A chromosome-level genome assembly for Chinese plum‘Wushancuili'reveals the molecular basis of its fruit color and susceptibility to rain-cracking

Chinese plum(Prunus salicina Lindl.)originates from China and makes a large contribution to the global production of plums.The P.salicina‘Wushancuili'has a green coloration and high fruit quality and is economically important in eliminating poverty and protecting ecology in the Yangtze River Three Gorges Reservoir.However,rain-induced cracking(rain-cracking,literally skin cracking caused by rain)is a limitation to‘Wushancuili'fruit production and causes severe losses.This study reported a high-quality‘Wushancuili'genome assembly consisting of a 302.17-Mb sequence with eight pseudo-chromosomes and a contig N50 of 23.59 Mb through the combination of Illumina sequencing,Pacific Biosciences HiFiⅢsequencing,and high-throughput chromosome conformation capture technology.A total of 25109 protein-coding genes are predicted and 54.17%of the genome is composed of repetitive sequences.‘Wushancuili'underwent a remarkable orthoselection during evolution.Gene identification revealed that loss-of-function in four core MYB10 genes results in the anthocyanin deficiency and absence of red color,revealing the green coloration due to the residual high chlorophyll in fruit skin.Besides,the occurrence of cracking is assumed to be closely associated with cell wall modification and frequently rain-induced pathogen enrichment through transcriptomic analysis.The loss of MYB10 genes might render fruit more susceptible to pathogen-mediated cracking by weakening the epidermal strength and reactive oxygen species(ROS)scavenging.Our findings provided fundamental knowledge regarding fruit coloration and rain-cracking and will facilitate genetic improvement and cultivation management in Chinese plums.

Genome-wide analysis of Panonychus citrimicro RNAs witha focus on potential insecticidal activity of 4 micro RNAs toeggs and nymphs

Panonychus citriMcGregor(Acari:Tetranychidae),a destructive citrus pest,causes considerable annual economic losses due to its short lifespan and rapid resistancedevelopment.MicroRNA(miRNA)-induced RNA interference is a promising approachfor pest control because of endogenous regulation of pest growth and development.Tosearch for miRNAs with potential insecticidal activity inP.citri,genome-wide analysis ofmiRNAs at different developmental stages was conducted,resulting in the identificationof 136 miRNAs,including 73 known and 63 novel miRNAs.A total of 17 isomiRNAsand 12 duplicated miRNAs were characterized.MiR-1 and miR-252-5p were identified asreference miRNAs forP.citriandTetranychus urticae.Based on differential expressionanalysis,treatments with miR-let-7a and miR-315 mimics and the miR-let-7a antagomirsignificantly reduced the egg hatch rate and resulted in abnormal egg development.Over-expression or downregulation of miR-34-5p and miR-305-5p through feeding significantlydecreased the adult eclosion rate and caused molting defects.The 4 miRNAs,miR-let-7a,miR-315,miR-34-5p,and miR-305-5p,had important regulatory functions and insectici-dal properties in egg hatching and adult eclosion.In general,these data advance our un-derstanding of miRNAs in mite biology,which can assist future studies on insect-specificmiRNA-based green pest control technology.

EjGASA6 promotes flowering and root elongation by enhancing gibberellin biosynthesis

The Gibberellic Acid-stimulated Arabidopsis(GASA)gene family is involved in the regulation of gene expression and plant growth,development,and stress responses.To investigate the function of loquat GASA genes in the growth and developmental regulation of plants,a loquat EjGASA6 gene homologous to Arabidopsis AtGASA6 was cloned.EjGASA6 expression was induced by gibberellin,and ectopic transgenic plants containing this gene exhibited earlier bloom and longer primary roots since these phenotypic characteristics are related to higher gibberellin content.Transcriptome analysis and qRT-PCR results showed that the expression levels of GA3ox1 and GA20ox1,which encode key enzymes in gibberellin biosynthesis,were significantly increased.Furthermore,we confirmed that EjGASA6 could promote the expression of GA20ox1 via the luciferase reporter system.Overall,our results suggest that EjGASA6 promotes blooming and main-root elongation by positively regulating gibberellin biosynthesis.These findings broaden our understanding of the role of GASAs in plant development and growth,and lay the groundwork for future research into the functions of EjGASA6 in regulating loquat growth and development.

Novel flavin-containing monooxygenase protein FMO1 interacts with CAT2 to negatively regulate drought tolerance through ROS homeostasis and ABA signaling pathway in tomato

Drought stress is themajor abiotic factor that can seriously affect plant growth and crop production.The functions of flavin-containing monooxygenases(FMOs)are known in animals.They addmolecular oxygen to lipophilic compounds or produce reactive oxygen species(ROS).However,little information on FMOs in plants is available.Here,we characterized a tomato drought-responsive gene that showed homology to FMO,and it was designated as FMO1.FMO1 was downregulated promptly by drought and ABA treatments.Transgenic functional analysis indicated that RNAi suppression of the expression of FMO1(FMO1-Ri)improved drought tolerance relative to wild-type(WT)plants,whereas overexpression of FMO1(FMO1-OE)reduced drought tolerance.The FMO1-Ri plants exhibited lower ABA accumulation,higher levels of antioxidant enzyme activities,and less ROS generation comparedwith theWTand FMO1-OE plants under drought stress.RNA-seq transcriptional analysis revealed the differential expression levels of many drought-responsive genes thatwere co-expressed with FMO1,including PP2Cs,PYLs,WRKY,and LEA.Using Y2H screening,we found that FMO1 physically interacted with catalase 2(CAT2),which is an antioxidant enzyme and confers drought resistance.Our findings suggest that tomato FMO1 negatively regulates tomato drought tolerance in the ABA-dependent pathway and modulates ROS homeostasis by directly binding to SlCAT2.

Intron-retained alternatively spliced VIN3-LIKE 2 gene from Chimonanthus praecox promotes flowering in transgenic Arabidopsis

Vernalization is a process of acquiring or accelerating the flowering ability by prolonged cold exposure.VERNALIZATION INSENSITIVE3(VIN3)is induced by chilling and is extremely important for the vernalization response in Arabidopsis thaliana.However,the issue of the induction of the VIN3-LIKE genes in wintersweet(Chimonanthus praecox)has been largely neglected.In the present study,we explored the molecular regulation of the PHD type finger protein-encoding gene CpVIL2 in relation to the growth and development of wintersweet in Arabidopsis.In wintersweet,quantitative real-time PCR(qRT-PCR)analysis showed that the relative expression of CpVIL2-As2i(intron-retained alternatively spliced in the second intron)was extremely higher in the pistils than in the other tissues.And the relative CpVIL2-As2i expression in flower buds(FBs)treated at 8°C was higher than that of FBs in December,2016 under natural conditions,which was not detected in non-flowering FBs at 16°C.In Arabidopsis,the expression patterns of the CpVIL2-As2i gene were detected at first in CpVIL2-As2i pro::GUS(β-glucuronidase)lines,with predominantly higher expression in flowers and inflorescence.Meanwhile,the hormone-induced expression profiles of the CpVIL2-As2i promoter were confirmed using exogenous induction by abscisic acid(ABA)and indole acetic acid(IAA)phytohormones,where the GUS enzyme activity obviously decreased compared with that of control.In comparison with Arabidopsis/Col-0,early flowering was detected in ectopic 35S::CpVIL2-As2i lines.Overall,these results demonstrated the function of the CpVIL2-As2i gene,at the same time,provided us with new insights into the molecular mechanisms of early flowering and complex regulatory networks of vernalization in wintersweet.

RNA interference in insects:the link between antiviral defense and pest control

RNA interference(RNAi)is a form of gene silencing triggered by double-stranded RNA(dsRNA)that operates in all eukaryotic cells.RNAi has been widely investigated in insects to determine the underlying molecular mechanism,to investigate its role in systemic antiviral defense,and to develop strategies for pest control.When insect cells are infected by viruses,viral dsRNA signatures trigger a local RNAi response to block viral replication and generate virus-derived DNA that confers systemic immunity.RNAi-based insect pest control involves the application of exogenous dsRNA targeting genes essential for insect development or survival,but the efficacy of this approach has limited potency in many pests through a combination of rapid dsRNA degradation,inefficient dsRNA uptake/processing,and ineffective RNAi machinery.This could be addressed by dsRNA screening and evaluation,focusing on dsRNA design and off-target management,as well as dsRNA production and delivery.This review summarizes recent progress to determine the role of RNAi in antiviral defense and as a pest control strategy in insects,addressing gaps between our fundamental understanding of the RNAi mechanism and the exploitation of RNAi-based pest control strategies.