Retrotransposons account for a large proportion of the genome and genomic variation, and play key roles in creating novel genes and diversifying the genome in many eukaryotic species. Although retrotransposons are abu...Retrotransposons account for a large proportion of the genome and genomic variation, and play key roles in creating novel genes and diversifying the genome in many eukaryotic species. Although retrotransposons are abundant in plants, their roles had been underestimated because of a lack of research. Here, we characterized a gibberellin Acid (GA)-insensitive dwarf mutant, 84133, in foxtail millet. Map-based cloning revealed a 5.5-kb Copia-like retrotransposon insertion in DWARF1 (D1), which encodes a DELLA protein. Transcriptional analysis showed that the Copia retrotransposon mediated the transcriptional reprogramming of D1 leading to a novel N-terminal-deleted truncated DELLA transcript that was putatively driven by Copia's LTR, namely D1-TT, and another chimeric transcript. The presence of D1-TT was confirmed by protein immunodetection analysis. Furthermore, D1-TT protein was resistant to GA3 treatment compared with the intact DELLA protein due to its inability to interact with the GA receptor, SiGID1. Overexpression of D1-TT in foxtail millet resulted in dwarf plants, confirming that it determines the dwarfism of 84133. Thus, our study documents a rare instance of long terminal repeat (LTR) retrotransposon-mediated transcriptional reprograming in the plant kingdom. These results shed light on the function of LTR retrotransposons in generating new gene functions and genetic diversity.展开更多
One possible mechanism suggested for somaclonal variation is the activation of transposable elements. The activation of retrotransposons by stresses and external changes is commonly observed in plants. In previous stu...One possible mechanism suggested for somaclonal variation is the activation of transposable elements. The activation of retrotransposons by stresses and external changes is commonly observed in plants. In previous study, we isolated the reverse transcriptase （RT） gene sequences of Ty 1-copia retrotransposons from tissue culture strawberry （Fragaria x ananassa） plant, but not the transcriptionally active sequence. For further understanding the relationship between retrotransposon and somaclonal varation, in this study, we isolated the transcriptionally active RT gene sequences from strawberry plants subjected to different abiotic stresses. These retrotransposons were activated by spraying strawberry leaves with 2 mmol L^-1 salicylic acid （SA）, 50 mmol L^-1 methyl jasmonate （MeJA）, 50 mmol L^-1 abscisic acid （ABA）, 50 mmol L^-1 2,4- dichlorophenoxyacetic acid （2,4-D） or by inducing callus growth in 2 types of MS media： first medium supplemented with 0.5 mg L^-1 6-benzylaminopurine （6-BA）, 0.5 mg L^-1 gibberellic acid （GA3）, 1.0 mg L^-1 thidiazuron （TDZ）, and 0.1 mg L^-1 2,4-D, and the second medium supplemented with 0.5 mg L^-1 6-BA, 0.5 mg L^-1 GA3, 2.0 mg L^-1 TDZ, and 0.02 mg L^-1 indole butyric acid （1BA）. Analysis of gene sequences of 17 RTs revealed that none of them contained stop codons and/or indels disrupting the reading frame. These different stress-origin transcriptionally active RTs were remarkably similar to each other- FATEXP2-8 and FATEYS9-7 showed 100% sequence identity. Analysis of pylogenetic of these transcriptionally active RTs and the RT sequences from genome showed that there were close phylogenetic relationships of most of the transcriptionally active RTs. The results of this study have contributed to the background information necessary for future studies for evaluating the relationship between retrotransposons and somaclonal variation.展开更多
Retrotransposon-like elements are major constituents of most eukaryotic genomes. For example, they account for roughly 90% of the wheat （Triticum aestivum） genome, Previous study on a wheat strain treated by low-ene...Retrotransposon-like elements are major constituents of most eukaryotic genomes. For example, they account for roughly 90% of the wheat （Triticum aestivum） genome, Previous study on a wheat strain treated by low-energy N^＋ ions indicated the variations in AFLP （Amplified Fragment Length Polymorphism ） markers, One such variation was caused by the re-activation of Tyl-copia-like retrotransposons, implying that the mutagenic effects of lowenergy ions might work through elevated activation of retrotransposons, In this paper an expression profile of Tyl-copia-like retrotransposons in wheat treated by low-energy N^＋ ions is reported, The reverse transcriptase （RT） domains of these retrotransposons were amplified by reverse-transcriptional polymerase chain reaction （RT-PCR） and sequentially cloned, 42 and 65 clones were obtained from the treated （CL） and control materials （CK）, respectively, Sequence analysis of each clone was performed by software. Phylogeny and classification were calculated responding to the sequences of the RT domains. All the results show that there is much difference in the RT domain between the control sample and the treated sample, Especially, the RT domains from the treated group encode significantly more functional ORF （open reading frames） than those from the control sample, This observation suggests that the treated sample has higher activation of retrotransposons, possibly as a consequence of low-energy ion beam irradiation, It also suggests that retrotransposons in the two groups impact the host gene expression in two different ways and carry out different functions in wheat cells.展开更多
A novel rice d1 mutant was identified using map-based cloning and comparative analysis of known dl mutants.The mutant[d1-a) shows a mild dwarf trait,which differs only slightly from the wildtype in plant height at the...A novel rice d1 mutant was identified using map-based cloning and comparative analysis of known dl mutants.The mutant[d1-a) shows a mild dwarf trait,which differs only slightly from the wildtype in plant height at the tillering stage.The dl-a mutant is different from other dl mutants.We found that it was interrupted by an Osr4 long terminal repeat(LTR)-retrotransposon,which resulted in the loss of exon 7 in the mutant D1 mRNA.A paralog of the D1 gene.D1-like,was revealed.D1-like is a truncated gene that might have resulted from recombination between retrotransposons.We identified 65 Osr4LTR-retrotransposons in Nipponbare,and found more LTR variants in contrast to coding DNA sequence(CDS) in the retrotransposons.We also identified five possible regulatory motifs in LTRs which may control the expression of the retrotransposons.In addition,we predicted six putative functional Osr4 retrotransposons that contain complete CDSs and all important elements.Osr4 retrotransposons were classified into 4 groups,and this type of retrotransposon only appears to be present in monocots.Members of group 1-1,which included all putative functional retrotransposons,showed a high similarity with each other.The retrotransposons were expressed in all tissues,at especially higher levels in some leaves and seeds.These findings imply that transpositions of group 1-1 members might have occurred frequently and recently.展开更多
Mutagenesis and retrotransposons have a close relationship, but little attention has been paid yet to the activity of retrotransposons produced by physical mutagens. The variation of retrotransposon WIS 2-1A activity ...Mutagenesis and retrotransposons have a close relationship, but little attention has been paid yet to the activity of retrotransposons produced by physical mutagens. The variation of retrotransposon WIS 2-1A activity in wheat （Triticum aestivum L.） embryos at three different growth times （30 h, 45 h and 60 h） was investigated after they had been treated with N^＋ implantation in a vacuum of 5× 10^-2 Pa and irradiation by ^60Coγ-ray respectively. For each of the three growth times the expression of WIS 2-1A showed almost entirely a same trend of downregulation, upregulation, then downregulation, and upregulation again with the increase in dose of N^＋ implantation, but the expression appeared irregular with the increase in irradiation of ^60Coγ-ray. In conclusion, the acutely activating effect of WIS 2-1A stimulated by vacuum and high dose N^＋ implantation within a shorter incubation time may provide a convenient tool to advance the research on mutagenic breeding and function genes.展开更多
In order to identify the variation and estimate the genetic diversity among the fig (Ficus carica L.) genotypes collected from Algeria and Turkey, the genetic relationships between 86 genotypes were investigated using...In order to identify the variation and estimate the genetic diversity among the fig (Ficus carica L.) genotypes collected from Algeria and Turkey, the genetic relationships between 86 genotypes were investigated using 23 inter primer binding sites (iPBS)-retrotransposon and 16 simple sequence repeat (SSR) primers. A total of 63 polymorphic bands for the iPBS-retrotransposon markers and 25 alleles for the SSR markers were identified with an average of 2.7 and 1.6 per primer, respectively. The average value of polymorphism information content (PIC) for the iPBS markers (0.73) was higher than that for the SSR markers (0.69). Applying the neighbor-joining method to the combined iPBS-retrotransposon and SSR data, the fig genotypes were clustered into two groups. The STRUCTURE software was used to determine the population structure. Among the genotypes studied, two populations (K = 2) were identified indicating a low diversity between the Algerian and Turkish varieties. Both types of markers were able to differentiate all the fig genotypes and were efficient in discriminating the closely related genotypes. Our data also showed that as a universal marker, iPBS-retrotransposon is a useful tool for the molecular characterization of fig genotypes.展开更多
Long terminal repeat （LTR） retrotransposons, one of the foremost types of transposons, continually change or modify gene function and reorganize the genome through bursts of dramatic proliferation. Many LTR-TEs pref...Long terminal repeat （LTR） retrotransposons, one of the foremost types of transposons, continually change or modify gene function and reorganize the genome through bursts of dramatic proliferation. Many LTR-TEs preferen-tially insert within other LTR-TEs, but the cause and evolutionary significance of these nested LTR-TEs are not well under-stood. In this study, a total of 1.52 Gb of Brassica sequence containing 2020 bacterial artificial chromosomes （BACs） was scanned, and six bacterial artificial chromosome （BAC） clones with extremely nested LTR-TEs （LTR-TEs density： 7.24/kb） were selected for further analysis. The majority of the LTR-TEs in four of the six BACs were found to be derived from the rapid proliferation of retrotransposons originating within the BAC regions, with only a few LTR-TEs originating from the proliferation and insertion of retrotransposons from outside the BAC regions approximately 5-23 Mya. LTR-TEs also pref-erably inserted into TA-rich repeat regions. Gene prediction by Genescan identified 207 genes in the 0.84Mb of total BAC sequences. Only a few genes （3/207） could be matched to the Brassica expressed sequence tag （EST） database, indicating that most genes were inactive after retrotransposon insertion. Five of the six BACs were putatively centromeric. Hence, nested LTR-TEs in centromere regions are rapidly duplicated, repeatedly inserted, and act to suppress activity of genes and to reshuffle the structure of the centromeric sequences. Our results suggest that LTR-TEs burst and proliferate on a local scale to create nested LTR-TE regions, and that these nested LTR-TEs play a role in the formation of centromeres.展开更多
Histone lysine methylation plays an important role in heterochromatin formation and reprogramming of gene expression. SET-domain-containing proteins are shown to have histone lysine methyltransferase activities. A lar...Histone lysine methylation plays an important role in heterochromatin formation and reprogramming of gene expression. SET-domain-containing proteins are shown to have histone lysine methyltransferase activities. A large number of SET-domain genes are identified in plant genomes. The function of most SET-domain genes is not known. In this work, we studied the 12 rice （Oryza sativa） homologs of Su（var）3-9, the histone H3 lysine 9 （H3K9） methyltransferase identified in Drosophila. Several rice SUVHs （i.e. SDG714, SDG727, and SDG710） were found to have an antagonistic func- tion to the histone H3K9 demethylase JMJ706, as down-regulation of these genes could partially complement the jmj706 phenotype and reduced histone H3K9 methylation. Down-regulation of a rice Su（var）3-9 homolog （SUVH）, namely SDG728, decreased H3K9 methylation and altered seed morphology. Overexpression of the gene increased H3K9 methylation. SDG728 and other SUVH genes were found to be involved in the repression of retrotransposons such as Tos17 and a Tyl-copia element. Analysis of histone methylation suggested that SDG728-mediated H3K9 methylation may play an important role in retrotransposon repression.展开更多
As the largest transposable element in the plant genome, retrotransposons are thought to be involved in citrus genetic instability and genome evolution, especially in sweet orange, which is prone to bud mutation. In t...As the largest transposable element in the plant genome, retrotransposons are thought to be involved in citrus genetic instability and genome evolution, especially in sweet orange, which is prone to bud mutation. In the present study, the presence of copia-like retrotransposons, their heterogeneity, genomic distribution, and transcriptional activities in Citrus were investigated in 12 sweet orange （Citrus sinensis Osbeck） cultivars using a PCR assay designed to detect copia-like reverse transcriptase （RT） sequences. Twelve amplification products from each cultivar were cloned and sequenced. The cloned sequences showed great heterogeneity, except “Dream” navel and “Hamlin”, both of which shared the same sequence. Frame shifting, termination, deletion, and substitution accounted for the heterogeneity of RT sequences. Southern blot hybridization using the RT1 clone from the “Cara Cara” navel as a probe showed that multiple copies were integrated throughout the sweet orange genomes, which made the retrotransposon possible an effective molecular marker to detect citrus evolution events and to reveal its relationship with bud mutation. No transcriptional activities of the retrotransposon were detected by RT-PCR and Northern analysis in the fruits and leaves of either “Cara Cara” or “Seike” navels.展开更多
The conserved domains of reverse transcriptase （RT） genes of Tyl-copia and Ty3-gypsy groups of long terminal repeat （LTR） retrotransposons were isolated from the Malus domestica genome using degenerate oligonucleo...The conserved domains of reverse transcriptase （RT） genes of Tyl-copia and Ty3-gypsy groups of long terminal repeat （LTR） retrotransposons were isolated from the Malus domestica genome using degenerate oligonucleotide primers. Sequence analysis showed that 45% of Ty1-copia and 63% of Ty3-gypsy RT sequences contained premature stop codons and/or indels disrupting the reading frame. High heterogeneity among RT sequences of both Ty1-copia and Ty3-gypsy group retrotransposons was observed, but Ty3-gypsy group retrotransposons in the apple genome are less heterogeneous than Ty1-copia elements. Retrotransposon copy number was estimated by dot blot hybridizations for Ty1-copia （-5000） and Ty3-gypsy （-26000）. All elements of the two types of LTR retrotransposons comprise approximately 38% of the M. domestica genome, with the Ty3-gypsy group contribution being higher （33.5%） than the Tyl.copia one （4.6%）. Transcription was not detected by reverse transcription-polymerase chain reaction for either Ty1-copia or Ty3-gypsy retrotransposons in the leaves of plants in vitro or in leaf explants cultured on medium supplemented with high concentration benzylaminopurine. This research reveals the differences in heterogeneity and copy number between Ty1-copia and Ty3-gypsy retrotransposons in the apple genome. Ty1-copia retrotransposon has higher heterogeneity than Ty3-gypsy retrotransposon, but the latter has a higher copy number, which implies that Ty3-gypsy retrotransposons may play a more important role in the apple genome evolution.展开更多
Eight intact LTR retrotransposons （Nbr1-Nbr8） have been previously characterized from the genome of Nosema bombycis, a eu- karyotic parasite with a compact and reduced genome. Here we describe six novel transcribed ...Eight intact LTR retrotransposons （Nbr1-Nbr8） have been previously characterized from the genome of Nosema bombycis, a eu- karyotic parasite with a compact and reduced genome. Here we describe six novel transcribed Nbr elements （Nbr9-Nbr14） identified through either cDNA library or RT-PCR. Like previously determined ones, all of them belong to the Ty3/Gypsy superfamily. Retrotransposon diversity and incomplete domains with insertions （Nbr12）, deletions （Nbrll） and in-frame stop codons in coding regions （Nbr9） were detected, suggesting that both defective and loss events of LTR retrotransposon have happened in N. bornbycis genome. Analysis of selection showed that strong purifying selection acts on all elements except Nbr11. This implies that selective pressure keeps both these Nbrs and their functions in genome. Interestingly, Nbrll is under positive selection and some positively selected codons were identified, indicating that new functionality might have evolved in the Nbrll retrotransposon. Unlike other transposable elements, Nbrll has integrated into a conserved syntenic block and probably resulted in the inversion of both flanking regions. This demonstrates that transposable element is an important factor for the reshuffling and evolution of their host genomes, and may be maintained under natural selection.展开更多
Transposable elements(TEs)constitute the most abundant portions of plant genomes and can dramatically shape host genomes during plant evolution.They also play important roles in crop domestication.However,whether TEs ...Transposable elements(TEs)constitute the most abundant portions of plant genomes and can dramatically shape host genomes during plant evolution.They also play important roles in crop domestication.However,whether TEs themselves are also selected during crop domestication has remained unknown.Here,we identify an active long terminal repeat(LTR)retrotransposon,HUO,as a potential target of selection during rice domestication and breeding.HUO is a low-copy-number LTR retrotransposon,and is active under natural growth conditions and transmitted through male gametogenesis,preferentially inserting into genomic regions capable of transcription.HUO exists in all wild rice accessions and about half of the archaeological rice grains(1200–7000 years ago)and landraces surveyed,but is absent in almost all modern varieties,indicating its gradual elimination during rice domestication and breeding.Further analyses showed that HUO is subjected to strict gene silencing through the RNA-directed DNA methylation pathway.Our results also suggest that multiple HUO copies may trigger genomic instability through altering genome-wide DNA methylation and small RNA biogenesis and changing global gene expression,resulting in decreased disease resistance and yield,coinciding with its elimination during rice breeding.Together,our study suggests that negative selection of an active retrotransposon might be important for genome stability during crop domestication and breeding.展开更多
Stochastic introgression of alien DNA may impose a genomic stress to the recipient genome. Herein, we report that apparent de novo genomic rearrangements in 10 of 13 selected endogenous, lowcopy, and potentially activ...Stochastic introgression of alien DNA may impose a genomic stress to the recipient genome. Herein, we report that apparent de novo genomic rearrangements in 10 of 13 selected endogenous, lowcopy, and potentially active long terminal repeat （LTR） retrotransposons occurred in one or more of three rice lines studied that were introgressed by wild rice （Zizania latifolia Griseb.）. For nine retrotransposons in which both the reverse-transcriptase （RT） region and the LTR region were available, largely concordant rearrangements occurred at both regions in five elements and at the RT region only in the remaining four elements. A marked proportion of the genomic changes was shared by two or all three introgression lines that were derived from a single F~ plant. This indicates that most of the genomic changes occurred at early developmental stages of the F~ somatic cells, which then gave rise to germline cells, and, hence, ensured inheritance of the changes to later generations. Possible causes and potential implications of the introgression-induced genomic rearrangements in LTR retrotransposons are discussed in the context of plant genome evolution and breeding.展开更多
Long terminal repeat retrotransposons(LTR-RTs) are a predominant group of plant transposable elements(TEs) that are an important component of plant genomes. A large number of LTR-RTs have been annotated in the genomes...Long terminal repeat retrotransposons(LTR-RTs) are a predominant group of plant transposable elements(TEs) that are an important component of plant genomes. A large number of LTR-RTs have been annotated in the genomes of the agronomically important oil and vegetable crops of the genus Brassica. Herein, full-length LTR-RTs in the genomes of Brassica and other closely related species were systematically analyzed. The full-length LTR-RT content varied greatly(from 0.43% to 23.4%) between different species, with Gypsy-like LTR-RTs constituting a primary group across these genomes. More importantly, many annotated LTR-RTs(from 10.03% to 33.25% of all detected LTR-RTs) were found to be enriched in localized hotspot regions. Furthermore, all of the analyzed species showed evidence of having experienced at least one round of a LTR-RT burst, with Raphanus sativus experiencing three or more. Moreover, these relatively ancient LTR-RT amplifications exhibited a clear expansion at specific time points. To gain a further understanding of this timing, Brassica rapa, B. oleracea, and R. sativus were examined for the presence of syntenic regions, but none were present. These findings indicate that these LTR-RT burst events were not inherited from a common ancestor,but instead were species-specific bursts that occurred after the divergence of Brassica species. This study further exemplifies the complexities of TE amplifications during the evolution of plant genomes and suggests that these LTR-RT bursts play an important role in genome expansion and divergence in Brassica species.展开更多
New classes of repetitive DNA elements were effectively identified by isolating small fragments of the elements from the wheat genome. A wheat A genome library was constructed from Triticum monococcum by degenerate cl...New classes of repetitive DNA elements were effectively identified by isolating small fragments of the elements from the wheat genome. A wheat A genome library was constructed from Triticum monococcum by degenerate cleavage with EcoO1091, the recognition sites of which consisted of 5＇-PuGGNCCPy-3＇ multi-sequences. Three novel repetitive sequences pTm6, pTm69 and pTm58 derived from the A genome were screened and tested for high copy number using a blotting approach, pTm6 showed identity with integrase domains of the barley Tyl-Copia-retrotransposon BARE-1 and pTm58 showed similarity to the barley Ty3-gypsy-like retrotransposon Romani. pTm69, however, constituted a tandem array with useful genomic specificities, but did not share any identity with known repetitive elements. This study also sought to isolate wheat D-genome-specific repetitive elements regardless of the level of methylation, by genomic subtraction. Total genomic DNA of Aegilops tauschii was cleaved into short fragments with a methylation-insensitive 4 bp cutter, Mbol, and then common DNA sequences between Ae. tauschii and Triticum turgidum were subtracted by annealing with excess T. turgidum genomic DNA. The D genome repetitive sequence pAt1 was isolated and used to identify an additional novel repetitive sequence family from wheat bacterial artificial chromosomes with a size range of 1 395-1 850 bp. The methods successfully led pathfinding of two unique repetitive families.展开更多
The dynamic activity of transposable elements(TEs)contributes to the vast diversity of genome size and architecture among plants.Here,we examined the genomic distribution and transposition activity of long terminal re...The dynamic activity of transposable elements(TEs)contributes to the vast diversity of genome size and architecture among plants.Here,we examined the genomic distribution and transposition activity of long terminal repeat retrotransposons(LTR-RTs)in Arabidopsis thaliana(Ath)and three of its relatives,Arabidopsis lyrata(Aly),Eutrema salsugineum(Esa),and Schrenkiella parvula(Spa),in Brassicaceae.Our analyses revealed the distinct evolutionary dynamics of Gypsy retrotransposons,which reflects the different patterns of genome size changes of the four species over the past million years.The rate of Gypsy transposition in Aly is approximately five times more rapid than that of Ath and Esa,suggesting an expanding Aly genome.Gypsy insertions in Esa are strictly confined to pericentromeric heterochromatin and associated with dramatic centromere expansion.In contrast,Gypsy insertions in Spa have been largely suppressed over the last million years,likely as a result of a combination of an inherent molecular mechanism of preferential DNA removal and purifying selection at Gypsy elements.Additionally,species-specific clades of Gypsy elements shaped the distinct genome architectures of Aly and Esa.展开更多
Tnt1 is an active retrotransposon originally identified in tobacco(Nicotiana tabacum L.)(Grandbastien et al.,1989),but its transposition activity could be activated through tissue culture in other plant species.The in...Tnt1 is an active retrotransposon originally identified in tobacco(Nicotiana tabacum L.)(Grandbastien et al.,1989),but its transposition activity could be activated through tissue culture in other plant species.The insertions are stable and inheritable in the progeny,which has made it a valuable and versatile tool for developing insertional mutagenesis libraries in several plant species.Here,we explored its utility for mutagenesis in cucumber(Cucumis sativus L.).T_3 Tnt1 transgenic cucumber plants were subjected to tissue culture to regenerate self-pollinated progeny.With PCR and analyses and Southern hybridization,we found regenerated plants maintained the original Tnt1 insertion and created new insertions suggesting characteristic re-transposition activity of Tnt1 during this process.Using genome walking,some flanking sequences of Tnt1 insertions were recovered in regenerated plants.The results demonstrated that Tnt1 could be stably inherited and re-transposable during tissue culture in cucumber and that it is feasible to use for developing an insertional mutagenesis library for cucumber.展开更多
Soybean(Glycine max) is an important legume crop that was domesticated in temperate regions.Soybean varieties from these regions generally mature early and exhibit extremely low yield when grown under inductive short-...Soybean(Glycine max) is an important legume crop that was domesticated in temperate regions.Soybean varieties from these regions generally mature early and exhibit extremely low yield when grown under inductive short-day(SD) conditions at low latitudes. The long-juvenile(LJ) trait, which is characterized by delayed flowering and maturity,and improved yield under SD conditions, allowed the cultivation of soybean to expand to lower latitudes. Two major loci control the LJ trait: J and E6. In the current study, positional cloning, sequence analysis, and transgenic complementation confirmed that E6 is a novel allele of J, the ortholog of Arabidopsis thaliana EARLY FLOWERING 3(ELF3). The mutant allele e6^(PG), which carries a Ty1/Copia-like retrotransposon insertion, does not suppress the legume-specific flowering repressor E1, allowing E1 to inhibit Flowering Locus T(FT) expression and thus delaying flowering and increasing yields under SD conditions. The e6^(PG)allele is a rare allele that has not been incorporated into modern breeding programs.The dysfunction of J might have greatly facilitated the adaptation of soybean to low latitudes. Our findings increase our understanding of the molecular mechanisms underlying the LJ trait and provide valuable resources for soybean breeding.展开更多
基金supported by the National Natural Science Foundation of China (31871634, 31500985)
文摘Retrotransposons account for a large proportion of the genome and genomic variation, and play key roles in creating novel genes and diversifying the genome in many eukaryotic species. Although retrotransposons are abundant in plants, their roles had been underestimated because of a lack of research. Here, we characterized a gibberellin Acid (GA)-insensitive dwarf mutant, 84133, in foxtail millet. Map-based cloning revealed a 5.5-kb Copia-like retrotransposon insertion in DWARF1 (D1), which encodes a DELLA protein. Transcriptional analysis showed that the Copia retrotransposon mediated the transcriptional reprogramming of D1 leading to a novel N-terminal-deleted truncated DELLA transcript that was putatively driven by Copia's LTR, namely D1-TT, and another chimeric transcript. The presence of D1-TT was confirmed by protein immunodetection analysis. Furthermore, D1-TT protein was resistant to GA3 treatment compared with the intact DELLA protein due to its inability to interact with the GA receptor, SiGID1. Overexpression of D1-TT in foxtail millet resulted in dwarf plants, confirming that it determines the dwarfism of 84133. Thus, our study documents a rare instance of long terminal repeat (LTR) retrotransposon-mediated transcriptional reprograming in the plant kingdom. These results shed light on the function of LTR retrotransposons in generating new gene functions and genetic diversity.
基金supported by the National Natural Sci-ence Foundation of China (30871689)the Program for New Century Excellent Talents in University, China(NCET-07-0565)Science Foundation from the Department of Education of Liaoning Province, China(20060772)
文摘One possible mechanism suggested for somaclonal variation is the activation of transposable elements. The activation of retrotransposons by stresses and external changes is commonly observed in plants. In previous study, we isolated the reverse transcriptase （RT） gene sequences of Ty 1-copia retrotransposons from tissue culture strawberry （Fragaria x ananassa） plant, but not the transcriptionally active sequence. For further understanding the relationship between retrotransposon and somaclonal varation, in this study, we isolated the transcriptionally active RT gene sequences from strawberry plants subjected to different abiotic stresses. These retrotransposons were activated by spraying strawberry leaves with 2 mmol L^-1 salicylic acid （SA）, 50 mmol L^-1 methyl jasmonate （MeJA）, 50 mmol L^-1 abscisic acid （ABA）, 50 mmol L^-1 2,4- dichlorophenoxyacetic acid （2,4-D） or by inducing callus growth in 2 types of MS media： first medium supplemented with 0.5 mg L^-1 6-benzylaminopurine （6-BA）, 0.5 mg L^-1 gibberellic acid （GA3）, 1.0 mg L^-1 thidiazuron （TDZ）, and 0.1 mg L^-1 2,4-D, and the second medium supplemented with 0.5 mg L^-1 6-BA, 0.5 mg L^-1 GA3, 2.0 mg L^-1 TDZ, and 0.02 mg L^-1 indole butyric acid （1BA）. Analysis of gene sequences of 17 RTs revealed that none of them contained stop codons and/or indels disrupting the reading frame. These different stress-origin transcriptionally active RTs were remarkably similar to each other- FATEXP2-8 and FATEYS9-7 showed 100% sequence identity. Analysis of pylogenetic of these transcriptionally active RTs and the RT sequences from genome showed that there were close phylogenetic relationships of most of the transcriptionally active RTs. The results of this study have contributed to the background information necessary for future studies for evaluating the relationship between retrotransposons and somaclonal variation.
基金supported by National Natural Science Foundation of China (No.10505018)Natural Science Foundation of Henan Province (No.511030400)
文摘Retrotransposon-like elements are major constituents of most eukaryotic genomes. For example, they account for roughly 90% of the wheat （Triticum aestivum） genome, Previous study on a wheat strain treated by low-energy N^＋ ions indicated the variations in AFLP （Amplified Fragment Length Polymorphism ） markers, One such variation was caused by the re-activation of Tyl-copia-like retrotransposons, implying that the mutagenic effects of lowenergy ions might work through elevated activation of retrotransposons, In this paper an expression profile of Tyl-copia-like retrotransposons in wheat treated by low-energy N^＋ ions is reported, The reverse transcriptase （RT） domains of these retrotransposons were amplified by reverse-transcriptional polymerase chain reaction （RT-PCR） and sequentially cloned, 42 and 65 clones were obtained from the treated （CL） and control materials （CK）, respectively, Sequence analysis of each clone was performed by software. Phylogeny and classification were calculated responding to the sequences of the RT domains. All the results show that there is much difference in the RT domain between the control sample and the treated sample, Especially, the RT domains from the treated group encode significantly more functional ORF （open reading frames） than those from the control sample, This observation suggests that the treated sample has higher activation of retrotransposons, possibly as a consequence of low-energy ion beam irradiation, It also suggests that retrotransposons in the two groups impact the host gene expression in two different ways and carry out different functions in wheat cells.
基金supported by National and Fujian Provincial Natural Science Foundation(No.31371557 and K55NI921A)Fujian Provincial Department of Education Project(K58MLV03A)a startup fund from Fujian Agriculture and Forestry University(http://www.fafu.edu.cn/)to X.S
文摘A novel rice d1 mutant was identified using map-based cloning and comparative analysis of known dl mutants.The mutant[d1-a) shows a mild dwarf trait,which differs only slightly from the wildtype in plant height at the tillering stage.The dl-a mutant is different from other dl mutants.We found that it was interrupted by an Osr4 long terminal repeat(LTR)-retrotransposon,which resulted in the loss of exon 7 in the mutant D1 mRNA.A paralog of the D1 gene.D1-like,was revealed.D1-like is a truncated gene that might have resulted from recombination between retrotransposons.We identified 65 Osr4LTR-retrotransposons in Nipponbare,and found more LTR variants in contrast to coding DNA sequence(CDS) in the retrotransposons.We also identified five possible regulatory motifs in LTRs which may control the expression of the retrotransposons.In addition,we predicted six putative functional Osr4 retrotransposons that contain complete CDSs and all important elements.Osr4 retrotransposons were classified into 4 groups,and this type of retrotransposon only appears to be present in monocots.Members of group 1-1,which included all putative functional retrotransposons,showed a high similarity with each other.The retrotransposons were expressed in all tissues,at especially higher levels in some leaves and seeds.These findings imply that transpositions of group 1-1 members might have occurred frequently and recently.
基金supported by National Natural Science Foundation of China(Nos.10505018,30800204)the Chinese Ministry of Agriculture(No.200803034)
文摘Mutagenesis and retrotransposons have a close relationship, but little attention has been paid yet to the activity of retrotransposons produced by physical mutagens. The variation of retrotransposon WIS 2-1A activity in wheat （Triticum aestivum L.） embryos at three different growth times （30 h, 45 h and 60 h） was investigated after they had been treated with N^＋ implantation in a vacuum of 5× 10^-2 Pa and irradiation by ^60Coγ-ray respectively. For each of the three growth times the expression of WIS 2-1A showed almost entirely a same trend of downregulation, upregulation, then downregulation, and upregulation again with the increase in dose of N^＋ implantation, but the expression appeared irregular with the increase in irradiation of ^60Coγ-ray. In conclusion, the acutely activating effect of WIS 2-1A stimulated by vacuum and high dose N^＋ implantation within a shorter incubation time may provide a convenient tool to advance the research on mutagenic breeding and function genes.
文摘In order to identify the variation and estimate the genetic diversity among the fig (Ficus carica L.) genotypes collected from Algeria and Turkey, the genetic relationships between 86 genotypes were investigated using 23 inter primer binding sites (iPBS)-retrotransposon and 16 simple sequence repeat (SSR) primers. A total of 63 polymorphic bands for the iPBS-retrotransposon markers and 25 alleles for the SSR markers were identified with an average of 2.7 and 1.6 per primer, respectively. The average value of polymorphism information content (PIC) for the iPBS markers (0.73) was higher than that for the SSR markers (0.69). Applying the neighbor-joining method to the combined iPBS-retrotransposon and SSR data, the fig genotypes were clustered into two groups. The STRUCTURE software was used to determine the population structure. Among the genotypes studied, two populations (K = 2) were identified indicating a low diversity between the Algerian and Turkish varieties. Both types of markers were able to differentiate all the fig genotypes and were efficient in discriminating the closely related genotypes. Our data also showed that as a universal marker, iPBS-retrotransposon is a useful tool for the molecular characterization of fig genotypes.
文摘Long terminal repeat （LTR） retrotransposons, one of the foremost types of transposons, continually change or modify gene function and reorganize the genome through bursts of dramatic proliferation. Many LTR-TEs preferen-tially insert within other LTR-TEs, but the cause and evolutionary significance of these nested LTR-TEs are not well under-stood. In this study, a total of 1.52 Gb of Brassica sequence containing 2020 bacterial artificial chromosomes （BACs） was scanned, and six bacterial artificial chromosome （BAC） clones with extremely nested LTR-TEs （LTR-TEs density： 7.24/kb） were selected for further analysis. The majority of the LTR-TEs in four of the six BACs were found to be derived from the rapid proliferation of retrotransposons originating within the BAC regions, with only a few LTR-TEs originating from the proliferation and insertion of retrotransposons from outside the BAC regions approximately 5-23 Mya. LTR-TEs also pref-erably inserted into TA-rich repeat regions. Gene prediction by Genescan identified 207 genes in the 0.84Mb of total BAC sequences. Only a few genes （3/207） could be matched to the Brassica expressed sequence tag （EST） database, indicating that most genes were inactive after retrotransposon insertion. Five of the six BACs were putatively centromeric. Hence, nested LTR-TEs in centromere regions are rapidly duplicated, repeatedly inserted, and act to suppress activity of genes and to reshuffle the structure of the centromeric sequences. Our results suggest that LTR-TEs burst and proliferate on a local scale to create nested LTR-TE regions, and that these nested LTR-TEs play a role in the formation of centromeres.
文摘Histone lysine methylation plays an important role in heterochromatin formation and reprogramming of gene expression. SET-domain-containing proteins are shown to have histone lysine methyltransferase activities. A large number of SET-domain genes are identified in plant genomes. The function of most SET-domain genes is not known. In this work, we studied the 12 rice （Oryza sativa） homologs of Su（var）3-9, the histone H3 lysine 9 （H3K9） methyltransferase identified in Drosophila. Several rice SUVHs （i.e. SDG714, SDG727, and SDG710） were found to have an antagonistic func- tion to the histone H3K9 demethylase JMJ706, as down-regulation of these genes could partially complement the jmj706 phenotype and reduced histone H3K9 methylation. Down-regulation of a rice Su（var）3-9 homolog （SUVH）, namely SDG728, decreased H3K9 methylation and altered seed morphology. Overexpression of the gene increased H3K9 methylation. SDG728 and other SUVH genes were found to be involved in the repression of retrotransposons such as Tos17 and a Tyl-copia element. Analysis of histone methylation suggested that SDG728-mediated H3K9 methylation may play an important role in retrotransposon repression.
基金the National Natural Science Foundation of China，国家科技攻关项目
文摘As the largest transposable element in the plant genome, retrotransposons are thought to be involved in citrus genetic instability and genome evolution, especially in sweet orange, which is prone to bud mutation. In the present study, the presence of copia-like retrotransposons, their heterogeneity, genomic distribution, and transcriptional activities in Citrus were investigated in 12 sweet orange （Citrus sinensis Osbeck） cultivars using a PCR assay designed to detect copia-like reverse transcriptase （RT） sequences. Twelve amplification products from each cultivar were cloned and sequenced. The cloned sequences showed great heterogeneity, except “Dream” navel and “Hamlin”, both of which shared the same sequence. Frame shifting, termination, deletion, and substitution accounted for the heterogeneity of RT sequences. Southern blot hybridization using the RT1 clone from the “Cara Cara” navel as a probe showed that multiple copies were integrated throughout the sweet orange genomes, which made the retrotransposon possible an effective molecular marker to detect citrus evolution events and to reveal its relationship with bud mutation. No transcriptional activities of the retrotransposon were detected by RT-PCR and Northern analysis in the fruits and leaves of either “Cara Cara” or “Seike” navels.
基金the National Natural Science Foundation of China (30500348)Specialized Research Fund for the Doctoral Program of Higher Education(20050157003)
文摘The conserved domains of reverse transcriptase （RT） genes of Tyl-copia and Ty3-gypsy groups of long terminal repeat （LTR） retrotransposons were isolated from the Malus domestica genome using degenerate oligonucleotide primers. Sequence analysis showed that 45% of Ty1-copia and 63% of Ty3-gypsy RT sequences contained premature stop codons and/or indels disrupting the reading frame. High heterogeneity among RT sequences of both Ty1-copia and Ty3-gypsy group retrotransposons was observed, but Ty3-gypsy group retrotransposons in the apple genome are less heterogeneous than Ty1-copia elements. Retrotransposon copy number was estimated by dot blot hybridizations for Ty1-copia （-5000） and Ty3-gypsy （-26000）. All elements of the two types of LTR retrotransposons comprise approximately 38% of the M. domestica genome, with the Ty3-gypsy group contribution being higher （33.5%） than the Tyl.copia one （4.6%）. Transcription was not detected by reverse transcription-polymerase chain reaction for either Ty1-copia or Ty3-gypsy retrotransposons in the leaves of plants in vitro or in leaf explants cultured on medium supplemented with high concentration benzylaminopurine. This research reveals the differences in heterogeneity and copy number between Ty1-copia and Ty3-gypsy retrotransposons in the apple genome. Ty1-copia retrotransposon has higher heterogeneity than Ty3-gypsy retrotransposon, but the latter has a higher copy number, which implies that Ty3-gypsy retrotransposons may play a more important role in the apple genome evolution.
基金supported by the National Basic Research Program of China(No.2005CB121000)the project of Chongqing Science & Technology Commission(CSTC,No.2006AA5019 and 2009BB1241)+1 种基金the Programme of Introducing Talents of Discipline to Universities(No.B07045)State Development Fund at Risk of Callus Silk(No.M012005-000Y-00070)
文摘Eight intact LTR retrotransposons （Nbr1-Nbr8） have been previously characterized from the genome of Nosema bombycis, a eu- karyotic parasite with a compact and reduced genome. Here we describe six novel transcribed Nbr elements （Nbr9-Nbr14） identified through either cDNA library or RT-PCR. Like previously determined ones, all of them belong to the Ty3/Gypsy superfamily. Retrotransposon diversity and incomplete domains with insertions （Nbr12）, deletions （Nbrll） and in-frame stop codons in coding regions （Nbr9） were detected, suggesting that both defective and loss events of LTR retrotransposon have happened in N. bornbycis genome. Analysis of selection showed that strong purifying selection acts on all elements except Nbr11. This implies that selective pressure keeps both these Nbrs and their functions in genome. Interestingly, Nbrll is under positive selection and some positively selected codons were identified, indicating that new functionality might have evolved in the Nbrll retrotransposon. Unlike other transposable elements, Nbrll has integrated into a conserved syntenic block and probably resulted in the inversion of both flanking regions. This demonstrates that transposable element is an important factor for the reshuffling and evolution of their host genomes, and may be maintained under natural selection.
基金grants from the Chinese Academy of Sciences(XDB27040201)the National Natural Science Foundation of China(91640114).
文摘Transposable elements(TEs)constitute the most abundant portions of plant genomes and can dramatically shape host genomes during plant evolution.They also play important roles in crop domestication.However,whether TEs themselves are also selected during crop domestication has remained unknown.Here,we identify an active long terminal repeat(LTR)retrotransposon,HUO,as a potential target of selection during rice domestication and breeding.HUO is a low-copy-number LTR retrotransposon,and is active under natural growth conditions and transmitted through male gametogenesis,preferentially inserting into genomic regions capable of transcription.HUO exists in all wild rice accessions and about half of the archaeological rice grains(1200–7000 years ago)and landraces surveyed,but is absent in almost all modern varieties,indicating its gradual elimination during rice domestication and breeding.Further analyses showed that HUO is subjected to strict gene silencing through the RNA-directed DNA methylation pathway.Our results also suggest that multiple HUO copies may trigger genomic instability through altering genome-wide DNA methylation and small RNA biogenesis and changing global gene expression,resulting in decreased disease resistance and yield,coinciding with its elimination during rice breeding.Together,our study suggests that negative selection of an active retrotransposon might be important for genome stability during crop domestication and breeding.
文摘Stochastic introgression of alien DNA may impose a genomic stress to the recipient genome. Herein, we report that apparent de novo genomic rearrangements in 10 of 13 selected endogenous, lowcopy, and potentially active long terminal repeat （LTR） retrotransposons occurred in one or more of three rice lines studied that were introgressed by wild rice （Zizania latifolia Griseb.）. For nine retrotransposons in which both the reverse-transcriptase （RT） region and the LTR region were available, largely concordant rearrangements occurred at both regions in five elements and at the RT region only in the remaining four elements. A marked proportion of the genomic changes was shared by two or all three introgression lines that were derived from a single F~ plant. This indicates that most of the genomic changes occurred at early developmental stages of the F~ somatic cells, which then gave rise to germline cells, and, hence, ensured inheritance of the changes to later generations. Possible causes and potential implications of the introgression-induced genomic rearrangements in LTR retrotransposons are discussed in the context of plant genome evolution and breeding.
基金supported by the National Natural Science Foundation of China (NSFC grants 31722048 and 31630068)the National Program on Key Research Project (2016YFD0100307)+2 种基金the Science and Technology Innovation Program of the Chinese Academy of Agricultural Sciencesthe Key Laboratory of Biology and Genetic Improvement of Horticultural Crops, Ministry of Agriculture, ChinaProspect of Shandong Seed Project, China (Shandong Gov. [2015] Reference No. 212)
文摘Long terminal repeat retrotransposons(LTR-RTs) are a predominant group of plant transposable elements(TEs) that are an important component of plant genomes. A large number of LTR-RTs have been annotated in the genomes of the agronomically important oil and vegetable crops of the genus Brassica. Herein, full-length LTR-RTs in the genomes of Brassica and other closely related species were systematically analyzed. The full-length LTR-RT content varied greatly(from 0.43% to 23.4%) between different species, with Gypsy-like LTR-RTs constituting a primary group across these genomes. More importantly, many annotated LTR-RTs(from 10.03% to 33.25% of all detected LTR-RTs) were found to be enriched in localized hotspot regions. Furthermore, all of the analyzed species showed evidence of having experienced at least one round of a LTR-RT burst, with Raphanus sativus experiencing three or more. Moreover, these relatively ancient LTR-RT amplifications exhibited a clear expansion at specific time points. To gain a further understanding of this timing, Brassica rapa, B. oleracea, and R. sativus were examined for the presence of syntenic regions, but none were present. These findings indicate that these LTR-RT burst events were not inherited from a common ancestor,but instead were species-specific bursts that occurred after the divergence of Brassica species. This study further exemplifies the complexities of TE amplifications during the evolution of plant genomes and suggests that these LTR-RT bursts play an important role in genome expansion and divergence in Brassica species.
基金Supported by Grants-in-Aid for Scientific Research (01760004 and 04760006) from the Japanese Ministry of Education,Culture,Sports,Science and Technology (MEXT)
文摘New classes of repetitive DNA elements were effectively identified by isolating small fragments of the elements from the wheat genome. A wheat A genome library was constructed from Triticum monococcum by degenerate cleavage with EcoO1091, the recognition sites of which consisted of 5＇-PuGGNCCPy-3＇ multi-sequences. Three novel repetitive sequences pTm6, pTm69 and pTm58 derived from the A genome were screened and tested for high copy number using a blotting approach, pTm6 showed identity with integrase domains of the barley Tyl-Copia-retrotransposon BARE-1 and pTm58 showed similarity to the barley Ty3-gypsy-like retrotransposon Romani. pTm69, however, constituted a tandem array with useful genomic specificities, but did not share any identity with known repetitive elements. This study also sought to isolate wheat D-genome-specific repetitive elements regardless of the level of methylation, by genomic subtraction. Total genomic DNA of Aegilops tauschii was cleaved into short fragments with a methylation-insensitive 4 bp cutter, Mbol, and then common DNA sequences between Ae. tauschii and Triticum turgidum were subtracted by annealing with excess T. turgidum genomic DNA. The D genome repetitive sequence pAt1 was isolated and used to identify an additional novel repetitive sequence family from wheat bacterial artificial chromosomes with a size range of 1 395-1 850 bp. The methods successfully led pathfinding of two unique repetitive families.
基金the“Innovation Project for the Postdoctoral Researchers”in Chinathe“Open Research Fund Program”of Beijing Key Lab of Plant Resource Research and Development,Beijing Technology and Business University。
文摘The dynamic activity of transposable elements(TEs)contributes to the vast diversity of genome size and architecture among plants.Here,we examined the genomic distribution and transposition activity of long terminal repeat retrotransposons(LTR-RTs)in Arabidopsis thaliana(Ath)and three of its relatives,Arabidopsis lyrata(Aly),Eutrema salsugineum(Esa),and Schrenkiella parvula(Spa),in Brassicaceae.Our analyses revealed the distinct evolutionary dynamics of Gypsy retrotransposons,which reflects the different patterns of genome size changes of the four species over the past million years.The rate of Gypsy transposition in Aly is approximately five times more rapid than that of Ath and Esa,suggesting an expanding Aly genome.Gypsy insertions in Esa are strictly confined to pericentromeric heterochromatin and associated with dramatic centromere expansion.In contrast,Gypsy insertions in Spa have been largely suppressed over the last million years,likely as a result of a combination of an inherent molecular mechanism of preferential DNA removal and purifying selection at Gypsy elements.Additionally,species-specific clades of Gypsy elements shaped the distinct genome architectures of Aly and Esa.
基金supported by the National Natural Science Foundation of China(No.31471156)the Agri-X Project of Shanghai Jiao Tong University(Agri-X2015002)the Shanghai Graduate Education and Innovation Program(Horticulture)
文摘Tnt1 is an active retrotransposon originally identified in tobacco(Nicotiana tabacum L.)(Grandbastien et al.,1989),but its transposition activity could be activated through tissue culture in other plant species.The insertions are stable and inheritable in the progeny,which has made it a valuable and versatile tool for developing insertional mutagenesis libraries in several plant species.Here,we explored its utility for mutagenesis in cucumber(Cucumis sativus L.).T_3 Tnt1 transgenic cucumber plants were subjected to tissue culture to regenerate self-pollinated progeny.With PCR and analyses and Southern hybridization,we found regenerated plants maintained the original Tnt1 insertion and created new insertions suggesting characteristic re-transposition activity of Tnt1 during this process.Using genome walking,some flanking sequences of Tnt1 insertions were recovered in regenerated plants.The results demonstrated that Tnt1 could be stably inherited and re-transposable during tissue culture in cucumber and that it is feasible to use for developing an insertional mutagenesis library for cucumber.
基金supported by the National Natural Science Foundation of China(31725021)funded by the Major Program of Guangdong Basic and Applied Research(2019B030302006)。
文摘Soybean(Glycine max) is an important legume crop that was domesticated in temperate regions.Soybean varieties from these regions generally mature early and exhibit extremely low yield when grown under inductive short-day(SD) conditions at low latitudes. The long-juvenile(LJ) trait, which is characterized by delayed flowering and maturity,and improved yield under SD conditions, allowed the cultivation of soybean to expand to lower latitudes. Two major loci control the LJ trait: J and E6. In the current study, positional cloning, sequence analysis, and transgenic complementation confirmed that E6 is a novel allele of J, the ortholog of Arabidopsis thaliana EARLY FLOWERING 3(ELF3). The mutant allele e6^(PG), which carries a Ty1/Copia-like retrotransposon insertion, does not suppress the legume-specific flowering repressor E1, allowing E1 to inhibit Flowering Locus T(FT) expression and thus delaying flowering and increasing yields under SD conditions. The e6^(PG)allele is a rare allele that has not been incorporated into modern breeding programs.The dysfunction of J might have greatly facilitated the adaptation of soybean to low latitudes. Our findings increase our understanding of the molecular mechanisms underlying the LJ trait and provide valuable resources for soybean breeding.
