Authorship：Corresponding author   Publishing type：Research paper (scientific journal)   Publisher：Elsevier BV  

DOI： 10.1016/j.aquabot.2020.103276

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Language：English   Publishing type：Research paper (scientific journal)   Publisher：JAPAN SOC HORTICULTURAL SCI  

The number of petals in a flower is one of the most important appearance qualities of ornamental flowers. In Eustoma, the number of petals fluctuates significantly and little is known about how it is controlled. We investigated the cultivating conditions that affect the number of petals in double flowers and tried to develop a technique for growing splendid corolla. High temperature in the reproductive phase reduces the number of petals. The transient treatment of high temperature just prior to the petal initiation stage is sufficient to control such a reduction. The measurement of flower bud growth showed that one week of temperature treatment is necessary to control the number of petals in a flower. The integration of our results demonstrated that both daytime and nighttime temperatures affected the number of petals and that the number of petals was clearly correlated with average daily temperature within the range of 20 degrees C < x < 25 degrees C. This phenomenon applies to various cultivars in Eustoma grandiflorum. We propose the greenhouse conditions necessary to achieve both high quality flowers and reduced energy consumption by considering the temperature and stages of flower development.

DOI： 10.2503/hortj.OKD-138

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Language：English   Publishing type：Research paper (scientific journal)   Publisher：JAPAN INT RESEARCH CENTER AGRICULTURAL SCIENCES  

Following the accident at the Fukushima Daiichi Nuclear Power Plant, agricultural fields in Fukushima Prefecture were subject to severe radioactive contamination. Given the fact that radiocesium is readily transferred along the food chain and potentially poses a radiation risk to humans, effective methods of lowering the transfer of radioactivity from soil to crops are needed. We examined the effect of neutralized schwertmannite (NS) produced in the disused Yanahara mine as a new agricultural material for reducing radiocesium uptake by crops. The application of NS to soil at 1% or 5% showed inhibitory effects on radiocesium accumulation in the harvests of four upland crops (sweet potato, radish, turnip, and Chinese cabbage) and rice. Substantial amounts of radiocesium were detected in brown rice and hulls following cultivation in soil with recommended levels of exchangeable potassium (&gt; 200 mg kg(-1)). Decreased levels of radiocesium were observed in both tissues following the application of 5% NS, suggesting additional effects of NS application in potassium-rich soil. Our results indicated the efficacy of NS application and suggest its practical use in minimizing the radioactivity in crops cultivated in radiocesium-contaminated soil.

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Authorship：Lead author,　Corresponding author   Language：English   Publishing type：Research paper (scientific journal)   Publisher：JAPAN INT RESEARCH CENTER AGRICULTURAL SCIENCES  

During arching cultivation of roses in autumn and winter, long-day treatment using fluorescent lamps placed above the base of the plants slightly increased the number of cut flowers and also tended to increase the cut flower length in the first year. To further investigate these effects, the light condition of assimilation shoots was modified by supplemental lighting using white light-emitting diodes (LEDs) placed above the assimilation shoots. Supplemental lighting at two different levels of photosynthetic photon flux density (PPFD), 100 and 250 mu mol m(-2) s(-1), increased the number of cut flowers from the middle portion of the assimilation shoots, and the total number and weight of cut flowers according to the light intensity. Irradiation at 250 mu mol m(-2) s(-1) PPFD also increased the number of cut flowers over 80 cm long and the length, weight and stem diameter of cut flowers over 60 cm long. Long-day treatment using fluorescent lamps did not affect the number of cut flowers in the second year. These results indicate that long-day treatment using fluorescent lamps can effectively increase the yield of cut rose flowers in some years, while supplemental lighting using white LEDs for assimilation shoots is a method of increasing it more strongly.

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Language：English   Publishing type：Research paper (scientific journal)   Publisher：OXFORD UNIV PRESS  

The whole-genome sequence of carnation (Dianthus caryophyllus L.) cv. 'Francesco' was determined using a combination of different new-generation multiplex sequencing platforms. The total length of the non-redundant sequences was 568 887 315 bp, consisting of 45 088 scaffolds, which covered 91% of the 622 Mb carnation genome estimated by k-mer analysis. The N50 values of contigs and scaffolds were 16 644 bp and 60 737 bp, respectively, and the longest scaffold was 1 287 144 bp. The average GC content of the contig sequences was 36%. A total of 1050, 13, 92 and 143 genes for tRNAs, rRNAs, snoRNA and miRNA, respectively, were identified in the assembled genomic sequences. For protein-encoding genes, 43 266 complete and partial gene structures excluding those in transposable elements were deduced. Gene coverage was similar to 98%, as deduced from the coverage of the core eukaryotic genes. Intensive characterization of the assigned carnation genes and comparison with those of other plant species revealed characteristic features of the carnation genome. The results of this study will serve as a valuable resource for fundamental and applied research of carnation, especially for breeding new carnation varieties.

DOI： 10.1093/dnares/dst053

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Language：English   Publishing type：Research paper (scientific journal)   Publisher：JAPAN SOC HORTICULTURAL SCI  

In senescing carnation (Dianthus caryophyllus L.) flowers, ethylene production begins in the gynoecium, and the resulting ethylene acts on petals, inducing autocatalytic ethylene production. We investigated the role of abscisic acid (ABA) in ethylene production in the gynoecium of flowers. First, cDNAs of major genes involved in ABA biosynthesis and signaling were cloned from carnation flower tissues. Then, changes in ABA content and gene expression of ABA biosynthesis and signaling in the ovary were examined using three cultivars, 'Light Pink Barbara (LPB)' and 'Excerea', whose cut flowers produce ethylene during senescence and have an ordinary vase-life of about one week, and 'Miracle Rouge', whose cut flowers produce no detectable ethylene during senescence and have a vase-life of about three weeks. ABA content in the ovary was 530-710 pmol.g(-1) fresh weight (FW) from Os 2 (early opening stage) to Os 6 (end of opening stage) in 'LPB', and at 200-380 pmol.g(-1) FW in 'Excerea' at the same stages; but 930 pmol.g(-1) FW at Ss 1 (early senescence stage). The ABA content remained at 70-160 pmol.g(-1) FW in 'Miracle Rouge'. The changes in ABA content were in parallel with the transcript levels of DcNCED1 (carnation gene for 9-cis-epoxycarotenoid dioxygenase). DcPYR1 (ABA receptor gene) transcript was 0.004-0.007 relative expression level (r.e.l.) in 'LPB' ovary at Os 1-Os 3, and 0.028 r.e.l. at Ss 1 (beginning of senescence stage). In 'Excerea' ovary,DcPYR1 transcript was 0.025-0.037 r.e.l. during flower opening and higher at Ss 1. By contrast, DcPYR1 transcript remained at 0.002-0.006 r.e.l. in 'Miracle Rouge' ovary during flower opening and senescence. The transcripts of DcACS1, the key gene for ethylene biosynthesis, were detected at Ss 1 in 'LPB', and at Ss 2 in 'Excerea', but not in 'Miracle Rouge' throughout flower opening and senescence stages. These findings suggest that ABA plays a causal role in inducing the expression of the DcACS1 gene in the gynoecium, leading to ethylene biosynthesis, and that both the ABA content and DcPYR1 expression must be above putative threshold levels for ABA to exert its action.

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Language：English   Publishing type：Research paper (scientific journal)   Publisher：JAPAN SOC HORTICULTURAL SCI  

The cuticle, composed of cutin and associated waxes, probably acts as a barrier against water evaporation from the epidermal surface of flower petals. Cuticle formation begins with the biosynthesis of very-long-chain fatty acids (VLCFAs), catalyzed by a fatty acid elongase complex in epidermal cells. In the present study, cDNAs were cloned and analyzed for three enzymes (DcKCR1,DcHCD1, and DcECR1). Combined with the previously obtained cDNA for DcKCS1, the present study completes the identification of cDNAs for the fatty acid elongase complex in 'Light Pink Barbara' carnation for the first time. DcKCS1 transcripts were accumulated at flower opening stage (Os) 2 through Os 6 (full opening stage) with slight changes, but decreased markedly at senescence stage (Ss) 2 and Ss 4. Also, transcripts for DcKCR1, DcHCD1, and DcECR1 were present in considerable amounts during flower opening stages from Os 2 to Os 6. These findings suggested that the expressions of four genes are active during flower opening stage, which is concomitant with the expansion growth in petals requiring rapid formation of a waxy cuticle. Cut flowers of 'Miracle Rouge' carnation have an extremely long vase-life of about three weeks. The cuticle layer on the epidermal cells of 'Miracle Rouge' petals was thinner than that of 'Light Pink Barbara' petals, and 'Miracle Rouge' flowers had a depressed expression of DcKCS1,DcKCR1, and DcHCD1 in petals. These findings suggested that the prolonged vase-life of 'Miracle Rouge' flowers is not related to cuticle formation.

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Language：English   Publishing type：Research paper (scientific journal)   Publisher：JAPAN SOC HORTICULTURAL SCI  

We cloned seven cDNAs coding for ubiquitin (polyubiquitin) (DcUbq1-7) from car-tuition petals: DcUbq1, 2, 3 encoded polyubiquitins consisting of five ubiquitin monomers; DcUbq4, three monomers and DcUbq5, 6, 7, a monomer. The 3'-UTR nucleotide sequences were separated into three groups; two were specific to DcUbq1 and DcUbq2, respectively, and the third was almost always common to other genes (DcUbq3-7). The transcript levels of DcUbq1 and DcUbq2 in petals fluctuated during flower opening, whereas those of DcUbq3-7 remained unchanged except for an increase in the last stage. On the other hand, during flower senescence, the transcript levels of DcUbq1 and DcUbq2 increased at later stages, and those of DcUbq3-7 remained almost constant during the process. Based on these findings, we suggest an association of ubiquitin gene expression with petal growth during flower opening and petal wilting during the senescence of carnation flowers through the degradation of specific proteins by the ubiquitin-proteasome system. Furthermore, we showed the successful use of DcUby3-7 transcripts as a normalizing standard in the determination of transcript levels of a target gene in senescing carnation petals, where massive degradation of RNA, such as actin mRNA and rRNA, usually occurs, causing inaccuracy in the estimation of transcript levels of interest.

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Language：English   Publishing type：Research paper (scientific journal)   Publisher：SPRINGER HEIDELBERG  

Ethylene has an inhibitory effect on flowering in a short-day (SD) plant chrysanthemum (Chrysanthemum morifolium Ramat.). In this study, we used a hexaploid chrysanthemum 'Sei-Marine' and found that its transgenic lines transformed with a mutated ethylene receptor gene mDG-ERS1(etr1-4), which conferred reduced ethylene sensitivity (J. Plant Biol. 51: 424-427, 2008), opened flowers earlier than the non-transformed control. We examined whether the accelerated flower induction in the transformant occurred through the enhanced expression of chrysanthemum genes homologous to FLOWERING LOCUS T (FT), a floral inducer gene in Arabidopsis. We cloned three cDNAs for FT homologs (CmFTL1, CmFTL2, and CmFTL3) from 'Sei-Marine'. CmFTL2 putatively encodes a non-functional gene product due to a frame shift caused by a 2 bp-deletion in the coding region. RT-PCR analysis revealed differential expression patterns of CmFTL genes in the transgenic and control lines, suggesting that these genes might be under the control of ethylene. CmFTL1/2 mRNA level was lower in a SD condition than a long-day (LD) condition. CmFTL3 mRNA accumulated abundantly under SD condition as compared with LD condition in the transgenic line. These results suggest the association of increased expression of CmFTL3 gene with the accelerated flowering in the transgenic line with reduced ethylene sensitivity.

DOI： 10.1007/s12374-012-0109-8

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Language：English   Publishing type：Research paper (scientific journal)   Publisher：JAPAN SOC HORTICULTURAL SCI  

The intron structures of two variants of 1-aminocyclopropane-1-carboxylate synthase (ACS) genes (DcACS1a and DcACS1b) in carnation (Dianthus caryophyllus) and genes homologous to them (ACS1 homologous genes) in other 110 Dianthus species (16 strains in total) were studied by comparing the sizes of the PCR amplificates and nucleotide sequence of the introns. All 16 sequenced homologous ACS1 genes, including DcACS1 genes themselves, had five exons and four introns. The exons had similar nucleotide sequences and consequently similar deduced amino-acid sequences.. The sizes of three introns (intron-1, -2, -3) were variable among the homologous genes, whereas that of the fourth intron (intron-4) was almost identical. The variation in introns was probably caused by the insertion (and deletion) of nucleotide fragments of given lengths. Interestingly, the 3'-UTR of DcACS1a was different from that of DcACS1b, and the latter was similar to other 14 ACS1 homologous genes. Moreover, the length of Thr repeat in the C-terminal region was long in DcACS1a protein but short in DcACS1b protein, and the latter resembled ACS1 homologous proteins in other Dianthus species. The present findings suggest that (1) the variation in intron structure between two variants of carnation DcACS1 is reminiscent of the variation that occurred universally in Dianthus species, (2) DcACS1a is probably a gene intrinsic to carnation, and (3) DcACS1b was acquired from another, as yet unknown, Dianthus species, in the course of breeding modern carnal ion cultivars.

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Language：English   Publishing type：Research paper (scientific journal)   Publisher：JAPAN SOC HORTICULTURAL SCI  

Flower opening in carnations (Dianthus caryophyllus L.) is the result of the enlargement of petal cells, which requires sugar metabolism. A cDNA encoding sucrose synthase (DcSUS1) was isolated from carnation petals as a candidate gene acting in the initial step of sugar metabolism in petal cells. DcSUS1 transcripts were detected abundantly in floral tissues of flowering carnation plants; the transcripts accumulated most in the petals and style followed by the ovary, whereas only small accumulation were found in stems, leaves, and calyces. Moreover, nearly constant accumulation of DcSUS1 transcripts was found in the petals during flower opening, fully open, and early senescence periods, whereas decreasing accumulation was detected in petals when senescence progressed. These findings suggested the involvement of DcSUS1 expression in petal cell growth during the opening of carnation flowers.

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Authorship：Lead author   Language：English   Publishing type：Research paper (scientific journal)   Publisher：SPRINGER  

Carnation (Dianthus caryophyllus) flowers exhibit climacteric ethylene production followed by petal wilting, a senescence symptom. DcACS1, which encodes 1-aminocyclopropane-1-carboxylate synthase (ACS), is a gene involved in this phenomenon. We determined the genomic DNA structure of DcACS1 by genomic PCR. In the genome of &apos;Light Pink Barbara&apos;, we found two distinct nucleotide sequences: one corresponding to the gene previously shown as DcACS1, designated here as DcACS1a, and the other novel one designated as DcACS1b. It was revealed that both DcACS1a and DcACS1b have five exons and four introns. These two genes had almost identical nucleotide sequences in exons, but not in some introns and 3&apos;-UTR. Analysis of transcript accumulation revealed that DcACS1b is expressed in senescing petals as well as DcACS1a. Genomic PCR analysis of 32 carnation cultivars showed that most cultivars have only DcACS1a and some have both DcACS1a and DcACS1b. Moreover, we found two DcACS1 orthologous genes with different nucleotide sequences from D. superbus var. longicalycinus, and designated them as DsuACS1a and DsuACS1b. Petals of D. superbus var. longicalycinus produced ethylene in response to exogenous ethylene, accompanying accumulation of DsuACS1 transcripts. These data suggest that climacteric ethylene production in flowers was genetically established before the cultivation of carnation.

DOI： 10.1007/s00299-010-0962-1

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Authorship：Lead author   Language：English   Publishing type：Research paper (scientific journal)   Publisher：OXFORD UNIV PRESS  

Growth of petal cells is a basis for expansion and morphogenesis (outward bending) of petals during opening of carnation flowers (Dianthus caryophyllus L.). Petal growth progressed through elongation in the early stage, expansion with outward bending in the middle stage, and expansion of the whole area in the late stage of flower opening. In the present study, four cDNAs encoding xyloglucan endotransglucosylase/hydrolase (XTH) (DcXTH1-DcXTH4) and three cDNAs encoding expansin (DcEXPA1-DcEXPA3) were cloned from petals of opening carnation flowers and characterized. Real-time reverse transcription-PCR analyses showed that transcript levels of XTH and expansin genes accumulated differently in floral and vegetative tissues of carnation plants with opening flowers, indicating regulated expression of these genes. DcXTH2 and DcXTH3 transcripts were detected in large quantities in petals as compared with other tissues. DcEXPA1 and DcEXPA2 transcripts were markedly accumulated in petals of opening flowers. The action of XTH in growing petal tissues was confirmed by in situ staining of xyloglucan endotransglucosylase (XET) activity using a rhodamine-labelled xyloglucan nonasaccharide as a substrate. Based on the present findings, it is suggested that two XTH genes (DcXTH2 and DcXTH3) and two expansin genes (DcEXPA1 and DcEXPA2) are associated with petal growth and development during carnation flower opening.

DOI： 10.1093/jxb/erq319

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Language：English   Publishing type：Research paper (scientific journal)   Publisher：WILEY-BLACKWELL  

P&gt;This comprehensive overview of the xyloglucan endotransglucosylase/hydrolase (XTH) family of genes and proteins in bryophytes, based on research using genomic resources that are newly available for the moss Physcomitrella patens, provides new insights into plant evolution. In angiosperms, the XTH genes are found in large multi-gene families, probably reflecting the diverse roles of individual XTHs in various cell types. As there are fewer cell types in P. patens than in angiosperms such as Arabidopsis and rice, it is tempting to deduce that there are fewer XTH family genes in bryophytes. However, the present study unexpectedly identified as many as 32 genes that potentially encode XTH family proteins in the genome of P. patens, constituting a fairly large multi-gene family that is comparable in size with those of Arabidopsis and rice. In situ localization of xyloglucan endotransglucosylase activity in this moss indicates that some P. patens XTH proteins exhibit biochemical functions similar to those found in angiosperms, and that their expression profiles are tissue-dependent. However, comparison of structural features of families of XTH genes between P. patens and angiosperms demonstrated the existence of several bryophyte-specific XTH genes with distinct structural and functional features that are not found in angiosperms. These bryophyte-specific XTH genes might have evolved to meet morphological and functional needs specific to the bryophyte. These findings raise interesting questions about the biological implications of the XTH family of proteins in non-seed plants.

DOI： 10.1111/j.1365-313X.2010.04351.x

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Authorship：Lead author   Language：English   Publishing type：Research paper (scientific journal)   Publisher：OXFORD UNIV PRESS  

Flower opening is an event accompanied by morphological changes in petals which include elongation, expansion, and outward-curving. Petal cell growth is a fundamental process that underlies such phenomena, but its molecular mechanism remains largely unknown. Suppression subtractive hybridization was performed between petals during the early elongation period (stage 1) and during the opening period (stage 5) in carnation flowers and a pair of subtraction libraries abundant in differentially expressed genes was constructed at each stage. 393 cDNA clones picked up by differential screening out of 1728 clones were sequenced and 235 different cDNA fragments were identified, among which 211 did not match any known nucleotide sequence of carnation genes in the databases. BLASTX search of nucleotide sequences revealed that putative functions of the translational products can be classified into several categories including transcription, signalling, cell wall modification, lipid metabolism, and transport. Open reading frames of 15 selected genes were successfully determined by rapid amplification of cDNA ends (RACE). Time-course analysis of these genes by real-time RT-PCR showed that transcript levels of several genes correlatively fluctuate in petals of opening carnation flowers, suggesting an association with the morphological changes by elongation or curving. Based on the results, it is suggested that the growth of carnation petals is controlled by co-ordinated gene expression during the progress of flower opening. In addition, the possible roles of some key genes in the initiation of cell growth, the construction of the cell wall and cuticle, and transport across membranes were discussed.

DOI： 10.1093/jxb/erq064

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Authorship：Lead author   Language：English   Publishing type：Research paper (scientific journal)   Publisher：SPRINGER  

Pondweed (Potamogeton distinctus A. Benn.), a monocot aquatic plant species, has turions, which are overwintering buds forming underground as an asexual reproductive organ. Turions not only survive for more than one month but also elongate under strict anoxia, maintaining high-energy charge by activation of fermentation. We cloned 82 cDNA fragments of genes, that are up-regulated during anoxic growth of pondweed turions, by suppression subtractive hybridization. The transcript levels of 44 genes were confirmed to be higher under anoxia than those in air by both Northern blot analysis and a semi-quantitative reverse transcription polymerase chain reaction (RT-PCR) method. A homology search for their nucleotide sequences revealed that some of them are highly homologous to known sequences of genes from other plants. They included alcohol dehydrogenase, pyruvate decarboxylase (PDC), glyceraldehyde-3-phosphate dehydrogenase (GAPDH), vacuolar H+-translocating pyrophosphatase and a plasma membrane intrinsic protein. Time courses of transcript accumulation of some genes under anoxia were different from those in air. The activity of PDC increased under anoxic conditions but the activities of GAPDH and pyrophosphatase remained constant after anoxic treatment. Anoxically up-regulated genes are possibly involved in physiological events to control energy production, pH regulation and cell growth under anoxia. These results suggest that transcriptional regulation of these genes serves as an essential part of survival and growth of pondweed turions under anoxia.

DOI： 10.1007/s00425-007-0537-8

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Language：English   Publishing type：Doctoral thesis  

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Authorship：Lead author   Language：English   Publishing type：Research paper (scientific journal)   Publisher：OXFORD UNIV PRESS  

Background and Aims Overwintering buds (turions) of the monocot aquatic pondweed species (Potamogeton distinctus) are highly tolerant to anoxic stress. Sucrose metabolism accompanied by enhanced activity of sucrose synthase (SuSy) operates actively during anaerobic elongation of pondweed turions. The aim of this study is to isolate SuSy genes from the turions and to investigate their transcriptional changes in response to anoxia and other stimuli.
Methods SuSy genes were isolated from pondweed turions by PCR methods and transcript levels of SuSy genes were examined in response to anoxia, sugars and plant hormones. In addition, the effects of anoxia on SuSy activity were examined both in the soluble fraction and in the microsomal fraction.
Key Results cDNAs of two SuSy genes (PdSUS1 and PdSUS2) were cloned from pondweed turions. The levels of PdSUS1 transcripts increased under anoxia but did not with sugar treatments. Anoxia-stimulated elongation of turions was further enhanced by 2,4-dichlorophenoxyacetic acid (2,4-D) and suppressed by treatments with sorbitol, 2-deoxyglucose (2-dGlc) and abscisic acid (ABA). The levels of PdSUS1 transcripts were increased by 2,4-D and decreased by sorbitol under anoxia. The levels of PdSUS2 transcripts were not significantly affected by anoxia and any other treatments. SuSy activity of turions under anoxia was enhanced in the soluble fraction, but not in the microsomal fraction.
Conclusions Up-regulation of PdSUS1 transcription under anoxia may not be attributed to sugar starvation under anoxia. A positive correlation between stem elongation and the level of PdSUS1 transcripts was observed in turions treated with anoxic conditions, 2,4-D and sorbitol. The increase in SuSy activity in the cytosol may contribute to sugar metabolism and sustain stem elongation under anoxia.

DOI： 10.1093/aob/mci220

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Authorship：Lead author   Language：Japanese  

CiNii Article

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Authorship：Lead author   Language：English   Publishing type：Research paper (scientific journal)  

DOI： 10.1023/A:1024585015697

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Language：Japanese  

CiNii Article

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Language：English   Publishing type：Research paper (scientific journal)   Publisher：OXFORD UNIV PRESS  

Stem segments prepared from pondweed (Potamo geton distinctus A. Benn.) turions (overwintering buds) elongate in anaerobic conditions, whereas there is almost no elongation in air. The anaerobic elongation was accompanied by a decrease in dry weights of stem segments, mainly due to consumption of storage starch in the amyloplasts of stem cells. On the other hand, total contents of amino acids increased in stem segments, in which contents of alanine, valine, leucine, and isoleucine increased, but contents of asparatic acid decreased. Moreover, contents of lactate in stem tissues increased at an early stage of anaerobic incubation. In tracer experiments with C-14-glucose, C-14 incorporation into stem tissues in anoxia was only half of that in normoxia. However, conversion of C-14 to ethanol occurred exclusively in anoxia. C-14-labelled metabolites were analysed by two-dimensional cellulose thin-layer chromatography. C-14 incorporation into sucrose and alanine was significantly increased in anoxia. The activity of alanine aminotransferase was enhanced by anoxia, suggesting that pyruvate is a precursor of alanine synthesis. The results suggest that pondweed turions produce energy necessary for anaerobic elongation by activating conversion of storage starch in the amyloplasts to ethanol, alanine and lactate.

DOI： 10.1093/jxb/erf036

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Event date： 2022.10.1

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Event date： 2022.9.14 - 2022.9.17

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Event date： 2020.12.15 - 2020.12.17

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