It is reported that the production of floral sexual phenotype in hexaploid monoecious persimmon (Diospyros kaki) is closely related to a pseudogene called OGI, and a short interspersed nuclear element (SINE)-like insertion (named Kali) in the OGI promoter leads to the gene silence. As a result, DNA methylation level of MeGI promoter determines the development of male or female flowers. However, the molecular mechanism in androecious D. kaki, which only bear male flowers, remains elusive. Here, real-time quantitative polymerase chain reaction (RT-qPCR), molecular cloning, and bisulfite PCR sequencing technique were carried out using 87 materials, including 56 androecious resources, 15 monoecious, and 16 gynoecious cultivars, to investigate the performance of OGI and MeGI on the specific androecious type of D. kaki in China. In conclusion, the Kali insertion was exactly located in the OGI promoter region, and the OGI gene and the Kali sequence were existing and conserved in androecious D. kaki. Meanwhile, we also demonstrated that the MeGI gene was widespread in our investigated samples. Ultimately, our result convincingly provided evidence that the low expression of OGI is probably ascribed to the presence of Kali displaying strong methylation in the OGI promoter, and low expression of MeGI, as well as high DNA methylation level, in the promoter was closely connected with the production of male flowers; this result was consistent with the monoecious persimmon model. Our findings provide predominant genetic aspects for investigation into androecious D. kaki, and future perfecting the sex-determining mechanisms in persimmon.
DNA methylation is an epigenetic mechanism with the potential to regulate gene expression and affect plant phenotypes. Both hybridization and genome doubling may affect the DNA methylation status of newly formed allopolyploid plants. Previous studies demonstrated that changes in cytosine methylation levels and patterns were different among individual hybrid plant, therefore, studies investigating the characteristics of variation in cytosine methylation status must be conducted at the population level to avoid sampling error. In the present study, an F1 hybrid diploid population and three allotriploid populations with different heterozygosity [originating from first-division restitution (FDR), second-division restitution (SDR), and post-meiotic restitution (PMR) 2n eggs of the same female parent] were used to investigate cytosine methylation inheritance and variation relative to their common parents using methylation-sensitive amplification polymorphism (MSAP). The variation in cytosine methylation in individuals in each population exhibited substantial differences, confirming the necessity of population epigenetics. The total methylation levels of the diploid population were significantly higher than in the parents, but those of the three allotriploid populations were significantly lower than in the parents, indicating that both hybridization and polyploidization contributed to cytosine methylation variation. The vast majority of methylated status could be inherited from the parents, and the average percentages of non-additive variation were 6.29, 3.27, 5.49 and 5.07% in the diploid, FDR, SDR and PMR progeny populations, respectively. This study lays a foundation for further research on population epigenetics in allopolyploids.
In this report, we compared transcriptomic differences between a synthetic Populus section Tacamahaca triploid driven by second-division restitution and its parents using a high-throughput RNA-seq method. A total of 4,080 genes were differentially expressed between the high-growth vigor allotriploids (SDR-H) and their parents, and 719 genes were non-additively expressed in SDR-H. Differences in gene expression between the allotriploid and male parent were more significant than those between the allotriploid and female parent, which may be caused by maternal effects. We observed 3,559 differentially expressed genes (DEGs) between the SDR-H and male parent. Notably, the genes were mainly involved in metabolic process, cell proliferation, DNA methylation, cell division, and meristem and developmental growth. Among the 1,056 DEGs between SDR-H and female parent, many genes were associated with metabolic process and carbon utilization. In addition, 1,789 DEGs between high- and low-growth vigor allotriploid were mainly associated with metabolic process, auxin poplar transport, and regulation of meristem growth. Our results indicated that the higher poplar ploidy level can generate extensive transcriptomic diversity compared with its parents. Overall, these results increased our understanding of the driving force for phenotypic variation and adaptation in allopolyploids driven by second-division restitution.
Polyploid breeding is important in Populus genetic improvement programs because polyploid trees generally display increased height growth compared to their diploid parents. However, the genetic mechanism underlying this phenomenon remains unknown. In the present study, apical bud transcriptomes of vigorous, fast growing Populus allotriploid progeny genotypes and their diploid parents were sequenced and analyzed. We found that these allotriploids exhibited extensive transcriptomic diversity. In total, 6020 differentially expressed genes (DEGs) were found when the allotriploid progeny and their parents were compared, among which 791 overlapped between the allotriploids and both parents. Many genes associated with cell differentiation and meristem development were preferentially expressed in apical buds of the fast growing Populus allotriploids compared to their diploid parents. In addition, many auxin-, gibberellin-, and jasmonic acid-related genes were also preferentially expressed in the allotriploids compared to their parents. Our findings show that allotriploidy can have considerable effects on duplicate gene expression in Populus. In particular we identified and considered DEGs that provide important clues for improving our mechanistic understanding of positive heterosis of vigor- and growth-related traits in Populus allotriploids.
The induction of nonreduced gametes, whether from first-division restitution (FDR), second-division restitution (SDR), or both, is an important approach for polyploidization. However, an accurate method for determining the genetic constitution of polyploid hybrids is not available. In this study, based on both flow cytometric analysis and somatic chromosome counting, 164 triploid hybrids of the female parent Populus pseudo-simonii × P. nigra ‘Zheyin3#’ (2n = 2x = 38, abbreviated ZY3) and male parent P. × beijingensis (2n = 2x = 38, abbreviated BJY) were produced by high-temperature treatment during ZY3 megasporogenesis. Using six simple sequence repeat (SSR) markers with low recombination frequencies to reduce the impact of recombination, we analyzed the allelic configurations of the obtained triploid hybrids. Although the allelic configurations were not always consistent at all six loci, by combining the allelic configurations at the six loci, we inferred that 40 triploids originated from FDR 2n eggs of ZY3, whereas the others originated from SDR 2n eggs. In conclusion, our study provides a novel and effective tool for analyzing 2n gametes and performing early selection to improve triploid poplar breeding programs.
Dioecy, a sexual system of single-sexual (gynoecious/androecious) individuals, is rare in flowering plants. This rarity may be a result of the frequent transition from dioecy into systems with co-sexual individuals.In this study, co-sexual expression (monoecy and hermaphroditic development), previously thought to be polyploid-specific in Diospyros species, was identified in the diploid D. oleifeara historically. We characterized potential genetic mechanisms that underlie the dissolution of dioecy to monoecy and andro(gyno)monoecy, based on multiscale genome-wide investigations of 150 accessions of Diospyros oleifera. We found all co-sexual plants, including monoecious and andro(gyno)monoecious individuals, possessed the male determinant gene OGI, implying the presence of genetic factors controlling gynoecia development in genetically male D. oleifera. Importantly, discrepancies in the OGI/MeGI module were found in diploid monoecious D. oleifera compared with polyploid monoecious D. kaki, including no Kali insertion on the promoter of OGI, no different abundance of smRNAs targeting MeGI (a counterpart of OGI), and no different expression of MeGI between female and male floral buds. On the contrary, in both single- and co-sexual plants, female function was expressed in the presence of a genome-wide decrease in methylation levels, along with sexually distinct regulatory networks of smRNAs and their targets. Furthermore, a genome-wide association study (GWAS) identified a genomic region and a DUF247 gene cluster strongly associated with the monoecious phenotype and several regions that may contribute to andromonoecy.Collectively, our findings demonstrate stable breakdown of the dioecious system in D. oleifera, presumably also a result of genomic features of the Y-linked region.
To explore the mechanisms of the color formation of different colored persimmon fruits, we used two different colored persimmon cultivars (yellow-peeled persimmon fruit ‘Zhongshi No.6’ and red-peeled persimmon fruit ‘Hongdenglong’) as materials to study the synthesis and accumulation of carotenoids in three stages (full green, color transition, and full yellow or red) using targeted metabolomic and transcriptomic methods. A total of 14 carotenoids and 30 carotenoid lipids were identified in the peel of the two persimmon cultivars. After analysis, it was found that the total carotenoid content of the red persimmon cultivar was higher than that of the yellow persimmon cultivar. The contents of lycopene, α-carotenoid, β-carotenoid, (E/Z)-phytoene, and β-cryptoxanthin are the main reasons for the difference in total carotenoid content between the two persimmon cultivars, especially lycopene. Twelve structural genes involved in the metabolism of carotenoids were also found in this study. In comprehensive metabolome and transcriptome analysis, we found that, between the two persimmon cultivars, lycopene was the key metabolite responsible for the color difference, and PSY, LCYE, and ZDS were the key genes that regulated the differences in lycopene accumulation. The results of this study provide us with new information regarding persimmon fruit synthesis and accumulation. In addition, they also provide a theoretical foundation for improving persimmon fruit germplasm.
Abstract PA-enhanced content causes astringency in persimmon fruit. PCNA persimmons can lose their astringency naturally and they become edible when still on the tree, which allows for conserves of physical and financial resources. C-PCNA persimmon originates in China. Its deastringency trait primarily depends on decreased PA biosynthesis and PA insolubilization at the late stage of fruit development. Although some genes and transcription factors that may be involved in the deastringency of C-PCNA persimmon have been reported, the expression patterns of these genes during the key deastringency stage are reported less. To investigate the variation in PA contents and the expression patterns of deastringency-related genes during typical C-PCNA persimmon ‘Xiaoguo-tianshi’ fruit development and ripening, PA content and transcriptional profiling were carried out at five late stages from 70 to 160 DAF. The combinational analysis phenotype, PA content, and DEG enrichment revealed that 120–140 DAF and 140–160 DAF were the critical phases for PA biosynthesis reduction and PA insolubilization, respectively. The expression of PA biosynthesis-associated genes indicated that the downregulation of the ANR gene at 140–160 DAF may be associated with PA biosynthesis and is decreased by inhibiting its precursor cis-flavan-3-ols. We also found that a decrease in acetaldehyde metabolism-associated ALDH genes and an increase in ADH and PDC genes might result in C-PCNA persimmon PA insolubilization. In addition, a few MYB-bHLH-WD40 (MBW) homologous transcription factors in persimmon might play important roles in persimmon PA accumulation. Furthermore, combined coexpression network analysis and phylogenetic analysis of MBW suggested that three putative transcription factors WD40 ( evm.TU.contig1.155 ), MYB ( evm.TU.contig8910.486 ) and bHLH ( evm.TU.contig1398.203 ), might connect and co-regulate both PA biosynthesis and its insolubilization in C-PCNA persimmon. The present study elucidated transcriptional insights into PA biosynthesis and insolubilization during the late development stages based on the C-PCNA D. kaki genome (unpublished). Thus, we focused on PA content variation and the expression patterns of genes involved in PA biosynthesis and insolubilization. Our work has provided additional evidence on previous knowledge and a basis for further exploration of the natural deastringency of C-PCNA persimmon.
Background: Polyploid breeding is a powerful approach for Populus genetic improvement because polyploid trees have valuable characteristics, including better timber quality and a higher degree of stress resistance compared with their full-sib diploids. However, the genetic mechanism underlying this phenomenon remains unknown. Keywords: iTRAQ, Populus, allotriploid, diploids, proteome, polyploid.
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