The present work initially identified the design parameters of a temporary immersion bioreactor to later scale it to a complete system for the </span><i><span style="font-family:Verdana;">i...The present work initially identified the design parameters of a temporary immersion bioreactor to later scale it to a complete system for the </span><i><span style="font-family:Verdana;">in vitro</span></i><span style="font-family:Verdana;"> multiplication of </span><i><span style="font-family:Verdana;">Ananas comosus</span></i><span style="font-family:Verdana;"> var. </span><span style="font-family:Verdana;">Trujillana</span><span style="font-family:Verdana;"> Red. Thus, a low-cost pneumatic temporary immersion bioreactor system was designed and built with 24 tanks of 2 L each. The automation of the system was designed and implemented by means of a timer circuit whose design parameters were: duration of the propagation process, which depends on the multiplication period of the crop and is an open variable, which means that the operator decides when to turn off the system;the duration of each dive, which for reasons of </span><span style="font-family:Verdana;">complexity</span><span style="font-family:Verdana;"> of the algorithm was standardized as one minute;immersion frequency, which was programmed for intervals of 1, 2, 3, 4, 5, 6, 7, 8 hours respectively and duration of aeration, which from a test run times of 0.20 were chosen, 30, 40, 50, 60, 70, and 80 seconds that correspond to the time of delivery of compressed air;additionally, the multiplication rate of </span><i><span style="font-family:Verdana;">Ananas comosus</span></i><span style="font-family:Verdana;"> var. </span><span style="font-family:Verdana;">Trujillana</span><span style="font-family:Verdana;"> Red in the immersion system which was 6.5 times per propagative unit inoculated in thirty days.展开更多
By employing temporary immersion bioreactor system(TIBs),we studied virus-free culture of seedlings from sugarcane varieties ROC16 and ROC22,from medium recipe,inoculation amount,sucrose concentration,and variety diff...By employing temporary immersion bioreactor system(TIBs),we studied virus-free culture of seedlings from sugarcane varieties ROC16 and ROC22,from medium recipe,inoculation amount,sucrose concentration,and variety difference. The results showed,using this method,that proliferation rate of ROC16 improved by 40 times,per flask generated about 800 plantlets; of ROC22 improved by 30 times,per flask generated about 400-600 plantlets. The results provided basis for using TIBs in rapid propagation of plantlets via tissue culture.展开更多
Although several studies have reported on the propagation of the Sagittaria sagittifolia to date, none of these methods have efficiently achieved the mass production of these plants. The present study aimed to investi...Although several studies have reported on the propagation of the Sagittaria sagittifolia to date, none of these methods have efficiently achieved the mass production of these plants. The present study aimed to investigate the propagation and growth of S. sagittifolia using a temporary immersion bioreactor system （TIBS） compared with conventional semi-solid and liquid culture. The effect of different immersion frequencies and immersion times together with supplementation of various plant growth regulators to the Murashige and Skoog （MS） medium was evaluated on shoot proliferation and plant growth. The results showed that the higher immersion frequency （every 6 h） and shorter immersion time （3 min and 10 rain） in medium containing 4 mg/L BA and 0.1 mg/L NAA produced the highest multiplication rate （23）, which are significantly higher than conventional semi-solid （3.6） and liquid （4.5） method, and the best plant growth parameter. While, the lower immersion frequency and longer immersion time （30 rain every 12 h and 60 min every 24 h） induced vitrification and pollution rate in shoot tips 16.6% and 19%, 42% and 37%, respectively. There is distinct decrease in pollution rate （8.3%） in TIBS （10 min every 6 h） compared with the conventional semi-solid and liquid cultures. Medium containing 4 mg/L BA and 0.5 mg/L NAA using 10 min immersion every 6 h showed satisfaction at the rooting stage, with high shoot proliferation rate （21.6）, 100% rooting and 94% plant survival. Therefore, applying TIBS in S. sagittifolia is an efficient method for scaling up the production of plantlets with high quality seedlings.展开更多
Agave guiengola Gentry is an endemic plant from a very small locality in Oaxaca,Mexico.Its conservation status is fragile and can rapidly worsen.Because of its scarcity,this agave has been used solely for ornamental p...Agave guiengola Gentry is an endemic plant from a very small locality in Oaxaca,Mexico.Its conservation status is fragile and can rapidly worsen.Because of its scarcity,this agave has been used solely for ornamental purposes,but it could have other uses if more plants were available.In vitro propagation by enhanced axillary sprouting from stem segments was attained using Murashige and Skoog Basal Medium(MS)as well as basal medium supplemented with cytokinins 6-Benzylaminopurine(BA)or 6-(γ,γ-Dimethylallylamino)purine(2iP).The best treatment for shoot induction in semisolid medium consisted in MS supplemented with 2 mg l^(–1) BA,obtaining a mean of 3.7 shoots per explant.Other interesting responses were observed,such as nodular callus induction using combinations of BA and 2,4-Dichlorophenoxyacetic acid(2,4-D);root induction without Plant Growth Regulators(PGR);and generation of shoot clusters.These clusters constituted an excellent explant for micropropagation in temporary immersion bioreactors,obtaining a propagation rate of 43 shoots per explant with 1 min immersion and 6 h immersion frequencies.All new plants rooted and survived the transfer to soil.This study developed an in vitro propagation scheme to produce individuals that can be used either for reforestation,economical purposes,or to carry out studies in this species to assess its full potential,avoiding exploitation from wild plants.展开更多
In this study, we employed a temporary immersion bioreactor(TIB)system for the micropropagation of strawberry seedlings. The TIB method and the conventional method for the micropropagation of strawberry seedlings were...In this study, we employed a temporary immersion bioreactor(TIB)system for the micropropagation of strawberry seedlings. The TIB method and the conventional method for the micropropagation of strawberry seedlings were compared in terms of explant propagation coefficient, seedling fresh weight,contents of anthocyanins and chlorophyll, and photosynthetic characteristics. The results showed that an inoculation density of 40 explants/L was suitable for the micropropagation of ’Benihope’ strawberry seedlings in the TIB. The propagation coefficient, fresh weight, photosynthetic rate, stomatal conductance, transpiration coefficient, and the contents of total anthocyanins and chlorophyll of seedlings micropropagated in the TIB were significantly higher than those of the seedlings micropropagated by the conventional method. In conclusion, the TIB method was superior to the conventional method in the micropropagation of strawberry seedlings.The negative reciprocity did not occur between anthocyanins and chlorophyll in strawberry seedlings cultured under forced ventilation such as TIB.展开更多
In Latin America the forestry of exotic species such as teak has been increasing in recent decades, due to their advantages in wood quality, rapid growth;and the relative ease of producing clones and their multiplicat...In Latin America the forestry of exotic species such as teak has been increasing in recent decades, due to their advantages in wood quality, rapid growth;and the relative ease of producing clones and their multiplication with respect to native species. Therefore, there is great interest in developing larger-scale propagation strategies that reduce costs and intensive manual labor. Culture in liquid media with temporary immersion and the semi-automation of the system has raised expectations for large-scale micropropagation. We report a protocol for teak, which reuses the primary explants in several culture cycles in semi-solid medium to produce nodal explants for the multiplication phase in temporary immersion bioreactors (RITA®). The control of factors such as cytokinin concentration, explants density, immersion frequencies and culture duration was analyzed. The number of shoots increased with 0.5 mg·l-1 of BA (6-Benzyladenine), alone or in combination with 0.5 mg·l-1 of Kinetin, with 2 daily immersions of 1 minute each;however, these shoots showed a high degree of hyperhydricity. When 0.05 mg·l-1 of BA was used with 1 immersion of 1 minute every 2 days, the hyperhydricity decreased. Although the number of shoots was lower, they showed good length to be used during multiplication and rooting ex vitro. Our results suggest that teak micropropagation can be simplified in two phases in vitro, the establishment and multiplication;followed by rooting ex vitro and acclimatization. This would imply a reduction in production costs, since most of the multiplication would take place in RITA®containers.展开更多
A new procedure for blueberry (Vaccinium corymbosum L.) micropropagation in programmed Temporary Immersion Bioreactors (TIBs based on two separate bottles) was developed for the commercial genotypes Biloxi, Sharp Blue...A new procedure for blueberry (Vaccinium corymbosum L.) micropropagation in programmed Temporary Immersion Bioreactors (TIBs based on two separate bottles) was developed for the commercial genotypes Biloxi, Sharp Blue and Brillita. Plant cultures were developed in a controlled environment with 0.4 MPa CO2 enrichment, sucrose-reduced medium, and light intensity of 60 mM m-2·s-1. Principal component analysis showed that component 1 (C1) grouped 64.08% of the total variability, while the first two components accounted for 86.97%. Representation of the principal components demonstrated three clusters corresponding with the blueberry genotypes, and within each cluster plants micropropagated in agar-base medium grouped separately from those plants multiplied in TIBs. Both plant number and total internodes traits (related to the productive efficiency) were demonstrated superior in blueberries propagated in TIBs. Additionally, when transferred to greenhouse conditions, blueberries propagated in TIBs showed higher adaptability and growing rates than those cultured by the conventional approach, altogether evidencing the occurrence of a photomixotrophic stage in the vitroplantlets cultured in TIBs.展开更多
文摘The present work initially identified the design parameters of a temporary immersion bioreactor to later scale it to a complete system for the </span><i><span style="font-family:Verdana;">in vitro</span></i><span style="font-family:Verdana;"> multiplication of </span><i><span style="font-family:Verdana;">Ananas comosus</span></i><span style="font-family:Verdana;"> var. </span><span style="font-family:Verdana;">Trujillana</span><span style="font-family:Verdana;"> Red. Thus, a low-cost pneumatic temporary immersion bioreactor system was designed and built with 24 tanks of 2 L each. The automation of the system was designed and implemented by means of a timer circuit whose design parameters were: duration of the propagation process, which depends on the multiplication period of the crop and is an open variable, which means that the operator decides when to turn off the system;the duration of each dive, which for reasons of </span><span style="font-family:Verdana;">complexity</span><span style="font-family:Verdana;"> of the algorithm was standardized as one minute;immersion frequency, which was programmed for intervals of 1, 2, 3, 4, 5, 6, 7, 8 hours respectively and duration of aeration, which from a test run times of 0.20 were chosen, 30, 40, 50, 60, 70, and 80 seconds that correspond to the time of delivery of compressed air;additionally, the multiplication rate of </span><i><span style="font-family:Verdana;">Ananas comosus</span></i><span style="font-family:Verdana;"> var. </span><span style="font-family:Verdana;">Trujillana</span><span style="font-family:Verdana;"> Red in the immersion system which was 6.5 times per propagative unit inoculated in thirty days.
基金Supported by Youth Science Foundation of Guangxi ( Guikeqing0832060)S&T Development Project from Guangxi Academy of Agricultural Sciences(2006006)~~
文摘By employing temporary immersion bioreactor system(TIBs),we studied virus-free culture of seedlings from sugarcane varieties ROC16 and ROC22,from medium recipe,inoculation amount,sucrose concentration,and variety difference. The results showed,using this method,that proliferation rate of ROC16 improved by 40 times,per flask generated about 800 plantlets; of ROC22 improved by 30 times,per flask generated about 400-600 plantlets. The results provided basis for using TIBs in rapid propagation of plantlets via tissue culture.
文摘Although several studies have reported on the propagation of the Sagittaria sagittifolia to date, none of these methods have efficiently achieved the mass production of these plants. The present study aimed to investigate the propagation and growth of S. sagittifolia using a temporary immersion bioreactor system （TIBS） compared with conventional semi-solid and liquid culture. The effect of different immersion frequencies and immersion times together with supplementation of various plant growth regulators to the Murashige and Skoog （MS） medium was evaluated on shoot proliferation and plant growth. The results showed that the higher immersion frequency （every 6 h） and shorter immersion time （3 min and 10 rain） in medium containing 4 mg/L BA and 0.1 mg/L NAA produced the highest multiplication rate （23）, which are significantly higher than conventional semi-solid （3.6） and liquid （4.5） method, and the best plant growth parameter. While, the lower immersion frequency and longer immersion time （30 rain every 12 h and 60 min every 24 h） induced vitrification and pollution rate in shoot tips 16.6% and 19%, 42% and 37%, respectively. There is distinct decrease in pollution rate （8.3%） in TIBS （10 min every 6 h） compared with the conventional semi-solid and liquid cultures. Medium containing 4 mg/L BA and 0.5 mg/L NAA using 10 min immersion every 6 h showed satisfaction at the rooting stage, with high shoot proliferation rate （21.6）, 100% rooting and 94% plant survival. Therefore, applying TIBS in S. sagittifolia is an efficient method for scaling up the production of plantlets with high quality seedlings.
基金This work was supported financially by project AGS-2015-02-01-267656,Fondo Mixto CONACYT-Gobierno del Estado de Aguascalientes,and project PIBT-18-2,Universidad Autónoma de Aguascalientes,México.
文摘Agave guiengola Gentry is an endemic plant from a very small locality in Oaxaca,Mexico.Its conservation status is fragile and can rapidly worsen.Because of its scarcity,this agave has been used solely for ornamental purposes,but it could have other uses if more plants were available.In vitro propagation by enhanced axillary sprouting from stem segments was attained using Murashige and Skoog Basal Medium(MS)as well as basal medium supplemented with cytokinins 6-Benzylaminopurine(BA)or 6-(γ,γ-Dimethylallylamino)purine(2iP).The best treatment for shoot induction in semisolid medium consisted in MS supplemented with 2 mg l^(–1) BA,obtaining a mean of 3.7 shoots per explant.Other interesting responses were observed,such as nodular callus induction using combinations of BA and 2,4-Dichlorophenoxyacetic acid(2,4-D);root induction without Plant Growth Regulators(PGR);and generation of shoot clusters.These clusters constituted an excellent explant for micropropagation in temporary immersion bioreactors,obtaining a propagation rate of 43 shoots per explant with 1 min immersion and 6 h immersion frequencies.All new plants rooted and survived the transfer to soil.This study developed an in vitro propagation scheme to produce individuals that can be used either for reforestation,economical purposes,or to carry out studies in this species to assess its full potential,avoiding exploitation from wild plants.
基金Supported by the Innovation Platform Open Fund of Education Department in Hunan Province (18K100)。
文摘In this study, we employed a temporary immersion bioreactor(TIB)system for the micropropagation of strawberry seedlings. The TIB method and the conventional method for the micropropagation of strawberry seedlings were compared in terms of explant propagation coefficient, seedling fresh weight,contents of anthocyanins and chlorophyll, and photosynthetic characteristics. The results showed that an inoculation density of 40 explants/L was suitable for the micropropagation of ’Benihope’ strawberry seedlings in the TIB. The propagation coefficient, fresh weight, photosynthetic rate, stomatal conductance, transpiration coefficient, and the contents of total anthocyanins and chlorophyll of seedlings micropropagated in the TIB were significantly higher than those of the seedlings micropropagated by the conventional method. In conclusion, the TIB method was superior to the conventional method in the micropropagation of strawberry seedlings.The negative reciprocity did not occur between anthocyanins and chlorophyll in strawberry seedlings cultured under forced ventilation such as TIB.
基金the support provided by the National Institute of Forest Science(NIFoS)of the Republic of South Korea
文摘In Latin America the forestry of exotic species such as teak has been increasing in recent decades, due to their advantages in wood quality, rapid growth;and the relative ease of producing clones and their multiplication with respect to native species. Therefore, there is great interest in developing larger-scale propagation strategies that reduce costs and intensive manual labor. Culture in liquid media with temporary immersion and the semi-automation of the system has raised expectations for large-scale micropropagation. We report a protocol for teak, which reuses the primary explants in several culture cycles in semi-solid medium to produce nodal explants for the multiplication phase in temporary immersion bioreactors (RITA®). The control of factors such as cytokinin concentration, explants density, immersion frequencies and culture duration was analyzed. The number of shoots increased with 0.5 mg·l-1 of BA (6-Benzyladenine), alone or in combination with 0.5 mg·l-1 of Kinetin, with 2 daily immersions of 1 minute each;however, these shoots showed a high degree of hyperhydricity. When 0.05 mg·l-1 of BA was used with 1 immersion of 1 minute every 2 days, the hyperhydricity decreased. Although the number of shoots was lower, they showed good length to be used during multiplication and rooting ex vitro. Our results suggest that teak micropropagation can be simplified in two phases in vitro, the establishment and multiplication;followed by rooting ex vitro and acclimatization. This would imply a reduction in production costs, since most of the multiplication would take place in RITA®containers.
文摘A new procedure for blueberry (Vaccinium corymbosum L.) micropropagation in programmed Temporary Immersion Bioreactors (TIBs based on two separate bottles) was developed for the commercial genotypes Biloxi, Sharp Blue and Brillita. Plant cultures were developed in a controlled environment with 0.4 MPa CO2 enrichment, sucrose-reduced medium, and light intensity of 60 mM m-2·s-1. Principal component analysis showed that component 1 (C1) grouped 64.08% of the total variability, while the first two components accounted for 86.97%. Representation of the principal components demonstrated three clusters corresponding with the blueberry genotypes, and within each cluster plants micropropagated in agar-base medium grouped separately from those plants multiplied in TIBs. Both plant number and total internodes traits (related to the productive efficiency) were demonstrated superior in blueberries propagated in TIBs. Additionally, when transferred to greenhouse conditions, blueberries propagated in TIBs showed higher adaptability and growing rates than those cultured by the conventional approach, altogether evidencing the occurrence of a photomixotrophic stage in the vitroplantlets cultured in TIBs.
