In plants,the lysine and histidine transporter(LHT)family represent a class of proteins that mediate the uptake,translocation,and utilization of amino acids.The tea plant(Camellia sinensis)is a perennial evergreen wit...In plants,the lysine and histidine transporter(LHT)family represent a class of proteins that mediate the uptake,translocation,and utilization of amino acids.The tea plant(Camellia sinensis)is a perennial evergreen with a relatively high level of amino acids.However,systematic identification and molecular characterization of the LHT gene family has rarely been reported in tea plants.In this study,22 CsLHTs were identified from the‘Shuchazao’genome and classified into two groups.The modeled three-dimensional structure and the conserved domains presented a high similarity among the LHTs proteins.Moreover,it was predicted that a few genes were conserved through the analysis of the physiochemical characters,structures and cis-elements in promoters.The expression patterns in tea plants revealed that CsLHT7 was mainly expressed in the roots,and CsLHT4 and CsLHT11 exhibited relatively high expression in both the roots and leaves.Moreover,the expression of all three genes could be induced by organic nitrogen.Additionally,heterogeneous expression of CsLHT4,CsLHT7 and CsLHT11 in Arabidopsis thaliana decreased the aerial parts biomass compared with that in WT plants while significantly increased the rosette biomass only for CsLHT11transgenic plants versus WT plants.Overall,our results provide fundamental information about CsLHTs and potential genes in N utilization for further analysis in tea plants.展开更多
Pre-harvest water deficit(PHWD)plays an important role in sugar accumulation of citrus fruit.However,the mechanism is not known well.Here,it was confirmed that PHWD promoted sucrose accumulation of citrus fruit,but ha...Pre-harvest water deficit(PHWD)plays an important role in sugar accumulation of citrus fruit.However,the mechanism is not known well.Here,it was confirmed that PHWD promoted sucrose accumulation of citrus fruit,but had limited effect on fructose,glucose and total acid.A sucrose transporter,Cs SUT1,which localizes to the plasma membrane,was demonstrated to function in sucrose transport induced by PHWD.Compared to wild-type,Cs SUT1 overexpression in citrus calli stimulated sucrose,fructose and glucose accumulation,while its silencing in juice sacs reduced sucrose accumulation.Increased sugar accumulation in transgenic lines enhanced plant drought tolerance,and resulted in decreased electrolyte leakage,malondialdehyde and hydrogen peroxide contents,as well as increased superoxide dismutase activity and proline contents.An abscisic acid(ABA)-responsive transcription factor,Cs ABF3,was found with a same expression pattern with Cs SUT1 under PHWD.Yeast one-hybrid,electrophoretic mobility shift assay and dual-luciferase assays all revealed that Cs ABF3 directly bound with the Cs SUT1 promoter by ABA responsive elements.When Cs ABF3 was overexpressed in citrus calli,the sucrose,fructose and glucose concentration increased correspondingly.Further,transgenic studies demonstrated that Cs ABF3 could affect sucrose accumulation by regulating Cs SUT1.Overall,this study revealed a regulation of Cs ABF3 promoting Cs SUT1 expression and sucrose accumulation in response to PHWD.Our results provide a detail insight into the quality formation of citrus fruit.展开更多
BACKGROUND Ankylosing spondylitis(AS)is a chronic immune-mediated inflammatory disease.The prevailing theory links AS onset to infections in susceptible individuals.Furthermore,infections may impair the immune respons...BACKGROUND Ankylosing spondylitis(AS)is a chronic immune-mediated inflammatory disease.The prevailing theory links AS onset to infections in susceptible individuals.Furthermore,infections may impair the immune responses.Numerous studies have investigated links between AS and various infections-bacterial,viral,fungal,and other microorganism infections.However,limited attention has been given to the association between AS and Clonorchis sinensis(C.sinensis)infection.CASE SUMMARY A 27-year-old male with a 10-yr history of AS presented to our hospital with inflammatory lower back pain as the primary manifestation.Ten years ago,the patient had achieved a stable condition after treatment with biological agents.However,he experienced a recurrence of lumbosacral pain with an unexplained cause 10 d before hospital admission.A lumbosacral magnetic resonance imaging(MRI)scan revealed bone marrow edema in the left sacroiliac joint,and laboratory indicators were elevated.Moreover,the presence of C.sinensis eggs was detected in the stool.The patient was prescribed praziquantel,resulting in the disap-pearance of C.sinensis eggs in subsequent routine stool tests and relief from lumbosacral pain.A follow-up MRI scan performed after 4 months revealed a reduction in bone marrow edema around the left sacroiliac joint.CONCLUSION C.sinensis infections could potentially trigger the exacerbation of AS.Clinicians should pay attention to investigating the presence of infections.INTRODUCTION Ankylosing spondylitis(AS)is a chronic inflammatory and rheumatic disease resulting from an imbalance between innate and acquired immune responses[1].While it can affect any part of the spine,its primary symptoms are persistent back pain and stiffness in the lower back and pelvis.The prevalence of AS per 10000 individuals is 23.8 in Europe,31.9 in North America,16.7 in Asia,10.2 in Latin America,and 7.4 in Africa[2].Infections commonly occur in the first 3 months and may act as potential triggers for the first symptoms of AS,often manifesting as gastrointestinal,urinary tract,and respiratory infections of microbiological origin[3,4].Clonorchis sinensis(C.sinensis)infection is a severe parasitic disease affecting millions globally,especially prevalent in China,South Korea,the Far East of Russia,and Vietnam,with an estimated 15 million cases[5].Transmission occurs through the consumption of undercooked freshwater fish containing metacercariae.Adult C.sinensis parasites then establish themselves within the human hepatobiliary system[6].C.sinensis infection triggers the activation of sphingosine 1-phosphate receptor 2,leading to the injury and fibrosis of the hepatobiliary[7].Recent research in a rat model found that C.sinensis infection increases the risk of hepatocellular carcinoma by stimulating hepatic progenitor cell proliferation[8].Complications of C.sinensis infection include cholestasis,cholangitis,biliary system fibrosis,and in severe cases,the development of cholangiocarcinoma[9].Consequently,the primary preventive measure is to abstain from consuming raw or undercooked freshwater fish.Praziquantel is the recommended and effective treatment for this infection[10].While there is existing literature on the coexistence of AS and parasitic infections,there is limited research specifically addressing the simultaneous presence of AS and C.sinensis infection.This case report details a rare scenario of AS coexisting with C.sinensis infection,underscoring the potential impact of C.sinensis infection on AS disease activity.展开更多
Citrus is the typical mycorrhizal fruit tree species establishing symbiosis with arbuscular mycorrhizal (AM) fungi. However, arbuscule development and senescence in colonized citrus roots, especially in response to dr...Citrus is the typical mycorrhizal fruit tree species establishing symbiosis with arbuscular mycorrhizal (AM) fungi. However, arbuscule development and senescence in colonized citrus roots, especially in response to drought stress, remain unclear, which is mainly due to the difficulty in clearing and staining lignified roots with the conventional method. Here, we improved the observation of colonized roots of citrus plants with the sectioning method, which enabled the clear observation of AM fungal structures. Furthermore, we investigated the effects of one week of drought stress on arbuscule development and senescence with the sectioning method. Microscopy observations indicated that drought stress significantly decreased mycorrhizal colonization (F%and M%) although it did not affect plant growth performance. Fluorescence probes (WGA 488 and/or Nile red) revealed that drought stress inhibited arbuscule development by increasing the percentage of arbuscules at the early stage and decreasing the percentages of arbuscules at the midterm and mature stages. Meanwhile, drought stress accelerated arbuscule senescence, which was characterized by the increased accumulation of neutral lipids. Overall, the sectioning method developed in this study enables the in-depth investigation of arbuscule status, and drought stress can inhibit arbuscule development but accelerate arbuscule senescence in the colonized roots of citrus plants. This study paves the way to elaborately dissecting the arbuscule dynamics in the roots of fruit tree species in response to diverse abiotic stresses.展开更多
Chinese Assam tea(Camellia sinensis var.assamica)is an important tea crop with a long history of cultivation in Yunnan,China.Despite its potential value as a genetic resource,its genetic diversity and domestication/br...Chinese Assam tea(Camellia sinensis var.assamica)is an important tea crop with a long history of cultivation in Yunnan,China.Despite its potential value as a genetic resource,its genetic diversity and domestication/breeding history remain unclear.To address this issue,we genotyped 469 ancient tea plant trees representing 26 C.sinensis var.assamica populations,plus two of its wild relatives(six and three populations of C.taliensis and C.crassicolumna,respectively)using 16 nuclear microsatellite loci.Results showed that Chinese Assam tea has a relatively high,but comparatively lower gene diversity(H_(S)=0.638)than the wild relative C.crassicolumna(H_S=0.658).Clustering in STRUCTURE indicated that Chinese Assam tea and its two wild relatives formed distinct genetic groups,with considerable interspecific introgression.The Chinese Assam tea accessions clustered into three gene pools,corresponding well with their geographic distribution.However,New Hybrids analysis indicated that 68.48%of ancient Chinese Assam tea plants from Xishuangbanna were genetic intermediates between the Puer and Lincang gene pools.In addition,10%of the ancient Chinese Assam tea individuals were found to be hybrids between Chinese Assam tea and C.taliensis.Our results suggest that Chinese Assam tea was domesticated separately in three gene pools(Puer,Lincang and Xishuangbanna)in the Mekong River valley and that the hybrids were subsequently selected during the domestication process.Although the domestication history of Chinese Assam tea in southwestern Yunnan remains complex,our results will help to identify valuable genetic resources that may be useful in future tea breeding programs.展开更多
One of the main diseases that adversely impacts the global citrus industry is citrus bacterial canker(CBC),caused by the bacteria Xanthomonas citri subsp.citri(Xcc).Response to CBC is a complex process,with both prote...One of the main diseases that adversely impacts the global citrus industry is citrus bacterial canker(CBC),caused by the bacteria Xanthomonas citri subsp.citri(Xcc).Response to CBC is a complex process,with both proteinDNA as well as protein–protein interactions for the regulatory network.To detect such interactions in CBC resistant regulation,a citrus high-throughput screening system with 203 CBC-inducible transcription factors(TFs),were developed.Screening the upstream regulators of target by yeast-one hybrid(Y1H)methods was also performed.A regulatory module of CBC resistance was identified based on this system.One TF(CsDOF5.8)was explored due to its interactions with the 1-kb promoter fragment of CsPrx25,a resistant gene of CBC involved in reactive oxygen species(ROS)homeostasis regulation.Electrophoretic mobility shift assay(EMSA),dual-LUC assays,as well as transient overexpression of CsDOF5.8,further validated the interactions and transcriptional regulation.The CsDOF5.8–CsPrx25 promoter interaction revealed a complex pathway that governs the regulation of CBC resistance via H2O2homeostasis.The high-throughput Y1H/Y2H screening system could be an efficient tool for studying regulatory pathways or network of CBC resistance regulation.In addition,it could highlight the potential of these candidate genes as targets for efforts to breed CBC-resistant citrus varieties.展开更多
This study investigates the nitrogen (N), phosphorous (P), and potassium (K) contents in raw biomasses of Camellia sinensis, Gliricidia sepium, and Musa acuminata. Therein, the highest N and P content was seen in Came...This study investigates the nitrogen (N), phosphorous (P), and potassium (K) contents in raw biomasses of Camellia sinensis, Gliricidia sepium, and Musa acuminata. Therein, the highest N and P content was seen in Camellia sinensis 116.80 ± 0.08 mg and 66.00 ± 0.14 mg respectively. The highest K content (106.80 ± 0.04 mg) was observed in Musa acuminata. Next, all three types of plant materials were allowed to decompose in water for 3 weeks, and a sample from each was analyzed for NPK after the 1st, 2nd, and 3rd week during decomposition. A significant increase in the release of N, P, and K by the Camellia sinensis to water (P Musa acuminate were not significantly changed (P > 0.05) over time. The ratio for N:P:K was calculated for raw biomass samples and decomposed samples to find the best fitting N:P:K ratio to apply to young tea plants as organic fertilizers. In addition to that, the microbial insight of these organic compounds was analyzed by observing how microbial population increased with decomposition by the enumeration of the total microbial count. A considerable increment in total microbial count was observed up to 3.28 × 10<sup>6</sup>, 1.21 × 10<sup>10</sup>, 2.18 × 10<sup>8</sup>, and 6.49 × 10<sup>7</sup> CFU/ml for Camellia sinensis, Gliricidia sepium, Musa accuminata (leaves), and Musa accuminata (trunk) respectively. The presence of phosphate solubilizing bacteria (PSB) and nitrogen solubilizing bacteria (NSB) throughout the decomposition period was confirmed by their growth on NBRIP and a modified nutrient medium that was specifically designed for the identification of ammonifiers respectively. Prepared fertilizer samples were applied to young tea plants that were grown in the Mawanella area in Sri Lanka (7°15'12.42"N 80°26'47.62"E) and according to the results, it is clear that fertilizer mixture 1 (N:P:K, 10:5:10, tea dust + Gliricidia + banana trunk) and fertilizer mixture 2 (N:P:K, 10:5:10, tea dust + Gliricidia + banana leaves) has the potential to increase the growth of young tea plants.展开更多
An efficient in vitro regeneration system by direct organogenesis from mature nodal and internodal stem segments ofNewhall navel orange (Citrus sinensis L. Osbeck) was developed. Illuminating conditions together with...An efficient in vitro regeneration system by direct organogenesis from mature nodal and internodal stem segments ofNewhall navel orange (Citrus sinensis L. Osbeck) was developed. Illuminating conditions together with plant growthregulators affected the adventitious bud regeneration frequency and efficiency. The initial 15 d darkness inoculation isbeneficial for the adventitious bud regeneration. The highest regeneration frequency (85.2%) and bud formationefficiency (3.7 per responsive internodal stem segment) were obtained in the media supplemented with 1.0 mg L-1 BAPand 0.5 mg L-1 NAA. ABA at 0.2 mg L-1 positively affected the bud formation efficiency, which amounted to 8.5 buds perinternodal segment in the presence of BAP at 1.0 mg L-1. The adventitious shoots successfully rooted and weretransferred to the soil.展开更多
Objective:To assess the larvicidal and irritant activities of the hexane extracts of leaves of Citrus sinensis(C.sinensis)against the early fourth instars and female adults of Aedes aegypti(Ae.aegypti).Methods:The lar...Objective:To assess the larvicidal and irritant activities of the hexane extracts of leaves of Citrus sinensis(C.sinensis)against the early fourth instars and female adults of Aedes aegypti(Ae.aegypti).Methods:The larvicidal potential of the prepared leaf extract was evaluated against early fourth instar larvae of Ae.aegypli using WHO protocol.The mortality counts were made after 24 h and LC_(50)and LG_(50)values were calculated.The efficacy of extract as mosquito irritant was assessed by contact irritancy assays.Extract-impregnated paper was placed on a glass plate over which a perspex funnel with a hole on the top was kept inverted.Single female adult,3-day old unfed/blood-fed,was released inside the funnel.After 3 min of acclimatization time,the time taken for the first take-off and total number of flights undertaken during 15 min were scored.Results:The citrus leaf extracts from hexane possessed moderate larvicidal efficiency against dengue vector.The bioassays resulted in an LC_(50)and LC_(90)value of 446.84 and 1370.96 ppm,respectively after 24 h of exposure.However,the extracts were proved to be remarkable irritant against adults Ae.aegypti,more pronounced effects being observed on blood-fed females than unfed females.The extract-impregnated paper was thus proved to be 7-11 times more irritable as compared with the control paper.Conclusions:The hexane extracts from C.sinensis leaves are proved to be reasonably larvicidal But remarkably irritant against dengue vector.Further studies are needed to identify the possible role of extract as adulticide,oviposition deterrent and ovicidal agent.The isolation of active ingredient from the extract could help in formulating strategies for mosquito control.展开更多
Citrus bacterial canker(CBC)results from Xanthomonas citri subsp.citri(Xcc)infection and poses a grave threat to citrus production.Class III peroxidases(CIII Prxs)are key proteins to the environmental adaptation of ci...Citrus bacterial canker(CBC)results from Xanthomonas citri subsp.citri(Xcc)infection and poses a grave threat to citrus production.Class III peroxidases(CIII Prxs)are key proteins to the environmental adaptation of citrus plants to a range of exogenous pathogens,but the role of CIII Prxs during plant resistance to CBC is poorly defined.Herein,we explored the role of CsPrx25 and its contribution to plant defenses in molecular detail.Based on the expression analysis,CsPrx25 was identified as an apoplast-localized protein that is differentially regulated by Xcc infection,salicylic acid,and methyl jasmone acid in the CBC-susceptible variety Wanjincheng(C.sinensis)and the CBC-resistant variety Calamondin(C.madurensis).Transgenic Wanjincheng plants overexpressing CsPrx25 were generated,and these transgenic plants exhibited significantly increased CBC resistance compared with the WT plants.In addition,the CsPrx25-overexpressing plants displayed altered reactive oxygen species(ROS)homeostasis accompanied by enhanced H_(2)O_(2) levels,which led to stronger hypersensitivity responses during Xcc infection.Moreover,the overexpression of CsPrx25 enhanced lignification as an apoplastic barrier for Xcc infection.Taken together,the results highlight how CsPrx25-mediated ROS homeostasis reconstruction and cell wall lignification can enhance the resistance of sweet orange to CBC.展开更多
Pathological hypertrophy(cell enlargement)plays an important role in the development of citrus canker,but its regulators are largely unknown.Although WRKY22 is known to be involved in pathogen-triggered immunity and p...Pathological hypertrophy(cell enlargement)plays an important role in the development of citrus canker,but its regulators are largely unknown.Although WRKY22 is known to be involved in pathogen-triggered immunity and positively regulates resistance to bacterial pathogens in Arabidopsis,rice and pepper,the CRISPR/Cas9-mediated partial knockout of CsWRKY22 improves resistance to Xanthomonas citri subsp.citri(Xcc)in Wanjincheng orange(Citrus sinensis Osbeck).Here,we demonstrate that CsWRKY22 is a nucleus-localized transcriptional activator.CsWRKY22-overexpressing plants exhibited dwarf phenotypes that had wrinkled and thickened leaves and were more sensitive to Xcc,whereas CsWRKY22-silenced plants showed no visible phenotype changes and were more resistant to Xcc.Microscopic observations revealed that the overexpression of CsWRKY22 increased cell size in the spongy mesophyll.Transcriptome analysis showed that cell growth-related pathways,such as the auxin and brassinosteroid hormonal signaling and cell wall organization and biogenesis pathways,were significantly upregulated upon CsWRKY22 overexpression.Interestingly,CsWRKY22 activated the expression of CsLOB1,which is a key gene regulating susceptibility to citrus canker.We further confirmed that CsWRKY22 bound directly to the W-boxes just upstream of the transcription start site of CsLOB1 in vivo and in vitro.We conclude that CsWRKY22 enhances susceptibility to citrus canker by promoting host hypertrophy and CsLOB1 expression.Thus,our study provides new insights into the mechanism regulating pathological hypertrophy and the function of WRKY22 in citrus.展开更多
Citrus fruits are rich in carotenoids.In the carotenoid biosynthetic pathway,lycopene β-cyclase（LCYb,EC：1.14.-.-） is a key regulatory enzyme in the catalysis of lycopene to β-carotene,an important dietary precurs...Citrus fruits are rich in carotenoids.In the carotenoid biosynthetic pathway,lycopene β-cyclase（LCYb,EC：1.14.-.-） is a key regulatory enzyme in the catalysis of lycopene to β-carotene,an important dietary precursor of vitamin A for human nutrition.Two closely related lycopene β-cyclase cDNAs,designated CsLCYb1 and CsLCYb2,were isolated from the pulp of orange fruits（Citrus sinensis）.The expression level of CsLCYb genes is lower in the flavedo and juice sacs of a lycopeneaccumulating genotype Cara Cara than that in common genotype Washington,and this might be correlated with lycopene accumulation in Cara Cara fruit.The CsLCYb1 efficiently converted lycopene into the bicyclic β-carotene in an Escherichia coli expression system,but the CsLCYb2 exhibited a lower enzyme activity and converted lycopene into the β-carotene and the monocyclic γ-carotene.In tomato transformation studies,expression of CsLCYb1 under the control of the cauliflower mosaic virus（CaMV） 35S constitutive promoter resulted in a virtually complete conversion of lycopene into β-carotene,and the ripe fruits displayed a bright orange colour.However,the CsLCYb2 transgenic tomato plants did not show an altered fruit colour during development and maturation.In fruits of the CsLCYb1 transgenic plants,most of the lycopene was converted into β-carotene with provitamin A levels reaching about 700 μg g-1DW.Unexpectedly,most transgenic tomatoes showed a reduction in total carotenoid accumulation,and this is consistent with the decrease in expression of endogenous carotenogenic genes in transgenic fruits.Collectively,these results suggested that the cloned CsLCYb1 and CsLCYb2 genes encoded two functional lycopene β-cyclases with different catalytic efficiency,and they may have potential for metabolite engineering toward altering pigmentation and enhancing nutritional value of food crops.展开更多
基金supported by the National Key Research and Development Program of China(Grant No.2018YFD1000600)the National Natural Science Foundation of China(Grant No.32070376)。
文摘In plants,the lysine and histidine transporter(LHT)family represent a class of proteins that mediate the uptake,translocation,and utilization of amino acids.The tea plant(Camellia sinensis)is a perennial evergreen with a relatively high level of amino acids.However,systematic identification and molecular characterization of the LHT gene family has rarely been reported in tea plants.In this study,22 CsLHTs were identified from the‘Shuchazao’genome and classified into two groups.The modeled three-dimensional structure and the conserved domains presented a high similarity among the LHTs proteins.Moreover,it was predicted that a few genes were conserved through the analysis of the physiochemical characters,structures and cis-elements in promoters.The expression patterns in tea plants revealed that CsLHT7 was mainly expressed in the roots,and CsLHT4 and CsLHT11 exhibited relatively high expression in both the roots and leaves.Moreover,the expression of all three genes could be induced by organic nitrogen.Additionally,heterogeneous expression of CsLHT4,CsLHT7 and CsLHT11 in Arabidopsis thaliana decreased the aerial parts biomass compared with that in WT plants while significantly increased the rosette biomass only for CsLHT11transgenic plants versus WT plants.Overall,our results provide fundamental information about CsLHTs and potential genes in N utilization for further analysis in tea plants.
基金supported by the National Natural Science Foundation of China(Grant No.32172520)the earmarked fund for China Agriculture Research System(Grant No.CARS-26)。
文摘Pre-harvest water deficit(PHWD)plays an important role in sugar accumulation of citrus fruit.However,the mechanism is not known well.Here,it was confirmed that PHWD promoted sucrose accumulation of citrus fruit,but had limited effect on fructose,glucose and total acid.A sucrose transporter,Cs SUT1,which localizes to the plasma membrane,was demonstrated to function in sucrose transport induced by PHWD.Compared to wild-type,Cs SUT1 overexpression in citrus calli stimulated sucrose,fructose and glucose accumulation,while its silencing in juice sacs reduced sucrose accumulation.Increased sugar accumulation in transgenic lines enhanced plant drought tolerance,and resulted in decreased electrolyte leakage,malondialdehyde and hydrogen peroxide contents,as well as increased superoxide dismutase activity and proline contents.An abscisic acid(ABA)-responsive transcription factor,Cs ABF3,was found with a same expression pattern with Cs SUT1 under PHWD.Yeast one-hybrid,electrophoretic mobility shift assay and dual-luciferase assays all revealed that Cs ABF3 directly bound with the Cs SUT1 promoter by ABA responsive elements.When Cs ABF3 was overexpressed in citrus calli,the sucrose,fructose and glucose concentration increased correspondingly.Further,transgenic studies demonstrated that Cs ABF3 could affect sucrose accumulation by regulating Cs SUT1.Overall,this study revealed a regulation of Cs ABF3 promoting Cs SUT1 expression and sucrose accumulation in response to PHWD.Our results provide a detail insight into the quality formation of citrus fruit.
基金Supported by Chongqing Municipal Health Commission's Key Discipline Construction Project of Traditional Chinese Medicine Bi Disease,Yu Traditional Chinese Medicine(2021),No.16.
文摘BACKGROUND Ankylosing spondylitis(AS)is a chronic immune-mediated inflammatory disease.The prevailing theory links AS onset to infections in susceptible individuals.Furthermore,infections may impair the immune responses.Numerous studies have investigated links between AS and various infections-bacterial,viral,fungal,and other microorganism infections.However,limited attention has been given to the association between AS and Clonorchis sinensis(C.sinensis)infection.CASE SUMMARY A 27-year-old male with a 10-yr history of AS presented to our hospital with inflammatory lower back pain as the primary manifestation.Ten years ago,the patient had achieved a stable condition after treatment with biological agents.However,he experienced a recurrence of lumbosacral pain with an unexplained cause 10 d before hospital admission.A lumbosacral magnetic resonance imaging(MRI)scan revealed bone marrow edema in the left sacroiliac joint,and laboratory indicators were elevated.Moreover,the presence of C.sinensis eggs was detected in the stool.The patient was prescribed praziquantel,resulting in the disap-pearance of C.sinensis eggs in subsequent routine stool tests and relief from lumbosacral pain.A follow-up MRI scan performed after 4 months revealed a reduction in bone marrow edema around the left sacroiliac joint.CONCLUSION C.sinensis infections could potentially trigger the exacerbation of AS.Clinicians should pay attention to investigating the presence of infections.INTRODUCTION Ankylosing spondylitis(AS)is a chronic inflammatory and rheumatic disease resulting from an imbalance between innate and acquired immune responses[1].While it can affect any part of the spine,its primary symptoms are persistent back pain and stiffness in the lower back and pelvis.The prevalence of AS per 10000 individuals is 23.8 in Europe,31.9 in North America,16.7 in Asia,10.2 in Latin America,and 7.4 in Africa[2].Infections commonly occur in the first 3 months and may act as potential triggers for the first symptoms of AS,often manifesting as gastrointestinal,urinary tract,and respiratory infections of microbiological origin[3,4].Clonorchis sinensis(C.sinensis)infection is a severe parasitic disease affecting millions globally,especially prevalent in China,South Korea,the Far East of Russia,and Vietnam,with an estimated 15 million cases[5].Transmission occurs through the consumption of undercooked freshwater fish containing metacercariae.Adult C.sinensis parasites then establish themselves within the human hepatobiliary system[6].C.sinensis infection triggers the activation of sphingosine 1-phosphate receptor 2,leading to the injury and fibrosis of the hepatobiliary[7].Recent research in a rat model found that C.sinensis infection increases the risk of hepatocellular carcinoma by stimulating hepatic progenitor cell proliferation[8].Complications of C.sinensis infection include cholestasis,cholangitis,biliary system fibrosis,and in severe cases,the development of cholangiocarcinoma[9].Consequently,the primary preventive measure is to abstain from consuming raw or undercooked freshwater fish.Praziquantel is the recommended and effective treatment for this infection[10].While there is existing literature on the coexistence of AS and parasitic infections,there is limited research specifically addressing the simultaneous presence of AS and C.sinensis infection.This case report details a rare scenario of AS coexisting with C.sinensis infection,underscoring the potential impact of C.sinensis infection on AS disease activity.
基金supported by grants from the Natural Science Foundation of China (Grant No.42077040)the open competition program of top ten critical priorities of Agricultural Science and Technology Innovation for the 14th Five-Year Plan of Guangdong Province (Grant Nos.2022SDZG09,2023SDZG09)+1 种基金the Natural Science Foundation of Guangdong (Grant No.2021B1515010868)the GDAS Project of Science and Technology Development(2021GDASYL-20210103023)。
文摘Citrus is the typical mycorrhizal fruit tree species establishing symbiosis with arbuscular mycorrhizal (AM) fungi. However, arbuscule development and senescence in colonized citrus roots, especially in response to drought stress, remain unclear, which is mainly due to the difficulty in clearing and staining lignified roots with the conventional method. Here, we improved the observation of colonized roots of citrus plants with the sectioning method, which enabled the clear observation of AM fungal structures. Furthermore, we investigated the effects of one week of drought stress on arbuscule development and senescence with the sectioning method. Microscopy observations indicated that drought stress significantly decreased mycorrhizal colonization (F%and M%) although it did not affect plant growth performance. Fluorescence probes (WGA 488 and/or Nile red) revealed that drought stress inhibited arbuscule development by increasing the percentage of arbuscules at the early stage and decreasing the percentages of arbuscules at the midterm and mature stages. Meanwhile, drought stress accelerated arbuscule senescence, which was characterized by the increased accumulation of neutral lipids. Overall, the sectioning method developed in this study enables the in-depth investigation of arbuscule status, and drought stress can inhibit arbuscule development but accelerate arbuscule senescence in the colonized roots of citrus plants. This study paves the way to elaborately dissecting the arbuscule dynamics in the roots of fruit tree species in response to diverse abiotic stresses.
基金supported by funds from the National Natural Science Foundation of China(31970363,31161140350)the Key Basic Research Program of Yunnan Province,China(202101BC070003)supported by the Scottish Government’s Rural and Environment Science and Analytical Services division。
文摘Chinese Assam tea(Camellia sinensis var.assamica)is an important tea crop with a long history of cultivation in Yunnan,China.Despite its potential value as a genetic resource,its genetic diversity and domestication/breeding history remain unclear.To address this issue,we genotyped 469 ancient tea plant trees representing 26 C.sinensis var.assamica populations,plus two of its wild relatives(six and three populations of C.taliensis and C.crassicolumna,respectively)using 16 nuclear microsatellite loci.Results showed that Chinese Assam tea has a relatively high,but comparatively lower gene diversity(H_(S)=0.638)than the wild relative C.crassicolumna(H_S=0.658).Clustering in STRUCTURE indicated that Chinese Assam tea and its two wild relatives formed distinct genetic groups,with considerable interspecific introgression.The Chinese Assam tea accessions clustered into three gene pools,corresponding well with their geographic distribution.However,New Hybrids analysis indicated that 68.48%of ancient Chinese Assam tea plants from Xishuangbanna were genetic intermediates between the Puer and Lincang gene pools.In addition,10%of the ancient Chinese Assam tea individuals were found to be hybrids between Chinese Assam tea and C.taliensis.Our results suggest that Chinese Assam tea was domesticated separately in three gene pools(Puer,Lincang and Xishuangbanna)in the Mekong River valley and that the hybrids were subsequently selected during the domestication process.Although the domestication history of Chinese Assam tea in southwestern Yunnan remains complex,our results will help to identify valuable genetic resources that may be useful in future tea breeding programs.
基金funded by the National Key Research and Development Program of China(2022YFD1201600)the earmarked fund for the China Agriculture Research System(CARS-26)+1 种基金the Fundamental Research Funds for the Central Universities,China(SWU-XDJH202308)the Science and Technology Research Program of Chongqing Municipal Education Commission,China(KJQN202001418)。
文摘One of the main diseases that adversely impacts the global citrus industry is citrus bacterial canker(CBC),caused by the bacteria Xanthomonas citri subsp.citri(Xcc).Response to CBC is a complex process,with both proteinDNA as well as protein–protein interactions for the regulatory network.To detect such interactions in CBC resistant regulation,a citrus high-throughput screening system with 203 CBC-inducible transcription factors(TFs),were developed.Screening the upstream regulators of target by yeast-one hybrid(Y1H)methods was also performed.A regulatory module of CBC resistance was identified based on this system.One TF(CsDOF5.8)was explored due to its interactions with the 1-kb promoter fragment of CsPrx25,a resistant gene of CBC involved in reactive oxygen species(ROS)homeostasis regulation.Electrophoretic mobility shift assay(EMSA),dual-LUC assays,as well as transient overexpression of CsDOF5.8,further validated the interactions and transcriptional regulation.The CsDOF5.8–CsPrx25 promoter interaction revealed a complex pathway that governs the regulation of CBC resistance via H2O2homeostasis.The high-throughput Y1H/Y2H screening system could be an efficient tool for studying regulatory pathways or network of CBC resistance regulation.In addition,it could highlight the potential of these candidate genes as targets for efforts to breed CBC-resistant citrus varieties.
文摘This study investigates the nitrogen (N), phosphorous (P), and potassium (K) contents in raw biomasses of Camellia sinensis, Gliricidia sepium, and Musa acuminata. Therein, the highest N and P content was seen in Camellia sinensis 116.80 ± 0.08 mg and 66.00 ± 0.14 mg respectively. The highest K content (106.80 ± 0.04 mg) was observed in Musa acuminata. Next, all three types of plant materials were allowed to decompose in water for 3 weeks, and a sample from each was analyzed for NPK after the 1st, 2nd, and 3rd week during decomposition. A significant increase in the release of N, P, and K by the Camellia sinensis to water (P Musa acuminate were not significantly changed (P > 0.05) over time. The ratio for N:P:K was calculated for raw biomass samples and decomposed samples to find the best fitting N:P:K ratio to apply to young tea plants as organic fertilizers. In addition to that, the microbial insight of these organic compounds was analyzed by observing how microbial population increased with decomposition by the enumeration of the total microbial count. A considerable increment in total microbial count was observed up to 3.28 × 10<sup>6</sup>, 1.21 × 10<sup>10</sup>, 2.18 × 10<sup>8</sup>, and 6.49 × 10<sup>7</sup> CFU/ml for Camellia sinensis, Gliricidia sepium, Musa accuminata (leaves), and Musa accuminata (trunk) respectively. The presence of phosphate solubilizing bacteria (PSB) and nitrogen solubilizing bacteria (NSB) throughout the decomposition period was confirmed by their growth on NBRIP and a modified nutrient medium that was specifically designed for the identification of ammonifiers respectively. Prepared fertilizer samples were applied to young tea plants that were grown in the Mawanella area in Sri Lanka (7°15'12.42"N 80°26'47.62"E) and according to the results, it is clear that fertilizer mixture 1 (N:P:K, 10:5:10, tea dust + Gliricidia + banana trunk) and fertilizer mixture 2 (N:P:K, 10:5:10, tea dust + Gliricidia + banana leaves) has the potential to increase the growth of young tea plants.
基金The authors acknowledge the financial support by the National Natural Science Foundation of China(002002)
文摘An efficient in vitro regeneration system by direct organogenesis from mature nodal and internodal stem segments ofNewhall navel orange (Citrus sinensis L. Osbeck) was developed. Illuminating conditions together with plant growthregulators affected the adventitious bud regeneration frequency and efficiency. The initial 15 d darkness inoculation isbeneficial for the adventitious bud regeneration. The highest regeneration frequency (85.2%) and bud formationefficiency (3.7 per responsive internodal stem segment) were obtained in the media supplemented with 1.0 mg L-1 BAPand 0.5 mg L-1 NAA. ABA at 0.2 mg L-1 positively affected the bud formation efficiency, which amounted to 8.5 buds perinternodal segment in the presence of BAP at 1.0 mg L-1. The adventitious shoots successfully rooted and weretransferred to the soil.
基金supported by University Grants Commission[grant No.F.35-74/2009(SR)]
文摘Objective:To assess the larvicidal and irritant activities of the hexane extracts of leaves of Citrus sinensis(C.sinensis)against the early fourth instars and female adults of Aedes aegypti(Ae.aegypti).Methods:The larvicidal potential of the prepared leaf extract was evaluated against early fourth instar larvae of Ae.aegypli using WHO protocol.The mortality counts were made after 24 h and LC_(50)and LG_(50)values were calculated.The efficacy of extract as mosquito irritant was assessed by contact irritancy assays.Extract-impregnated paper was placed on a glass plate over which a perspex funnel with a hole on the top was kept inverted.Single female adult,3-day old unfed/blood-fed,was released inside the funnel.After 3 min of acclimatization time,the time taken for the first take-off and total number of flights undertaken during 15 min were scored.Results:The citrus leaf extracts from hexane possessed moderate larvicidal efficiency against dengue vector.The bioassays resulted in an LC_(50)and LC_(90)value of 446.84 and 1370.96 ppm,respectively after 24 h of exposure.However,the extracts were proved to be remarkable irritant against adults Ae.aegypti,more pronounced effects being observed on blood-fed females than unfed females.The extract-impregnated paper was thus proved to be 7-11 times more irritable as compared with the control paper.Conclusions:The hexane extracts from C.sinensis leaves are proved to be reasonably larvicidal But remarkably irritant against dengue vector.Further studies are needed to identify the possible role of extract as adulticide,oviposition deterrent and ovicidal agent.The isolation of active ingredient from the extract could help in formulating strategies for mosquito control.
基金The National Key Research and Development Program of China(2018YFD1000306)the Natural Science Foundation of Chongqing(cstc2020jcyj-msxmX1064)+2 种基金the Fundamental Research Funds for the Central Universities(SWU115025)the Earmarked Funds for the China Agriculture Research System(CARS-26)the Key Project of Guangxi Science and Technology(GuiKeAA18118046-6)funded this study.
文摘Citrus bacterial canker(CBC)results from Xanthomonas citri subsp.citri(Xcc)infection and poses a grave threat to citrus production.Class III peroxidases(CIII Prxs)are key proteins to the environmental adaptation of citrus plants to a range of exogenous pathogens,but the role of CIII Prxs during plant resistance to CBC is poorly defined.Herein,we explored the role of CsPrx25 and its contribution to plant defenses in molecular detail.Based on the expression analysis,CsPrx25 was identified as an apoplast-localized protein that is differentially regulated by Xcc infection,salicylic acid,and methyl jasmone acid in the CBC-susceptible variety Wanjincheng(C.sinensis)and the CBC-resistant variety Calamondin(C.madurensis).Transgenic Wanjincheng plants overexpressing CsPrx25 were generated,and these transgenic plants exhibited significantly increased CBC resistance compared with the WT plants.In addition,the CsPrx25-overexpressing plants displayed altered reactive oxygen species(ROS)homeostasis accompanied by enhanced H_(2)O_(2) levels,which led to stronger hypersensitivity responses during Xcc infection.Moreover,the overexpression of CsPrx25 enhanced lignification as an apoplastic barrier for Xcc infection.Taken together,the results highlight how CsPrx25-mediated ROS homeostasis reconstruction and cell wall lignification can enhance the resistance of sweet orange to CBC.
基金the National Key Research and Development Program of China(2018YFD1000300,to S.C.)the National Natural Sciences Foundation of China(31972393,to X.Z.)+2 种基金the Natural Science Foundation Project of Chongqing City(cstc2019jcyj-bshX0024 to Q.L.)the Fundamental Research Funds for the Central Universities(XDJK2019C027,to Q.L.)the Earmarked Fund for China Agriculture Research System(CARS-26,to S.C.).
文摘Pathological hypertrophy(cell enlargement)plays an important role in the development of citrus canker,but its regulators are largely unknown.Although WRKY22 is known to be involved in pathogen-triggered immunity and positively regulates resistance to bacterial pathogens in Arabidopsis,rice and pepper,the CRISPR/Cas9-mediated partial knockout of CsWRKY22 improves resistance to Xanthomonas citri subsp.citri(Xcc)in Wanjincheng orange(Citrus sinensis Osbeck).Here,we demonstrate that CsWRKY22 is a nucleus-localized transcriptional activator.CsWRKY22-overexpressing plants exhibited dwarf phenotypes that had wrinkled and thickened leaves and were more sensitive to Xcc,whereas CsWRKY22-silenced plants showed no visible phenotype changes and were more resistant to Xcc.Microscopic observations revealed that the overexpression of CsWRKY22 increased cell size in the spongy mesophyll.Transcriptome analysis showed that cell growth-related pathways,such as the auxin and brassinosteroid hormonal signaling and cell wall organization and biogenesis pathways,were significantly upregulated upon CsWRKY22 overexpression.Interestingly,CsWRKY22 activated the expression of CsLOB1,which is a key gene regulating susceptibility to citrus canker.We further confirmed that CsWRKY22 bound directly to the W-boxes just upstream of the transcription start site of CsLOB1 in vivo and in vitro.We conclude that CsWRKY22 enhances susceptibility to citrus canker by promoting host hypertrophy and CsLOB1 expression.Thus,our study provides new insights into the mechanism regulating pathological hypertrophy and the function of WRKY22 in citrus.
基金supported by the National Basic Research Program of China (973 Program, 2011CB100600)the National Natural Science Foundation of China (30771482, 30921002)
文摘Citrus fruits are rich in carotenoids.In the carotenoid biosynthetic pathway,lycopene β-cyclase（LCYb,EC：1.14.-.-） is a key regulatory enzyme in the catalysis of lycopene to β-carotene,an important dietary precursor of vitamin A for human nutrition.Two closely related lycopene β-cyclase cDNAs,designated CsLCYb1 and CsLCYb2,were isolated from the pulp of orange fruits（Citrus sinensis）.The expression level of CsLCYb genes is lower in the flavedo and juice sacs of a lycopeneaccumulating genotype Cara Cara than that in common genotype Washington,and this might be correlated with lycopene accumulation in Cara Cara fruit.The CsLCYb1 efficiently converted lycopene into the bicyclic β-carotene in an Escherichia coli expression system,but the CsLCYb2 exhibited a lower enzyme activity and converted lycopene into the β-carotene and the monocyclic γ-carotene.In tomato transformation studies,expression of CsLCYb1 under the control of the cauliflower mosaic virus（CaMV） 35S constitutive promoter resulted in a virtually complete conversion of lycopene into β-carotene,and the ripe fruits displayed a bright orange colour.However,the CsLCYb2 transgenic tomato plants did not show an altered fruit colour during development and maturation.In fruits of the CsLCYb1 transgenic plants,most of the lycopene was converted into β-carotene with provitamin A levels reaching about 700 μg g-1DW.Unexpectedly,most transgenic tomatoes showed a reduction in total carotenoid accumulation,and this is consistent with the decrease in expression of endogenous carotenogenic genes in transgenic fruits.Collectively,these results suggested that the cloned CsLCYb1 and CsLCYb2 genes encoded two functional lycopene β-cyclases with different catalytic efficiency,and they may have potential for metabolite engineering toward altering pigmentation and enhancing nutritional value of food crops.
