solve the chilling stress of mango fruit for a higher value.This study addresses chilling stress modulation by investigating the effects of melatonin treatment on CI,proline metabolism,and related gene expressions of ‘Keitt’ mango during cold storage after dipped in 0 (control),0.1 (MT1),and 0.2 mmol L^(-1)(MT2) melatonin solution for 30 min.The results revealed that melatonin treatment in MT1 significantly reduced CI development and increased proline content in mango fruit during cold storage compared to the control.These changes were along with increases in the activity of critical enzymes as well as the expression of encoding genes involved in proline biosynthesis,such as pyrroline-5-carboxylate synthetase (P5CS),pyrroline-5-carboxylate reductase (P5CR),ornithine D-aminotransferase (OAT),P5CS2,P5CR2,and OAT3.Additionally,proline dehydrogenase (PDH) activity and the expression of the PDH3 gene associated with proline dehydrogenation were lower in MT1-treated mangoes than the controlled group.Thus,melatonin treatment has regulated proline metabolism resulting in the accumulation of proline,subsequently contributing to enhancing the chilling tolerance of‘Keitt’mango fruit.

Down-regulation of MeMYB2 leads to anthocyanin accumulation and increases chilling tolerance in cassava(Manihot esculenta Crantz)

Chilling-induced accumulation of reactive oxygen species(ROS) is harmful to plants,which usually produce anthocyanins to scavenge ROS as protection from chilling stress.As a tropical crop,cassava is hypersensitive to chilling,but the biochemical basis of this hypersensitivity remains unclear.We previously generated Me MYB2-RNAi transgenic cassava with increased chilling tolerance.Here we report that Me MYB2-RNAi transgenic cassava accumulated less ROS but more cyanidin-3-O-glucoside than the wild type under early chilling stress.Under this stress,the anthocyanin biosynthesis pathway was more active in Me MYB2-RNAi lines than in the wild type,and several genes involved in the pathway,including Me TT8,were up-regulated by Me MYB2-RNAi in the transgenic cassava.Me MYB2 bound to the Me TT8 promoter and blocked its expression under both normal and chilling conditions,thereby inhibiting anthocyanin accumulation.Me TT8 was shown to bind to the promoter of Dihydroflavonol 4-reductase(Me DFR-2)and increased Me DFR-2 expression.Me MYB2 appears to act as an inhibitor of chilling-induced anthocyanin accumulation during the rapid response of cassava to chilling stress.

Insight into the spoilage heterogeneity of meat-borne bacteria isolates with high-producing collagenase

Chilled chicken is inevitably contaminated by microorganisms during slaughtering and processing,resulting in spoilage.Cutting parts of chilled chicken,especially wings,feet,and other skin-on products,are abundant in collagen,which may be the primary target for degradation by spoilage microorganisms.In this work,a total of 17 isolates of spoilage bacteria that could secrete both collagenase and lipase were determined by raw-chicken juice agar(RJA)method,and the results showed that 7 strains of Serratia,Aeromonas,and Pseudomonas could significantly decompose the collagen ingredients.The gelatin zymography showed that Serratia liquefaciens(F5)and(G7)had apparent degradation bands around 50 kDa,and Aeromonas veronii(G8)and Aeromonas salmonicida(H8)had a band around.65 and 95 kDa,respectively.The lipase and collagenase activities were detected isolate-by-isolate,with F5 showing the highest collagenase activity.For spoilage ability on meat in situ,F5 performed strongest in spoilage ability,indicated by the total viable counts,total volatile basic nitrogen content,sensory scores,lipase,and collagenase activity.This study provides a theoretical basis for spoilage heterogeneity of strains with high-producing collagenase in meat.

Research Progress and Outlook of Low Temperature Chilling Injury in Northeast China

Focused on the major agro-meteorological disaster in northeast China area—low temperature chilling injury,the research progresses of low temperature chilling injury in northeast China were reviewed systematically.The basic concepts which included the connotation and extension of chilling damage,the circulation characteristics and effect factors which formed summer low temperature in northeast China,the prediction,forecast and defense of low temperature chilling injury were summarized and done the outlook.

Effect of Polyamine Priming on Chilling Tolerance of Lolium perenne during Seed Imbibition

[Objective] This study aimed to investigate the effect of potyamine priming on physiological and biochemical variations of Lolium perenne embryos and seed germination. [Method] With annual Lolium perenne （Diamond T and Grazing-8000） as experimental materials, after priming with 0.5 mmol/L putrescine （Put）, spermidine （Spd） and spermine （Spm） for 24 h and chilling imbibition at 5 ℃ for 12, 24, 36 and 48 h, the effect of Put, Spd and Spm priming on chilling tolerance and germination ability of annual Lolium perenne seeds during imbibition was investigated. [Result] Put, Spd and Spm priming improved the activities of peroxidase （POD）, catalase （CAT）, superoxide dismutase （SOD） and ascorbate peroxidase （APX） and content of soluble protein content under low temperature stress, significantly in-creased the germination rate, and shortened the average germination duration. After chilling imbibition for 48 h, compared with the control, the average germination rate of annual Lolium perenne seeds was improved by 15.5% and 12.0% after Put, Spd and Spm priming, and the average germination duration was shortened by 1.21 and 1.14 d. During seed imbibition, the chilling tolerance of Grazing-8000 was stronger than that of Diamond T. Overall, Put, Spd and Spm treatment could increase the chilling tolerance of annual Lolium perenne seeds during imbibition, and improve the germination ability of seeds under low temperature stress. [Conclusion] Results of this study provided theoretical basis for the application of seed priming technology in the production of annual ryegrass.

Analysis and Evaluation Indicator Selection of Chilling Tolerance of Different Cotton Genotypes

[Objectivc] This study aimed to investigate the chilling tolerance of seedlings of different cotton genotypes and screen appropriate indicators for assess- ing chilling tolerance, to establish reliable mathematical evaluation model for chilling tolerance of cotton, thus providing theoretical basis for breeding and promoting new chilling-tolerant cotton germplasms and large-scale evaluation of chilling tolerance of cotton varieties. [Method] Fifteen cotton varieties （lines） were used as experimental materials. The photosynthetic gas exchange parameters, chlorophyll fluorescence ki- netic parameters, chlorophyll content, relative soluble sugar content, malonaldehyde content, relative proiine content, relative conductivity and other 12 physiological indi- cators of seedling leaves under low temperature treatment （5 ℃, 12 h） and recovery treatment （25 ℃. 24 h） were determined; based on the chilling tolerance coefficient （CTC） of various individual indicators, the comprehensive evaluation of chilling toler- ance was conducled by using principal component analysis, hierarchical cluster anal- ysis and stepwise regression analysis. [Result] The results showed that the 12 indi- vidual physiological indicators could be classified into 7 independent comprehensive components by principal component analysis; 15 cotton varieties （lines） were clus- tered into three categories by using membership function method and hierarchical cluster analysis; the mathematical model for evaluating chilling tolerance of cotton seedlings was established： D =0.275 -0.244Fo1 ＋0.206Fv/Fm1＋0.326g,%-0.056SS ＋ 0.225MDA＋O.O38REC （FF=0.995）, and the evaluation accuracy of the equation was higher than 94.25%,0. Six identification indicators closely related to chilling tolerance were screened, including Fo,, Fv/Fm1, Seedling leaves of cotton varieties （lines） gs2, SS, MDA, and REC. [Conclusion] with high chilling tolerance are less dam- aged under low temperature stress, and are able to maintain relatively high photo- synthetic electron transport capacity and high stomatal conductance after recovery treatment, which is contributed to gas exchange and recovery of photosynthetic ca- pacity. Determination of the six indicators under the same stress condition can be adopted for rapid identification and prediction of the chilling tolerance of other cotton varieties, which provides basis for the breeding, promotion, identification and screen- ing of chilling tolerant germplasms.

AtpH Is a Chilling-Repressed Gene in Rice

A chilling-repressed cDNA fragment was cloned from rice ( Oryza sativa L.) seedlings by reverse transcription-polymerase chain reaction (RT-PCR) differential display. Database comparison of the cDNA sequence is identical to atpH encoded by rice chloroplast genome. Northern hybridization showed that its RNA transcript was reduced greatly on the first day and fully inhibited after 2 d of chilling treatment.

Peculiarity of transcriptional and H3K27me3 dynamics during peach bud dormancy

Bud dormancy facilitates the survival of meristems under harsh environmental conditions.To elucidate how molecular responses to chilling accumulation controlling dormancy in peach buds,chromatin immunoprecipitation sequencing to identify the H3K27me3 modifications and RNA sequencing of two peach cultivars with pronounced differences in chilling requirement were carried out,the results showed that genes associated with abscisic acid and gibberellic acid signal pathways play key roles in dormancy regulation.The results demonstrated that peach flower bud differentiation occurred continuously in both cultivars during chilling accumulation,which was correlated with the transcript abundance of key genes involved in phytohormone metabolism and flower bud development under adverse conditions.The more increased strength in high chillingrequirement cultivar along with the chilling accumulation at the genome-wide level.The function of the dormancy-associated MADS-box gene PpDAM6 was identified,which is involved in leaf bud break in peach and flower development in transgenic Nicotiana tabacum(NC89).In addition,PpDAM6 was positively regulated by PpCBF,and the genes of putative dormancy-related and associated with metabolic pathways were proposed.Taken together,these results constituted a theoretical basis for elucidating the regulation of peach bud dormancy transition.

Study on Chilling Requirement of Different Fruit Branch of a Sweet Cherry Cultivar ‘Hongmanao'

With ＇Hongmanao＇, a sweet cherry cultivar from central Shanxi Province, as the experimental material, the chilling requirements of different types of fruit branches （including mixed fruit branch, long fruit branch, medium fruit branch, short fruit branch and blossom fruit branch） were studied by the 0-7.2 ℃ model in a solar greenhouse during 2012-2013. In the greenhouse, the fruit branches of ＇Hongmanao＇ were cultured in water in a thermostatic environment. The results showed that the chilling requirement of leaf buds of ＇Hongmanao＇ was lower than that of its flower buds; and the chilling requirements of the five types of fruit branches ranked as blossom fruit branch＇s〈short fruit branch＇s〈medium fruit branch＇s〈long fruit branch＇s〈mixed fruit branch＇s,

Smart Colorimetric Corn Starch Films Combined with Anthocyanin-Loaded Glutenin-Carboxymethyl Chitosan Nanocomplexes for Freshness Monitoring of Chilled Pork

In this study,intelligent,pH-responsive colorimetric films were prepared by encapsulating anthocyanins in nanocomplexes prepared from glutenin and carboxymethyl chitosan.These nanocomplexes were added to a corn starch matrix and used in the freshness monitoring of chilled pork.The effects of anthocyanin-loaded nanocomplexes on the physical,structural,and functional characteristics of the films were investigated.The addition of anthocyanin-loaded nanocomplexes increased the tensile strength,elongation at break,hydrophobicity,and light transmittance of the films while decreasing their water vapor permeability.This is because new hydrogen bonds are formed between the film components,resulting in a more homogeneous and dense structure.The colorimetric film has a significant color response to pH changes.These films were used in experiments on the freshness of chilled pork,in which the pH changes with changing freshness states.The results show that the colorimetric film can monitor changes in the freshness of chilled pork in real time,where orange,pink,and green represent the fresh,secondary fresh,and putrefied states of pork,respectively.Therefore,the intelligent colorimetric film developed in this study has good application potential in the food industry.

Effect of Delay Chilling on μ-calpain Activities and the Tenderness of Bovine M. longissimus

μ-calpain activities and shear force values of bovine M. longissimus from Chinese Yellow crossbred bulls were analyzed,and the effect of delay chilling on μ-calpain activities and the tenderness of beef during postmortem aging were studied. The results showed that delay chilling significantly improved μ-calpain activities (P<0.05) and enhanced the tenderness of bovine M. longissimus during earlier aging periods compared with conventional chilling. But in later aging periods,delay chilling weakened the effect on the tenderness of beef because of premature consumption of μ-calpain. The experiment results confirmed that delay chilling improved the rate of postmortem aging of beef and remarkably enhanced the tenderness of beef through the effect of delay chilling on μ-calpain activities.

Temperature Trends and Accumulation of Chill Hours, Chill Units, and Chill Portions in South Carolina

There is considerable concern about the potential impact of climate change on agriculture, such as the accumulation of chilling hours needed to break the dormancy of many perennial plants, like fruit trees. Therefore, this study aimed to determine if there had been a significant change in air temperatures and chill hours, chill units, and chill portion accumulation in South Carolina over the last two decades. Two decades of daily maximum (Tmax) and minimum (Tmin) air temperature records were obtained from weather stations in thirty-one counties in South Carolina. Hourly temperature data, reconstructed from the daily data, were used to calculate the daily and annual chill hours, chill units, and chill portions accumulation using four different chill models for each location and year. The chill models included the T(t) °C model, the 0°C °C model, the Utah model, and the Dynamic model. For each county, regression analyses were conducted to evaluate the historical trends. Despite year-to-year variability, the tendency was a statistically significant (α = 0.05) increase in air temperature, averaging 0.089°C per year for 20 out of 31 counties in South Carolina. The other 11 counties had no significant change in temperature. The average temperature increase in the 31 counties was 0.072°C per year. The temperature increase resulted in a decrease in annual chill accumulation during the fall to spring, averaging 17.7 chill hours, 8.6 chill hours, 17.0 chill units, and 0.40 chill portions per year calculated with the T(t) °C, 0°C °C, Utah, and Dynamic models, respectively. However, whether this decrease in chill values was statistically significant or not depended on the chill model used. This study did not investigate the cause of the observed historical trends in temperature and chill accumulation. Still, if the trends continue, they could significantly impact the future of the temperate fruit tree industry in the state.

Methyl Jasmonate Reduces Chilling Injury and Maintains Postharvest Quality in Peaches

Zhaohui peaches(Prunus persica Batsch)were treated with 0(CK), 1, 10 or 100 μmol L-1 methyl jasmonate(MeJA)vapor at 20℃ for 24 h before stored at 0℃ for 35 d. The untreated fruits showed chilling injury(CD symptoms after 4 wk of storage, as indicated by increased fruit firmness and reduced ex-tractable juice, which is referred as leatheriness. Treatment with 1 and 10 μmol L-1 MeJA promoted normal ripening and softening, maintained higher levels of extractable juice, titratable acidity, pectinesterase(PE)and polygalacturonase(PC)activities, inhibited increases in fruit fresh weight loss, decay incidence, electrolyte leakage and MDA content, and improved color development, thereby preventing chilling injury symptoms development and maintaining edible quality. MeJA treatment also delayed the climacteric rise in respiratory rate, but promoted ethylene production during the later period of cold storage, suggesting that ethylene may involve in CI development of peaches. These results indicate that 1 and 10 μmol L-1 MeJA treatments could be used to reduce CI development and decay incidence in peaches.

Water relations and an expression analysis of plasma membrane intrinsic proteins in sensitive and tolerant rice during chilling and recovery

A symptom of chilling injury is development of water deficit in shoots, resulting from an imbalance of water transport and transpiration. In this work, two rice varieties （Oryza sativa L. var. Wasetoitsu and Somewake） seedlings were chilled at 7 ℃, followed by recovery at 28 ℃. Based on the growth phenotype and electrolyte leakage tests, Somewake was shown to be a chilling-tolerant variety, and Wasetoitsu a chilling-sensitive one. The chilling stress reduced markedly the relative water content （RWC） of leaves, accumulative transpiration and osmotic root hydraulic conductivity （Lp） in both varieties. But when retumed to 28 ℃, the water relation balance of Somewake recovered better. The mRNA expression profile of all the 11 plasma membrane intrinsic proteins （PIPs）, a subgroup of aquaporins, was subsequently determined by real-time reverse transcription （RT）-PCR with TaqMan-minor grove binder （MGB） probes derived from rice var. Nipponbare during chilling treatment and recovery. Most of the PIP genes was down-regulated at the low temperature, and recovered at the warm temperature. The relative expression of some PIPs in both Somewake and Wasetoitsu decreased in parallel during the chilling. However during the recovery, the relative expression of OsPIP1;1, OsPIP2;1, OsPIP2;7 in shoots and OsPIP1：1, OsPIP2：1 in roots were significantly higher in Somewake than Wasetoitsu. This supports the role of PIPs in re-establishing water balance after chilling conditions. We discuss the diversified roles played by members of the aquaporin PIP subfamily in plant chilling tolerance depending on aquaporin isoforms, plant tissue and the stage of chilling duration.