Journal of plant physiology最新文献

{"title":"Chitosan-thiourea and their derivatives: Applications and action mechanisms for imparting drought tolerance","authors":"","doi":"10.1016/j.jplph.2024.154365","DOIUrl":"10.1016/j.jplph.2024.154365","url":null,"abstract":"<div><div>The increasing abiotic stresses from changing global climatic conditions, including drought, extreme temperatures, salinity, storms, pollutants, and floods, impend crop cultivation and sustainability. To mitigate these effects, numerous synthetic and non-synthetic chemicals or plant growth regulators are in practice. Chitosan, a natural organic substance rich in nitrogen and carbon, and thiourea, a synthetic plant growth regulator containing sulfur and nitrogen, have garnered significant interest for their roles in enhancing plant stress tolerance. Despite extensive use, the precise mechanisms of their actions remain unclear. Towards this endeavor, the present review examines how chitosan and thiourea contribute to stress tolerance in crop plants, particularly under drought conditions, to improve production and sustainability. It also explores thiourea's potential as a hydrogen sulfide (H₂S) donor and the possible applications of thiolated chitosan derivatives and chitosan-thiourea combinations, emphasizing their biological functions and benefits for sustainable agriculture.</div></div>","PeriodicalId":16808,"journal":{"name":"Journal of plant physiology","volume":null,"pages":null},"PeriodicalIF":4.0,"publicationDate":"2024-10-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142391382","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Changes in the anthocyanin pathway related to phenolic compounds and gene expression in skin and pulp of cv. 'Istrska belica' (Olea europaea L.) during ripening","authors":"","doi":"10.1016/j.jplph.2024.154364","DOIUrl":"10.1016/j.jplph.2024.154364","url":null,"abstract":"<div><div>The purpose of research was to study in detail the dynamics of the anthocyanin pathway during the ripening of olives, comprising the relative gene expression of nine enzymes and the contents of twelve phenolic compounds. The analyses were conducted on cv. 'Istrska belica' at seven maturity stages, separately in the pulp and the skin. Most phenolic compounds showed a higher content in the skin than in the pulp. Results showed that the accumulation of dihidroquercetin and dihydromyricetin started at the latest maturity stages. The most abundant phenolics evaluated in the current study present in both tissues were cyanidin-3-<em>O</em>-rutinoside and delphinidin-3-<em>O</em>-glucoside, both presented at all maturity stages, even when colour was not yet visible in the skin or pulp. Gene expression of enzymes revealed tissue-specific regulation during ripening. Genes expressions for phenylalanine ammonia lyase, chalcone synthase, chalcone isomerase, flavonoid 3-hydroxylase and flavonoid 3′-hydroxylase showed higher levels in the skin than in the pulp, and an upregulation during ripening in both tissues. Anthocyanidin synthase was the only gene with the highest expression at the beginning of ripening, with extreme decrease between second and third maturity stage, which suggests that the enzyme is mainly synthesized at the beginning of ripening and that enzyme activation starts at latest maturity stages. Our research contributes to a better understanding of the dynamics of phenolic accumulation and the relative gene expression of enzymes involved in the anthocyanin pathway in reveals tissue-specific changes during olive fruit ripening. The previous results are also supported by physical changes, which are reflected in a statistical increase in fruit weight, a decrease in fruit firmness and also by changes in appearance observed during ripening. Understanding the accumulation of anthocyanins could, through further study, help to improve the quality of the fruit and therefore the quality of olive products.</div></div>","PeriodicalId":16808,"journal":{"name":"Journal of plant physiology","volume":null,"pages":null},"PeriodicalIF":4.0,"publicationDate":"2024-09-28","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142375594","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"KAR1-dormancy release in Avena fatua caryopses includes increased AfMAN gene expression and ENDO-β-MANNANASE activity in the coleorhiza and radicle","authors":"","doi":"10.1016/j.jplph.2024.154363","DOIUrl":"10.1016/j.jplph.2024.154363","url":null,"abstract":"<div><div>KAR₁, at very low concentration (3x10⁻⁹ M) released dormancy in <em>Avena fatua</em> caryopses, which was expressed in almost complete emergence of coleorhiza (CE) and radicle (RE) just after three days of germination. The dormancy-releasing effect of KAR₁ was associated with an increased activity of ENDO-β-MANNANASE (MAN; EC 3.2.1.78) (hydrolase and transglycosylase) in coleorhiza and radicle before RE. The <em>MAN</em> genes, <em>MAN</em>1, <em>MAN2</em>, <em>MAN</em>3, <em>MAN</em>4, and <em>MAN5</em> were for the first time identified in the genome of <em>A. fatua</em>. KAR₁ induced expression of <em>AfMAN1-3</em> and <em>AfMAN5</em> in the coleorhiza and <em>AfMAN2</em> and <em>AfMAN3</em> in the radicle during caryopses germination. The increase in transcripts in the coleorhiza of <em>AfMAN1,5</em> after 8 h and <em>AfMAN3,5</em> after 12 h germination in the presence of KAR₁ is probably responsible for the increase in MAN activity determined after 18 h before RE. KAR₁ also increased <em>AfMAN</em>3 expression in radicle after 12 h which probably caused the increased MAN activity after 18 h. Therefore, release of caryopses dormancy by KAR₁ involves increasing expression of <em>MAN</em> genes and MAN activity both in the coleorhiza and radicle, which might facilitate the passage of the radicle through the coleorhiza. The work provides the first data on the contribution of MAN, present in coleorhiza and radicle, in the dormancy release of caryopses by KAR₁.</div></div>","PeriodicalId":16808,"journal":{"name":"Journal of plant physiology","volume":null,"pages":null},"PeriodicalIF":4.0,"publicationDate":"2024-09-26","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142375595","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Identification of BpEXP family genes and functional characterization of the BpEXPA1 gene in the stems development of Betula platyphylla","authors":"","doi":"10.1016/j.jplph.2024.154361","DOIUrl":"10.1016/j.jplph.2024.154361","url":null,"abstract":"<div><div>Expansins (EXPs) are unique plant cell wall proteins with the ability to induce cell wall expansion and play potential roles in xylem development. In the present study, a total of 25 <em>BpEXP</em> genes were identified in <em>Betula platyphylla</em>. Results of bioinformatics analysis described that BpEXP gene family was highly conserved in the process of evolution. All these genes were clustered into four groups, EXPA (Expansin A), EXPB (Expansin B), EXLA (Expansin-like A) and EXLB (Expansin-like B), according to phylogenetic analysis and BpEXPA1 was highly homologous to PttEXP1 and PttEXP2. The results of RT-qPCR showed that <em>BpEXPA1</em> was expressed higher in stems and preferentially expressed in the first internodes, followed by apical buds and the third internodes, promoter expression analysis with GUS assay demonstrated that it was expressed in developing xylem, suggesting that <em>BpEXPA1</em> might be involved in the development of the primary stems of birch. Overexpression of <em>BpEXPA1</em> can promote cortex cell expansion and then enlarge the cortex cell area and layer, however inhibit the secondary cell wall deposition and result in the thinner cell wall and larger lumens of xylem fiber in transgenic plants. This study will provide information for investigating the regulation mechanism of <em>BpEXP</em> family genes and gene resources for birch genetics improvement.</div></div>","PeriodicalId":16808,"journal":{"name":"Journal of plant physiology","volume":null,"pages":null},"PeriodicalIF":4.0,"publicationDate":"2024-09-21","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142324190","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Comparative analysis of element and hormone content in zygotic embryos of Pinus elliottii and P. elliottii × P. caribaea","authors":"","doi":"10.1016/j.jplph.2024.154359","DOIUrl":"10.1016/j.jplph.2024.154359","url":null,"abstract":"<div><div>Somatic embryogenesis is a crucial method for achieving clonal forestry in conifers. Understanding the development of zygotic embryos is essential not only for enhancing the efficiency and quality of somatic embryogenesis, but also for advancing forestry breeding programs. This study investigated dynamic changes of element and hormone contents during ZE development of <em>Pinus elliottii</em> and its hybrid <em>P. elliottii</em> × <em>P. caribaea</em>. Significant differences in embryo development speed among different clones were observed. Elemental analysis was conducted using inductively coupled plasma mass spectrometry (ICP-MS) and identified 68 elements, including major, minor, and beneficial elements. In both species, the contents of potassium (K), calcium (Ca), iron (Fe), boron (B) and five beneficial elements decreased during early ZE development, while phosphorus (P) and copper (Cu) increased. Significantly higher levels of K, Ca and Fe at the initial stage, and sulfur (S) and nickel (Ni) decreased at later stages were detected in <em>P. elliottii</em> than in the hybrid. For the other elements, except for very few significant differences at certain stages, most differences between the two species did not reach a significant level. The contents of endogenous hormones were determined and different accumulation patterns were detected in most hormones between the two species, except abscisic acid (ABA) which simultaneously decreased with developments by stage 8. Significant differences were found in indole-3-acetic acid (IAA) contents at most stages between species, while higher levels of total cytokinin (CK) at each stage were detected in the hybrid in comparison with those in <em>P. elliottii</em>. As a result, lower IAA to CK ratios in the hybrid than in <em>P. elliottii</em>. Methyl jasmonate (JA-me) and gibberellin A3 (GA3) contents showed a similar pattern and exhibited an M-shaped fluctuation in the hybrid. Furthermore, JA-me, GA3, gibberellin A4 (GA4) and brassinolide (BR) showed significantly higher levels in the hybrid than in <em>P. elliottii</em>. K-means clustering and correlation analyses were used to explore relationships between elements and hormones during embryo development, revealing complex interplay in both species. These data indicate different requirement in element and hormone contents for embryogenesis and suggest species-specific media composition for each step in somatic embryogenesis. The findings provide insights into their developmental processes and informing future research and applications in somatic embryogenesis and forestry breeding.</div></div>","PeriodicalId":16808,"journal":{"name":"Journal of plant physiology","volume":null,"pages":null},"PeriodicalIF":4.0,"publicationDate":"2024-09-19","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142324191","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Diethyl ether anaesthesia does not block local touch response in Arabidopsis thaliana","authors":"","doi":"10.1016/j.jplph.2024.154358","DOIUrl":"10.1016/j.jplph.2024.154358","url":null,"abstract":"<div><div>Plants can sense and respond to non-damaging mechanical stimulation such as touch, rain, or wind. Mechanical stimulation induces an increase of cytosolic calcium ([Ca²⁺]_cyt), accumulation of phytohormones from the group of jasmonates (JAs) and activation of gene expression, which can be JAs-dependent or JAs-independent. Response to touch shares similar properties with reactions to stresses such as wounding or pathogen attack, and regular mechanical stimulation leads to changes in growth and development called thigmomorphogenesis. Previous studies showed that well-known seismonastic plants such as Venus flytrap (<em>Dionaea muscipula</em>) or sensitive plant (<em>Mimosa pudica</em>) lost their touch-induced motive responses during exposure to general volatile anaesthetic (GVA) diethyl ether. Here, we investigated the effect of diethyl ether anaesthesia on touch response in <em>Arabidopsis thaliana</em>. We monitored [Ca²⁺]_cyt level, accumulation of JAs and expression of touch-responsive genes. Our results showed that none of the investigated responses was affected by diethyl ether. However, diethyl ether alone increased [Ca²⁺]_cyt and modulated JAs-independent touch-responsive genes, thus partially activating touch response non-specifically. Together with our previous studies, we concluded that GVA diethyl ether cannot block the local rise of [Ca²⁺]_cyt but only its systemic propagation dependent on GLUTAMATE LIKE RECEPTOR 3s (GLR3s) channels.</div></div>","PeriodicalId":16808,"journal":{"name":"Journal of plant physiology","volume":null,"pages":null},"PeriodicalIF":4.0,"publicationDate":"2024-09-19","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142324192","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Effects of LED polarized and vortex light on growth and photosynthetic characteristics of pepper (Capsicum annuum L.)","authors":"","doi":"10.1016/j.jplph.2024.154360","DOIUrl":"10.1016/j.jplph.2024.154360","url":null,"abstract":"<div><div>Most studies currently focus on traditional illuminant regulating plant growth, while less attention has been given to the LED internal luminescence. This study examined how polarized and vortex light affect the growth and photosynthetic traits of pepper plants, with LED light used as the control. The findings indicated that circular polarized light significantly increased the aboveground biomass of pepper. Additionally, both polarized and vortex light treatment significantly influenced the root development of pepper. In comparison to the control group, the chlorophyll content was highest under circular polarized light, while the Pn, Sc, Tr, and Ci values were highest under linear polarized light, and the enzyme activity of Rubisco was increased. Circular polarized light notably increased the activities of POD, CAT, and SOD, the activity of SOD reached its peak under the left vortex light. Moreover, the content of MDA was observed to be the lowest under linear and right vortex light treatments. The expressions of key genes for chlorophyll synthesis (<em>CaHEMA1</em> and <em>CaCAO</em>) and antioxidant enzyme synthesis (<em>CaPOD, CaSOD</em>, and <em>CaMDHAR</em>) were significantly altered under varying polarized light conditions, The latter genes, which play crucial roles in antioxidant enzyme activity, also showed significant variations in response to different polarized light treatments. In conclusion, polarized light significantly impacts the growth of pepper and is anticipated to be utilized for plant growth, setting the stage for future research in this area.</div></div>","PeriodicalId":16808,"journal":{"name":"Journal of plant physiology","volume":null,"pages":null},"PeriodicalIF":4.0,"publicationDate":"2024-09-19","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142348897","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Phloretin inhibits the growth of Arabidopsis shoots by inducing chloroplast damage and programmed cell death","authors":"","doi":"10.1016/j.jplph.2024.154354","DOIUrl":"10.1016/j.jplph.2024.154354","url":null,"abstract":"<div><div>Phloretin is a key secondary metabolite produced by apple trees. Known for its strong antioxidant properties, this dihydrochalcone has been extensively studied in animals but less so in plants. Recently, we identified phloretin as a phytotoxic allelochemical that inhibits growth in the model plant Arabidopsis by disrupting auxin metabolism and distribution in the roots. In this study, we found that phloretin significantly hinders the growth of Arabidopsis seedlings' aerial parts after a short-term treatment (10 days) and causes their decay after long-term exposure (28 days). These effects result from ultrastructural damage in the mesophyll cells of the leaves, including chloroplast displacement and swelling, lesions, and alterations in thylakoid and cell wall organization. Interestingly, phloretin-treated plants showed a decrease in malondialdehyde levels and antioxidant enzyme activities, while hydrogen peroxide and proline levels remained unchanged. This suggests that phloretin-induced chlorosis and seedling decay are not due to oxidative stress but rather to severe chloroplast structural damage, leading to inefficient photosynthesis, starch degradation, starvation, and activation of micro- and macroautophagic processes for self-preservation. Ultimately, these processes result in programmed cell death. These new insights into the phytotoxic effects of phloretin on Arabidopsis shoots could pave the way for future research into phloretin as a potential multitarget bioherbicide and enhance our understanding of autoallelopathy in apple trees.</div></div>","PeriodicalId":16808,"journal":{"name":"Journal of plant physiology","volume":null,"pages":null},"PeriodicalIF":4.0,"publicationDate":"2024-09-19","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142328195","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Inducible tolerance to low Ca:Mg in serpentine ecotype of Erythranthe guttata","authors":"","doi":"10.1016/j.jplph.2024.154355","DOIUrl":"10.1016/j.jplph.2024.154355","url":null,"abstract":"<div><div>In serpentine soils, the low level of calcium relative to magnesium (Ca:Mg) is detrimental to the growth of most plant species. Ecotypic variation in <em>Erythranthe guttata</em> allows for some populations to maintain high photosynthetic rates and biomass despite low Ca:Mg. In this study, the mechanism of tolerance was investigated by treating hydroponically grown plants with either high (1.0) or low (0.02) Ca:Mg growth solutions and assaying excised leaf discs for rates of photosynthesis and disc expansion, and for starch, Ca²⁺ and Mg²⁺ ion concentrations. Low Ca:Mg in the assay solutions reduced both photosynthesis and leaf disc expansion after one week of treatment. However, serpentine tissues show stable photosynthetic rates after one week and a recovery in leaf tissue expansion after two weeks exposure to low Ca:Mg conditions. Values for non-serpentine tissues continued to decline. Increased growth of low Ca:Mg treated discs supplied with exogenous sucrose suggests that growth in serpentine-exposed tissues is limited by availability of carbon products from photosynthesis. Serpentine leaves had higher vacuole Mg concentrations than non-serpentine leaves after three weeks of treatment with low Ca:Mg. The combination of elevated starch concentrations, reduced growth and lower vacuolar Mg concentrations in leaves of non-serpentine plants grown in low Ca:Mg indicate an inefficient use of carbon resources and starch degradation as an observed response to Mg toxicity. Together, these results suggest that serpentine <em>E. guttata</em> exhibits an inducible tolerance to low Ca:Mg through gradual compartmentalization of magnesium to maintain the production and metabolism of photosynthates necessary for growth.</div></div>","PeriodicalId":16808,"journal":{"name":"Journal of plant physiology","volume":null,"pages":null},"PeriodicalIF":4.0,"publicationDate":"2024-09-18","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142357689","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Hydrogen sulfide alleviates cadmium stress in germinating carrot seeds by promoting the accumulation of proline","authors":"","doi":"10.1016/j.jplph.2024.154357","DOIUrl":"10.1016/j.jplph.2024.154357","url":null,"abstract":"<div><div>Carrot (<em>Daucus carota</em> L.), a widely cultivated economically vegetable from the <em>Apiaceae</em> family, is grown globally. However, carrots can be adversely impacted by cadmium (Cd) pollution in the soil due to its propensity to accumulate in the fleshy root, thus impeding carrot growth and posing health hazards to consumers. Given the potential of hydrogen sulfide (H₂S) to improve plant resistance against Cd stress, we treated germinating carrot seeds with varying concentrations of sodium hydrosulfide (NaHS), aiming to alleviate the toxic impacts of Cd stress on carrot seed germination. The results revealed that carrot seeds treated with a concentration of 0.25 mM NaHS displayed better seed germination-associated characteristics compared to seeds treated with NaHS concentrations of 0.1 mM and 0.5 mM. Further investigation revealed a rise in the expression levels of <em>L-cysteine desulfhydrase</em> and <em>D-cysteine desulfhydrase</em>, along with enhanced activity of L-cysteine desulfhydrase and D-cysteine desulfhydrase among the NaHS treatment group, thereby leading to H₂S accumulation. Moreover, NaHS treatment triggered the expression of <em>pyrroline-5-carboxylate synthase</em> and <em>pyrroline-5-carboxylate reductase</em> and promoted the accumulation of endogenous proline, while the contents of soluble sugar and soluble protein increased correspondingly. Interestingly, since the application of exogenous proline did not influence the accumulation of endogenous H₂S, suggesting that H₂S served as the upstream regulator of proline. Histochemical staining and biochemical indices revealed that NaHS treatment led to elevated antioxidant enzyme activity, alongside a suppression of superoxide anion and hydrogen peroxide generation. Furthermore, high performance liquid chromatography analysis revealed that NaHS treatment reduced Cd²⁺ uptake, thereby promoting germination rate, seed vitality, and hypocotyl length of carrot seeds under Cd stress. Overall, our findings shed light on the application of NaHS to enhance carrot resistance against Cd stress and lay a foundation for exploring the regulatory role of H₂S in plants responding to Cd stress.</div></div>","PeriodicalId":16808,"journal":{"name":"Journal of plant physiology","volume":null,"pages":null},"PeriodicalIF":4.0,"publicationDate":"2024-09-18","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142310964","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}