CsABF3-activated CsSUT1 pathway is implicated in pre-harvest water deficit inducing sucrose accumulation in citrus fruit

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.

Eureka lemon zinc finger protein ClDOF3.4 interacts with citrus yellow vein clearing virus coat protein to inhibit viral infection

Citrus yellow vein clearing virus(CYVCV)is a new citrus virus that has become an important factor restricting the development of China’s citrus industry,and the CYVCV coat protein(CP)is associated with viral pathogenicity.In this study,the Eureka lemon zinc finger protein(ZFP)ClDOF3.4 was shown to interact with CYVCV CP in vivo and in vitro.Transient expression of ClDOF3.4 in Eureka lemon induced the expression of salicylic acid(SA)-related and hypersensitive response marker genes,and triggered a reactive oxygen species burst,ion leakage necrosis,and the accumulation of free SA.Furthermore,the CYVCV titer in ClDOF3.4 transgenic Eureka lemon plants was approximately 69.4%that in control plants 6 mon after inoculation,with only mild leaf chlorotic spots observed in those transgenic plants.Taken together,the results indicate that ClDOF3.4 not only interacts with CP but also induces an immune response in Eureka lemon by inducing the SA pathways.This is the first report that ZFP is involved in the immune response of a citrus viral disease,which provides a basis for further study of the molecular mechanism of CYVCV infection.

Improved observation of colonized roots reveals the regulation of arbuscule development and senescence by drought stress in the arbuscular mycorrhizae of citrus

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.

High-throughput screening system of citrus bacterial cankerassociated transcription factors and its application to the regulation of citrus canker resistance

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.

航天搭载后福橘(Citrus reticulata Blanco)茎尖无性系有丝分裂行为和细胞超微结构的变化

福橘(Citrus reticulata Blanco)茎尖经‘实践八号’育种卫星搭载后,经组织培养获得其再生植株,以此为材料,结合常规压片法与超薄切片技术,研究航天搭载对福橘当代无性系细胞有丝分裂行为及细胞超微结构的影响。结果表明:航天搭载对福橘茎尖具有诱变作用,所得13个诱变无性系中,有丝分裂过程存在不同程度的异常,其中‘08004’无性系植株的有丝分裂畸变率明显高于其他无性系:中期染色体数目异常畸变率为0.34%,落后染色体畸变率为0.69%,微核畸变率为0.86%;并且有丝分裂过程中纺锤体结构发生异常,可见C-形(0.17%)或S-形纺锤体(1.20%),细胞发生多极分裂(1.03%)。绝大多数航天搭载当代无性系的细胞超微结构正常。‘08004’无性系植株叶肉细胞超微结构呈现细胞程序性死亡(PCD)典型特征:细胞核内染色质凝集;叶绿体类囊体结构解体;出现自体吞噬小体;细胞液泡化并发生质壁分离,最终形成凋亡小体。PCD过程中,线粒体保持其结构完整性,直至PCD末期才发生结构变化。

农杆菌对蕉柑(Citrusreticulata var.tankan Hayata)胚性细胞的附着研究

农杆菌能够附着蕉柑的胚性细胞。农杆菌对蕉柑胚性细胞的附着曲线表明 ,农杆菌与蕉柑胚性细胞之间的附着是一个动态过程。农杆菌在蕉柑胚性细胞上的附着量受共培养介质中乙酰丁香酮 (As)和表面活性物质影响较小 。

Phenolic and flavonoid contents of mandarin (Citrus reticulata Blanco) fruit tissues and their antioxidant capacity as evaluated by DPPH and ABTS methods

The total phenolic and flavonoid contents in the fruit tissues （peels, pulp residues, seeds, and juices） of 19 citrus genotypes belonged to Citrus reticulata Blanco were evaluated and their antioxidant capacity was tested by 2,2-diphenyl-l-picrylhydra- zyl radicals （DPPH） method and 2,2＇-azino-bis（3-ethylbenzthiozoline-6）-sulphonic acid （ABTS） method. The total phenolic and flavonoid contents, and their antioxidant capacity varied in different citrus fruit tissues. Generally, the peel had both the highest average of total phenolics （27.18 mg gallic acid equivalent （GAE） g^-1 DW） and total flavonoids （38.97 mg rutin equivalent （RE） g^-1 DW）. The highest antioxidant capacity was also the average of DPPH value （21.92 mg vitamin C equiv- alent antioxidant capacity （VCEAC） g^-1 DW） and average of ABTS value （78.70 mg VCEAC g-1 DW） in peel. The correlation coefficient between the total phenolics and their antioxidant capacity of different citrus fruits tissues ranged from 0.079 to 0.792, and from -0.150 to 0.664 for the total flavonoids. The antioxidant capacity of fruit tissues were correlated with the total phenoilc content and flavonoid content except in case of the peel. In addition, the total phenolic content and antioxidant capacity varied in different citrus genotypes. Manju and Karamandarin were better genotypes with higher antioxidation and the phenolic content, however Shagan was the poorest genotype with lower antioxidation and the phenolic content.

Multiomics-based dissection of citrus flavonoid metabolism using a Citrus reticulata × Poncirus trifoliata population

Deciphering the genetic basis of plant secondary metabolism will provide useful insights for genetic improvement and enhance our fundamental understanding of plant biological processes.Although citrus plants are among the most important fruit crops worldwide,the genetic basis of secondary metabolism in these plants is largely unknown.Here,we use a high-density linkage map to dissect large-scale flavonoid metabolic traits measured in different tissues(young leaf,old leaf,mature pericarp,and mature pulp)of an F_(1) pseudo-testcross citrus population.We detected 80 flavonoids in this population and identified 138 quantitative trait loci(QTLs)for 57 flavonoids in these four tissues.Based on transcriptional profiling and functional annotation,twenty-one candidate genes were identified,and one gene encoding flavanone 3-hydroxylase(F3H)was functionally verified to result in naturally occurring variation in dihydrokaempferol content through genetic variations in its promoter and coding regions.The abundant data resources collected for diverse citrus germplasms here lay the foundation for complete characterization of the citrus flavonoid biosynthetic pathway and will thereby promote efficient utilization of metabolites in citrus quality improvement.

Digital Gene Expression Analysis of Huanglongbing Affected Mandarins(Citrus reticulata Blanco)In Response to Thermotherapy

Huanglongbing(HLB),which is mainly associated with“Candidatus Liberibacter asiaticus”(CLas),is currently threatening citrus production around the world.This destructive century-old disease results in callose deposition in phloem plasmodesmata and starch overaccumulation in leaf chloroplasts.Heat treatment with high humidity has proven to be effective in eliminating the CLas titers in potted citrus trees.This study explored the curative mechanisms and impact of heat treatment(from 32 to 48°C with 75%±5%relative humidity,14h in light and 10h in dark for four days)on mandarin trees(Citrus reticulata Blanco)that were severely infected by CLas using Solexa/Illumina’s digital gene expression(DGE)profiling.A total of 838 differentially expressed genes(DEGs)were identified,of which 510 were downregulated.Proteinprotein interaction analysis of the DEGs indicated that the thermotherapy of HLB trees downregulated the circadian clock related genes in chloroplasts and upregulated the RNA-editing process in the mitochondria,characterized by 141 genes encoding pentatricopeptide repeatcontaining proteins(PPR)being all upregulated.Furthermore,the expression of phenylpropanoid and flavonoid biosynthesis genes,and auxininduced genes were suppressed.The defense response pathway of the thermotherapy treated diseased trees was apparently disrupted,as indicated by the downregulation of LRR receptor-like serine/threonine-protein kinase FLS2 gene.This study improves our understanding of the curative mechanisms of heat treatment on HLB trees and CLas biological regulation.Importantly,this provides new insights on the involvement of RNA modification by PPR genes and changes in the circadian clock after the heat stress treatment of CLas-infected citrus.

Manual thinning increases fruit size and sugar content of Citrus reticulata Blanco and affects hormone synthesis and sugar transporter activity

Manual fruit thinning(MFT)in fruit trees has been previously shown to increase fruit size and enhance fruit quality,but the effect of MFT on Ponkan(Citrus reticulata Blanco)and the underlying mechanisms remain poorly understood.In this study,efforts were made to elucidate how MFT influences the fruit quality of Ponkan.The results showed that MFT substantially increased fruit size and elevated fruit total soluble solids in comparison with the fruit from the unthinned trees(used as control).Expression analyses demonstrated that m RNA abundance of three important sugar transporter genes,including CrSUT1,CrSTP1 and CrTMT1,was evidently elevated in the flesh of thinned fruit when compared with those of the control.In addition,MFT prominently up-regulated the transcript levels of various auxin and gibberellin(GA)biosynthesis and signaling genes,including CrYUC6,CrAUX/IAA,CrGA20ox1 and CrGA3ox1.Concurrently,the contents of endogenous IAA and GA3,measured at 90 d after fruit thinning,were notably elevated in the fruit from trees with the thinning treatment relative to the control,although no difference was detected in the two groups before the thinning manipulation.Taken together,these results indicate that manual fruit thinning could greatly improve fruit quality,which may be attributed to promoting fruit expansion due to the increased auxin levels and expediting sugar accumulation through the up-regulation of sugar transporter genes.

Genome-wide analysis of cytochrome P450 genes in Citrus clementina and characterization of a CYP gene encoding f lavonoid 3′-hydroxylase

Cytochrome P450s(CYPs)are the largest family of enzymes in plant and play multifarious roles in development and defense but the available information about the CYP superfamily in citrus is very limited.Here we provide a comprehensive genome-wide analysis of the CYP superfamily in Citrus clementina genome,identifying 301 CYP genes grouped into ten clans and 49 families.The characteristics of both gene structures and motif compositions strongly supported the reliability of the phylogenetic relationship.Duplication analysis indicated that tandem duplication was the major driving force of expansion for this superfamily.Promoter analysis revealed numerous cis-acting elements related to various responsiveness.RNA-seq data elucidated their expression patterns in citrus fruit peel both during development and in response to UV-B.Furthermore,we characterize a UV-B-induced CYP gene(Ciclev10019637m,designated CitF3′H)as a f lavonoid 3-hydroxylase for the first time.CitF3 H catalyzed numerous f lavonoids and favored naringenin in yeast assays.Virusinduced silencing of CitF3′ H in citrus seedlings significantly reduced the levels of 3′-hydroxylated f lavonoids and their derivatives.These results together with the endoplasmic reticulum-localization of CitF3 H in plant suggest that this enzyme is responsible for the biosynthesis of 3-hydroxylated f lavonoids in citrus.Taken together,our findings provide extensive information about the CYP superfamily in citrus and contribute to further functional verification.

Transcriptomic analysis reveals the defense mechanisms of citrus infested with Diaphorina citri

Huanglongbing(HLB) is a devastating disease that has led to an acute crisis for growers of citrus, one of the world's most important fruit crops. The phloem-feeding Asian citrus psyllid(ACP), Diaphorina citri, is the main pest at the new shoot stage and is the only natural vector of HLB pathogenic bacteria. Little is known about how plants perceive and defend themselves from this destructive pest. Here, we characterized changes in the expression of various genes in citrus plants that were continuously infested by D. citri for different durations(12, 24, and 48 h). A total of 5 219 differentially expressed genes(DEGs) and 643 common DEGs were identified across all time points. Several pathways related to defense were activated, such as peroxisome, alpha-linolenic acid metabolism, and phenylpropanoid and terpenoid biosynthesis, and some pathways related to growth and signal transduction were suppressed in response to D. citri infestation. The expression of genes including kinases(CML44, CIPK6, and XTH6), phytohormones(SAMT, LOX6, and NPR3), transcription factors(bHLH162, WRKY70, and WRKY40), and secondary metabolite synthesis-related genes(PAL, 4CL2, UGT74B1 and CYP82G1) was significantly altered in response to D. citri infestation. The findings of this study greatly enhance our understanding of the mechanisms underlying the defense response of citrus plants to D. citri infestation at the molecular level. Functional characterization of the candidate defense-related genes identified in this study will aid the molecular breeding of insect-resistant citrus varieties.

Mapping of QTLs and candidate genes associated with multiple phenotypic traits for Huanglongbing tolerance in citrus

Huanglongbing(HLB)is the most devastating disease for citrus worldwide.Candidatus Liberibacter asiaticus(C Las),vectored by Asian citrus psyllid(ACP,Diaphorina citri Kuwayama),is the most common pathogen causing the disease.Commercial citrus varieties are highly susceptible to HLB,whereas trifoliate orange(Poncirus trifoliata)is considered highly tolerant to HLB.An F1 segregating population and their parent trifoliate orange and sweet orange,which had been exposed to intense HLB pressure for three years,was evaluated for disease symptoms,ACP colonization,C Las titer and tree vigor repeatedly for two to three years.Trifoliate orange and sweet orange showed significant differences for most of the phenotypic traits,and the F1 population exhibited a large variation.A high-density SNP-based genetic map with 1402 markers was constructed for trifoliate orange,which exhibited high synteny and high coverage of its reference genome.A total of 26 quantitative trait locus(QTLs)were identified in four linkage groups LG-t6,LG-t7,LG-t8 and LG-t9,of which four QTL clusters exhibit a clear co-localization of QTLs associated with different traits.Through genome-wide analysis of gene expression in response to C Las infection in‘Flying Dragon’and‘Larger-Flower DPI-50-7’trifoliate orange,85 differentially expressed genes were found located within the QTL clusters.Among them,seven genes were classified as defense or immunity protein which exhibited the highest transcriptional change after C Las infection.Our results indicate a quantitative genetic nature of HLB tolerance and identified candidate genes that should be valuable for searching for genetic solutions to HLB through breeding or genetic engineering.

An efficient transient gene expression system for protein subcellular localization assay and genome editing in citrus protoplasts

Protoplast has been widely used in biotechnologies to circumvent the breeding obstacles in citrus, including long juvenility, polyembryony, and male/female sterility. The protoplast-based transient gene expression system is a powerful tool for gene functional characterization and CRISPR/Cas9 genome editing in higher plants, but it has not been widely used in citrus. In this study, the polyethylene glycol(PEG)-mediated method was optimized for citrus callus protoplast transfection, with an improved transfection efficiency of 68.4%. Consequently, the efficiency of protein subcellular localization assay was increased to 65.8%, through transient expression of the target gene in protoplasts that stably express the fluorescent organelle marker protein. The gene editing frequencies in citrus callus protoplasts reached 14.2% after transient expression of CRISPR/Cas9 constructs. We demonstrated that the intronic polycistronic tRNAgRNA(inPTG) genome editing construct was functional in both the protoplast transient expression system and epicotyl stable transformation system in citrus. With this optimized protoplast transient expression system, we improved the efficiency of protein subcellular localization assay and developed the genome editing system in callus protoplasts, which provides an approach for prompt test of CRISPR vectors.

Transcription factor CsTT8 promotes fruit coloration by positively regulating the methylerythritol 4-phosphate pathway and carotenoid biosynthesis pathway in citrus (Citrus spp.)

Carotenoids directly influence citrus fruit color and nutritional value,which is critical to consumer acceptance.Elucidating the potential molecular mechanism underlying carotenoid metabolism is of great importance for improving fruit quality.Despite the well-established carotenoid biosynthetic pathways,the molecular regulatory mechanism underlying carotenoid metabolism remains poorly understood.Our previous studies have reported that the Myc-type basic helix-loop-helix(bHLH)transcription factor(TF)regulates citrus proanthocyanidin biosynthesis.Transgenic analyses further showed that overexpression of CsTT8 could significantly promote carotenoid accumulation in transgenic citrus calli,but its regulatory mechanism is still unclear.In the present study,we found that overexpression of CsTT8 enhances carotenoid content in citrus fruit and calli by increasing the expression of CsDXR,CsHDS,CsHDR,CsPDS,CsLCYE,CsZEP,and CsNCED2,which was accompanied by changes in the contents of abscisic acid and gibberellin.The in vitro and in vivo assays indicated that CsTT8 directly bound to the promoters of CsDXR,CsHDS,and CsHDR,the keymetabolic enzymes of the methylerythritol 4-phosphate(MEP)pathway,thus providing precursors for carotenoid biosynthesis and transcriptionally activating the expression of these three genes.In addition,CsTT8 activated the promoters of four key carotenoid biosynthesis pathway genes,CsPDS,CsLCYE,CsZEP,and CsNCED2,directly promoting carotenoid biosynthesis.This study reveals a novel network of carotenoid metabolism regulated by CsTT8.Our findings will contribute to manipulating carotenoid metabolic engineering to improve the quality of citrus fruit and other crops.

An Efficient Regeneration System for Native Orange (<i>Citrus reticulata</i>) through <i>In-Vitro</i>Culture Technique

Citrus reticulata (Mandarin Orange), commonly known as “Sweet Orange”, is one of the most difficult plants to improve through traditional breeding approaches as it poses various biological limitations that greatly hinder the cultivar improvement. In the present study, using the fresh seed of native orange as explant, an efficient, reproducible, regeneration method was developed through in vitro organogenesis. Mature, healthy and dehusked seeds were treated with Murashige and Skoog, (MS) media containing 3% sucrose, 0.7% agar supplemented with different concentrations and combinations of phytohormones. The highest calli initiation (93.3% ± 0.5%) responses were observed on MS medium supplemented with 2,4-dichlorophenoxyacetic acid (2,4-D) at 3.0 mg/L followed by 2,4-D at 3.5 mg/L (86.7% ± 1.75%) in this experiment. Maximum shoot regeneration (86.7% ± 3.35%) responses were reported using MS medium supplemented with the combination of 6-benzylaminopurine (BAP) at 3.0 mg/L and 1-naphthaleneacetic acid (NAA) at 2.0 mg/L. MS medium supplemented with NAA at 1.0 mg/L showed the best rooting (80% ± 2.89%) response in comparison to (70% ± 5.20%) indole-3-butyric acid (IBA) at 1.0 mg/L. The regenerated plantlets were acclimatized in pots containing sterile garden soil mixture to examine their response in natural conditions.

Plantlets Regeneration via Somatic Embryogenesis from the Nucellus Tissues of Kinnow Mandarin (Citrus reticulata L.)

Studies were initiated to explore the role of nucellus tissues and growth regulators in plantlets regeneration via somatic embryogenesis of Kinnow mandarin [Citrus reticulata L. (Blanco)]. Nucellus tissues were cultured on MS medium supplemented with different concentrations and combinations of auxins, cytokinins and malt extract for primary callus induction. The best response for primary callus induction (90%) was obtained when MS medium was supplemented with 5 mg/l 2,4-D and 500 mg/l malt extract. Best results for embryogenic callus induction (80%) were obtained in C8 medium. The induction of somatic embryos was highest when MS medium was supplemented with 1 mg/l BAP and maturation of somatic embryos occurred when MS medium was supplemented with 5 mg/l 2,4-D and 1 mg/l BAP. Maximum plantlets were regenerated (92%) from the somatic embryos on half strength MS medium with no hormones. The plantlets were successfully acclimatized in different potting mixtures and highest survival rate (100%) was achieved in potting mixture containing sand and peat moss (2:1).

Protective effect of Citrus reticulata peel extract against potassium dichromate-induced reproductive toxicity in rats

Objective:To study the protective effect of Citrus reticulata peel extract against potassium dichromate-induced reproductive impairment in male Wistar rats.Methods:A total of 35 male Wistar rats were equally divided into 5 groups.The control group orally received an equivalent volume of saline(1 mL);group 2 was given potassium dichromate at 10 mg/kg/body weight(b.w.)orally for eight weeks;group 3 and 4 orally received Citrus reticulata peel extract at 200 and 400 mg/kg/b.w.respectively,followed by treatment with potassium dichromate.Group 5 was administered with 400 mg/kg/b.w.Citrus reticulata peel extract only.The parameters including oxidative stress(malondialdehyde,antioxidant enzymes),sex hormones(testosterone,luteinizing hormone,follicle-stimulating hormone)and inflammation indexes(tumor necrosis factor-alpha,inter leukin 6)were measured.Sperm characteristics(sperm morphology,motility and count)were also determined.Results:The decrease of male sex hormones concentration in plasma(testosterone,luteinizing hormone),testicular antioxidant parameters(reduced glutathione,catalase,superoxide dismutase,carnitine),sperm motility and sperm count induced by potassium dichromate was mitigated by Citrus reticulata peel extract.Moreover,the peel extract alleviated the increase of sperm abnormalities,follicle-stimulating hormone,testicular malondialdehyde,tumor necrosis factor-alpha and nitric oxide levels caused by potassium dichromate.The sections of rats treated with the potassium dichromate were characterized by some histopathological changes as disorganization of spermatogonial cell layers and deformation of Leydig cells.A positive immune-reaction for p53 was observed in testicular cells of the rats treated with potassium dichromate.The extract of Citrus reticulata peel improved the testicular histology and decreased p53 positive immune-reaction in potassium dichromate administered rats.Conclusions:Citrus reticulata peel extract can alleviate the reproductive toxicity triggered by potassium dichromatevia its antioxidant and anti-inflammatory properties.Citrus reticulata peel extract may act as a new natural prophylactically agent for the treatment of potassium dichromate-induced testicular damage.

Effects of Storage and Preservation of Postharvest Gonggan Mandarin(Citrus reticulata)Fruit Immersion in Salicylic Acid Combined with Calcium Chloride

[Objectives]To investigate the effects and mechanism of fruit immersion treatment with salicylic acid(SA),calcium chloride(CaCl_(2))solutions at different concentrations or their combinations on post-harvest preservation of Gonggan Mandarin(Citrus reticulata)fruit.[Methods]13 experiments(2 storage temperatures,3 control treatments,3 concentrations of SA solution,4 concentrations of CaCl_(2) solution and 3 combinations of SA+CaCl_(2) solution for fruit immersion treatment)were conducted for 90 d.Gonggan fruit weight loss rate,rotting rate,disease index,disease type,soluble solids and organic acids content,and the differences in the activity of six enzymes(APX,PPO,CHI,CAT,SOD,PAL)related to fruit disease resistance were detected regularly.[Results]With the increase of storage days,the weight loss rate,rotting rate and disease index increased,the content of soluble solids and organic acids decreased,the activity of APX,PPO,CHI and SOD increased,and the activity of CAT and PAL decreased.However,the increase or decrease of fruit loss at low temperature was the smallest,and the increase or decrease of fruit at natural room temperature was the largest.Gonggan fruit cleaning before storage is an effective preservation and fresh-keeping technology,but its preservation and fresh-keeping effect is far less than that of fruit immersion by SA,CaCl_(2) and SA+CaCl_(2) compound solution.The effect of fruit immersion by SA solution was greater than that of fruit immersion by CaCl_(2) solution;the best concentration of SA alone was 6.0 mmol/L,which was not significantly different from 3.0 mmol/L,but the effect of 9.0 mmol/L decreased;the best concentration of CaCl_(2) alone was 6.0 mmol/L,which was significantly better than 3.0 and 9.0 mmol/L;the optimal concentration combination of SA+CaCl_(2) compound solution for fruit immersion treatment was 6.0 mmol/L SA+3.0 mmol/L CaCl_(2).[Conclusions]The fruit immersion by SA,CaCl_(2) and their compound solutions induced the increase of activity of six enzymes related to disease resistance in Gonggan fruit,and PPO and CHI were judged to be two key enzymes based on response intensity.

Omics analyses in citrus reveal a possible role of RNA translation pathways and Unfolded Protein Response regulators in the tolerance to combined drought, high irradiance, and heat stress

Environmental changes derived from global warming and human activities increase the intensity and frequency of stressful conditions for plants.Multiple abiotic factors acting simultaneously enhance stress pressure and drastically reduce plant growth,yield,and survival.Stress combination causes a specific stress situation that induces a particular plant response different to the sum of responses to the individual stresses.Here,by comparing transcriptomic and proteomic profiles to different abiotic stress combinations in two citrus genotypes,Carrizo citrange(Citrus sinensis×Poncirus trifoliata)and Cleopatra mandarin(Citrus reshni),with contrasting tolerance to different abiotic stresses,we revealed key responses to the triple combination of heat stress,high irradiance and drought.The specific transcriptomic response to this stress combination in Carrizo was directed to regulate RNA metabolic pathways and translation processes,potentially conferring an advantage with respect to Cleopatra.In addition,we found endoplasmic reticulum stress response as common to all individual and combined stress conditions in both genotypes and identified the accumulation of specific groups of heat shock proteins(HSPs),such as small HSPs and HSP70s,and regulators of the unfolded protein response,BiP2 and PDIL2-2,as possible factors involved in citrus tolerance to triple stress combination.Taken together,our findings provide new insights into the acclimation process of citrus plants to multiple stress combination,necessary for increasing crop tolerance to the changing climatic conditions.