A highly efficient and simple micropropagation system for Phalaenopsis was developed using elongated protocorm-like bodies (ePLBs) obtained by skotomorphogenesis. When normal protocorm-like bodies (nPLBs) without growing point excision were cultured under different light conditions (dark: 0 mmol·m-2·s-1, low light: 2 mmol·m-2·s-1, high light: 80 mmol·m-2·s-1, of photosynthetic photon flux density respectively), PLB proliferation efficiency was higher under dark than high light and low light conditions. In addition, shoot formation percentage was lower under dark conditions (8.0%) than under low light (66.0%) and high light conditions (68.2%) and few PLBs developed shoots during culture under dark conditions. The Secondary ePLBs obtained after culturing under dark conditions were approximately twice as long as nPLBs. After acclimation under low light conditions for 2 weeks, the ePLBs were transferred to high light conditions after making a partial incision in their apical parts. Under high light conditions, a large number of secondary PLBs were obtained from ePLBs, 6 times as many as from nPLBs treated with the same partial incisions. The findings of this study showed that culturing PLBs in dark conditions suppresses shoot formation that might interfere with PLB proliferation, and that a large number of secondary PLBs could be obtained from these ePLBs compared with nPLBs after exposure to high light intensity.
{"title":"Development of a Highly Efficient and Simple Micropropagation System for Phalaenopsis Using Elongated Protocorm-like Bodies Induced by Skotomorphogenesis under Dark Conditions","authors":"Shinichi Enoki, Y. Takahara","doi":"10.2503/JJSHS1.CH-083","DOIUrl":"https://doi.org/10.2503/JJSHS1.CH-083","url":null,"abstract":"A highly efficient and simple micropropagation system for Phalaenopsis was developed using elongated protocorm-like bodies (ePLBs) obtained by skotomorphogenesis. When normal protocorm-like bodies (nPLBs) without growing point excision were cultured under different light conditions (dark: 0 mmol·m-2·s-1, low light: 2 mmol·m-2·s-1, high light: 80 mmol·m-2·s-1, of photosynthetic photon flux density respectively), PLB proliferation efficiency was higher under dark than high light and low light conditions. In addition, shoot formation percentage was lower under dark conditions (8.0%) than under low light (66.0%) and high light conditions (68.2%) and few PLBs developed shoots during culture under dark conditions. The Secondary ePLBs obtained after culturing under dark conditions were approximately twice as long as nPLBs. After acclimation under low light conditions for 2 weeks, the ePLBs were transferred to high light conditions after making a partial incision in their apical parts. Under high light conditions, a large number of secondary PLBs were obtained from ePLBs, 6 times as many as from nPLBs treated with the same partial incisions. The findings of this study showed that culturing PLBs in dark conditions suppresses shoot formation that might interfere with PLB proliferation, and that a large number of secondary PLBs could be obtained from these ePLBs compared with nPLBs after exposure to high light intensity.","PeriodicalId":17343,"journal":{"name":"Journal of The Japanese Society for Horticultural Science","volume":null,"pages":null},"PeriodicalIF":0.0,"publicationDate":"2014-04-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://sci-hub-pdf.com/10.2503/JJSHS1.CH-083","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"69158020","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
A recently popular Japanese yellow-green-skin table grape, ‘Shine Muscat’ (Vitis labruscana Bailey × V. vinifera L.), has the problem of berry skin browning, which occurs at the maturation stage just before harvest. Tiny reddish-brown blotches appear on the surface of berries and considerably decrease the grape’s market value. Although the mechanisms and factors for browning are unknown, we hypothesized the involvement of polyphenol compounds and their oxidation reactions. In this study, the gene expressions of polyphenol oxidase (PPO), stilbene synthase (STS), and chalcone synthase (CHS), which are key enzymatic genes related to the metabolic pathway for polyphenols, were analyzed during berry maturation to examine the molecular basis for browning. Skin browning occurred on several berries in a bunch of ‘Shine Muscat’ from 80 days after full bloom (DAFB), after which the number of berries with skin browning increased, and the browned area spread on the berry surfaces with maturation. Increases in the expression of VvPPO2, VvSTS type B, and VvCHS1 were associated with skin browning, and the trans-resveratrol content also increased in the browning skin, suggesting that biosynthesis and metabolic pathways for phenolic compounds were activated at the time of browning. In terms of VvPPO genes, specific up-regulation of VvPPO2 expression was observed compared with the VvPPO1 gene. The promoter sequence of VvPPO2 contains more Myb binding motifs and W-box motifs than does VvPPO1. The specific up-regulation of VvPPO2 gene expression will play a crucial role in understanding and managing the skin-browning mechanism in the grape berries of ‘Shine Muscat’.
{"title":"Skin Browning and Expression of PPO, STS, and CHS Genes in the Grape Berries of ‘Shine Muscat’","authors":"Yuka Suehiro, K. Mochida, H. Itamura, T. Esumi","doi":"10.2503/JJSHS1.CH-095","DOIUrl":"https://doi.org/10.2503/JJSHS1.CH-095","url":null,"abstract":"A recently popular Japanese yellow-green-skin table grape, ‘Shine Muscat’ (Vitis labruscana Bailey × V. vinifera L.), has the problem of berry skin browning, which occurs at the maturation stage just before harvest. Tiny reddish-brown blotches appear on the surface of berries and considerably decrease the grape’s market value. Although the mechanisms and factors for browning are unknown, we hypothesized the involvement of polyphenol compounds and their oxidation reactions. In this study, the gene expressions of polyphenol oxidase (PPO), stilbene synthase (STS), and chalcone synthase (CHS), which are key enzymatic genes related to the metabolic pathway for polyphenols, were analyzed during berry maturation to examine the molecular basis for browning. Skin browning occurred on several berries in a bunch of ‘Shine Muscat’ from 80 days after full bloom (DAFB), after which the number of berries with skin browning increased, and the browned area spread on the berry surfaces with maturation. Increases in the expression of VvPPO2, VvSTS type B, and VvCHS1 were associated with skin browning, and the trans-resveratrol content also increased in the browning skin, suggesting that biosynthesis and metabolic pathways for phenolic compounds were activated at the time of browning. In terms of VvPPO genes, specific up-regulation of VvPPO2 expression was observed compared with the VvPPO1 gene. The promoter sequence of VvPPO2 contains more Myb binding motifs and W-box motifs than does VvPPO1. The specific up-regulation of VvPPO2 gene expression will play a crucial role in understanding and managing the skin-browning mechanism in the grape berries of ‘Shine Muscat’.","PeriodicalId":17343,"journal":{"name":"Journal of The Japanese Society for Horticultural Science","volume":null,"pages":null},"PeriodicalIF":0.0,"publicationDate":"2014-04-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://sci-hub-pdf.com/10.2503/JJSHS1.CH-095","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"69158905","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
The infection efficiency of Agrobacterium to cotyledon explants of Japanese apricot ( Prunus mume ) was markedly improved by sonication treatment. The use of sGFP(S65T) reporter gene in this study enabled direct observation of transgene expression, thus allowing the evaluation of Agrobacterium infection efficiency. Immature cotyledons of different cultivars and different developmental stages were subjected to sonication treatment of durations ranging from 10 sec to 2 min. When early-stage immature cotyledons of ‘Nanko’ were subjected to sonication treatment, the transient GFP expression frequency after co-cultivation was nearly 100% and GFP fluorescence was distributed over almost the entire cotyledon surface. In contrast, when the inoculation was carried out according to the standard dipping method, transient GFP expression frequency was less than 10% and GFP fluorescence was observed only in spots. The highest frequency of somatic embryogenesis (SEG) with GFP expression was obtained from 40 sec sonication treatment of May 14 ‘Nanko’ immature cotyledons. However, with younger cotyledons, which have a higher SEG frequency, 20 sec of sonication treatment was found to be sufficient to increase the infection frequency.
{"title":"Improving Infection Efficiency of Agrobacterium to Immature Cotyledon Explants of Japanese Apricot (Prunus mume) by Sonication Treatment","authors":"M. Gao-Takai, R. Tao","doi":"10.2503/JJSHS1.CH-085","DOIUrl":"https://doi.org/10.2503/JJSHS1.CH-085","url":null,"abstract":"The infection efficiency of Agrobacterium to cotyledon explants of Japanese apricot ( Prunus mume ) was markedly improved by sonication treatment. The use of sGFP(S65T) reporter gene in this study enabled direct observation of transgene expression, thus allowing the evaluation of Agrobacterium infection efficiency. Immature cotyledons of different cultivars and different developmental stages were subjected to sonication treatment of durations ranging from 10 sec to 2 min. When early-stage immature cotyledons of ‘Nanko’ were subjected to sonication treatment, the transient GFP expression frequency after co-cultivation was nearly 100% and GFP fluorescence was distributed over almost the entire cotyledon surface. In contrast, when the inoculation was carried out according to the standard dipping method, transient GFP expression frequency was less than 10% and GFP fluorescence was observed only in spots. The highest frequency of somatic embryogenesis (SEG) with GFP expression was obtained from 40 sec sonication treatment of May 14 ‘Nanko’ immature cotyledons. However, with younger cotyledons, which have a higher SEG frequency, 20 sec of sonication treatment was found to be sufficient to increase the infection frequency.","PeriodicalId":17343,"journal":{"name":"Journal of The Japanese Society for Horticultural Science","volume":null,"pages":null},"PeriodicalIF":0.0,"publicationDate":"2014-04-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://sci-hub-pdf.com/10.2503/JJSHS1.CH-085","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"69158500","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Y. Mochizuki, Y. Iwasaki, Mitsutoshi Fuke, I. Ogiwara
. Nagahata, H. and A. Kuroda. 2004. Varietal differences in dry matter production and nitrogen absorption among early-maturing rice varieties under an artificial high temperature condition. The Hokuriku Crop Sci. 39: 78–80 (In Japanese). Nagashima, Y. and M. Sada. 1982. Root development in strawberry forcing culture. Bull. Shizuoka Agr. Exp. Stn. 27: 31–36
{"title":"Analysis of a High-yielding Strawberry (Fragaria ×ananassa Duch.) Cultivar ‘Benihoppe’ with Focus on Root Dry Matter and Activity","authors":"Y. Mochizuki, Y. Iwasaki, Mitsutoshi Fuke, I. Ogiwara","doi":"10.2503/JJSHS1.CH-088","DOIUrl":"https://doi.org/10.2503/JJSHS1.CH-088","url":null,"abstract":". Nagahata, H. and A. Kuroda. 2004. Varietal differences in dry matter production and nitrogen absorption among early-maturing rice varieties under an artificial high temperature condition. The Hokuriku Crop Sci. 39: 78–80 (In Japanese). Nagashima, Y. and M. Sada. 1982. Root development in strawberry forcing culture. Bull. Shizuoka Agr. Exp. Stn. 27: 31–36","PeriodicalId":17343,"journal":{"name":"Journal of The Japanese Society for Horticultural Science","volume":null,"pages":null},"PeriodicalIF":0.0,"publicationDate":"2014-04-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"69158761","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
The MADS-box gene family is one of the largest transcription factor gene families in plants and is necessary at various developmental stages. Many studies on flower development show that especially MIKCc-type MADSbox genes are essential for proper floral organ development. We identified and characterized MIKCc-type MADS-box genes expressed in Eustoma grandiflorum flowers. Twenty-three genes were identified and grouped into 10 clades, which were characterized by conserved specific motifs. Phylogenetic analysis indicated the diversification of AG/PLE, AP3/DEF, PI/GLO, and SEP clades and the occurrence of recent gene duplication events. The floral organ-specific expression patterns were partly diversified within the gene members of AP3/DEF and SEP clades, while they were conserved in AG/PLE and PI/GLO clades. These results suggest that genes with conserved expression as well as those with diversified expression contribute to specifying floral organ identity in E. grandiflorum.
{"title":"Conservation and Diversification of Floral Homeotic MADS-box Genes in Eustoma grandiflorum","authors":"M. Ishimori, S. Kawabata","doi":"10.2503/JJSHS1.CH-098","DOIUrl":"https://doi.org/10.2503/JJSHS1.CH-098","url":null,"abstract":"The MADS-box gene family is one of the largest transcription factor gene families in plants and is necessary at various developmental stages. Many studies on flower development show that especially MIKCc-type MADSbox genes are essential for proper floral organ development. We identified and characterized MIKCc-type MADS-box genes expressed in Eustoma grandiflorum flowers. Twenty-three genes were identified and grouped into 10 clades, which were characterized by conserved specific motifs. Phylogenetic analysis indicated the diversification of AG/PLE, AP3/DEF, PI/GLO, and SEP clades and the occurrence of recent gene duplication events. The floral organ-specific expression patterns were partly diversified within the gene members of AP3/DEF and SEP clades, while they were conserved in AG/PLE and PI/GLO clades. These results suggest that genes with conserved expression as well as those with diversified expression contribute to specifying floral organ identity in E. grandiflorum.","PeriodicalId":17343,"journal":{"name":"Journal of The Japanese Society for Horticultural Science","volume":null,"pages":null},"PeriodicalIF":0.0,"publicationDate":"2014-04-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://sci-hub-pdf.com/10.2503/JJSHS1.CH-098","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"69158963","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Solanaceae, Rosaceae, and Plantaginaceae exhibit the S-RNase-based gametophytic self-incompatibility (GSI) system. This type of GSI is controlled by a single polymorphic locus ( S locus) containing the pistil S determinant gene, S-ribonuclease ( S-RNase ), and the pollen S determinant, the S locus F-box gene ( SFB/SLF ). In addition to these determinant genes, non-S factors, called modifier genes, are required for the GSI reaction. Here, we conducted large-scale transcriptome analysis of unpollinated, self-pollinated, and cross-pollinated pistils of Japanese apricot ( Prunus mume Sieb. et Zucc. cv. Nanko) to capture all of the molecular events induced by the GSI reaction in Prunus , using next-generation sequencing technologies. We obtained 40,061 unigenes from 77,521,310 reads from pollinated and unpollinated pistils and pollen grains. Among these unigenes, 29,985 and 27,898 unigene sequences showed at least one hit against the NCBI nr and TAIR10 protein databases, respective - ly, in BLASTX searches using an E-value cutoff of 1e-6. Digital expression analysis showed that 8,907 and 10,190 unigenes were expressed at significantly different levels between unpollinated (UP) and cross-pollinated (CP) pistils and between UP and self-pollinated (SP) pistils, respectively. The expression of 4,348 unigenes in both CP and SP pollination was commonly and significantly different from that in UP, while the expression of 4,559 and 5,842 unigenes in CP and SP, respectively, was specifically and significantly different from UP. The expression of 2,227 unigenes was up-regulated both in CP and SP compared with UP. Genes supposedly involved in S-RNase-based GSI were included among the unigenes up-regulated by pollination, while no unigenes homologous to the solanaceous pistil modifiers HT-B or 120K were included among the unigenes up-regulated by pollination or in the whole unpollinated/pollinated pistil transcriptome. We discuss the distinct molecular mechanism of S-RNase-based GSI in Prunus .
{"title":"Transcriptome Analysis of Self- and Cross-pollinated Pistils of Japanese Apricot (Prunus mume Sieb. et Zucc.)","authors":"T. Habu, R. Tao","doi":"10.2503/JJSHS1.CH-086","DOIUrl":"https://doi.org/10.2503/JJSHS1.CH-086","url":null,"abstract":"Solanaceae, Rosaceae, and Plantaginaceae exhibit the S-RNase-based gametophytic self-incompatibility (GSI) system. This type of GSI is controlled by a single polymorphic locus ( S locus) containing the pistil S determinant gene, S-ribonuclease ( S-RNase ), and the pollen S determinant, the S locus F-box gene ( SFB/SLF ). In addition to these determinant genes, non-S factors, called modifier genes, are required for the GSI reaction. Here, we conducted large-scale transcriptome analysis of unpollinated, self-pollinated, and cross-pollinated pistils of Japanese apricot ( Prunus mume Sieb. et Zucc. cv. Nanko) to capture all of the molecular events induced by the GSI reaction in Prunus , using next-generation sequencing technologies. We obtained 40,061 unigenes from 77,521,310 reads from pollinated and unpollinated pistils and pollen grains. Among these unigenes, 29,985 and 27,898 unigene sequences showed at least one hit against the NCBI nr and TAIR10 protein databases, respective - ly, in BLASTX searches using an E-value cutoff of 1e-6. Digital expression analysis showed that 8,907 and 10,190 unigenes were expressed at significantly different levels between unpollinated (UP) and cross-pollinated (CP) pistils and between UP and self-pollinated (SP) pistils, respectively. The expression of 4,348 unigenes in both CP and SP pollination was commonly and significantly different from that in UP, while the expression of 4,559 and 5,842 unigenes in CP and SP, respectively, was specifically and significantly different from UP. The expression of 2,227 unigenes was up-regulated both in CP and SP compared with UP. Genes supposedly involved in S-RNase-based GSI were included among the unigenes up-regulated by pollination, while no unigenes homologous to the solanaceous pistil modifiers HT-B or 120K were included among the unigenes up-regulated by pollination or in the whole unpollinated/pollinated pistil transcriptome. We discuss the distinct molecular mechanism of S-RNase-based GSI in Prunus .","PeriodicalId":17343,"journal":{"name":"Journal of The Japanese Society for Horticultural Science","volume":null,"pages":null},"PeriodicalIF":0.0,"publicationDate":"2014-04-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://sci-hub-pdf.com/10.2503/JJSHS1.CH-086","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"69158642","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
A. Kalisz, S. Cebula, P. Siwek, A. Sękara, A. Grabowska, J. Gil
The aim of the research was to determine the impact of row covers on the growth, yield, and selected nutritional compounds contents in Chinese cabbage (Brassica rapa L. Pekinensis Group), cultivated in a field with or without non-woven fleece (17 g·m -2 ). Application of row covers accelerated the growth and development of the plants due to more favorable microclimatic conditions than in an open field. All tested biometrical parameters of the rosettes, determined after removing covers, were considerably higher than in the uncovered control. Therefore, direct covers could be successfully used for promote the growth of Chinese cabbage rosettes after transplanting. Total and marketable yields of covered plants were higher by 36% and 91%, respectively, than in the control. No external flower stalks were observed in either treatment, but about 50% of control heads had internal bolters. Row covering could be an effective prevention method against Chinese cabbage bolting in spring production in Central Europe. Laboratory analysis performed directly after removing covers showed higher contents of L-ascorbic acid, chlorophylls, and carotenoids in the plants in an open field. However, in the subsequent few weeks, such differences between treatments decreased and at harvest time, the level of these compounds was similar. Mature heads of control Chinese cabbage contained significantly more soluble sugars, crude fiber and thiocyanates than covered plants. The models were proposed to predict changes in fresh and dry weight as a function of time.
本研究旨在确定行盖对在有无无纺羊毛(17 g·m -2)条件下栽培的大白菜(Brassica rapa L. Pekinensis Group)生长、产量和部分营养物质含量的影响。由于小气候条件比露天条件更有利，行盖的施用加速了植物的生长发育。除去盖后测定的莲座的所有生物特征参数都明显高于未盖的对照组。因此，直接覆盖可以成功地促进大白菜莲座移栽后的生长。覆盖植株的总产量和可销售产量分别比对照高36%和91%。两种处理均未观察到外花茎，但约50%的对照头有内栓。行覆盖是中欧地区春季大白菜抽苔的有效防治措施。在去除覆盖物后直接进行的实验室分析显示，在开阔的田野中，植物中l -抗坏血酸、叶绿素和类胡萝卜素的含量较高。然而，在随后的几周内，处理之间的这种差异减少了，在收获季节，这些化合物的水平相似。对照大白菜成熟穗中可溶性糖、粗纤维和硫氰酸盐含量显著高于覆盖植株。提出了预测鲜重和干重随时间变化的模型。
{"title":"Effects of Row Covers Using Non-woven Fleece on the Yields, Rate of Bolting, and Quality of Heading Chinese Cabbage in Early Spring Cultivation","authors":"A. Kalisz, S. Cebula, P. Siwek, A. Sękara, A. Grabowska, J. Gil","doi":"10.2503/JJSHS1.CH-064","DOIUrl":"https://doi.org/10.2503/JJSHS1.CH-064","url":null,"abstract":"The aim of the research was to determine the impact of row covers on the growth, yield, and selected nutritional compounds contents in Chinese cabbage (Brassica rapa L. Pekinensis Group), cultivated in a field with or without non-woven fleece (17 g·m -2 ). Application of row covers accelerated the growth and development of the plants due to more favorable microclimatic conditions than in an open field. All tested biometrical parameters of the rosettes, determined after removing covers, were considerably higher than in the uncovered control. Therefore, direct covers could be successfully used for promote the growth of Chinese cabbage rosettes after transplanting. Total and marketable yields of covered plants were higher by 36% and 91%, respectively, than in the control. No external flower stalks were observed in either treatment, but about 50% of control heads had internal bolters. Row covering could be an effective prevention method against Chinese cabbage bolting in spring production in Central Europe. Laboratory analysis performed directly after removing covers showed higher contents of L-ascorbic acid, chlorophylls, and carotenoids in the plants in an open field. However, in the subsequent few weeks, such differences between treatments decreased and at harvest time, the level of these compounds was similar. Mature heads of control Chinese cabbage contained significantly more soluble sugars, crude fiber and thiocyanates than covered plants. The models were proposed to predict changes in fresh and dry weight as a function of time.","PeriodicalId":17343,"journal":{"name":"Journal of The Japanese Society for Horticultural Science","volume":null,"pages":null},"PeriodicalIF":0.0,"publicationDate":"2014-04-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://sci-hub-pdf.com/10.2503/JJSHS1.CH-064","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"69158272","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
T. Kawai, Ayako Gonoi, Michiya Nitta, M. Kaido, N. Yamagishi, N. Yoshikawa, R. Tao
Apple latent spherical virus (ALSV) vectors have been shown to effectively induce stable virus-induced gene silencing (VIGS) in a wide range of plant species, including rosaceous fruit tree species such as apple (Malus × domestica Borkh.), pear (Pyrus communis L.), and Japanese pear (P. pyrifolia Nakai). In this study, we attempted to develop a VIGS-based gene evaluation system for two Prunus fruit tree species, apricot and Japanese apricot, using ALSV vectors. A partial sequence of the P. armeniaca PHYTOENE DESATURASE (ParPDS) gene was cloned and ligated into the T-DNA region of a binary vector, pBICAL2, designed based on RNA2 of ALSV. The resultant pBICAL2-ParPDS was introduced into a disarmed Agrobacterium strain, EHA105. pBICAL1, a binary plasmid for the expression of ALSV RNA1 in plants, was also introduced into EHA105. Leaves of Nicotiana benthamiana were infected with pBICAL1/EHA105 and pBICAL2-ParPDS/EHA105 simultaneously to produce and amplify recombinant ALSV particles. The amplified ParPDS-ALSV in N. benthamiana was isolated and infected into the cotyledons of apricot and Japanese apricot seedlings by particle bombardment. Although our attempts to infect wild and recombinant ALSVs into Japanese apricot seedlings were unsuccessful, uniform discoloration of the upper leaves, a typical phenotype of PDS knock down, was observed several weeks after inoculation in apricot seedlings. We discuss the possible use of this VIGS-based gene evaluation system in Prunus.
苹果潜伏球形病毒(ALSV)载体已被证明能在广泛的植物物种中有效诱导稳定的病毒诱导基因沉默(VIGS)，包括玫瑰科果树物种，如苹果(Malus × domestica Borkh.)、梨(Pyrus communis L.)和日本梨(P. pyrifolia Nakai)。本研究尝试以ALSV为载体，建立基于vigs的杏、杏两种李属果树的基因评价体系。本文克隆了亚美尼亚植物烯去饱和酶(ParPDS)基因的部分序列，并将其连接到基于ALSV RNA2设计的二元载体pBICAL2的T-DNA区。将得到的pBICAL2-ParPDS导入解除武装的农杆菌菌株EHA105中。pBICAL1是在植物中表达ALSV RNA1的双质粒，也被引入EHA105中。利用pBICAL1/EHA105和pBICAL2-ParPDS/EHA105同时侵染本烟叶片，产生并扩增重组ALSV颗粒。利用粒子轰击的方法，分离出本拟白僵菌ParPDS-ALSV扩增体，并将其侵染到杏和杏苗子叶中。虽然我们尝试将野生和重组ALSVs感染日本杏幼苗都没有成功，但在接种杏幼苗几周后，观察到PDS敲低的典型表型，即上部叶片均匀变色。讨论了基于vigs的李树基因评价系统的应用前景。
{"title":"Virus-induced Gene Silencing in Apricot (Prunus armeniaca L.) and Japanese Apricot (P. mume Siebold & Zucc.) with the Apple Latent Spherical Virus Vector System","authors":"T. Kawai, Ayako Gonoi, Michiya Nitta, M. Kaido, N. Yamagishi, N. Yoshikawa, R. Tao","doi":"10.2503/JJSHS1.CH-091","DOIUrl":"https://doi.org/10.2503/JJSHS1.CH-091","url":null,"abstract":"Apple latent spherical virus (ALSV) vectors have been shown to effectively induce stable virus-induced gene silencing (VIGS) in a wide range of plant species, including rosaceous fruit tree species such as apple (Malus × domestica Borkh.), pear (Pyrus communis L.), and Japanese pear (P. pyrifolia Nakai). In this study, we attempted to develop a VIGS-based gene evaluation system for two Prunus fruit tree species, apricot and Japanese apricot, using ALSV vectors. A partial sequence of the P. armeniaca PHYTOENE DESATURASE (ParPDS) gene was cloned and ligated into the T-DNA region of a binary vector, pBICAL2, designed based on RNA2 of ALSV. The resultant pBICAL2-ParPDS was introduced into a disarmed Agrobacterium strain, EHA105. pBICAL1, a binary plasmid for the expression of ALSV RNA1 in plants, was also introduced into EHA105. Leaves of Nicotiana benthamiana were infected with pBICAL1/EHA105 and pBICAL2-ParPDS/EHA105 simultaneously to produce and amplify recombinant ALSV particles. The amplified ParPDS-ALSV in N. benthamiana was isolated and infected into the cotyledons of apricot and Japanese apricot seedlings by particle bombardment. Although our attempts to infect wild and recombinant ALSVs into Japanese apricot seedlings were unsuccessful, uniform discoloration of the upper leaves, a typical phenotype of PDS knock down, was observed several weeks after inoculation in apricot seedlings. We discuss the possible use of this VIGS-based gene evaluation system in Prunus.","PeriodicalId":17343,"journal":{"name":"Journal of The Japanese Society for Horticultural Science","volume":null,"pages":null},"PeriodicalIF":0.0,"publicationDate":"2014-01-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://sci-hub-pdf.com/10.2503/JJSHS1.CH-091","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"69158774","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Yasuhisa Tsuchida, H. Yakushiji, Takaaki Oe, K. Negoro, N. Gato, Tatsuya Kotani, Onishi Yuriko, T. Kobata, M. Tamura
1Japanese Apricot Laboratory, Wakayama Fruit Tree Experiment Station, Minabe, Wakayama 645-0021, Japan 2Grape and Persimmon Research Division, NARO Institute of Fruit Tree Science, Higashi-Hiroshima 739-2494, Japan 3Food Science Research Laboratory, Nakano BC Co. Ltd., Kainan 642-0034, Japan 4R & D Department, Kobata Research Institute Limited., Gojo, Nara 637-0071, Japan 5NARO Institute of National Food Research Institute, Tsukuba 305-8642, Japan
{"title":"Differences in Cell-wall Polysaccharide Degradation during Softening Process in Two Cultivars of Japanese Apricot Fruits","authors":"Yasuhisa Tsuchida, H. Yakushiji, Takaaki Oe, K. Negoro, N. Gato, Tatsuya Kotani, Onishi Yuriko, T. Kobata, M. Tamura","doi":"10.2503/JJSHS1.CH-067","DOIUrl":"https://doi.org/10.2503/JJSHS1.CH-067","url":null,"abstract":"1Japanese Apricot Laboratory, Wakayama Fruit Tree Experiment Station, Minabe, Wakayama 645-0021, Japan 2Grape and Persimmon Research Division, NARO Institute of Fruit Tree Science, Higashi-Hiroshima 739-2494, Japan 3Food Science Research Laboratory, Nakano BC Co. Ltd., Kainan 642-0034, Japan 4R & D Department, Kobata Research Institute Limited., Gojo, Nara 637-0071, Japan 5NARO Institute of National Food Research Institute, Tsukuba 305-8642, Japan","PeriodicalId":17343,"journal":{"name":"Journal of The Japanese Society for Horticultural Science","volume":null,"pages":null},"PeriodicalIF":0.0,"publicationDate":"2014-01-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://sci-hub-pdf.com/10.2503/JJSHS1.CH-067","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"69158294","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
D. Mizuta, A. Nakatsuka, T. Ban, I. Miyajima, N. Kobayashi
We compared anthocyanidin composition patterns and the expression of anthocyanin biosynthesis genes in Rhododendron kiusianum, R. kaempferi, and their natural hybrids from the Kirishima mountain mass. Compared with the habitat, phenotypic characteristics including tree height and flower color did not change in the transplanted individuals after cutting propagation. According to flower color measurements, R. kiusianum and R. kaempferi belonged to purple and red series, respectively, and their natural hybrids belonged to either the red or purple series. HPLC analysis showed that the petals of most R. kiusianum and natural hybrids contained both cyanidin and delphinidin series pigments, while the petals of R. kaempferi contained only cyanidin series pigments. However, one R. kiusianum individual in the purple series contained only cyanidin series pigments and one natural hybrid individual in the red series contained both cyanidin and delphinidin series pigments. These individuals were thought to be influenced by co-pigmentation or a lack thereof, respectively. All samples expressed F3′H, DFR, and ANS genes in real-time quantitative RT-PCR, and the F3′5′H gene was always expressed in samples containing delphinidin series pigments. These results suggest that the expression of F3′5′H is essential for R. kiusianun and its natural hybrids to produce delphinidin series pigments. This study showed that interspecific hybridization between wild species with purple series flowers and wild species with red series flowers varied the pigment composition and anthocyanin biosynthesis-related gene expression in the natural hybrids, suggesting that it caused the flower color variation in the wild populations in the Kirishima mountains.
{"title":"Pigment composition patterns and expression of anthocyanin biosynthesis genes in Rhododendron kiusianum, R. kaempferi, and their natural hybrids on Kirishima mountain mass, Japan","authors":"D. Mizuta, A. Nakatsuka, T. Ban, I. Miyajima, N. Kobayashi","doi":"10.2503/JJSHS1.CH-087","DOIUrl":"https://doi.org/10.2503/JJSHS1.CH-087","url":null,"abstract":"We compared anthocyanidin composition patterns and the expression of anthocyanin biosynthesis genes in Rhododendron kiusianum, R. kaempferi, and their natural hybrids from the Kirishima mountain mass. Compared with the habitat, phenotypic characteristics including tree height and flower color did not change in the transplanted individuals after cutting propagation. According to flower color measurements, R. kiusianum and R. kaempferi belonged to purple and red series, respectively, and their natural hybrids belonged to either the red or purple series. HPLC analysis showed that the petals of most R. kiusianum and natural hybrids contained both cyanidin and delphinidin series pigments, while the petals of R. kaempferi contained only cyanidin series pigments. However, one R. kiusianum individual in the purple series contained only cyanidin series pigments and one natural hybrid individual in the red series contained both cyanidin and delphinidin series pigments. These individuals were thought to be influenced by co-pigmentation or a lack thereof, respectively. All samples expressed F3′H, DFR, and ANS genes in real-time quantitative RT-PCR, and the F3′5′H gene was always expressed in samples containing delphinidin series pigments. These results suggest that the expression of F3′5′H is essential for R. kiusianun and its natural hybrids to produce delphinidin series pigments. This study showed that interspecific hybridization between wild species with purple series flowers and wild species with red series flowers varied the pigment composition and anthocyanin biosynthesis-related gene expression in the natural hybrids, suggesting that it caused the flower color variation in the wild populations in the Kirishima mountains.","PeriodicalId":17343,"journal":{"name":"Journal of The Japanese Society for Horticultural Science","volume":null,"pages":null},"PeriodicalIF":0.0,"publicationDate":"2014-01-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://sci-hub-pdf.com/10.2503/JJSHS1.CH-087","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"69158700","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
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