Plant-Microbe Interactions under the Extreme Habitats and Their Potential Applications.

Plant-microbe associations define a key interaction and have significant ecological and biotechnological perspectives. In recent times, plant-associated microbes from extreme environments have been extensively explored for their multifaceted benefits to plants and the environment, thereby gaining momentum in global research. Plant-associated extremophiles highlight ubiquitous occurrences, inhabiting extreme habitats and exhibiting enormous diversity. The remarkable capacity of extremophiles to exist in extreme environmental conditions is attributed to the evolution of adaptive mechanisms in these microbes at genetic and physiological levels. In addition, the plant-associated extremophiles have a major impact in promoting plant growth and development and conferring stress tolerance to the host plant, thereby contributing immensely to plant adaptation and survival in extreme conditions. Considering the major impact of plant-associated extremophiles from a socio-economic perspective, the article discusses their significance in emerging biotechnologies with a key focus on their ecological role and dynamic interaction with plants. Through this article, the authors aim to discuss and understand the favorable impact and dynamics of plant-associated extremophiles and their biotechnological utilities.

Silencing of the phytoene desaturase (PDS) gene affects the expression of fruit-ripening genes in tomatoes.

BACKGROUND: Past research has shown that virus-induced phytoene desaturase (PDS) gene silencing via agroinjection in the attached and detached fruit of tomato plants results in a pale-yellow fruit phenotype. Although the PDS gene is often used as a marker for gene silencing in tomatoes, little is known about the role of PDS in fruit ripening. In this study, we investigated whether the pepper PDS gene silenced endogenous PDS genes in the fruit of two tomato cultivars, Dotaerang Plus and Legend Summer. RESULTS: We found that the pepper PDS gene successfully silenced endogenous PDS in tomato fruit at a silencing frequency of 100% for both cultivars. A pale-yellow silenced area was observed over virtually the entire surface of individual fruit due to the transcriptional reduction in phytoene desaturase (PDS), zeta-carotene (ZDS), prolycopene isomerase (CrtlSO), and beta-carotene hydroxylase (CrtR-b2), which are the carotenoid biosynthesis genes responsible for the red coloration in tomatoes. PDS silencing also affected the expression levels of the fruit-ripening genes Tomato AGAMOUS-LIKE1 (TAGL1), RIPENING INHIBITOR (RIN), pectin esterase gene (PE), lipoxygenase (LOX), FRUITFULL1/FRUITFUL2 (FUL1/FUL2), and the ethylene biosynthesis and response genes 1-aminocyclopropane-1-carboxylate oxidase 1 and 3 (ACO1 and ACO3) and ethylene-responsive genes (E4 and E8). CONCLUSION: These results suggest that PDS is a positive regulator of ripening in tomato fruit, which must be considered when using it as a marker for virus-induced gene silencing (VIGS) experiments in order to avoid fruit-ripening side effects.

Tomato seeds pretreated with Antifreeze protein type I (AFP I) promotes the germination under cold stress by regulating the genes involved in germination process.

This study was conducted to investigate the involvement of antifreeze proteins (AFPs; type I and III) in the germination mechanism of tomato seeds under low temperature stress. Germination of the seeds grown at a room temperature (25°C) was observed on 5 days after sowing (DAS), while all seeds exposed to a low temperature started to germinate at 16 days after sowing (DAS). However, in comparison with control seeds (0 µg/l), seeds treated with AFP I (100, 300, or 500 µg/l) germinated earlier and at a higher percentage until 20 DAS, and seeds treated with 100 µg/l AFP I showed the highest percentage of germination. Surprisingly, AFP III did not significantly increase germination, and the rate was lower among 500 µg/l AFP III-treated seeds compared with control seeds (0 µg/l). The transcription levels of the plasma membrane-associated H+-ATPase gene and antioxidant-related superoxide dismutase (SOD) and catalase 1 (CAT1) genes were analyzed, and the transcription levels of the genes in the seeds grown at 25°C were relatively low. For low temperature-treated seeds, H+-ATPase in control seeds (0 µg/l) was higher compared with that in AFP I-treated seeds and was lower compared with that in AFP III-treated seeds. The expression levels of the antioxidant-related genes (SOD and CAT1) were lower in AFP I-treated seeds than in control seeds (0 µg/l); however, they were higher in AFP III-treated seeds than in control seeds (0 µg/l). Overall, compared with AFP III, AFP I may potentially function as a cold-protective agent by modulating the genes associated with seed germination.

The role of antifreeze proteins in the regulation of genes involved in the response of Hosta capitata to cold.

Cold temperatures are a major source of stress for plants and negatively impact crop yield. A possible way to protect plants is to treat them with antifreeze proteins (AFPs). Here, we investigated whether fish AFPs can shield the rare ornamental species Hosta capitata from low-temperature stress. We elucidated the expression patterns of the cold-inducible genes C-repeat binding factor 1 (CBF1) and dehydrin 1 (DHN1), as well as the antioxidant genes superoxide dismutase (SOD) and catalase (CAT). All were upregulated at low temperature (4 °C). With increasing exposure time, CBF1 and DHN1 expression generally rose (except CBF1 at 48 h). In contrast, SOD and CAT expression gradually declined from 6 to 48 h. Depending on exposure duration, AFP regulation of gene transcription varied with concentration. However, compared with other concentrations, 100 µg/L AFP reduced CBF1 and DHN1 expression and increased SOD and CAT expression in plants, regardless of exposure time. Both AFP I and III were likely to be most effective at protecting plants against cold stress at concentrations of 100 µg/L. Their involvement in H. capitata cold-stress treatment occurred through regulating the expression of important stress-response genes.

Differential expression of anthocyanin structural genes and transcription factors determines coloration patterns in gerbera flowers.

We investigated the expression of anthocyanin structural genes and transcription factors (TFs) associated with varying anthocyanin content during different developmental stages (S1-S4) of the gerbera cultivars 'Nathasha' and 'Rosalin'. Accumulation of anthocyanin started at S1 and reached a maximum at S3 in both cultivars. Enhancement of anthocyanin content in 'Nathasha' was associated with upregulation of ANS and MYB10, whereas in 'Rosalin', upregulation was associated with CHS1, MYB10, and MYC1. Low-temperature exposure (6 °C) enhanced anthocyanin content to a greater extent than that at 22 °C via stronger upregulation of CHS1 and MYB10 in 'Nathasha' and CHS1 in 'Rosalin', irrespective of flower developmental stage. However, differences in anthocyanin content between the two cultivars were found to be influenced by the expression levels of all structural genes and TFs, irrespective of flower developmental stage and temperature conditions. We suggest that differences in the regulation mechanisms of anthocyanin biosynthesis and coloration pattern between 'Nathasha' and 'Rosalin' are related to differences in the expression patterns of structural genes and TFs; however, further functional studies of the key genes in anthocyanin biosynthesis are needed.

Hymenobacter jeollabukensis sp. nov., isolated from soil.

A Gram-stain-negative, non-motile, rod-shaped, aerobic bacterial strain, designated 1-3-3-8T, was isolated from soil and characterized taxonomically using a polyphasic approach. Comparative 16S rRNA gene sequence analysis showed that strain 1-3-3-8T belongs to the family Cytophagaceae of phylum Bacteroidetes and is most closely related to Hymenobacter paludis KBP-30T (96.8% similarity), Hymenobacter ocellatus Myx2105T (96.8%), Hymenobacter coalescens WW84T (95.6%), and Hymenobacter deserti ZLB-3T (95.4%). The G + C content of the genomic DNA of strain 1-3-3-8T was 63.6 mol%. The isolate contained C15:0 iso (28.4%), summed feature 4 (C17:1 anteiso B/C17:1 iso I; 18.9%), and C15:0 anteiso (17.6%) as major fatty acids, MK-7 as the predominant respiratory quinone, and sym-homospermidine as the predominant polyamine. The major polar lipids were phosphatidylethanolamine and an unidentified lipid. The phenotypic and chemotaxonomic data supported the affiliation of strain 1-3-3-8T with the genus Hymenobacter. The DNA-DNA relatedness between strain 1-3-3-8T and H. paludis KCTC 32237T and H. ocellatus DSM 11117T were 24.5 and 27.4% respectively, clearly showing that the isolate is not related to them at the species level. Overall, the novel strain could be differentiated from its phylogenetic neighbors on the basis of several phenotypic, genotypic, and chemotaxonomic features. Therefore, strain 1-3-3-8T represents a novel species of the genus Hymenobacter, for which the name Hymenobacter jeollabukensis sp. nov. has been proposed. The type strain is 1-3-3-8T (= KCTC 52741T = JCM 32192T).

Hymenobacter terrigena sp. nov., isolated from soil.

A Gram-stain-negative, non-motile, non-spore-forming, rodshaped, aerobic bacterial strain, designated S1-2-2-5T, was isolated from the Jeollabuk-do province, Republic of Korea, and was characterized taxonomically using a polyphasic approach. Comparative 16S rRNA gene sequence analysis showed that strain S1-2-2-5T belonged to the family Cytophagaceae in phylum Bacteroidetes, and was most closely related to Hymenobacter terrae DG7AT (98.2%), Hymenobacter rubidus DG7BT (98.0%), Hymenobacter soli PB17T (97.7%), Hymenobacter daeguensis 16F3Y-2T (97.2%) and Hymenobacter saemangeumensis GSR0100T (97.0%). The G + C content of the genomic DNA of strain S1-2-2-5T was 59.4 mol%. The detection of menaquinone MK-7 as the predominant respiratory quinone, a fatty acid profile with summed feature 3 (C16:1ω7c/C16:1ω6c; 32.0%), C15:0 iso (19.0%), and C15:0 anteiso (15.0%) as the major components, and a polar lipid profile with phosphatidylethanolamine as the major component supported the affiliation of strain S1-2-2-5T to the genus Hymenobacter. The DNA-DNA relatedness between strain S1-2-2-5T and H. terrae KCTC 32554T, H. rubidus KCTC 32553T, H. soli KCTC 12607T, H. daeguensis KCTC 52537T, and H. saemangeumensis KACC 16452T were 49.5, 48.2, 34.1, 28.1, and 31.8% respectively, clearly showing that the isolate is not related to them at the species level. Strain S1-2-2-5T could be clearly differentiated from its closest neighbors on the basis of its phenotypic, genotypic and chemotaxonomic features. Therefore, strain S1-2-2-5T represents a novel species of the genus Hymenobacter, for which the name Hymenobacter terrigena sp. nov. is proposed. The type strain is S1-2-2-5T (= KCTC 52737T = JCM 32195T).

Transcriptional control of anthocyanin biosynthesis genes and transcription factors associated with flower coloration patterns in Gerbera hybrida.

We analyzed the expression of anthocyanin biosynthesis genes and transcription factors (TFs) in the Gerbera hybrida cultivars 'Bintang' and 'Alliance' that exhibit different coloration patterns. Differential expression of biosynthesis genes and TFs was associated with variable anthocyanin content at different flower developmental stages (S1-S3) in both cultivars; higher anthocyanin content was correlated with higher levels of gene expression. Exposure to different temperatures (6 and 22 °C) also resulted in different anthocyanin content levels: the lower temperature (6 °C) enhanced anthocyanin content compared to the higher temperature (22 °C). However, the increased anthocyanin content of 'Bintang' compared to 'Alliance' was the result of higher levels of expression of all detected genes, regardless of flower stage and temperature conditions. Therefore, we conclude that transcriptional control of the detected genes is associated with the mechanisms of anthocyanin biosynthesis and coloration patterns in gerberas; however, further studies of the key genes are needed.

Larkinella roseus sp. nov., a species of the family Cytophagaceae isolated from beach soil.

The taxonomic position of bacterial strain, designated 15J16-1T3AT, recovered from a soil sample was established using a polyphasic approach. Phylogenic analysis based on the 16S rRNA gene sequence showed that strain 15J16-1T3AT belonged to the family Cytophagaceae, phylum Bacteroidetes, and was most closely related to 'Larkinella harenae' 15J9-9 (95.9% similarity), Larkinella ripae 15J11-11T (95.6%), Larkinella bovis M2TB15T (94.7%), Larkinella arboricola Z0532T (93.9%), and Larkinella insperata LMG 22510T (93.5%). Cells were rod-shaped, Gram-stain-negative, aerobic, and nonmotile. The isolate grew on NA, R2A, TSA, but not on LB agar. The strain was able to grow at temperature range from 10°C to 30°C with an optimum at 25°C and pH 6-8. Menaquinone MK-7 was the predominant respiratory quinone. The major cellular fatty acids comprised C16:1ω5c (48.6%) and C15:0 iso (24.1%). Phosphatidylethanolamine, phosphatidylserine, and an unidentified lipid were the major polar lipids. The G + C content of the genomic DNA was 49.5 mol%. Strain 15J16-1T3AT could be distinguished from its closest phylogenetic neighbors based on its phenotypic, genotypic, and chemotaxonomic features. Therefore, the isolate is considered to represent a novel species in the genus Larkinella, for which the name Larkinella roseus sp. nov. is proposed. The type strain is 15J16-1T3AT (= KCTC 52004T = JCM 31991T).

Anthocyanin mutants of Japanese and common morning glories exhibit normal proanthocyanidin accumulation in seed coats.

Anthocyanin and proanthocyanidin biosynthesis pathways are believed to overlap. This study examined proanthocyanidin accumulation in seed coats of morning glories (Ipomoea nil and I. purpurea) carrying mutations in CHS-D, CHI, and ANS genes encoding chalcone synthase, chalcone isomerase, and anthocyanidin synthase, respectively. Chemical staining revealed that mutants accumulate proanthocyanidin normally. Thus, the tested genes are not essential to proanthocyanidin biosynthesis, but are essential to anthocyanin biosynthesis in flowers and stems. Based on the results and the I. nil draft genome sequence, the genes involved in proanthocyanidin biosynthesis, including a new copy of the flavanone 3-hydroxylase gene could be predicted. Moreover, the genome has no homologs for known enzymes involved in producing flavan-3-ols, the starter and extension units of proanthocyanidin. These results suggested that I. nil produces flavan-3-ols through an undiscovered biosynthesis pathway. To characterize proanthocyanidin pigmentation further, we conducted mutant screening using a large I. nil population. We discovered that the brown mutant lines (exhibiting brown seeds and normal anthocyanin pigmentation) do not accumulate proanthocyanidin in their seed coats. Thus, the brown mutation should be useful for further investigations into the various mechanisms controlling anthocyanin and proanthocyanidin pathways.

Overexpression of snapdragon Delila (Del) gene in tobacco enhances anthocyanin accumulation and abiotic stress tolerance.

BACKGROUND: Rosea1 (Ros1) and Delila (Del) co-expression controls anthocyanin accumulation in snapdragon flowers, while their overexpression in tomato strongly induces anthocyanin accumulation. However, little data exist on how Del expression alone influences anthocyanin accumulation. RESULTS: In tobacco (Nicotiana tabacum 'Xanthi'), Del expression enhanced leaf and flower anthocyanin production through regulating NtCHS, NtCHI, NtF3H, NtDFR, and NtANS transcript levels. Transgenic lines displayed different anthocyanin colors (e.g., pale red: T0-P, red: T0-R, and strong red: T0-S), resulting from varying levels of biosynthetic gene transcripts. Under salt stress, the T2 generation had higher total polyphenol content, radical (DPPH, ABTS) scavenging activities, antioxidant-related gene expression, as well as overall greater salt and drought tolerance than wild type (WT). CONCLUSION: We propose that Del overexpression elevates transcript levels of anthocyanin biosynthetic and antioxidant-related genes, leading to enhanced anthocyanin production and antioxidant activity. The resultant increase of anthocyanin and antioxidant activity improves abiotic stress tolerance.

Genome sequence and analysis of the Japanese morning glory Ipomoea nil.

Ipomoea is the largest genus in the family Convolvulaceae. Ipomoea nil (Japanese morning glory) has been utilized as a model plant to study the genetic basis of floricultural traits, with over 1,500 mutant lines. In the present study, we have utilized second- and third-generation-sequencing platforms, and have reported a draft genome of I. nil with a scaffold N50 of 2.88 Mb (contig N50 of 1.87 Mb), covering 98% of the 750 Mb genome. Scaffolds covering 91.42% of the assembly are anchored to 15 pseudo-chromosomes. The draft genome has enabled the identification and cataloguing of the Tpn1 family transposons, known as the major mutagen of I. nil, and analysing the dwarf gene, CONTRACTED, located on the genetic map published in 1956. Comparative genomics has suggested that a whole genome duplication in Convolvulaceae, distinct from the recent Solanaceae event, has occurred after the divergence of the two sister families.

Expression of RsMYB1 in chrysanthemum regulates key anthocyanin biosynthetic genes

Background Several MYB genes belonging to R2R3 MYB transcription factors have been used in several plant species to enhance anthocyanin production, and have shown various expression or regulation patterns. This study focused on the effect of ectopic expression of an RsMYB1 isolated from radish (Raphanus sativa) on chrysanthemum cv. ‘Shinma'. Results The RT-PCR results confirmed that RsMYB1 regulated the expression of three key biosynthetic genes (CmF3H, CmDFR, and CmANS) that are responsible for anthocyanin production in transgenic chrysanthemum, but were not detected in the non-transgenic line. In all transgenic plants, higher expression levels of key biosynthetic genes were observed in flowers than in leaves. However, the presence of RsMYB1 in chrysanthemum did not affect any morphological characteristics, such as plant height, leaf shape or size, and number of flowers. Furthermore, no anthocyanin accumulation was visually observed in the leaves and floral tissue of any of the transgenic lines, which was further confirmed by anthocyanin content estimation. Conclusion To our knowledge, this is the first time the role of an MYB transcription factor in anthocyanin production has been investigated in chrysanthemum.

Activation and epigenetic regulation of DNA transposon nDart1 in rice.

A large part of the rice genome is composed of transposons. Since active excision/reintegration of these mobile elements may result in harmful genetic changes, many transposons are maintained in a genetically or epigenetically inactivated state. However, some non-autonomous DNA transposons of the nDart1-3 subgroup, including nDart1-0, actively transpose in specific rice lines, such as pyl-v which carries an active autonomous element, aDart1-27, on chromosome 6. Although nDart1-3 subgroup elements show considerable sequence identity, they display different excision frequencies. The most active element, nDart1-0, had a low cytosine methylation status. The aDart1-27 sequence showed conservation between pyl-stb (pyl-v derivative line) and Nipponbare, which both lack autonomous activity for transposition of nDart1-3 subgroup elements. In pyl-v plants, the promoter region of the aDart1-27 transposase gene was more hypomethylated than in other rice lines. Treatment with the methylation inhibitor 5-azacytidine (5-azaC) induced transposition of nDart1-3 subgroup elements in both pyl-stb and Nipponbare plants; the new insertion sites were frequently located in genic regions. 5-AzaC treatment principally induced expression of Dart1-34 transposase rather than the other 38 aDart1-related elements in both pyl-stb and Nipponbare treatment groups. Our observations show that transposition of nDart1-3 subgroup elements in the nDart1/aDart1 tagging system is correlated with the level of DNA methylation. Our system does not cause somaclonal variation due to an absence of transformed plants, offers the possibility of large-scale screening in the field and can identify dominant mutants. We therefore propose that this tagging system provides a valuable addition to the tools available for rice functional genomics.

A WD40-repeat protein controls proanthocyanidin and phytomelanin pigmentation in the seed coats of the Japanese morning glory.

The protein complex composed of the transcriptional regulators containing R2R3-MYB domains, bHLH domains, and WDR in plants controls various epidermal traits, including anthocyanin and proanthocyanidin pigmentation, trichome and root hair formation, and vacuolar pH. In the Japanese morning glory (Ipomoea nil), InMYB1 having R2R3-MYB domains and InWDR1 containing WDR were shown to regulate anthocyanin pigmentation in flowers, and InWDR1 was reported to control dark-brown pigmentation and trichome formation on seed coats. Here, we report that the seed pigments of I. nil mainly comprise proanthocyanidins and phytomelanins and that these pigments are drastically reduced in the ivory seed coats of an InWDR1 mutant. In addition, a transgenic plant of the InWDR1 mutant carrying the active InWDR1 gene produced dark-brown seeds, further confirming that InWDR1 regulates seed pigmentation. Early steps in anthocyanin and proanthocyanidin biosynthetic pathways are thought to be common. In the InWDR1 mutant, none of the structural genes for anthocyanin biosynthesis that showed reduced expression in the white flowers were down-regulated in the ivory seeds, which suggests that InWDR1 may activate different sets of the structural genes for anthocyanin biosynthesis in flowers and proanthocyanidin production in seeds. As in the flowers, however, we noticed that the expression of InbHLH2 encoding a bHLH regulator was down-regulated in the seeds of the InWDR1 mutant. We discuss the implications of these results with respect to the proanthocyanidin biosynthesis in the seed coats.

Identification of r mutations conferring white flowers in the Japanese morning glory (Ipomoea nil).

The wild-type Japanese morning glory [Ipomoea nil (L.) Roth.] exhibits blue flowers with red stems, and spontaneous r mutants display white flowers with green stems. We have identified two r mutations, r1-1 and r1-2, that are caused by insertions of Tpn1-related DNA transposable elements, Tpn3 (5.6 kb) and Tpn6 (4.7 kb), respectively, into a unique intron of the CHS-D gene, which is responsible for flower and stem pigmentation. Both Tpn3 and Tpn6, which belong to the En/Spm or CACTA superfamily, are nonautonomous elements lacking transposase genes but containing unrelated cellular DNA segments including exons and introns. Interestingly, r1-2 contains an additional 4-bp insertion at the Tpn3 integration site in r1-1, presumably a footprint caused by the excision of Tpn3. The results strengthen the previous notion that Tpn1 and its relatives are major spontaneous mutagens for generating various floriculturally important traits in I. nil. Since I. nil has an extensive history of genetic studies, molecular identification of classical spontaneous mutations would also facilitate reinterpretation of the abundant classical genetic data available.

A bHLH regulatory gene in the common morning glory, Ipomoea purpurea, controls anthocyanin biosynthesis in flowers, proanthocyanidin and phytomelanin pigmentation in seeds, and seed trichome formation.

The transcriptional regulators for anthocyanin pigmentation include proteins containing R2R3-MYB domains, bHLH domains and conserved WD40 repeats, and their interactions determine the set of genes to be expressed. Spontaneous ivory seed (ivs) mutants of Ipomoea purpurea displaying pale pigmented flowers and ivory seeds are caused by insertions of DNA transposons into the bHLH2 gene that encodes a bHLH transcriptional regulator. A partial reduction in the expression of all structural genes encoding enzymes for anthocyanin biosynthesis was observed in the young flower buds of these ivs mutants. The DFR-B and ANS transcripts were completely abolished in the ivs seed coats, whereas the early biosynthetic genes for flavonol biosynthesis remained active. The production and accumulation of both proanthocyanidin and phytomelanin pigments in the ivory seed coats were drastically reduced. Moreover, the unbranched trichomes in the ivory seeds were smaller in size and fewer in number than those in the wild-type dark-brown seeds, and the surface of the epidermis without trichomes in the dark-brown seeds looked rougher, due to the protruding tangential walls, than that in the ivory seeds. Although the I. purpurea bHLH2 gene is the most closely related to the petunia AN1 gene, whose mutation is known to confer white flowers and to be deficient in acidification of their vacuoles, the vacuolar alkalization in the epidermal flower limbs of I. purpurea ivs mutants appears to occur normally. These results are discussed with regard to the function of bHLH transcriptional regulators controlling flower and seed pigmentation as well as other epidermal traits.

Spontaneous mutations caused by a Helitron transposon, Hel-It1, in morning glory, Ipomoea tricolor.

Helitrons are newcomers among eukaryotic DNA transposons and have originally been identified by computational analysis in the genomes of Arabidopsis, rice and nematode. They are distinguished from other transposons in their structural features, and their proposed transposition mechanisms are involved in rolling circle replication. Computer-predicted autonomous Helitrons with conserved terminal sequences 5'-TC and CTRR-3' are presumed to encode a putative transposase, Rep/Hel-TPase, which contains a characteristic nuclease/ligase domain for the replication-initiation protein (Rep) and a DNA helicase domain (Hel). Plant Helitrons are thought to encode an additional transposase, RPA-TPase, which is related to the largest subunit of the replication protein A (RPA70). Although Helitrons are found in diverse genomes, neither an autonomous element nor a transposition event has been reported. Here we show that a spontaneous pearly-s mutant of Ipomoea tricolor cv. Pearly Gates, exhibiting white flowers and isolated in approximately 1940, has an 11.5-kbp novel Helitron, named Hel-It1, integrated into the DFR-B gene for anthocyanin pigmentation. Hel-It1 shows the predicted plant Helitron structure for an autonomous element with the conserved termini and carrying the two putative transposase genes, Rep/Hel-TPase and RPA-TPase, which contain a nonsense and a frameshift mutation, respectively. Hel-It1-related elements are scattered in the Ipomoea genome, and only a fraction of the pearly-s plants were found to carry Hel-It1 at another insertion site. The pearly-s mutant appears to bear an autonomous element and to express the wild-type RPA-TPase transcripts. The structures of a putative autonomous element and its transposase genes are discussed.

Genetics and epigenetics in flower pigmentation associated with transposable elements in morning glories.

Among the genus Ipomoea, three morning glories, I. nil (the Japanese morning glory), I. purpurea (the common morning glory), and I. tricolor, were domesticated well for floricultural plants, and many spontaneous mutants displaying various flower pigmentation patterns were isolated. Most of these spontaneous mutations were found to be caused by the insertion of DNA transposable elements in the genes for the anthocyanin pigmentation in flowers, and many of them exhibited variegated flowers, such as white flowers with pigmented spots and sectors. Here, we describe the historical background of the mutants displaying variegated flowers and review the genetic and epigenetic regulation in flower pigmentation associated with transposable elements of these morning glories. The flecked, speckled, r-1, and purple mutations in I. nil were caused by insertions of Tpn1 and its relatives in the En/Spm superfamily, Tpn2, Tpn3, and Tpn4, into the genes for anthocyanin coloration in flowers, i.e., DFR-B, CHI, CHS-D, and InNHX1, respectively. Similarly, the flaked and pink mutants of I. purpurea have distantly related elements, Tip100 and Tip201, in the Ac/Ds superfamily inserted into the CHS-D and F3'H genes, respectively. The flower variegation patterns can be determined by the frequency and timing of the excision of these transposons, and their stable insertions produce plain color flowers without generating pigmented spots or sectors; furthermore, both genetic and epigenetic regulation appeared to play important roles in determining the frequency and timing of the excision of the transposons. However, flower variegation is not always associated with the excision of an integrated DNA transposon from one of the genes for anthocyanin pigmentation. The mutant Flying Saucers of I. tricolor displaying variegated flowers was found to have the transposon ItMULE1 inserted into the DFR-B promoter region, but no excision of ItMULE1 from the DFR-B could be detected in the variegated flower lines. The instable pearly-vrg allele in cv. Flying Saucers is likely to be an epiallele because the DNA methylation in the DFR-B promoter appeared to be associated with flower pigmentation.

Genetics and epigenetics in flower pigmentation associated with transposable elements in morning glories.

Among the genus Ipomoea, three morning glories, I. nil the Japanese morning glory), I. purpurea (the common morning glory), and I. tricolor, were domesticated well for floricultural plants, and many spontaneous mutants displaying various flower pigmentation patterns were isolated. Most of these spontaneous mutations were found to be caused by the insertion of DNA transposable elements in the genes for the anthocyanin pigmentation in flowers, and many of them exhibited variegated flowers, such as white flowers with pigmented spots and sectors. Here, we describe the historical background of the mutants displaying variegated flowers and review the genetic and epigenetic regulation in flower pigmentation associated with transposable elements of these morning glories. The flecked, speckled, r-1, and purple mutations in I. nil were caused by insertions of Tpnl and its relatives in the En/Spm superfamily, Tpn2, Tpn3, and Tpn4, into the genes for anthocyanin coloration in flowers,i.e., DFR-B, CHI, CHS-D, and InNHXI, respectively. Similarly, the flaked and pink mutants of I. purpurea have distantly related elements, Tip100 and Tip201, in the Ac/Ds superfamily inserted into the CHS-D and F3'H genes, respectively. The flower variegation patterns can be determined by the frequency and timing of the excision of these transposons, and their stable insertions produce plain color flowers without generating pigmented spots or sectors; furthermore, both genetic and epigenetic regulation appeared to play important roles in determining the frequency and timing of the excision of the transposons. However, flower variegation is not always associated with the excision of an integrated DNA transposon from one of the genes for anthocyanin pigmentation. The mutant Flying Saucers of I. tricolor displaying variegated flowers was found to have the transposon ItMULE inserted into the DFR-B promoter region, but no excision of ITMULEL from the DFR-B could be detected in the variegated flower lines. The instable pearly-vrg allele in cv. Flying Saucers is likely to be an epiallele because the DNA methylation in the DFR-B promoter appeared to be associated with flower pigmentation.