掲載種別：研究論文（学術雑誌）   出版者・発行元：Japanese Society for Horticultural Science  

DOI： 10.2503/hrj.22.303

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DOI： 10.1101/2022.12.29.522208

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出版者・発行元：Cold Spring Harbor Laboratory  

Summary

Multiplexed inter-simple sequence repeats genotyping by sequencing (MIG-seq) is an next-generation sequencing library construction method developed for the analysis of DNA in ecology. Although MIG-seq can generate libraries from low-quality DNA, few polymorphisms can be obtained in species with small genomes. In this study, we developed degenerate oligonucleotide primer MIG-seq (dpMIG-seq) as an effective polymorphism discovery method that allows for variation in the number of polymorphisms while retaining the advantages of MIG-seq, including independence from DNA quality. In dpMIG-seq, a proportion of the simple sequence repeats in the primer sequence of the first PCR in MIG-seq was changed to degenerate oligonucleotides to enable annealing to a wider range of sequences. In tests of several crop species other than wheat, the number of loci that could be sequenced using dpMIG-seq with a data volume of 0.3 gigabases (Gb) was increased compared with that sequenced using MIG-seq. In wheat, the number of polymorphisms obtained via dpMIG-seq was higher than that obtained via MIG-seq when a data volume of about ≥2 Gb was obtained. In dpMIG-seq, different loci could be sequenced by changing the positions of the degenerate oligonucleotides. By applying dpMIG-seq, we constructed a linkage map consisting of 5,142 markers for the rice inter-subspecies F₂ population, and we detected quantitative trait loci for heading date in the regions where known heading-related genes were located. Overall, our results show that dpMIG-seq is a useful tool for the genetic analysis of crop species.

DOI： 10.1101/2022.08.25.504752

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記述言語：英語   掲載種別：研究論文（学術雑誌）  

In the evolutionary history of plants, variation in cis-regulatory elements (CREs) resulting in diversification of gene expression has played a central role in driving the evolution of lineage-specific traits. However, it is difficult to predict expression behaviors from CRE patterns to properly harness them, mainly because the biological processes are complex. In this study, we used cistrome datasets and explainable convolutional neural network (CNN) frameworks to predict genome-wide expression patterns in tomato (Solanum lycopersicum) fruit from the DNA sequences in gene regulatory regions. By fixing the effects of trans-acting factors using single cell-type spatiotemporal transcriptome data for the response variables, we developed a prediction model for crucial expression patterns in the initiation of tomato fruit ripening. Feature visualization of the CNNs identified nucleotide residues critical to the objective expression pattern in each gene, and their effects were validated experimentally in ripening tomato fruit. This cis-decoding framework will not only contribute to the understanding of the regulatory networks derived from CREs and transcription factor interactions, but also provides a flexible means of designing alleles for optimized expression.

DOI： 10.1093/plcell/koac079

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記述言語：英語   掲載種別：研究論文（学術雑誌）   出版者・発行元：International Society for Horticultural Science (ISHS)  

DOI： 10.17660/actahortic.2022.1338.49

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記述言語：英語   掲載種別：研究論文（学術雑誌）  

In flowering plants, different lineages have independently transitioned from the ancestral hermaphroditic state into and out of various sexual systems1. Polyploidizations are often associated with this plasticity in sexual systems2,3. Persimmons (the genus Diospyros) have evolved dioecy via lineage-specific palaeoploidizations. More recently, hexaploid D. kaki has established monoecy and also exhibits reversions from male to hermaphrodite flowers in response to natural environmental signals (natural hermaphroditism, NH), or to artificial cytokinin treatment (artificial hermaphroditism, AH). We sought to identify the molecular pathways underlying these polyploid-specific reversions to hermaphroditism. Co-expression network analyses identified regulatory pathways specific to NH or AH transitions. Surprisingly, the two pathways appeared to be antagonistic, with abscisic acid and cytokinin signalling for NH and AH, respectively. Among the genes common to both pathways leading to hermaphroditic flowers, we identified a small-Myb RADIALIS-like gene, named DkRAD, which is specifically activated in hexaploid D. kaki. Consistently, ectopic overexpression of DkRAD in two model plants resulted in hypergrowth of the gynoecium. These results suggest that production of hermaphrodite flowers via polyploidization depends on DkRAD activation, which is not associated with a loss-of-function within the existing sex determination pathway, but rather represents a new path to (or reinvention of) hermaphroditism.

DOI： 10.1038/s41477-022-01107-z

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掲載種別：研究論文（学術雑誌）   出版者・発行元：Elsevier {BV}  

DOI： 10.1016/j.postharvbio.2021.111787

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掲載種別：研究論文（学術雑誌）   出版者・発行元：Oxford University Press (OUP)  

Abstract

MIG-seq (Multiplexed inter-simple sequence repeats genotyping by sequencing) has been developed as a low cost genotyping technology, although the number of polymorphisms obtained is assumed to be minimal, resulting in the low application of this technique to analyses of agricultural plants. We applied MIG-seq to 12 plant species that include various crops and investigated the relationship between genome size and the number of bases that can be stably sequenced. The genome size and the number of loci, which can be sequenced by MIG-seq, are positively correlated. This is due to the linkage between genome size and the number of simple sequence repeats (SSRs) through the genome. The applicability of MIG-seq to population structure analysis, linkage mapping, and quantitative trait loci (QTL) analysis in wheat, which has a relatively large genome, was further evaluated. The results of population structure analysis for tetraploid wheat showed the differences among collection sites and subspecies, which agreed with previous findings. Additionally, in wheat biparental mapping populations, over 3,000 SNPs/indels with low deficiency were detected using MIG-seq, and the QTL analysis was able to detect recognized flowering-related genes. These results revealed the effectiveness of MIG-seq for genomic analysis of agricultural plants with large genomes, including wheat.

DOI： 10.1093/dnares/dsac011

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その他リンク： https://academic.oup.com/dnaresearch/article-pdf/29/2/dsac011/43439343/dsac011.pdf

掲載種別：研究論文（学術雑誌）  

In contrast to the progress in the research on physiological disorders relating to shelf life in fruit crops, it has been difficult to non-destructively predict their occurrence. Recent high-tech instruments have gradually enabled non-destructive predictions for various disorders in some crops, while there are still issues in terms of efficiency and costs. Here, we propose application of a deep neural network (or simply deep learning) to simple RGB images to predict a severe fruit disorder in persimmon, rapid over-softening. With 1,080 RGB images of ‘Soshu’ persimmon fruits, three convolutional neural networks (CNN) were examined to predict rapid over-softened fruits with a binary classification and the date to fruit softening. All of the examined CNN models worked successfully for binary classification of the rapid over-softened fruits and the controls with > 80% accuracy using multiple criteria. Furthermore, the prediction values (or confidence) in the binary classification were correlated to the date to fruit softening. Although the features for classification by deep learning have been thought to be in a black box by conventional standards, recent feature visualization methods (or “explainable” deep learning) has allowed identification of the relevant regions in the original images. We applied Grad-CAM, Guided backpropagation, and layer-wise relevance propagation (LRP), to find early symptoms for CNNs classification of rapid over-softened fruits. The focus on the relevant regions tended to be on color unevenness on the surface of the fruit, especially in the peripheral regions. These results suggest that deep learning frameworks could potentially provide new insights into early physiological symptoms of which researchers are unaware.

DOI： 10.2503/hortj.UTD-323

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記述言語：英語   掲載種別：研究論文（学術雑誌）  

Peel degreening is the most conspicuous aspect of fruit ripening in many citrus fruits because of its importance for marketability. In this study, peel degreening in response to propylene (an ethylene analog) and at varying storage temperatures was characterized in Satsuma mandarin (Citrus unshiu Marc.) fruit. Propylene treatment triggered rapid peel degreening (within 4-6 days), indicated by an increase in the citrus color index (CCI) and chlorophyll loss. Peel degreening was also observed in fruit at 10°C and 15°C after 28-42 days, with gradual CCI increase and chlorophyll reduction. However, fruit at 5°C, 20°C, and 25°C remained green, and no substantial changes in peel CCI and chlorophyll content were recorded during the 42-day storage duration. The transcriptomes of peels of fruit treated with propylene for 4 days and those stored at varying temperatures for 28 days were then analyzed by RNA-Seq. We identified three categories of differentially expressed genes that were regulated by (i) propylene (and by analogy, ethylene) alone, (ii) low temperature (5°C, 10°C, or 15°C vs. 25°C) alone, and (iii) either propylene or low temperature. Gene-encoding proteins associated with chlorophyll degradation (such as CuSGR1, CuNOL, CuACD2, CuCAB2, and CuLHCB2) and a transcription factor (CuERF114) were differentially expressed by propylene or low temperature. To further examine temperature-induced pathways, we also monitored gene expression during on-tree fruit maturation vs. postharvest. The onset of on-tree peel degreening coincided with autumnal drops in field temperatures, and it was accompanied by differential expression of low temperature-regulated genes. On the contrary, genes that were exclusively regulated by propylene (such as CuCOPT1 and CuPOX-A2) displayed insignificant expression changes during on-tree peel degreening. These findings indicate that low temperatures could be involved in the fruit ripening-related peel degreening independently of ethylene.

DOI： 10.3389/fpls.2022.918226

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記述言語：英語   掲載種別：研究論文（学術雑誌）  

DOI： 10.1016/j.postharvbio.2020.111436

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記述言語：英語   掲載種別：研究論文（学術雑誌）  

DOI： 10.3389/fpls.2021.638321

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掲載種別：研究論文（学術雑誌）   出版者・発行元：Japanese Society for Horticultural Science  

DOI： 10.2503/hrj.20.455

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記述言語：英語   掲載種別：研究論文（学術雑誌）  

DOI： 10.3389/fpls.2020.567249

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担当区分：最終著者,　責任著者   記述言語：英語   掲載種別：研究論文（学術雑誌）  

DOI： 10.1093/jxb/eraa206

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その他リンク： http://academic.oup.com/jxb/article-pdf/71/16/4778/33573305/eraa206.pdf

記述言語：英語   掲載種別：研究論文（学術雑誌）  

DOI： 10.1093/dnares/dsaa012

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記述言語：日本語   掲載種別：研究論文（学術雑誌）   出版者・発行元：一般社団法人 園芸学会  

モモ（Prunus persica（L.）Batsch）の肉質は，収穫後の貯蔵および消費者の嗜好性の観点から重要な形質である．溶質モモの果実は典型的なクライマクテリック型の成熟を示し，成熟後期にエチレン生成の急増と果実硬度の急速な低下がみられる．一方で，硬肉モモではエチレンがほとんど生成されず，樹上や収穫後において硬度が高く維持される．本研究では，‘川中島白桃’の放任受粉の実生から選抜されたと言われている‘桃水’が硬肉性を持つことを明らかにした．‘桃水’は棚持ち期間が長く，肉質がサクサクしているが，そのエチレン生成や軟化特性は明らかにされていない．そこで，エチレンのアナログであるプロピレンを外生処理した‘桃水’果実のエチレン生成および果実軟化の様相を調査した．無処理の果実では，収穫時期や栽培地に関わらずエチレンはほとんど検出されず，果実硬度が高く維持された．プロピレン処理により果実硬度は著しく低下したが，その間もエチレン生成はほとんど検出されなかった．モモの硬肉性の原因遺伝子として同定されたPpYUC11遺伝子の遺伝子型調査により，PpYUC11遺伝子の5′隣接領域におけるトランスポゾン様配列の挿入および第1イントロン内のSSRのいずれについても，‘桃水’の遺伝子型は他の硬肉品種の遺伝子型と同じであることが明らかになった．これらの結果から，‘桃水’はエチレン生成および果実軟化が抑制された硬肉モモであることが示唆された．

DOI： 10.2503/hrj.19.61

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その他リンク： https://www.jstage.jst.go.jp/article/hrj/19/1/19_61/_pdf

担当区分：最終著者,　責任著者   記述言語：英語   掲載種別：研究論文（学術雑誌）  

DOI： 10.3389/fpls.2020.554158

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担当区分：責任著者   記述言語：英語   掲載種別：研究論文（学術雑誌）  

DOI： 10.1016/j.postharvbio.2019.110949

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担当区分：筆頭著者   記述言語：英語   掲載種別：研究論文（学術雑誌）  

DOI： 10.1094/MPMI-04-19-0093-A

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記述言語：英語   掲載種別：研究論文（学術雑誌）  

DOI： 10.3389/fpls.2019.00888

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掲載種別：研究論文（学術雑誌）  

DOI： 10.1016/j.postharvbio.2018.08.017

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記述言語：英語   掲載種別：研究論文（学術雑誌）   出版者・発行元：OXFORD UNIV PRESS  

Parthenocarpy, a process in which fruit set occurs without fertilization, leads to the production of seedless fruit. A number of floral homeotic mutants with abnormal stamen development exhibit parthenocarpic fruit set. Flower development is thought to repress ovary growth before anthesis. However, the mechanism of parthenocarpic fruit development caused by aberrant flower formation is poorly understood. To investigate the molecular mechanism of parthenocarpic fruit development in floral homeotic mutants, we performed functional analysis of Tomato APETALA3 (TAP3) by loss-of-function approaches. Organ-specific promoter was used to induce organ-specific loss of function in stamen and ovary/fruit. We observed increased cell expansion in tap3 mutants and TAP3-RNAi lines during parthenocarpic fruit growth. These were predominantly accompanied by the up-regulation of GA biosynthesis genes, including SlGA20ox1, SlGA20ox2, and SlGA20ox3, as well as reduced expression of the GA-inactivating gene SlGA2ox1 and the auxin signaling gene SlARF7 involved in a crosstalk between GA and auxin. These transcriptional profiles are in agreement with the GA levels in these lines. These results suggest th

DOI： 10.1093/pcp/pcy184

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掲載種別：研究論文（国際会議プロシーディングス）  

DOI： 10.17660/ActaHortic.2018.1218.71

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記述言語：英語   掲載種別：研究論文（学術雑誌）  

DOI： 10.2503/hortj.OKD-140

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掲載種別：研究論文（学術雑誌）  

DOI： 10.1186/s12870-018-1264-y

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掲載種別：研究論文（学術雑誌）  

DOI： 10.2503/hortj.OKD-035

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記述言語：英語   掲載種別：研究論文（学術雑誌）  

DOI： 10.2503/hortj.OKD-094

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記述言語：英語   掲載種別：研究論文（学術雑誌）  

DOI： 10.2503/hortj.OKD-028

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記述言語：英語   掲載種別：研究論文（学術雑誌）  

DOI： 10.1093/pcp/pcw157

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記述言語：英語   掲載種別：研究論文（学術雑誌）  

DOI： 10.2503/hortj.MI-066

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記述言語：英語   掲載種別：研究論文（学術雑誌）   出版者・発行元：ELSEVIER SCIENCE BV  

We investigated the physiological effects of short-term postharvest near infrared (NIR) radiation on relative transpiration rates, stomatal apertures, and reactive oxygen species (ROS) levels in guard cells on excised young lettuce leaves and on transpiration of leaf lettuce at commercial maturity. When the young leaves were radiated by NIR of wavelengths longer than 850 nm at 100 mu mol m(-2) s(-1) for short duration (10-60) min, relative transpiration rates during subsequent storage were reduced, but not by visible light radiation and by longer radiation (180 min) of NIR. The reduction in transpiration rates by the short-term NIR radiation was greater at 10 degrees C than at 25 degrees C under both dark and light conditions during subsequent storage. The short-term NIR radiation enhanced stomatal closure and ROS accumulation in guard cells of young lettuce leaves. These results indicate that the suppression of transpiration by short-term NIR radiation is likely to be mediated through stomatal closure due to NIR-induced ROS accumulation. The reduction of transpiration by short-term NIR radiation was obtained not only in excised young leaves but also in leaf lettuce at commercial maturity, resulting in keeping freshness. The short-term NIR radiation could be an additional means to extend shelf life of leaf vegetables. (C) 2015 Elsevier B.V. All rights reserved.

DOI： 10.1016/j.postharvbio.2015.05.010

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担当区分：筆頭著者,　責任著者   記述言語：英語   掲載種別：研究論文（学術雑誌）   出版者・発行元：JAPAN SOC HORTICULTURAL SCI  

Two types of floral morph, called thrum and pin, occur in heterostylous species. The thrum and pin flowers have shorter and longer styles, respectively. Heterostyly in Linum has been studied since Darwin's era. The floral morph, self-incompatibility, and related phenotypes have been well characterized using a natural population, but genetic analysis using a segregated population has not been reported, and the mode of inheritance of heterostyly in Linum remains to be investigated. We prepared a segregated population by crossing thrum and pin flowers of Linum grandiflorum Desf. and investigated style and stamen lengths. On the basis of the style to stamen length ratio, the population could be divided into thrum and pin clearly at a ratio of 1:1. Style length of pins was 1.6 times longer than that of thrums. To investigate the factor regulating the difference in style length, we further measured the style cell length of thrums and pins. The style cell length of pins was longer than that of thrums, whose rate was comparable to the rate of the style length ratio between thrum and pin flowers. These findings indicate that the floral morph of heterostyly in Linum is controlled by a single diallelic locus, and that a difference in the cell expansion rate caused thrum and pin morphs in the style, such as in a typical heterostylous species. PCR genotyping showed that TSS1, an S candidate gene reported previously, cosegregated completely with the thrum phenotype, indicating strong linkage between the S locus and TSS1. Furthermore, three flower colors (red, pink, and white) were observed at a 1:2:1 ratio, and no white thrum flowers or red pin flowers were found in this population. These flower color phenotypes could also be controlled by a diallelic locus, whose two alleles (R and r) would be incompletely dominant, and therefore, may be linked to the S locus.

DOI： 10.2503/hortj.MI-045

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掲載種別：研究論文（学術雑誌）  

DOI： 10.2503/hrj.14.61

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記述言語：日本語   出版者・発行元：園芸学会  

エダマメは主に夏季に栽培，収穫され気温が高いことから，収穫後に品質低下しやすい．本研究では'ふくら'と'湯あがり娘'を用い，種々の貯蔵温度，形態（"枝葉付き"，"枝付き"，"もぎ莢"）が収穫後のエダマメの品質に及ぼす影響を経時的に調査した．いずれの品種でも，15°C以上の貯蔵では糖含量は3時間後に収穫時の約80％以下に，遊離アミノ酸含量は6時間後に収穫時の約80％以下に減少した．常温近くで貯蔵すると3時間という非常に短い間に品質が低下することが明らかになった．しかし，10°C以下の貯蔵条件は，外観保持効果に優れ，糖含量は10時間後でも収穫時の約80％以上，遊離アミノ酸含量は24時間後でも収穫時の約80％以上保持した．低温貯蔵は品質低下抑制に非常に有効であった．枝葉付き貯蔵にも一定の品質保持効果が見られたが，貯蔵温度と時間の影響の方が明らかに大きかった．これらの結果から，エダマメを高品質に保つためには，収穫後3時間以内に10°C以下の温度に管理することが重要と考えられた．

DOI： 10.2503/hrj.14.297

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その他リンク： http://agriknowledge.affrc.go.jp/RN/2010892824

記述言語：日本語   出版者・発行元：日本食品工学会  

エダマメを凍結すると細胞が損傷し，食感品質が低下する．デンプンを多く含むため，食感に影響する要素として糊化度に注目し，凍結時の糊化度が物性に及ぼす影響を，'味緑'と'湯あがり娘'を用いて調査した．<br>加熱時間を変えて糊化度の異なるエダマメを数種類作成し，凍結した．流水解凍した後，再度加熱し，ほぼ完全に糊化させた．物性を測定した結果，凍結時の糊化度の影響を受けて検体間で異なる値を示した．糊化度55～83％で凍結したエダマメは，解凍後の荷重，CI値が高く，ほぼ完全に糊化させても値は大きく低下せず，他糊化度で凍結したもの比べて高い値を示した．しかし，糊化度41％以下で凍結したエダマメは，解凍後高い荷重，CI値を示すが，加熱によりほぼ完全に糊化されることで値が大きく低下した．また，糊化度92％以上で凍結したエダマメは，解凍後の時点で低い荷重，CI値を示し，ほぼ完全糊化の加熱により値が若干低下した．

DOI： 10.11301/jsfe.16.63

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その他リンク： http://agriknowledge.affrc.go.jp/RN/2010891565

担当区分：筆頭著者   記述言語：英語   掲載種別：研究論文（学術雑誌）   出版者・発行元：ELSEVIER SCIENCE BV  

Double flowers induced by virus-induced gene silencing (VIGS) of two C-class MADS-box genes, pMADS3 and FBP6, were investigated in four cultivars of Petunia hybrida. In flowers induced by either pMADS3-VIGS or FBP6-VIGS, only small changes in commercial appearance were recognized regardless of cultivar, whereas in those induced by pMADS3/FBP6-VIGS, complete conversion of stamens into petaloid tissues and marked enlargement of upper limb-like tissues were observed, resulting in a decorative appearance in all the four cultivars. In whorl 4, cultivar-dependent conversion of carpels into new flowers was noted in pMADS3/FBP6-VIGS flowers. Of the four cultivars, only 'Mambo Purple' exhibited the development of new flowers instead of carpels and the emergence of ectopic new flowers from the axil of whorl 3 organs, which created an ornamental appearance with a high commercial value. Investigation of large and small petaloid stamens induced by pMADS3/FBP6-VIGS and pMADS3-VIGS, respectively, revealed only small differences in cell size compared to the large difference in total surface area. Quantitative RT-PCR analysis of SQUAMOSA/AP1/FRU type A-class genes, FBP29, PFG, and FBP26, showed no close relationship between the expression of those genes and petaloid stamen size. The suppressed C-class gene function at the late stage of flower development has little influence on the final size of petaloid tissue. (C) 2014 Elsevier IV. All rights reserved.

DOI： 10.1016/j.scienta.2014.07.029

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記述言語：英語   掲載種別：研究論文（学術雑誌）   出版者・発行元：WILEY-BLACKWELL  

Heterostylous species have two types of flowers, thrum and pin morphs, and these are controlled by a single diallelic locus designated the S locus; fertilization between these two types of flowers is successful. The S gene and the molecular mechanism by which it operates remain to be uncovered, although heterostyly has been studied since the time of Darwin. We compared transcripts and proteins of the thrum and pin flowers of heterostylous flax (Linum grandiflorum) to characterize the molecular differences between them and to elucidate the molecular machinery of heterostyly. Twelve floral morph-related genes were eventually isolated by an integrated study of subtraction and 2D-PAGE analyses, and four genes, TSS1, LgAP1, LgMYB21 and LgSKS1, were predicted to be related to heterostyly. TSS1, a thrum style-specific gene, showed some features suitable for the S gene. Although its biological function is unclear, TSS1 was expressed only in the thrum style and is probably linked to the S locus. LgMYB21, another thrum style gene, would be involved in floral morphogenesis. LgMYB21 was highly expressed in the thrum style, which is shorter than the pin style, and its overexpression in Arabidopsis reduced pistil length. Furthermore, a comparison of transcript and protein accumulations showed no differences in the mRNA accumulation of some thrum-specific proteins, including LgSKS1, suggesting that these are regulated by floral morph-specific post-transcriptional regulation. The Linum S locus regulates not only S specificity but also many floral phenotypes. Dynamic regulation of transcripts and proteins would be necessary for the pleiotropic function of the Linum S locus.

DOI： 10.1111/j.1365-313X.2011.04792.x

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記述言語：英語   掲載種別：研究論文（学術雑誌）   出版者・発行元：OXFORD UNIV PRESS  

Fruit ripening in response to treatments with propylene, 1-methycyclopropene (1-MCP), and low temperature was characterized in 'Sanuki Gold' kiwifruit, Actinidia chinensis Planch. Propylene treatment immediately induced rapid fruit softening, increased AC-PG (polygalacturonase) and AC-EXP (expansin) mRNA accumulation, and stimulated an increase in the soluble solid concentration (SSC) and a decrease in titratable acidity (TA). After 3 d exposure to propylene, ethylene production and AC-PL (pectate lyase) mRNA accumulation were observed. 1-MCP treatment after 24 h exposure to propylene eliminated AC-PG mRNA accumulation and suppressed continued changes in SSC and TA. Application of 1-MCP at the start of the treatment, followed by continuous propylene exposure, markedly delayed fruit softening, and the expression of the cell wall-modifying genes, and changes in the SSC and TA, indicating that kiwifruit become insensitive to ethylene at least for 3 d following 1-MCP exposure. Surprisingly, significant fruit softening, mRNA accumulation of AC-PG, AC-PL, and AC-EXP, and decreased TA were observed without ethylene production in intact fruit stored at low temperature for 1 month, but not in fruit stored at room temperature. Repeated 1-MCP treatments (twice a week) failed to inhibit the changes that occurred in low temperature storage. These observations indicate that low temperature modulates the ripening of kiwifruit in an ethylene-independent manner, suggesting that kiwifruit ripening is inducible by either ethylene or low temperature signals.

DOI： 10.1093/jxb/err324

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記述言語：英語   出版者・発行元：AMER SOC PLANT BIOLOGISTS  

DOI： 10.1105/tpc.111.231260

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記述言語：英語   掲載種別：研究論文（学術雑誌）   出版者・発行元：WILEY-BLACKWELL  

The S-RNase-based gametophytic self-incompatibility (SI) of Rosaceae, Solanaceae, and Plantaginaceae is controlled by at least two tightly linked genes located at the complex S locus; the highly polymorphic S-RNase for pistil specificity and the F-box gene (SFB/SLF) for pollen. Self-incompatibility in Prunus (Rosaceae) is considered to represent a self recognition by a single factor system, because loss-of-function of SFB is associated with self-compatibility, and allelic divergence of SFB is high and comparable to that of S-RNase. In contrast, Petunia (Solanaceae) exhibits non-self recognition by multiple factors. However, the distribution of self recognition and non-self recognition SI systems in different taxa is not clear. In addition, in non-self recognition systems, a loss-of-function phenotype of pollen S is unknown. Here we analyze the divergence of SFBB genes, the multiple pollen S candidates, of a rosaceous plant Japanese pear (Pyrus pyrifolia) and show that intrahaplotypic divergence is high and comparable to the allelic diversity of S-RNase while interhaplotypic divergence is very low. Next, we analyzed loss-of-function of the SFBB1 type gene. Genetic analysis showed that pollen with the mutant haplotype S4sm lacking SFBB1-S4 is rejected by pistils with an otherwise compatible S1 while it is accepted by other non-self pistils. We found that the S5 haplotype encodes a truncated SFBB1 protein, even though S5 pollen is accepted normally by pistils with S1 and other non-self haplotypes. These findings suggest that Japanese pear has a non-self recognition by multiple factors SI system, although it is a species of Rosaceae to which Prunus also belongs.

DOI： 10.1111/j.1365-313X.2011.04752.x

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記述言語：英語   掲載種別：研究論文（学術雑誌）   出版者・発行元：JAPAN SOC HORTICULTURAL SCI  

The effects of modified atmosphere (MA) storage and application of 1-methylcyclopropene (1-MCP) at harvest on the storability and quality of 'Sanuki Gold' kiwifruit harvested at two different maturity stages were investigated. MA storage in both fruit harvested early at 136 days after pollination (DAP) or late at 154 DAP delayed flesh softening, increase in soluble solid concentrations (SSC), decrease in titratable acids (TA), and reduction in fruit flesh color index compared to air stored fruit, suggesting that MA storage is effective in prolonging 'Sanuki Gold' kiwifruit storage life. Further, MA stored fruit did not attain full ripening flesh firmness and SSC thresholds even after 4 months of storage under MA conditions, suggesting that early harvested 'Sanuki Gold' kiwifruit can be stored for 4 months in MA. Fruit from both harvesting maturity stages stored under air conditions achieved maximum SSC (18%) values during storage, suggesting that two weeks early harvesting did not compromise edible quality characteristics. Only late harvested fruit treated with 1-MCP and stored in MA showed slight inhibitory effect specific to fruit softening during the first and second month of storage, suggesting that 1-MCP may have some limited ripening inhibitory effect during storage of 'Sanuki Gold' kiwifruit.

DOI： 10.2503/jjshs1.80.372

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記述言語：英語   掲載種別：研究論文（学術雑誌）   出版者・発行元：ELSEVIER SCIENCE BV  

Ethylene biosynthesis in kiwiftuit, Actinidia chinensis &apos;Sanuki Gold&apos; was characterized using propylene, an ethylene analog, and 1-methylcyclopropene (1-MCP), an inhibitor of ethylene perception. In fruit harvested between a young stage (66 days after pollination) (DAP) and an early commercial harvesting stage (143 DAP), 2 days of exposure to propylene were sufficient to initiate ethylene biosynthesis while in fruit harvested at commercial harvesting stage (154 DAP), 4 days of propylene treatment were required. This observation suggests that response of ethylene biosynthesis to propylene treatment in kiwifruit declined with fruit maturity. Propylene treatment resulted in up-regulated expression of AC-ACO1, AC-ACO2, AC-SAM1 and AC-SAM2, prior to the induction of AC-ACS1 and ethylene production, confirming that AC-ACS1 is the rate limiting step in ethylene biosynthesis in kiwiftuit. Treatment of fruit with more than 5 mu L L(-1) of 1-MCP after the induction of ethylene production subsequently suppressed ethylene production and expression of ethylene biosynthesis genes. Treatment of fruit with 1-MCP at harvest followed with propylene treatment delayed the induction of ethylene production and AC-ACS1 expression for 5 days. These observations suggest that in ripening kiwifruit, ethylene biosynthesis is regulated by positive feedback mechanism and that 1-MCP treatment at harvest effectively delays ethylene production by 5 days. (C) 2009 Elsevier B.V. All rights reserved.

DOI： 10.1016/j.postharvbio.2009.08.007

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記述言語：英語   掲載種別：研究論文（学術雑誌）   出版者・発行元：OXFORD UNIV PRESS  

Gametophytic self-incompatibility (GSI) in Solanaceae, Rosaceae, and Plantaginaceae is controlled by a multiallelic S-locus. The specificities of pistil and pollen are controlled by separate S-locus genes, S-RNase and SLF/SFB, respectively. Although the S-specificity is determined by the S-locus genes, factors located outside the S-locus are also required for expression of GSI. HT-B is one of the pistil non-S-factors identified in Nicotiana and Solanum, and encodes a small asparagine/aspartate-rich extracellular protein with unknown biochemical function. Here, HT-B was cloned from Petunia and characterized. The structural features and expression pattern of Petunia HT-B were very similar to those of Nicotiana and Solanum. Unlike other solanaceous species, expression of HT-B was also observed in self-compatible Petunia species. RNA interference (RNAi)-mediated suppression of Petunia HT-B resulted in partial breakdown of GSI. Quantitative analysis of the HT-B mRNA accumulation in the transgenics showed that a 100-fold reduction is not sufficient and a &gt; 1000-fold reduction is required to achieve partial breakdown of GSI.

DOI： 10.1093/jxb/erp005

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記述言語：日本語   出版者・発行元：日本食品保蔵科学会  

セイヨウナシ&#039;ラ・フランス&#039;果実は成熟時に爆発的なエチレン生成と急激な果肉軟化を示す典型的なクライマクテリック型果実である。一方, 同じナシ属に属するチュウゴクナシ品種&#039;ヤーリー&#039;果実は成熟中に多量のエチレンを生成するが軟化しない。本研究では果実軟化の制御にかかわる因子を同定するために, これらナシ果実の細胞壁タンパク質のプロテオーム解析を行った。さまざまな成熟ステージの&#039;ラ・フランス&#039;細胞壁タンパク質を二次元電気泳動で分離したところ, 成熟に伴い変化する19個のタンパク質スポットを得た。これらのうち, エチレン作用阻害剤である1-MCP処理による軟化抑制を行うと, 15個のタンパク質の蓄積がプレクライマクテリック期と同等になった。また, 19個のタンパク質のうち8個が&#039;ヤーリー&#039;果実では成熟に伴って変動せず, 果肉硬度と同じ挙動を示す結果となった。このうち1-MCPにより変動し, &#039;ヤーリー&#039;では変化しなかったのは2個のスポットである。これら2個のタンパク質の蓄積パターンは果肉軟化様相と同じ挙動を示しており, 軟化の鍵因子の有力な候補と考えられた。

DOI： 10.5891/jafps.34.127

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記述言語：英語   掲載種別：研究論文（学術雑誌）   出版者・発行元：ELSEVIER SCIENCE BV  

We investigated the differences in capability to produce ripening-associated ethylene between climacteric ('Yali', 'Xinqingli', and 'Zhuzuili') and non-climacteric ('Hongli', 'Yuanbali', and 'Hongxiaoli') Chinese pear (Pyrus bretschneideri Rehder) fruit varieties. Three ACS (PbACS1, PbACS2, and PbACS3), two ACO (PbACO1 and PbACO2), and three MADS-box (PbMADS1, PbMADS2, and PbMADS3) genes were cloned from ripening fruit. Fruit were harvested at the mature stage and treated with 5000 mu L L-1 propylene for 4 days. Ethylene production was induced by propylene in the climacteric but not in non-climacteric type fruit. In the ripening climacteric fruit, PbACS1 and PbACO1 transcript accumulation accompanied ethylene production but the accumulation of other ACS and ACO mRNAs was not detected. In 'Yali' fruit, 1-MCP exposure prior to propylene treatment completely inhibited the expression of these genes, while exposure after the commencement of ethylene production weakened their expression. Transcripts of PbACO1 accumulated in response to propylene treatment even in non-climacteric fruit but this accumulation was eliminated after the termination of propylene treatment. In response to wounding, transcripts of PbACS2, PbACS3, and PbACO2 genes accumulated in both climacteric and non-climacteric fruit, but accumulation of PbACS1 and PbACO1 mRNAs was not detected. In the Southern analysis of PbACS1, HindIII digests of genomic DNA showed 8.3, 3.5 and 2.9 kb bands. The 2.9 kb band was detected only in climacteric varieties while the 3.5 kb band was detected in both climacteric and non-climacteric varieties except in 'Yali'. These results suggest that Chinese pears may have two copies of the ACSI gene, in which PbACS1A could be linked to the varietal differences in the capability to produce ripening-associated ethylene. There was no correlation between the expression patterns of the three MADS-box genes cloned and the differences in ripening-associated ethylene production among the Chinese pear varieties. (C) 2006 Elsevier B.V. All rights reserved.

DOI： 10.1016/j.postharvbio.2006.12.010

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記述言語：英語   掲載種別：研究論文（学術雑誌）   出版者・発行元：GENETICS  

Although recent findings suggest that the F-box genes SFB/SLF control pollen-part S specificity in the S-RNase-based gametophytic self-incompatibility (GSI) system, how these genes operate in the system is unknown, and functional variation of pollen S genes in different species has been reported. Here, we analyzed the S locus of two species of Maloideae: apple (Malus domestica) and Japanese pear (Pyrus pyrifolia). The sequencing of a 317-kb region of the apple S-9 haplotype revealed two similar F-box genes. Homologous sequences were isolated from different haplotypies of apple and Japanese pear, and they were found to be polymorphic genes derived from the Slocus. Since each S haplotype contains two or three related genes, the genes were named SFBB for S locus F-box brothers. The SFBB genes are specifically expressed in pollen, and variable regions of the SFBB genes are under positive selection. In a style-specific mutant S haplotype of Japanese pear, the SFBB genes are retained. Apart from their multiplicity, SFBB genes meet the expected characteristics of pollen S. The unique multiplicity of SFBB genes as the pollen S candidate is discussed in the context of mechanistic variation in the S-RNase-based GSI system.

DOI： 10.1534/genetics.106.068858

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記述言語：英語   掲載種別：研究論文（学術雑誌）   出版者・発行元：JAPAN SOC HORTICULTURAL SCI  

Substantial decreases in cell wall bound galactosyl and arabinosyl residues are two of the most evident cell wall compositional changes that occur during fruit ripening. The roles of P-galactosidase (P-Gal) and alpha-L-arabinofuranosidase (alpha-Af), the enzymes responsible for these respective losses, were investigated and compared in European (Pyrus communis L. 'La France') and Chinese (Pyrus bretschneideri Rehd. 'Yali') pear fruits which exhibit different softening characteristics during ripening. The increase in the activities of P-Gal and a-Af during ripening in both types of pear fruit correlated well with an increase in climacteric ethylene production, and a concomitant decrease in flesh firmness in 'La France' fruit. However, there was no noticeable decrease in 'Yali' flesh firmness even after 28 days of storage at room temperature. In both fruit types, enzyme activity and the accumulation of transcripts hybridizing with PpGAL1, PpGAL4, PpARF2, and PcARF1 increased with fruit ripening. Increases in gene expression and enzyme activities in 'Yali' fruit with no detectable softening during ripening indicate that P-Gal and a-Af may not mediate difference in fruit softening between two pears, but that they could play some role(s) in cell wall changes, perhaps in cooperation with other cell wall-modifying enzymes such as polygalacturonase.

DOI： 10.2503/jjshs.76.85

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記述言語：英語   掲載種別：研究論文（学術雑誌）   出版者・発行元：OXFORD UNIV PRESS  

Cell wall disassembly in ripening fruit is highly complex, involving the dismantling of multiple polysaccharide networks by diverse families of wall-modifying proteins. While it has been reported in several species that multiple members of each such family are expressed in the same fruit tissue, it is not clear whether this reflects functional redundancy, with protein isozymes from a single enzyme class performing similar roles and contributing equally to wall degradation, or whether they have discrete functions, with some isoforms playing a predominant role. Experiments reported here sought to distinguish between cell wall-related processes in ripening melon that were softening-associated and softening-independent. Cell wall polysaccharide depolymerization and the expression of wall metabolism-related genes were examined in transgenic melon (Cucumis melo var. cantalupensis Naud.) fruit with suppressed expression of the 1-aminocyclopropane-1-carboxylate oxidase (ACO) gene and fruits treated with ethylene and 1-methylcyclopropene (1-MCP). Softening was completely inhibited in the transgenic fruit but was restored by treatment with exogenous ethylene. Moreover, post-harvest application of 1-MCP after the onset of ripening completely halted subsequent softening, suggesting that melon fruit softening is ethylene-dependent. Size exclusion chromatography of cell wall polysaccharides, from the transgenic fruits, with or without exogenous ethylene, indicated that the depolymerization of both pectins and xyloglucans was also ethylene dependent. However, northern analyses of a diverse range of cell wall-related genes, including those for polygalacturonases, xyloglucan endotransglucosylase/hydrolases, expansin, and beta-galactosidases, identified specific genes within single families that could be categorized as ethylene-dependent, ethylene-independent, or partially ethylene-dependent. These results support the hypothesis that while individual cell wall-modifying proteins from each family contribute to cell wall disassembly that accompanies fruit softening, other closely related family members are regulated in an ethylene-independent manner and apparently do not directly participate in fruit softening.

DOI： 10.1093/jxb/erl283

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記述言語：英語   掲載種別：研究論文（学術雑誌）   出版者・発行元：ELSEVIER SCIENCE BV  

We investigated the role of beta-galactosidase (EC 3.2.1.23; beta-Gal) in 'La France' pear (Pyrus communis L.) fruit growth and softening at both biochemical and molecular levels. Northern hybridization was carried out using probes prepared from Japanese pear beta-galactosidase (PpGAL) cDNA clones, designated PpGAL1, PpGAL2, PpGAL3, PpGAL4, PpGAL5, PpGAL6 and PpGAL7. P-Galactosidase activity and accumulation of the transcripts hybridizing with the PpGAL genes were investigated during fruit growth and in relation to fruit softening and in response to propylene and 1-methylcyclopropene (1-MCP) treatments during postharvest ripening. beta-Galactosidase activity was highest in very young fruit, decreased to low levels in later stages of fruit expansion and then increased during postharvest ripening. The increase in beta-Gal activity during fruit ripening paralleled the decrease in fruit firmness. Transcripts hybridizing with PpGAL1 and PpGAL4 were not detected during fruit growth but were detected in the ripening fruit. On the contrary, the abundance of transcripts hybridizing with PpGAL2, PpGAL3, PpGAL5, PpGAL6 and PpGAL7 was highest in the early stages of fruit development, decreased towards fruit maturity and except for PpGAL2 were detected in the ripening fruit, albeit at low levels. Propylene treatment resulted in increased ethylene production, decreased fruit firmness and increased beta-Gal activity. Besides, propylene stimulated the accumulation of transcripts hybridizing with PpGAL1 and PpGAL4 and suppressed the accumulation of transcripts hybridizing with PpGAL6 and PpGAL7 and had no effect on transcripts hybridizing with PpGAL2, PpGAL3 and PpGAL5. The converse was true for fruit treated with 1-MCP, although the inhibition of the accumulation of transcripts hybridizing with PpGAL1 and PpGAL4 by 1-MCP was not complete. These results indicate that PpGAL1 and PpGAL4 play a crucial role in 'La France' pear fruit softening and the increase in their expression at the onset of fruit ripening is in part due to up-regulation by ethylene. The decrease in transcripts hybridizing with PpGAL6 and PpGAL7 in ripe fruit may in part be due to down-regulation by ethylene whereas that of transcripts hybridizing with PpGAL2, PpGAL3 and PpGAL5 may be due to another mechanism(s) unrelated to ethylene action. The divergent members of the P-Gal gene family in 'La France' pear fruit, therefore, have differential developmental and hormonal regulation characteristics. (c) 2005 Published by Elsevier B.V.

DOI： 10.1016/j.postharvbio.2005.02.002

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記述言語：英語   掲載種別：研究論文（学術雑誌）   出版者・発行元：SPRINGER  

Gametophytic self-incompatibility (GSI) in sweet cherry (Prunus avium, Rosaceae) is controlled by the multi-allelic S-locus. This complex locus contains two linked genes, an S-RNase that controls pistil specificity, and an F-box gene named SFB (S-haplotype-specific F-box protein gene) that is the putative determinant of pollen specificity in Prunus species. The S-locus is considered to be a region of repressed recombination, as recombination has not been observed. Recombination between the pistil-S and pollen-S determinant genes would result in non-functional S-haplotypes and loss of self-incompatibility. With the recent identification of multiple SFB alleles in sweet cherry, along with the existing S-RNase alleles, it is now possible to estimate the linkage distance and quantify the physical distance between the two GSI specificity genes in multiple sweet cherry S-haplotypes. A recombinational analysis between S-RNase and SFB was performed for four sweet cherry S-haplotypes (S-2, S-3, S-4, S-6) using F-1 progeny from reciprocal crosses between 'Emperor Francis' (S-3 S-4) and 'NY 54' (S-2 S-6). For SFB genotyping, allele-specific primer sets were designed from the sequence of the SFB coding region. The S-RNase and SFB genotypes of 511 progeny, representing the outcomes of 1,022 meioses, were determined. All four S-haplotypes individually segregated according to the expected Mendelian ratio of 1:1. The S-RNase and SFB loci were completely linked as no recombinant individuals were identified, thus maintaining the co-evolved allele specificities for the pistil and putative pollen determinant. The physical distance between S-RNase and SFB in six sweet cherry S-haplotypes (S1-S6) was determined using PCR with genomic clones as the template. The relative order and transcriptional orientation of S-RNase and SFB were conserved across the six S-haplotypes. However, the physical distance between these two genes varied widely, ranging from 380 bp to approximately 40 kb. This study represents the first large-scale recombinational analysis of the S-locus region in the Rosaceae, serving as the starting point for future comparative analyses of physical distances, linkage distances, and sequence diversity among Prunus S-haplotypes.

DOI： 10.1007/s00497-004-0240-x

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記述言語：英語   掲載種別：研究論文（学術雑誌）   出版者・発行元：SPRINGER-VERLAG  

The gene SFB encodes an F-box protein that has appropriate S-haplotype-specific variation to be the pollen determinant in the S-RNase-based gametophytic self-incompatibility (GSI) reaction in Prunus (Rosaceae). To further characterize Prunus SFB, we cloned and sequenced four additional alleles from sweet cherry (P. avium), SFB1, SFB2, SFB4, and SFB5. These four alleles showed haplotype-specific sequence diversity similar to the other nine SFB alleles that have been cloned. In an amino acid alignment of Prunus SFBs, including the four newly cloned alleles, 121 out of the 384 sites were conserved and an additional 65 sites had only conservative replacements. Amino acid identity among the SFBs ranged from 66.0% to 82.5%. Based on normed variability indices (NVI), 34 of the non-conserved sites were considered to be highly variable. Most of the variable sites were located at the C-terminal region. A window-averaged plot of NVI indicated that there were two variable and two hypervariable regions. These variable and hypervariable regions appeared to be hydrophilic or at least not strongly hydrophobic, which suggests that these regions may be exposed on the surface and function in the allele specificity of the GSI reaction. Evidence of positive selection was detected using maximum likelihood methods with sites under positive selection concentrated in the variable and hypervariable regions.

DOI： 10.1007/s00497-003-0200-x

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記述言語：英語   掲載種別：研究論文（学術雑誌）  

Japanese apricot ( Prunus mume) exhibits the S-RNase-based gametophytic self-incompatibility system as do other self-incompatible Prunus species. This report identifies the S haplotype-specific F-box protein gene ( SFB), a candidate gene for pollen- S, of Japanese apricot, which leads to the development of a molecular typing system for S-haplotype in this fruit species. Both 5'- and 3'-RACE (rapid amplification of cDNA ends) were performed with SFB gene-specific oligonucleotide primers to clone Pm-SFB(1) and Pm-SFB(7) of 'Nanko ( S(1) S(7))'. As in the case of SFB of other Prunus species, Pm-SFB(1) and Pm-SFB(7) showed a high level of S-haplotype-specific sequence polymorphism and their expression was specific to pollen. Genomic DNA-blot analyses of 11 Japanese apricot cultivars with the Pm-SFB probes under low stringency conditions yielded RFLP bands specific to the S(1)- to S(8)-haplotypes as well as a self-compatible S(f)-haplotype. A practical usage of SFB as a molecular marker for S-haplotypes and self-compatibility in Japanese apricot is discussed.

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記述言語：英語   掲載種別：研究論文（学術雑誌）  

This study describes a novel F-box protein gene in the S-locus of sour cherry (Prunus cerasus) and sweet cherry (P. avium). The gene showed an S-haplotype-specific sequence polymorphism and the expression was specific to pollen. Genomic DNA blot analysis of eight sweet cherry cultivars with the probe for the F-box protein gene under low stringency conditions yielded RFLP bands specific to the S-haplotypes of each cultivar. We discuss the possibility of the gene for the F-box protein being a candidate for the male determinant of gametophytic self-incompatibility in PRUNUS:

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DOI： 10.1105/tpc.009290

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記述言語：英語   掲載種別：研究論文（学術雑誌）  

The genomic clone encoding the pistil-specific thaumatin/PR5-like protein (PsTL1) was isolated from Japanese pear (Pyrus serotina). Sequence analysis showed that the genomic clone contained the 5'-flanking sequence of 2.4 kb, the 3'-flanking sequence of 648 bp and the coding region interrupted by a intron of 351 bp. A sequence motif conserved in some pistil self-incompatibility gene promoters of solanaceous and brassicaceous species was located in the 5'-flanking region of the PsTL1 gene. The 2.4 kb 5'-flanking region was fused to the GUS coding sequence and transferred to tobacco. Transgenic tobacco showed GUS activity in pistil and, at low level, in anther, but not in other floral organs and leaf. Histochemical analysis localized GUS activity to stigma, transmitting tissue, anther and pollen of transgenic tobacco.

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掲載種別：研究論文（学術雑誌）  

In almond, gametophytic self-incompatibility is controlled by a single multiallelic locus (S-locus). In styles, the products of S-alleles are ribonucleases, the S-RNases. Cultivated almond in California have four predominant S-alleles (Sa, Sb, S(c), S(d)). We previously reported the cDNA cloning of three of these alleles, namely Sb, S(c) and S(d). In this paper we report the cloning and DNA sequence analysis of the Sa allele. The Sa-RNase displays approximately 55% similarity at the amino-acid level with other almond S-RNases (Sb, S(c), and S(d)) and this similarity was lower than that observed among the Sb, S(c) and S(d)-RNases. Using the cDNA sequence, a PCR-based identification system using genomic DNA was developed for each of the S-RNase alleles. Five almond cultivars with known self-incompatibility (SI) geno-types were analyzed. Common sequences among four S-alleles were used to create four primers, which, when used as sets, amplify DNA bands of unique size that corresponded to each of the four almond S-alleles; Sa (602 bp), Sb (1083 bp), S(c) (221 bp) and S(d) (343 bp). All PCR products obtained from genomic DNA isolated from the five almond cultivars were cloned and their DNA sequence obtained. The nucleotide sequence of these genomic DNA fragments matched the corresponding S-allele cDNA sequence in every case. The amplified products obtained for the Sa- and Sb-alleles were both longer than that expected for the coding region, revealing the presence of an intron of 84 bp in the Sa-allele and 556 bp in the Sb-allele. Both introns are present within the site of the hypervariable region common in S-RNases from the Rosaceae family and which may be important for S specificity. The exon portions of the genomic DNA sequences were completely consistent with the cDNA sequence of the corresponding S-allele. A useful application of these primers would be to identify the S-genotype of progeny in a breeding program, new varieties in an almond nursery, or new grower selections at the seedling stage.

DOI： 10.1007/s001220051489

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掲載種別：研究論文（学術雑誌）  

Genomic sequences of the self-incompatibility genes, the S-RNase genes, from two rosaceous species, Japanese pear and apple, were characterized. Genomic Southern blot and sequencing of a 4.5-kb genomic clone showed that the S4-RNase gene of Japanese pear is surrounded by repetitive sequences as in the case of the S-RNase genes of solanaceous species. The flanking regions of the S2- and S(f)-RNase genes of apple were also cloned and sequenced. The 5' flanking regions of the three alleles bore no similarity with those of the solanaceous S-RNase genes, although the position and sequence of the putative TATA box were conserved. The putative promoter regions of the Japanese pear S4- and apple S(f)-RNase genes shared a stretch of about 200 bp with 80% sequence identity. However, this sequence was not present in the S2-RNase gene of apple, and thus it may reflect a close relationship between the S4- and S(f)-RNase genes rather than a cis-element important in regulating gene expression. Despite the uniform pattern of expression of the rosaceous S-RNase genes, sequence motifs conserved in the 5' flanking regions of the three alleles were not found, implying that the cis-element controlling pistil specific gene expression also locates at the intragenic region or upstream of the analyzed promoter region.

DOI： 10.1016/S0378-1119(98)00105-X

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掲載種別：研究論文（学術雑誌）  

cDNAs encoding three S-RNases of almond (Prunus dulcis), which belongs to the family Rosaceae, were cloned and sequenced. The comparison of amino acid sequences between the S-RNases of almond and those of other rosaceous species showed that the amino acid sequences of the rosaceous S-RNases are highly divergent, and intra-subfamilial similarities are higher than inter-subfamilial similarities. Twelve amino acid sequences of the rosaceous S-RNases were aligned to characterize their primary structural features. In spite of their high level of diversification, the rosaceous S-RNases were found to have five conserved regions, C1, C2, C3, C5, and RC4 which is Rosaceae-specific conserved region. Many variable sites fall into one region, named RHV. RHV is located at a similar position to that of the hypervariable region a (HVa) of the solanaceous S-RNases, and is assumed to be involved in recognizing S-specificity of pollen. On the other hand, the region corresponding to another solanaceous hypervariable region (HVb) was not variable in the rosaceous S-RNases. In the phylogenetic tree of the T2/S type RNase, the rosaceous S-RNase fall into two subfamily-specific groups (Amygdaloideae and Maloideae). The results of sequence comparisons and phylogenetic analysis imply that the present S-RNases of Rosaceae have diverged again relatively recently, after the divergence of subfamilies.

DOI： 10.1007/s004380050894

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記述言語：日本語   出版者・発行元：園芸学会  

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記述言語：日本語  

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記述言語：英語   出版者・発行元：Japanese Society for Horticultural Science  

Substantial decreases in cell wall bound galactosyl and arabinosyl residues are two of the most evident cell wall compositional changes that occur during fruit ripening. The roles of β-galactosidase (β-Gal) and α-L-arabinofuranosidase (α-Af), the enzymes responsible for these respective losses, were investigated and compared in European (Pyrus communis L. 'La France') and Chinese (Pyrus bretschneideri Rehd. 'Yali') pear fruits which exhibit different softening characteristics during ripening. The increase in the activities of β-Gal and α-Af during ripening in both types of pear fruit correlated well with an increase in climacteric ethylene production, and a concomitant decrease in flesh firmness in 'La France' fruit. However, there was no noticeable decrease in 'Yali' flesh firmness even after 28 days of storage at room temperature. In both fruit types, enzyme activity and the accumulation of transcripts hybridizing with PpGAL1, PpGAL4, PpARF2, and PcARF1 increased with fruit ripening. Increases in gene expression and enzyme activities in 'Yali' fruit with no detectable softening during ripening indicate that β-Gal and α-Af may not mediate difference in fruit softening between two pears, but that they could play some role(s) in cell wall changes, perhaps in cooperation with other cell wall-modifying enzymes such as polygalacturonase.

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その他リンク： http://dl.ndl.go.jp/info:ndljp/pid/10471864

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記述言語：英語  

S4′ is a pollen-part mutant in sweet cherry (Prunus avium L.) that is extensively used to develop self-compatible cultivars. The S 4′-haplotype is known to have a functional stylar component and a nonfunctional pollen component. The pollen component in sweet cherry necessary for the specificity of the pollen reaction is believed to be an S-haplotype specific F-box protein gene, called SFB. This study describes two molecular markers that distinguish between SFB4 and SFB 4′ by taking advantage of a four base pair deletion in the mutant allele. The resulting polymerase chain reaction (PCR) products can either be separated directly on a polyacrylamide gel or they can be subjected to restriction enzyme digestion and the different sized products can be visualized on an agarose gel. The latter technique utilizes restriction sites created in the PCR products from the SFB4′ allele, but not the SFB 4 allele. Because the primer sets created differential restriction sites, these primer sets were termed dCAPS (derived cleaved amplified polymorphism sequence) markers. These molecular assays can be used to verify self-compatibility conferred by the S4′-haplotype.

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記述言語：英語   出版者・発行元：Japanese Society of Plant Physiologists  

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