Light spectrum plays an important role in regulating the growth and development of in vitro cultured potato(Solanum tuberosum L.) plantlets. The status of potato plantlets at the end of in vitro stage influences the m...Light spectrum plays an important role in regulating the growth and development of in vitro cultured potato(Solanum tuberosum L.) plantlets. The status of potato plantlets at the end of in vitro stage influences the minituber production after transplanting. With 100 μmol m^-2s^-1 total photosynthetic photon flux density(PPFD), a light spectrum study of 100% red light emitting diodes(LEDs) light spectrum(RR), 100% blue LEDs light spectrum(BB), 65% red+35% blue LEDs light spectrum(RB), and 45% red+35% blue+20% green LEDs light spectrum(RBG) providing illumination at the in vitro cultured stage of potato plantlets for 4 weeks using fluorescent lamp as control(CK) was performed to investigate the effects of LEDs light spectrum on the growth, leaf anatomy, and chloroplast ultrastructure of potato plantlets in vitro as well as the minituber yield after 2 months transplanting in the greenhouse. Compared to CK, RB and RBG promoted the growth of potato plantlets in vitro with increased stem diameter, plantlet fresh weight, plantlet dry weight, and health index. Furthermore, BB induced the greatest stem diameter as well as the highest health index in potato plantlets in vitro. Root activity, soluble protein, and free amino acid were also significantly enhanced by BB, whereas carbohydrates were improved by RR. In addition, thickness of leaf, palisade parenchyma and spongy parenchyma was significantly increased by BB and RBG. Chloroplasts under BB and RBG showed well-developed grana thylakoid and stroma thylakoid. Unexpectedly, distinct upper epidermis with greatest thickness was induced and palisade parenchyma and spongy parenchyma were arranged neatly in RR. After transplanting in the greenhouse for 2 months, potato plantlets in vitro from BB, RB, and RBG produced high percentage of large size tuber. BB improved fresh and dry weights of the biggest tuber but decreased tuber number per plantlet. In addition, RBG increased tuber number as well as tuber fresh and dry weight slightly. Our results suggested monochromatic blue LEDs as well as combined red, blue or/and green LEDs light spectrum were superior to fluorescent lamp spectrum in micro-propagation of potato plantlets. Therefore, the application of RBG was suitable;BB and RB could be used as alternatives.展开更多
Plant in vitro organogenesis is well-controlled and thus provides an ideal system for plant propagation and studying mechanisms of plant development. However, the data on systematic in vitro organogenesis from leaf ex...Plant in vitro organogenesis is well-controlled and thus provides an ideal system for plant propagation and studying mechanisms of plant development. However, the data on systematic in vitro organogenesis from leaf explant with various concentrations and combinations of hormones are limited. Arabidopsis is a very useful model plant species for many aspects of plant biological study. Here, we reported a simple, fast and efficient one-step process for evaluating leaf explant-derived in vitro Arabidopsis organogenesis involving the application of various concentrations and combinations of exogenous hormones. The central portion of the fourth rosette leaf was harvested from the 21-days-old seedling and cultured in vitro on the media containing 216 combinations of exogenous hormones. Different types of organs, including roots, shoots, inflorescences, and leaf-like organs were initiated from leaf explants. Several optimal experimental combinations were selected. A hormone combination, 1.00 μM NAA + 10.00 μM ZT, was considered as the most efficient one for adventitious shoot regeneration. And for adventitious root regeneration, six hormone combinations, [(NAA + ZT: 1.00 + 0.10 μM;10.00 + 0.01 μM;20.00 + 0.10 μM;20.00 + 1.00 μM) and (NAA + 6-BA: 10.00 + 0.10 μM;20.00 + 10.00 μM)], were thought to be the best ones. Further, both auxin and cytokinin ratios and concentrations were crucial for efficient in vitro organogenesis. Our study provides the important information for hormone-regulated organogenesis.展开更多
The specificities of tissue culture of wheat greatly limit the use of chloroplast transformation technologies in this crop. One limitation in wheat tissue culture is that it is difficult to regenerate plantlets from l...The specificities of tissue culture of wheat greatly limit the use of chloroplast transformation technologies in this crop. One limitation in wheat tissue culture is that it is difficult to regenerate plantlets from leaf tissue explants of regenerated plantlets, resulting in difficulty in obtaining homoplastic plants via multiple rounds of antibiotic selection of chloroplast transformants. Thus, a repeated in vitro regeneration system from leaf tissues was studied in this research. Our results showed that 2 mm leaf basal segments of the 4 cm high leaves from regenerated plantlets can give the best callus induction at present study. The best callus induction medium was Murashige and Skoog (MS) basal medium supplemented with 2 mg/L 2,4-dichlorophenoxyacetic acid and 1 mg/L naphthalenacetic acid, which gave a callus induction rate of up to 87.2%. The optimal differentiation medium was MS basal medium supplemented with 10 mg/L silver nitrate and 1 mg/L 2,3,5-triiodobenzoic acid, which gave a regeneration rate up to 33.7% for the wheat lines tested. This is the first report showing that leaf basal segments of in vitro regenerated plantlets can be used for regeneration of wheat. The establishment of a repetitive regeneration system should pave the way for the development of chloroplast transformation and the plant regeneration systems starting from leaf material of in vitro regenerated wheat and other cereal crops.展开更多
基金supported by the National High-Tech R&D Program of China (2013 AA 103005)
文摘Light spectrum plays an important role in regulating the growth and development of in vitro cultured potato(Solanum tuberosum L.) plantlets. The status of potato plantlets at the end of in vitro stage influences the minituber production after transplanting. With 100 μmol m^-2s^-1 total photosynthetic photon flux density(PPFD), a light spectrum study of 100% red light emitting diodes(LEDs) light spectrum(RR), 100% blue LEDs light spectrum(BB), 65% red+35% blue LEDs light spectrum(RB), and 45% red+35% blue+20% green LEDs light spectrum(RBG) providing illumination at the in vitro cultured stage of potato plantlets for 4 weeks using fluorescent lamp as control(CK) was performed to investigate the effects of LEDs light spectrum on the growth, leaf anatomy, and chloroplast ultrastructure of potato plantlets in vitro as well as the minituber yield after 2 months transplanting in the greenhouse. Compared to CK, RB and RBG promoted the growth of potato plantlets in vitro with increased stem diameter, plantlet fresh weight, plantlet dry weight, and health index. Furthermore, BB induced the greatest stem diameter as well as the highest health index in potato plantlets in vitro. Root activity, soluble protein, and free amino acid were also significantly enhanced by BB, whereas carbohydrates were improved by RR. In addition, thickness of leaf, palisade parenchyma and spongy parenchyma was significantly increased by BB and RBG. Chloroplasts under BB and RBG showed well-developed grana thylakoid and stroma thylakoid. Unexpectedly, distinct upper epidermis with greatest thickness was induced and palisade parenchyma and spongy parenchyma were arranged neatly in RR. After transplanting in the greenhouse for 2 months, potato plantlets in vitro from BB, RB, and RBG produced high percentage of large size tuber. BB improved fresh and dry weights of the biggest tuber but decreased tuber number per plantlet. In addition, RBG increased tuber number as well as tuber fresh and dry weight slightly. Our results suggested monochromatic blue LEDs as well as combined red, blue or/and green LEDs light spectrum were superior to fluorescent lamp spectrum in micro-propagation of potato plantlets. Therefore, the application of RBG was suitable;BB and RB could be used as alternatives.
文摘Plant in vitro organogenesis is well-controlled and thus provides an ideal system for plant propagation and studying mechanisms of plant development. However, the data on systematic in vitro organogenesis from leaf explant with various concentrations and combinations of hormones are limited. Arabidopsis is a very useful model plant species for many aspects of plant biological study. Here, we reported a simple, fast and efficient one-step process for evaluating leaf explant-derived in vitro Arabidopsis organogenesis involving the application of various concentrations and combinations of exogenous hormones. The central portion of the fourth rosette leaf was harvested from the 21-days-old seedling and cultured in vitro on the media containing 216 combinations of exogenous hormones. Different types of organs, including roots, shoots, inflorescences, and leaf-like organs were initiated from leaf explants. Several optimal experimental combinations were selected. A hormone combination, 1.00 μM NAA + 10.00 μM ZT, was considered as the most efficient one for adventitious shoot regeneration. And for adventitious root regeneration, six hormone combinations, [(NAA + ZT: 1.00 + 0.10 μM;10.00 + 0.01 μM;20.00 + 0.10 μM;20.00 + 1.00 μM) and (NAA + 6-BA: 10.00 + 0.10 μM;20.00 + 10.00 μM)], were thought to be the best ones. Further, both auxin and cytokinin ratios and concentrations were crucial for efficient in vitro organogenesis. Our study provides the important information for hormone-regulated organogenesis.
文摘The specificities of tissue culture of wheat greatly limit the use of chloroplast transformation technologies in this crop. One limitation in wheat tissue culture is that it is difficult to regenerate plantlets from leaf tissue explants of regenerated plantlets, resulting in difficulty in obtaining homoplastic plants via multiple rounds of antibiotic selection of chloroplast transformants. Thus, a repeated in vitro regeneration system from leaf tissues was studied in this research. Our results showed that 2 mm leaf basal segments of the 4 cm high leaves from regenerated plantlets can give the best callus induction at present study. The best callus induction medium was Murashige and Skoog (MS) basal medium supplemented with 2 mg/L 2,4-dichlorophenoxyacetic acid and 1 mg/L naphthalenacetic acid, which gave a callus induction rate of up to 87.2%. The optimal differentiation medium was MS basal medium supplemented with 10 mg/L silver nitrate and 1 mg/L 2,3,5-triiodobenzoic acid, which gave a regeneration rate up to 33.7% for the wheat lines tested. This is the first report showing that leaf basal segments of in vitro regenerated plantlets can be used for regeneration of wheat. The establishment of a repetitive regeneration system should pave the way for the development of chloroplast transformation and the plant regeneration systems starting from leaf material of in vitro regenerated wheat and other cereal crops.
