Pub Date : 2019-09-25DOI: 10.5511/plantbiotechnology.19.0807a
Toshiya Inamura, Manami Nakazawa, Mitsuyo Ishibe, M. Otani, M. Nakano
The liliaceous perennial plants, Tricyrtis spp., have recently become popular as ornamental plants for pot and garden uses. In order to broaden the variability in plant form, flower form and flower color of Tricyrtis spp., intersectional hybridization was examined between four T. formosana cultivars or T. hirta var. albescens (sect. Hirtae) and T. macranthopsis (sect. Brachycyrtis). After cross-pollination, ovary enlargement was observed only when T. macranthopsis was used as a pollen parent. Ovules with placental tissues were excised from enlarged ovaries and cultured on half-strength MS medium without plant growth regulators. From five cross-combinations, 31 ovule culture-derived plantlets were obtained and 20 of them were confirmed to be intersectional hybrids by flow cytometry and inter-simple sequence repeat analyses. Almost all hybrids grew well and produced flowers 1-2 years after transplantation to the greenhouse. Hybrids had semi-cascade-type shoots, which was intermediate between T. formosana cultivars and T. hirta var. albescens (erect-type shoots) and T. macranthopsis (cascade-type shoots). They produced flowers with novel forms and colors compared with the corresponding parents, and some were horticulturally attractive. The results obtained in the present study indicate the validity of intersectional hybridization via ovule culture for breeding of Tricyrtis spp.
{"title":"Production and characterization of intersectional hybrids between Tricyrtis sect. Brachycyrtis and sect. Hirtae via ovule culture.","authors":"Toshiya Inamura, Manami Nakazawa, Mitsuyo Ishibe, M. Otani, M. Nakano","doi":"10.5511/plantbiotechnology.19.0807a","DOIUrl":"https://doi.org/10.5511/plantbiotechnology.19.0807a","url":null,"abstract":"The liliaceous perennial plants, Tricyrtis spp., have recently become popular as ornamental plants for pot and garden uses. In order to broaden the variability in plant form, flower form and flower color of Tricyrtis spp., intersectional hybridization was examined between four T. formosana cultivars or T. hirta var. albescens (sect. Hirtae) and T. macranthopsis (sect. Brachycyrtis). After cross-pollination, ovary enlargement was observed only when T. macranthopsis was used as a pollen parent. Ovules with placental tissues were excised from enlarged ovaries and cultured on half-strength MS medium without plant growth regulators. From five cross-combinations, 31 ovule culture-derived plantlets were obtained and 20 of them were confirmed to be intersectional hybrids by flow cytometry and inter-simple sequence repeat analyses. Almost all hybrids grew well and produced flowers 1-2 years after transplantation to the greenhouse. Hybrids had semi-cascade-type shoots, which was intermediate between T. formosana cultivars and T. hirta var. albescens (erect-type shoots) and T. macranthopsis (cascade-type shoots). They produced flowers with novel forms and colors compared with the corresponding parents, and some were horticulturally attractive. The results obtained in the present study indicate the validity of intersectional hybridization via ovule culture for breeding of Tricyrtis spp.","PeriodicalId":20411,"journal":{"name":"Plant Biotechnology","volume":"36 3 1","pages":"175-180"},"PeriodicalIF":1.6,"publicationDate":"2019-09-25","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://sci-hub-pdf.com/10.5511/plantbiotechnology.19.0807a","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"46730816","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Pub Date : 2019-09-25DOI: 10.5511/plantbiotechnology.19.0506a
Yunfeng Li, Xiao-Qin Zeng, Hui Zhuang, Huan Chen, Ting Zhang, Jun Zhang, Hao Zheng, Jun Tang, Hong-Lei Wang, Suxian Ren, Y. Ling, G. He
In cereal crops, the grain number per panicle and the grain yield are greatly affected by the number of florets in a spikelet. In wild-type rice, a spikelet only produces one fertile floret and beneath the floret are a pair of sterile lemmas and a pair of rudimentary glumes. This study characterized a rice spikelet mutant nonstop glumes 2 (nsg2). In the nsg2 mutant, both the sterile lemmas and rudimentary glumes were elongated, and part of sterile lemma looked like a lemma in appearance, shape and size. Detailed histological analysis and qPCR analysis revealed that the sterile lemmas in the nsg2 mutant had homeotically transformed into lemma-like organs. This phenotype indicates that NSG2 is involved in the regulation of spikelet development and supports the long-held view that sterile lemmas were derived from the lemmas of the two lateral florets. This implies that the rice spikelet has the potential to be restored to the "three florets spikelet", which may have existed in its ancestors. Genetic analysis reveals that the nsg2 trait is controlled by a single recessive gene. The NSG2 gene was finally mapped between markers R-20 and R-39 on chromosome 7 with a physical region of 180 kb. The two MYB family factors LOC_Os07g44030 and LOC_Os07g44090 might be involved in the development of the spikelet and floral organ, and they were considered as candidate genes of NSG2. These results provide a foundation for map-based cloning and function analysis of NSG2, as well as evidence to support "three-florets spikelet" breeding in rice.
{"title":"Characterization and fine mapping of nonstop glumes 2 (nsg2) mutant in rice (Oryza sativa L.).","authors":"Yunfeng Li, Xiao-Qin Zeng, Hui Zhuang, Huan Chen, Ting Zhang, Jun Zhang, Hao Zheng, Jun Tang, Hong-Lei Wang, Suxian Ren, Y. Ling, G. He","doi":"10.5511/plantbiotechnology.19.0506a","DOIUrl":"https://doi.org/10.5511/plantbiotechnology.19.0506a","url":null,"abstract":"In cereal crops, the grain number per panicle and the grain yield are greatly affected by the number of florets in a spikelet. In wild-type rice, a spikelet only produces one fertile floret and beneath the floret are a pair of sterile lemmas and a pair of rudimentary glumes. This study characterized a rice spikelet mutant nonstop glumes 2 (nsg2). In the nsg2 mutant, both the sterile lemmas and rudimentary glumes were elongated, and part of sterile lemma looked like a lemma in appearance, shape and size. Detailed histological analysis and qPCR analysis revealed that the sterile lemmas in the nsg2 mutant had homeotically transformed into lemma-like organs. This phenotype indicates that NSG2 is involved in the regulation of spikelet development and supports the long-held view that sterile lemmas were derived from the lemmas of the two lateral florets. This implies that the rice spikelet has the potential to be restored to the \"three florets spikelet\", which may have existed in its ancestors. Genetic analysis reveals that the nsg2 trait is controlled by a single recessive gene. The NSG2 gene was finally mapped between markers R-20 and R-39 on chromosome 7 with a physical region of 180 kb. The two MYB family factors LOC_Os07g44030 and LOC_Os07g44090 might be involved in the development of the spikelet and floral organ, and they were considered as candidate genes of NSG2. These results provide a foundation for map-based cloning and function analysis of NSG2, as well as evidence to support \"three-florets spikelet\" breeding in rice.","PeriodicalId":20411,"journal":{"name":"Plant Biotechnology","volume":"36 3 1","pages":"125-134"},"PeriodicalIF":1.6,"publicationDate":"2019-09-25","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://sci-hub-pdf.com/10.5511/plantbiotechnology.19.0506a","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"49060690","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Pub Date : 2019-09-25DOI: 10.5511/plantbiotechnology.19.0531a
Yuki Kanesaka, Masaaki Okada, S. Ito, T. Oyama
The rapid assessment of gene function is crucial in biological research. The CRISPR/Cas9 system is widely used as a tool for targeted gene editing in many organisms including plants. Previously, we established a transient gene expression system for investigating cellular circadian rhythms in duckweed. In this system, circadian reporters and clock gene effectors-such as overexpressors, RNA interference (RNAi), and CRISPR/Cas9-were introduced into duckweed cells using a particle bombardment method. In the present study, we applied the CRISPR/Cas9 system at a single cell level to Arabidopsis thaliana, a model organism in plant biology. To evaluate the mutation induction efficiency of the system, we monitored single-cell bioluminescence after application of the CRISPR/Cas9 system targeting the ELF3 gene, which is essential for robust circadian rhythmicity. We evaluated the mutation induction efficiency by determining the proportion of cells with impaired circadian rhythms. Three single guide RNAs (sgRNAs) were designed, and the proportion of arrhythmic cells following their use ranged from 32 to 91%. A comparison of the mutation induction efficiencies of diploid and tetraploid Arabidopsis suggested that endoreduplication had a slight effect on efficiency. Taken together, our results demonstrate that the transiently introduced CRISPR/Cas9 system is useful for rapidly assessing the physiological function of target genes in Arabidopsis cells.
{"title":"Monitoring single-cell bioluminescence of Arabidopsis leaves to quantitatively evaluate the efficiency of a transiently introduced CRISPR/Cas9 system targeting the circadian clock gene ELF3.","authors":"Yuki Kanesaka, Masaaki Okada, S. Ito, T. Oyama","doi":"10.5511/plantbiotechnology.19.0531a","DOIUrl":"https://doi.org/10.5511/plantbiotechnology.19.0531a","url":null,"abstract":"The rapid assessment of gene function is crucial in biological research. The CRISPR/Cas9 system is widely used as a tool for targeted gene editing in many organisms including plants. Previously, we established a transient gene expression system for investigating cellular circadian rhythms in duckweed. In this system, circadian reporters and clock gene effectors-such as overexpressors, RNA interference (RNAi), and CRISPR/Cas9-were introduced into duckweed cells using a particle bombardment method. In the present study, we applied the CRISPR/Cas9 system at a single cell level to Arabidopsis thaliana, a model organism in plant biology. To evaluate the mutation induction efficiency of the system, we monitored single-cell bioluminescence after application of the CRISPR/Cas9 system targeting the ELF3 gene, which is essential for robust circadian rhythmicity. We evaluated the mutation induction efficiency by determining the proportion of cells with impaired circadian rhythms. Three single guide RNAs (sgRNAs) were designed, and the proportion of arrhythmic cells following their use ranged from 32 to 91%. A comparison of the mutation induction efficiencies of diploid and tetraploid Arabidopsis suggested that endoreduplication had a slight effect on efficiency. Taken together, our results demonstrate that the transiently introduced CRISPR/Cas9 system is useful for rapidly assessing the physiological function of target genes in Arabidopsis cells.","PeriodicalId":20411,"journal":{"name":"Plant Biotechnology","volume":"104 5","pages":"187-193"},"PeriodicalIF":1.6,"publicationDate":"2019-09-25","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://sci-hub-pdf.com/10.5511/plantbiotechnology.19.0531a","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"41297367","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Pub Date : 2019-06-25DOI: 10.5511/PLANTBIOTECHNOLOGY.19.0322A
Ichiro Kasajima
Plant colors such as 'green leaf' and 'red apple' are often described based on human sense, even in scientific papers. On the other hand, colors are measured based on colorimetric principles in some papers, especially in the studies of horticultural plants. The science of color measurements ('colorimetry') is not included in any of the popular lectures in schools and universities, thus the principles of color measurements would not be understood by most researchers. The present review will overview the principles of colorimetry, and will introduce colorimetric methods which can be used for scientific measurement of plant colors. That is to say, the reflection spectrum of visible light (380-780 nm) is measured at 5-nm intervals on the surface of leaves or petals in 'Spectrometric Color Measurement' (SCM). The spectral data is multiplied with RGB or XYZ color matching functions and integrated to obtain RGB or XYZ intensities. Alternatively, approximate RGB values are directly obtained in 'Photographic Color Measurement' (PCM). RGB/XYZ intensities are further calculated to obtain 'hue', 'saturation', and 'lightness', the three factors of colors. Colorimetric insights into genetic regulations (such as MYB gene) and physiological regulations (such as alexandrite effect) of plant colors are also described.
{"title":"Measuring plant colors.","authors":"Ichiro Kasajima","doi":"10.5511/PLANTBIOTECHNOLOGY.19.0322A","DOIUrl":"https://doi.org/10.5511/PLANTBIOTECHNOLOGY.19.0322A","url":null,"abstract":"Plant colors such as 'green leaf' and 'red apple' are often described based on human sense, even in scientific papers. On the other hand, colors are measured based on colorimetric principles in some papers, especially in the studies of horticultural plants. The science of color measurements ('colorimetry') is not included in any of the popular lectures in schools and universities, thus the principles of color measurements would not be understood by most researchers. The present review will overview the principles of colorimetry, and will introduce colorimetric methods which can be used for scientific measurement of plant colors. That is to say, the reflection spectrum of visible light (380-780 nm) is measured at 5-nm intervals on the surface of leaves or petals in 'Spectrometric Color Measurement' (SCM). The spectral data is multiplied with RGB or XYZ color matching functions and integrated to obtain RGB or XYZ intensities. Alternatively, approximate RGB values are directly obtained in 'Photographic Color Measurement' (PCM). RGB/XYZ intensities are further calculated to obtain 'hue', 'saturation', and 'lightness', the three factors of colors. Colorimetric insights into genetic regulations (such as MYB gene) and physiological regulations (such as alexandrite effect) of plant colors are also described.","PeriodicalId":20411,"journal":{"name":"Plant Biotechnology","volume":"36 2 1","pages":"63-75"},"PeriodicalIF":1.6,"publicationDate":"2019-06-25","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://sci-hub-pdf.com/10.5511/PLANTBIOTECHNOLOGY.19.0322A","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"43576237","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Pub Date : 2019-06-25DOI: 10.5511/PLANTBIOTECHNOLOGY.19.0328A
Yongming Luo, Shoki Aoyama, Y. Fukao, Y. Chiba, Takeo Sato, J. Yamaguchi
As major components of the ubiquitin system, ubiquitin ligases mediate the transfer of ubiquitin to specific target substrates, thereby playing important roles in regulating a wide range of cellular processes. The Arabidopsis Tóxicos en Levadura (ATL) family is a group of plant-specific RING-type ubiquitin ligases with N-terminal transmembrane-like domains. To date, 91 ATL isoforms have been identified in the Arabidopsis genome, with some reported to regulate plant responses to environmental stresses. However, the functions of most ATLs remain unclear. This study showed that ATL8 is a sugar starvation response gene and that ATL8 expression was significantly increased by sugar starvation conditions but repressed by exogenous sugar supply. The ATL8 protein was found to possess ubiquitin ligase activity in vitro and to localize to membrane-bound compartments in plant cells. In addition, Starch Synthase 4 was identified as a putative interactor with ATL8, suggesting that ATL8 may be involved in modulating starch accumulation in response to sugar availability. These findings suggest that ATL8 functions as a membrane-localized ubiquitin ligase likely to be involved in the adaptation of Arabidopsis plants to sugar starvation stress.
作为泛素系统的主要组成部分，泛素连接酶介导泛素向特定靶底物的转移，从而在调节广泛的细胞过程中发挥重要作用。拟南芥Tóxicos en Levadura（ATL）家族是一组具有N-末端跨膜样结构域的植物特异性RING型泛素连接酶。到目前为止，在拟南芥基因组中已经鉴定出91种ATL异构体，其中一些被报道可以调节植物对环境胁迫的反应。然而，大多数ATL的功能仍不清楚。本研究表明ATL8是一个糖饥饿反应基因，ATL8的表达在糖饥饿条件下显著增加，但受到外源糖供应的抑制。ATL8蛋白在体外具有泛素连接酶活性，并定位于植物细胞中的膜结合区。此外，淀粉合成酶4被鉴定为与ATL8的假定相互作用因子，表明ATL8可能参与调节淀粉积累以响应糖的可用性。这些发现表明，ATL8作为一种膜定位的泛素连接酶发挥作用，可能参与拟南芥植物对糖饥饿胁迫的适应。
{"title":"Involvement of the membrane-localized ubiquitin ligase ATL8 in sugar starvation response in Arabidopsis.","authors":"Yongming Luo, Shoki Aoyama, Y. Fukao, Y. Chiba, Takeo Sato, J. Yamaguchi","doi":"10.5511/PLANTBIOTECHNOLOGY.19.0328A","DOIUrl":"https://doi.org/10.5511/PLANTBIOTECHNOLOGY.19.0328A","url":null,"abstract":"As major components of the ubiquitin system, ubiquitin ligases mediate the transfer of ubiquitin to specific target substrates, thereby playing important roles in regulating a wide range of cellular processes. The Arabidopsis Tóxicos en Levadura (ATL) family is a group of plant-specific RING-type ubiquitin ligases with N-terminal transmembrane-like domains. To date, 91 ATL isoforms have been identified in the Arabidopsis genome, with some reported to regulate plant responses to environmental stresses. However, the functions of most ATLs remain unclear. This study showed that ATL8 is a sugar starvation response gene and that ATL8 expression was significantly increased by sugar starvation conditions but repressed by exogenous sugar supply. The ATL8 protein was found to possess ubiquitin ligase activity in vitro and to localize to membrane-bound compartments in plant cells. In addition, Starch Synthase 4 was identified as a putative interactor with ATL8, suggesting that ATL8 may be involved in modulating starch accumulation in response to sugar availability. These findings suggest that ATL8 functions as a membrane-localized ubiquitin ligase likely to be involved in the adaptation of Arabidopsis plants to sugar starvation stress.","PeriodicalId":20411,"journal":{"name":"Plant Biotechnology","volume":"36 2 1","pages":"107-112"},"PeriodicalIF":1.6,"publicationDate":"2019-06-25","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://sci-hub-pdf.com/10.5511/PLANTBIOTECHNOLOGY.19.0328A","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"47498134","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Pub Date : 2019-06-25DOI: 10.5511/PLANTBIOTECHNOLOGY.19.0220B
Takuya Suzaki, Mai Tsuda, H. Ezura, B. Day, K. Miura
Transient protein expression is an effective tool to rapidly unravel novel gene functions, such as transcriptional activity of promoters and sub-cellular localization of proteins. However, transient expression is not applicable to some species and varieties because of insufficient expression levels. We recently developed one of the strongest agroinfiltration-based transient protein expression systems for plant cells, termed 'Tsukuba system.' About 4 mg/g fresh weight of protein expression in Nicotiana benthamiana was obtained using this system. The vector pBYR2HS, which contains a geminiviral replication system and a double terminator, can be used in various plant species and varieties, including lettuces, eggplants, tomatoes, hot peppers, and orchids. In this study, we assessed the applicability of the Tsukuba system to several species of legumes, including Lotus japonicus, soybean Glycine max, and common bean Phaseolus vulgaris. The GFP protein was transiently expressed in the seedpods of all examined legume species, however, protein expression in leaves was observed only in P. vulgaris. Taken together, our system is an effective tool to examine gene function rapidly in several legume species based on transient protein expression.
{"title":"Agroinfiltration-based efficient transient protein expression in leguminous plants.","authors":"Takuya Suzaki, Mai Tsuda, H. Ezura, B. Day, K. Miura","doi":"10.5511/PLANTBIOTECHNOLOGY.19.0220B","DOIUrl":"https://doi.org/10.5511/PLANTBIOTECHNOLOGY.19.0220B","url":null,"abstract":"Transient protein expression is an effective tool to rapidly unravel novel gene functions, such as transcriptional activity of promoters and sub-cellular localization of proteins. However, transient expression is not applicable to some species and varieties because of insufficient expression levels. We recently developed one of the strongest agroinfiltration-based transient protein expression systems for plant cells, termed 'Tsukuba system.' About 4 mg/g fresh weight of protein expression in Nicotiana benthamiana was obtained using this system. The vector pBYR2HS, which contains a geminiviral replication system and a double terminator, can be used in various plant species and varieties, including lettuces, eggplants, tomatoes, hot peppers, and orchids. In this study, we assessed the applicability of the Tsukuba system to several species of legumes, including Lotus japonicus, soybean Glycine max, and common bean Phaseolus vulgaris. The GFP protein was transiently expressed in the seedpods of all examined legume species, however, protein expression in leaves was observed only in P. vulgaris. Taken together, our system is an effective tool to examine gene function rapidly in several legume species based on transient protein expression.","PeriodicalId":20411,"journal":{"name":"Plant Biotechnology","volume":"36 2 1","pages":"119-123"},"PeriodicalIF":1.6,"publicationDate":"2019-06-25","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://sci-hub-pdf.com/10.5511/PLANTBIOTECHNOLOGY.19.0220B","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"44198260","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Pub Date : 2019-06-25DOI: 10.5511/PLANTBIOTECHNOLOGY.19.0417A
Shigeru Hanamata, Jumpei Sawada, B. Toh, Seijiro Ono, K. Ogawa, Togo Fukunaga, K. Nonomura, Takamitsu Kurusu, K. Kuchitsu
We have previously shown that autophagy is required for post meiotic anther development including programmed cell death-mediated degradation of the tapetum and pollen maturation in rice. However, the spatiotemporal dynamics of autophagy in the tapetum remain poorly understood. We here established an in vivo imaging technique to analyze the dynamics of autophagy in rice tapetum cells by expressing green fluorescent protein-tagged AtATG8, a marker for autophagosomes. 3D-imaging analysis revealed that the number of autophagosomes/autophagy-related structures is extremely low at the tetrad stage (stage 8), and autophagy is dramatically induced at the uninucleate stages (stage 9-10) throughout the tapetal cells during anther development. The present monitoring system for autophagy offers a powerful tool to analyze the regulation of autophagy in rice tapetal cells during pollen maturation.
{"title":"Monitoring autophagy in rice tapetal cells during pollen maturation.","authors":"Shigeru Hanamata, Jumpei Sawada, B. Toh, Seijiro Ono, K. Ogawa, Togo Fukunaga, K. Nonomura, Takamitsu Kurusu, K. Kuchitsu","doi":"10.5511/PLANTBIOTECHNOLOGY.19.0417A","DOIUrl":"https://doi.org/10.5511/PLANTBIOTECHNOLOGY.19.0417A","url":null,"abstract":"We have previously shown that autophagy is required for post meiotic anther development including programmed cell death-mediated degradation of the tapetum and pollen maturation in rice. However, the spatiotemporal dynamics of autophagy in the tapetum remain poorly understood. We here established an in vivo imaging technique to analyze the dynamics of autophagy in rice tapetum cells by expressing green fluorescent protein-tagged AtATG8, a marker for autophagosomes. 3D-imaging analysis revealed that the number of autophagosomes/autophagy-related structures is extremely low at the tetrad stage (stage 8), and autophagy is dramatically induced at the uninucleate stages (stage 9-10) throughout the tapetal cells during anther development. The present monitoring system for autophagy offers a powerful tool to analyze the regulation of autophagy in rice tapetal cells during pollen maturation.","PeriodicalId":20411,"journal":{"name":"Plant Biotechnology","volume":"36 2 1","pages":"99-105"},"PeriodicalIF":1.6,"publicationDate":"2019-06-25","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://sci-hub-pdf.com/10.5511/PLANTBIOTECHNOLOGY.19.0417A","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"44087552","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Pub Date : 2019-06-25DOI: 10.5511/PLANTBIOTECHNOLOGY.19.0317A
B. Gutiérrez, M. M. Cobo, M. Orellana, J. Vega, V. Arahana, Viviana Jaramillo, M. Torres
The development of in vitro propagation methods can improve the current commercial use and conservation of plants like naranjilla (Solanum quitoense), a distinctive Andean crop and key emerging agricultural product. In the present study, we report in vitro culture protocols for naranjilla apical buds, hypocotyls and petioles. In apical bud culture, MS medium supplemented with 0.10 mg l-1 1-naphtaleneacetic acid (NAA) produced longer plantlets with greater number of leaves. Hypocotyl culture yielded higher number of shoots when using older explants in MS medium supplemented with different combinations of NAA, 6-benzylaminopurine (BAP) and gibberellic acid (GA3). Petiole culture produced a significantly higher number of shoots per explant, with more abundant and bigger leaves, when using MS medium supplemented with 0.02 mg l-1 NAA, 4.50 mg l-1 BAP and 1.00 mg l-1 GA3. A factorial analysis reveals that the interaction between GA3 and NAA/BAP plays an important role in shoot regeneration. These results provide new tools for the in vitro regeneration of naranjilla plants, improving on previously reported protocols for this species by using alternative explant types and regeneration protocols.
{"title":"Micropropagation of Solanum quitoense var. quitoense by apical bud, petiole and hypocotyl culture.","authors":"B. Gutiérrez, M. M. Cobo, M. Orellana, J. Vega, V. Arahana, Viviana Jaramillo, M. Torres","doi":"10.5511/PLANTBIOTECHNOLOGY.19.0317A","DOIUrl":"https://doi.org/10.5511/PLANTBIOTECHNOLOGY.19.0317A","url":null,"abstract":"The development of in vitro propagation methods can improve the current commercial use and conservation of plants like naranjilla (Solanum quitoense), a distinctive Andean crop and key emerging agricultural product. In the present study, we report in vitro culture protocols for naranjilla apical buds, hypocotyls and petioles. In apical bud culture, MS medium supplemented with 0.10 mg l-1 1-naphtaleneacetic acid (NAA) produced longer plantlets with greater number of leaves. Hypocotyl culture yielded higher number of shoots when using older explants in MS medium supplemented with different combinations of NAA, 6-benzylaminopurine (BAP) and gibberellic acid (GA3). Petiole culture produced a significantly higher number of shoots per explant, with more abundant and bigger leaves, when using MS medium supplemented with 0.02 mg l-1 NAA, 4.50 mg l-1 BAP and 1.00 mg l-1 GA3. A factorial analysis reveals that the interaction between GA3 and NAA/BAP plays an important role in shoot regeneration. These results provide new tools for the in vitro regeneration of naranjilla plants, improving on previously reported protocols for this species by using alternative explant types and regeneration protocols.","PeriodicalId":20411,"journal":{"name":"Plant Biotechnology","volume":"36 2 1","pages":"91-97"},"PeriodicalIF":1.6,"publicationDate":"2019-06-25","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://sci-hub-pdf.com/10.5511/PLANTBIOTECHNOLOGY.19.0317A","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"47572531","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Rice flo2 mutation produces grains showing a reduced amount of storage proteins. Using Nipponbare and the flo2 mutant, we created rice transformants that showed defective production of major allergen proteins RA14 and RA33 (14-16 kDa and 33 kDa allergen proteins, respectively) by RNAi introduction. The knock-down transformant generated using Nipponbare showed greatly reduced accumulation of both allergen proteins, normal growth, and production of a sufficient amount of normal-shaped seeds. F1 seeds were obtained by crossing between the transformants containing RNAi genes to RA14 and RA33, and showed decreased accumulation of both proteins. However, a peculiar phenotype was observed in the flo2 transformants that lacked accumulation of RA14 or RA33. They showed significantly reduced fertility. A wrinkled grain feature was found on the transformant lacking accumulation of RA14. F1 seeds obtained by crossing these transformants showed significantly lower fertility. F2 seeds showed decreases in both allergen proteins but morphological abnormality with small and severely wrinkled features. These results indicated that it is hard to obtain any transformant lacking accumulation of these allergen proteins using the flo2 mutant, whereas a knock-down transformant of both allergen protein genes was obtained when a wild-type Nipponbare was used as a host. These facts strongly suggest that RA14 and RA33 have some roles in rice seeds.
{"title":"Aberrant endosperm formation caused by reduced production of major allergen proteins in a rice flo2 mutant that confers low-protein accumulation in grains.","authors":"Hiroshi Teramura, Kyoko Kondo, Masato Suzuki, Hiroaki Kobayashi, Tsutomu Furukawa, Hiroaki Kusano, H. Shimada","doi":"10.5511/PLANTBIOTECHNOLOGY.19.0312A","DOIUrl":"https://doi.org/10.5511/PLANTBIOTECHNOLOGY.19.0312A","url":null,"abstract":"Rice flo2 mutation produces grains showing a reduced amount of storage proteins. Using Nipponbare and the flo2 mutant, we created rice transformants that showed defective production of major allergen proteins RA14 and RA33 (14-16 kDa and 33 kDa allergen proteins, respectively) by RNAi introduction. The knock-down transformant generated using Nipponbare showed greatly reduced accumulation of both allergen proteins, normal growth, and production of a sufficient amount of normal-shaped seeds. F1 seeds were obtained by crossing between the transformants containing RNAi genes to RA14 and RA33, and showed decreased accumulation of both proteins. However, a peculiar phenotype was observed in the flo2 transformants that lacked accumulation of RA14 or RA33. They showed significantly reduced fertility. A wrinkled grain feature was found on the transformant lacking accumulation of RA14. F1 seeds obtained by crossing these transformants showed significantly lower fertility. F2 seeds showed decreases in both allergen proteins but morphological abnormality with small and severely wrinkled features. These results indicated that it is hard to obtain any transformant lacking accumulation of these allergen proteins using the flo2 mutant, whereas a knock-down transformant of both allergen protein genes was obtained when a wild-type Nipponbare was used as a host. These facts strongly suggest that RA14 and RA33 have some roles in rice seeds.","PeriodicalId":20411,"journal":{"name":"Plant Biotechnology","volume":"36 2 1","pages":"85-90"},"PeriodicalIF":1.6,"publicationDate":"2019-06-25","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://sci-hub-pdf.com/10.5511/PLANTBIOTECHNOLOGY.19.0312A","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"45611009","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
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