Temporary Immersion Bioreactor System Research Articles

Bulbil initiation: a comprehensive review on resources, development, and utilisation, with emphasis on molecular mechanisms, advanced technologies, and future prospects.

Bulbil is an important asexual reproductive structure of bulbil plants. It mainly grows in leaf axils, leaf forks, tubers and the upper and near ground ends of flower stems of plants. They play a significant role in the reproduction of numerous herbaceous plant species by serving as agents of plant propagation, energy reserves, and survival mechanisms in adverse environmental conditions. Despite extensive research on bulbil-plants regarding their resources, development mechanisms, and utilisation, a comprehensive review of bulbil is lacking, hindering progress in exploiting bulbil resources. This paper provides a systematic overview of bulbil research, including bulbil-plant resources, identification of development stages and maturity of bulbils, cellular and molecular mechanisms of bulbil development, factors influencing bulbil development, gene research related to bulbil development, multi-bulbil phenomenon and its significance, medicinal value of bulbils, breeding value of bulbils, and the application of plant tissue culture technology in bulbil production. The application value of the Temporary Immersion Bioreactor System (TIBS) and Terahertz (THz) in bulbil breeding is also discussed, offering a comprehensive blueprint for further bulbil resource development. Additionally, additive, seven areas that require attention are proposed: (1) Utilization of modern network technologies, such as plant recognition apps or websites, to collect and identify bulbous plant resources efficiently and extensively; (2) Further research on cell and tissue structures that influence bulb cell development; (3) Investigation of the network regulatory relationship between genes, proteins, metabolites, and epigenetics in bulbil development; (4) Exploration of the potential utilization value of multiple sprouts, including medicinal, ecological, and horticultural applications; (5) Innovation and optimization of the plant tissue culture system for bulbils; (6) Comprehensive application research of TIBS for large-scale expansion of bulbil production; (7) To find out the common share genetics between bulbils and flowers.

Enhancing dendrobine production in Dendrobium nobile through mono-culturing of endophytic fungi, Trichoderma longibrachiatum (MD33) in a temporary immersion bioreactor system.

Dendrobine, a valuable alkaloid found in Dendrobium nobile, possesses significant pharmaceutical potential. In this study, we explored innovative approaches to enhance dendrobine production by utilizing endophytic fungi in a Temporary Immersion Bioreactor System (TIBS, Nanjing BioFunction Co. Ltd., China) and traditional test bottles. Dendrobine was unequivocally identified and characterised in D. nobile co-culture seedlings through UHPLC analysis and LC-MS qTOF analysis, supported by reference standards. The CGTB (control group) and EGTB (experimental group) 12-month-old D. nobile seedlings exhibited similar peak retention times at 7.6±0.1 minutes, with dendrobine identified as C16H25NO2 (molecular weight 264.195). The EGTB, co-cultured with Trichoderma longibrachiatum (MD33), displayed a 2.6-fold dendrobine increase (1804.23 ng/ml) compared to the CGTB (685.95 ng/ml). Furthermore, a bioanalytical approach was applied to investigate the mono-culture of T. longibrachiatum MD33 with or without D. nobile seedlings in test bottles. The newly developed UHPLC-MS method allowed for dendrobine identification at a retention time of 7.6±0.1 minutes for control and 7.6±0.1 minutes for co-culture. Additionally, we explored TIBS to enhance dendrobine production. Co-culturing D. nobile seedlings with Trichoderma longibrachiatum (MD33) in the TIBS system led to a substantial 9.7-fold dendrobine increase (4415.77 ng/ml) compared to the control (454.01 ng/ml) after just 7 days. The comparative analysis of dendrobine concentration between EGTB and EGTIBS highlighted the remarkable potential of TIBS for optimizing dendrobine production. Future research may focus on scaling up the TIBS approach for commercial dendrobine production and investigating the underlying mechanisms for enhanced dendrobine biosynthesis in D. nobile. The structural elucidation of dendrobine was achieved through 1H and 13C NMR spectroscopy, revealing a complex array of proton environments and distinct carbon environments, providing essential insights for the comprehensive characterization of the compound. These findings hold promise for pharmaceutical and industrial applications of dendrobine and underline the role of endophytic fungi in enhancing secondary metabolite production in medicinal plants.

Improving biomass and dendrobine-type total alkaloids (DTTAs) production of Dendrobium nobile through combining Temporary Immersion Bioreactor System (TIBS) with endophyte MD33 elicitation

Improving biomass and dendrobine-type total alkaloids (DTTAs) production of Dendrobium nobile through combining Temporary Immersion Bioreactor System (TIBS) with endophyte MD33 elicitation

Micropropagation of iraca palm (Carludovica palmata Ruiz y Pav) using a temporary immersion system

Carludovica palmata is a Neotropical plant with a promising potential for the agroindustry. It is an important source of fibers used to manufacture handmade goods. Colombia is one of the leading countries in the production of this species; however, it lacks efficient techniques for its propagation. We developed a protocol for massive micropropagation of C. palmata using a temporary immersion bioreactor (TIB) system. Immersion frequency, immersion time, culture medium volume, and explant density were evaluated using a split-split-plot design. The variables evaluated were the number and length of shoots and roots, the number of leaves, and the dry weight of shoots. The performance of three micropropagation systems, TIB, semi-solid medium, and liquid medium, was evaluated using a generalized randomized block design. Murashige and Skoog (MS) culture medium with 1.0 mg L−1 of 6-benzylaminopurine, 0.5 mg L−1 of 1-naphthaleneacetic acid, and 20 g L−1 of sucrose added to it was used. The results obtained show a frequency of 12 h with an immersion time of 1 min improved the length of shoots, roots, and number of leaves, and dry weight. A volume of 20 mL/explant was found to be an adequate condition to increase the number and length of shoots, the number of leaves, and the dry weight. According to the optimization module, the factor levels that will maximize the evaluated variables are a 1-minute immersion of the tissues in the culture medium with a frequency of 12 hours each time, with 175 mL of culture medium per bioreactor and density of 10 explants. In conclusion, TIB was shown to be efficient for massive micropropagation of C. palmata compared with conventional methods.

An Efficient Method of Pennisetum × advena ‘Rubrum’ Plantlets Production Using the Temporary Immersion Bioreactor Systems and Agar Cultures

The aim of this study is to develop an efficient method for micropropagation of Pennisetum × advena 'Rubrum'. Agar cultures containing Murashige and Skoog (MS) medium supplemented with 6-benzyl-amino-purine (BAP) in various concentrations (0.5 mg/L to 2 mg/L) and a temporary immersion bioreactor system (TIS) using liquid medium MS with an addition of 1 mg/L BAP were tested. Rooting was performed using ½ MS medium supplemented with different auxin combinations (indole-3-butyric acid IBA and α-naphthalene acetic acid NAA) and activated charcoal. The TIS method was found to be the most efficient, producing 36.9 new plants within four weeks. The resulting plantlets were thin and bright green in color, with no signs of hyperhydricity. The most suitable agar medium yielded 19.5 new plants within eight weeks. For rooting, ½ MS supplemented with 0.5 mg/L IBA and 0.5 mg/L NAA exhibited an 84% rooting rate, whereas the addition of activated charcoal inhibited rooting.

Constant and Intermittent Contact with the Volatile Organic Compounds of Serendipita indica Alleviate Salt Stress In Vitro Ocimum basilicum L.

Serendipita indica is a plant growth-promoting fungus. It is a natural soil dweller that can colonize the roots of a wide range of plants, including cultivated crops. S. indica has been reported to improve plant nutrient uptake and increase stress tolerance when inoculated into the soil. The present study was undertaken to study the effect of volatile organic compounds (VOCs) of S. indica on salt-stressed Ocimum basilicum ‘Fin vert’ in vitro, either in a culture vessel with a semi-solid medium or via a modified temporary immersion bioreactor system (SETIS). For all salt concentrations, VOCs of S. indica significantly improved plant growth in both semi-solid medium and SETIS bioreactors. This resulted in heavier and taller plants, more shoots per plant, and longer roots. This was even observed for the control without salt. At 9 g/L NaCl, plants with Serendipita were able to give longer roots than those without (1.2 cm vs. 0.0 and 1.7 cm vs. 1.7 cm) in the semi-solid medium and SETIS, respectively. Nevertheless, the VOCs were not able to make the plant salt tolerant to this high concentration. The increase in total phenolic and flavonoid content and radical scavenging suggest that the antioxidant defense system is triggered by the S. indica VOCs. In the semi-solid system, without VOCs, 1 g/L NaCl led to an increase in total chlorophyll content (TCC) and a significant decrease in TCC was further measured only at 6 g/L NaCl or more. However, when VOCs were added, the bleaching effect of the salt was partially restored, even at 6 and 9 g/L NaCl. A significant decrease in TCC was also measured in the SETIS system at 6 g/L NaCl or more and treatment with VOC did not make any difference. An exception was 9 g/L, where the VOC-treated plants produced more than three times more chlorophyll than the non-treated plants. These findings will encourage the application of Serendipita indica for stress reduction. In addition, the proposed original adaptation of a temporary immersion system will be instrumental to investigate stress reduction associated with volatile compounds and better understand their mechanism of action.

Establishment of Direct Organogenesis Protocol for Arachis hypogaea cv. Virginia in Liquid Medium by Temporary Immersion System (TIS)

Peanuts (Arachis hypogaea L.) are a rich source of herbal oil, proteins, minerals, vitamins, fibers, essential amino acids, as well as bioactive compounds, and are thus widely used for human nutrition and animal feed, and for prevention from certain diseases. However, the in vitro regeneration response of the species is generally low, and it also displays a significant variability among its varieties. Thus, the development of advanced protocols and approaches for the in vitro propagation of peanut is still of immense importance. A recently developed in vitro propagation technique, TIS; Temporary Immersion Bioreactor System, provides a new approach for the mass propagation of plants. Accordingly, the present study provides an efficient de novo regeneration protocol for Arachis hypogaea L. cv. Virginia by using a TIS. Different concentrations of cytokinins, i.e., benzyladenine (BA) or thidiazuron (TDZ), were tested with several combinations of dry and medium immersion periods of TIS, corresponding to a total of 8, 12, 16, 24, 32, 36, 48, 64, 72, and 96 min daily immersions for the induction of direct organogenesis. The study exhibited that an MS medium added to 110 µM BA or 10 µM TDZ are the most appropriate medium formulations in TIS, when applied for 16 min every 16 h. The application of optimized procedures to cv. NC7 and two valuable Turkish autochthonous varieties, 7 × 77 and Com74, is also reported. To the best of our knowledge, the present study draws attention also for being the first study in which a TIS was used for peanuts.

Stipagrostis pennata (Trin.) De Winter Artificial Seed Production and Seedlings Multiplication in Temporary Immersion Bioreactors

This study was conducted to develop the protocol for artificial seed production of Stipagrostis pennata (Trin.) De Winter via somatic embryo encapsulation as well as test a temporary bioreactor system for germination and seedling growth. Embryogenic calli were encapsulated using sodium alginate and calcium chloride and then sowed in the Murashige and Skoog (MS) germination medium in in vitro cultures. The experiments were conducted as a factorial based on a completely randomized design with three replications. The treatments include three concentrations of sodium alginate (1.5%, 2.5%, and 3.5%), two ion exchange times (20 and 30 min), and two artificial seed germination media (hormone-free MS and MS supplemented with zeatin riboside and L-proline). Germination percentage and number of days needed until the beginning of germination were studied. The highest percentage of artificial seed germination was obtained when 2.5% sodium alginate was used for 30 min (ion exchange time) and when the seeds were placed on the MS germination medium supplemented with zeatin riboside and L-proline. The results of the analysis of variance in the temporary immersion bioreactor system showed that the main effects observed on the seedling growth were associated with different growth hormones in culture media and the number of feeding cycles. Experimental results also indicated that the total protein analyses of zygotic seedlings and seedlings originating from the synthetic seeds showed no statistically significant differences between these samples.

In Vitro Multiplication of Eucalyptus pilularis and Eucalyptus grandis x E. urophylla (Urograndis Eucalypt): Effect of Light Quality in Temporary Immersion Bioreactor

Light quality is an important factor for in vitro development of explants in a bioreactor system. Based on the need to optimize, this study aimed to evaluate the in vitro multiplication of Eucalyptus pilularis and urograndis eucalypt using different light quality in a temporary immersion bioreactor (TIB) system. Different spectral qualities on in vitro multiplication were evaluated using three light sources, namely fluorescent lamp (white light), red and blue. Shoot length, number of buds, fresh weight per explant, vigor and hyperhydricity were evaluated according to an established scoring scale at 30 days. The results showed that fluorescent white light was the most appropriate for use in the in vitro multiplication of E. pilularis, and blue light for the urograndis eucalypt clone resulting in a greater shoot length (1.46 cm; 2.41 cm), number of buds (2.25; 10.20), vigor (1.8; 1.3) and fresh weight per explant (86.9 mg; 449 mg). The results can be applied to optimize clonal microplant production on a commercial scale.

Temporary Immersion Bioreactor System as an Efficient Method for Mass Production of In Vitro Plants in Horticulture and Medicinal Plants

A temporary immersion system (TIS) bioreactor has been used as an efficient and cost-effective method for the in vitro propagation of many plant species. In the current study, the applicability of a TIS bioreactor for plantlet regeneration Chrysanthemum morifolium Ramat., Fragaria × ananassa Duch., and Cnidium officinale Makino was studied. Shoot length, a number of leaves per regenerated shoot, fresh, and dry biomass of plantlets were optimal with the TIS compared to semi-solid and liquid immersion cultures. The leaf area in cryshanthmum, strawberry, and C. afficinale were 2.87 cm2, 3.51 cm2, and 1.43 cm2, respectively, in the plants regenerated by TIS. The photosynthetic pigments were highest in strawberry plants grown in TIS bioreactor culture, and there was no significant difference between semi-solid and liquid culture while the highest values were obtained in C. officinale maintained in semi-solid culture. The chrysanthemum and strawberry plants showed a 100% acclimatization rate in all culture systems. C. officinale plants showed the highest survival rate at 96.9%, which were regenerated in the TIS. TIS bioreactor culture, thus, provides a convenient method that could be adopted for commercial in vitro propagation of chrysanthemum, strawberry and C. officinale plants.

Improving large-scale biomass and total alkaloid production of Dendrobium nobile Lindl. using a temporary immersion bioreactor system and MeJA elicitation

BackgroundDendrobium nobile Lindl. is an important pharmacopeial plant with medicinal and ornamental value. This study sought to provide a technical means for the large-scale production of total alkaloid in D. nobile. Seedlings were cultured in vitro using a temporary immersion bioreactor system (TIBS). The four tested immersion frequencies (min/h; 5/2, 5/4, 5/6, and 5/8) influenced the production of biomass and total alkaloid content. In addition, to compare the effects of different concentrations of the phytohormone methyl jasmonate (MeJA) and treatment time on biomass and total alkaloid accumulation, MeJA was added to the TIBS medium after 50 days. Finally, total alkaloid production in semi-solid system (SSS), TIBS, and TIBS combined with the MeJA system (TIBS-MeJA) were compared.ResultsThe best immersion frequency was found to be 5/6 (5 min every 6 h), which ensured appropriate levels of biomass and total alkaloid content in plantlets. The alkaloid content and production level of seedlings were the highest after treatment with 10 μM MeJA separately for 20 and 30 days using TIBS. The maximum content (7.41 mg/g DW) and production level (361.24 mg/L) of total alkaloid on use of TIBS-MeJA were 2.32- and 4.69-fold, respectively, higher in terms of content, and 2.07- and 10.49-fold, respectively, higher in terms of production level than those on using of TIBS (3.20 mg/g DW, 174.34 mg/L) and SSS (1.58 mg/g DW, 34.44 mg/L).ConclusionsOur results show TIBS-MeJA is suitable for large-scale production of total alkaloid in in vitro seedlings. Therefore, this study provides a technical means for the large-scale production of total alkaloid in D. nobile.

Design and development of temporary immersion bioreactor system controlled by microcontroller

This research aims to develop, design and construct a temporary immersion bioreactor (TIB) controlled by microcontroller. The designed TIB can control the time for plant feeding and the carbon dioxide (CO2) concentration via application. The TIB consists of 2 sets of the plant tissue culture containers. The TIB can define the feeding and the CO2 concentration control times up to 10 and 4 time periods per day, respectively. The system feeds the plants and controls the CO2 concentration during the defined time periods. The test results of the feeding time control illustrated that the TIB could set the feeding times conveniently and quickly. Moreover, the system could properly work following the set times. For the test results of the CO2 concentration control, it was found that the TIB could control the CO2 concentration in the containers during the set operating time. It took about 7, 28, 30, 36 and 21 minutes after the system started working to adjust the CO2 concentration in order to be at the set levels of <750, 1,500, 2,000, 2,500 and 3,000 ppm, respectively. The CO2 concentration control system could properly work with error less than 10%.

Temporary immersion bioreactor system for propagation by somatic embryogenesis of hybrid larch (Larix × eurolepis Henry)

Somatic embryogenesis (SE) has high potential for large-scale clonal propagation of conifers. Different types of bioreactor cultures have been tested for the conifer SE process where the temporary immersion bioreactors (TIBs) have proved to be useful across the different developmental steps of the SE process. In the present study the use of TIBs was tested for hybrid larch (Larix × eurolepis Henry). The results showed two-fold increases in both fresh weight (FW) of pro-embryogenic masses (PEMs) and yield of cotyledonary embryos in the TIBs compared to solid medium in plates. For the germination phase, the highest number of roots per plant, the root length and height of plants were also obtained in the TIBs. The results show that the TIB system can be successfully used to support scale up of plant production in all steps of the SE process from proliferation to germination of hybrid larch (Larix × eurolepis Henry).

A novel temporary immersion bioreactor system for large scale multiplication of banana (Rasthali AAB\u2014Silk)

Musa sp. cultivar Rasthali (Silk AAB) is a choice variety of the Asian sub-continent. Its production and sustenance are threatened by Fusarium wilt, which affects the livelihoods of small and marginal farmers. The use of quality planting material is one of the strategies to manage the disease. Availability of quality planting material for varieties other than Grand Naine is limited. Large-scale micropropagation using existing technologies is laborious and expensive. Temporary immersion bioreactor system is emerging as a potential advancement in the micropropagation industry. In this study, a cost-effective temporary immersion bioreactor (TIB) system has been developed and an efficient micropropagation method has been standardized. Explants cultured in TIB with 250 ml of culture medium in a 2-min immersion frequency of 6 h were found to be efficient for shoot proliferation and rooting. Its efficacy has been compared with the semisolid culture method. At the end of the 6th subculture, 1496 ± 110 shoots per explant were obtained in TIB. Chlorophyll, carotenoid, stomatal index, and the number of closed stomata were examined to determine the physiological functions of the plants grown in TIB and compared with semisolid grown plantlets. Plantlets grown in TIB were genetically stable and were confirmed using inter-simple sequence repeat (ISSR) markers. The multiplication of shoots in TIB was 2.7-fold higher than the semisolid culture method, which is suitable for large-scale production of planting material for commercial applications.

Development of an in vitro Microtuberization and Temporary Immersion Bioreactor System to Evaluate Heat Stress Tolerance in Potatoes (Solanum tuberosum L.).

High temperature (heat) stress reduces tuber yield and quality of potatoes. Screening potatoes for heat tolerance is increasingly important, considering the climate change scenario and expansion of potatoes to countries where heat stress is an issue. In vitro screening for tolerance to abiotic stresses offers several advantages, including quick evaluation of numerous genotypes (clones) in reduced space, controlled environmental conditions (temperature and photoperiod), and free from confounding variables inherent to greenhouse and field conditions. In this study, we explored the feasibility of using a temporary immersion bioreactor system for heat tolerance screening of potatoes. We determined the best hormone-free microtuberizing media for this system (MSG with 8% sucrose) to enhance microtuber number and size. Comparisons of microtubers produced at 30°C as heat treatment, with 16°C as normal condition, allowed to identify heat tolerant and susceptible potato clones. The use of bioreactors allowed distinguishing well-formed (non-deformed) from deformed microtubers. Heat stress increased the total biomass of plant tissues in all the clones. However, the effect of heat stress on microtuber number and weight varied among the clones. Incubation at 30°C decreased the weight and number of non-deformed microtubers in all the clones except for Reveille Russet in which the weight of non-deformed microtubers was significantly increased and the count of non-deformed microtubers was not affected. The potato variety Reveille Russet, which was selected under high-temperature field conditions in Texas, had many non-deformed microtubers per explant and the highest microtuber weight among four clones evaluated under heat stress. We described a faster and reliable in vitro microtuberization system for abiotic stress tolerance screening, identified Reveille Russet as a promising heat-tolerant potato variety, and confirmed Russet Burbank and Atlantic as susceptible heat-tolerant checks.

Micropropagation of banana (Musa spp) using temporary immersion bioreactor system

Banana is an important crop in the tropics which possess the potential for commercial production in Nigeria. Large scale production requires large volume of planting materials which may be difficult to obtain using conventional methods of propagation. Temporary immersion bioreactor system (TIBs) is a cost effective method for micropropagation of plants. The present study was carried out to develop an efficient method for rapid multiplication of banana using temporary immersion bioreactor system (TIBs). Banana microshoots were regenerated from young suckers obtained from field grown plants using conventional plant tissue culture. Microshoots of 2cm length were used as explants for multiplication in temporary immersion bioreactor system. Ten (10) explants were cultured in bioreactor bottles containing Murashinge and Skoog (MS) liquid media supplemented with different concentrations of 6-bezylaminopurine (BAP) with or without 250mg/L Activated Charcoal (AC). Results showed that explants cultured in media supplemented with 2 mg/L or 1mg/L BAP without AC gave the highest shoot multiplication rate of 900% and 800%, respectively compared to hormone free media. Production of competent plants (plants ready for ex vitro establisment) were however, influenced by the presence of AC and the highest percentage of competent plants (80%) were produced when media was fortified with 1mg/L BAP+ 250mg AC. Regenerated plants were successfully established in the field and were morphologically normal and fertile.

Micropropagation of pineapple (Ananas comosus L. Var. Smooth cayenne) in temporary immersion bioreactor system (TIPS)

Pineapple is an important edible fruit in the family Bromeliaceae popularly grown in the tropical and subtropical countries. Commercial prodution of pineapple requires large volume of planting materials which could not easily be obtained using conventional method of propagation. A protocol for mass propagation of pineapple (Ananas comosus L. var.smooth cayenne) using temporary immersion bioreactor system has been developed.The protocol involves four immersion cycles in Murashinge and Skoog (MS) media fortified with 1mg/L or 2mg/L 6-Benzylaminopurine (BAP) with or without 0.25g/L activated charcoal (AC). The highest multiplication rate (120 -130 plants/bottle) was obtained when media was fortified with 1mg/l or 2mg/L BAP alone. The presence of activated charcoal (AC) promoted root morphogenesis, resulting in significant increase in roots formation in BAP suplemented media. A combination of BAP with AC significantly increased the number of competent plants(20 – 30 plants/bottle) after four weeks of culture in temporary immersion system. The system is recommended for rapid and efficient micropropagation of pineapple.

In-vitro development of Nauclea diderrichii (de Willd. &amp; Th. Dur) Merrin liquid-M Smedia supplemented with benzyl amino purine (BAP) and naphthalene acetic acid (NAA)

The growth of plantlets in Temporary Immersion Bioreactor system (TIBs) relies on initial successful liquid phase transition process. The response of N. diderrichii explants was assessed in liquid-M Smedia with a view to mass produce its seedlings using TIBs. Seven treatments consisting (A) 0.0/0.0, (B) 0.0/0.1, (C) 0.1/0.0, (D) 0.2/0.1, (E) 0.3/0.2, (F) 0.4/0.3 and (G) 0.5/0.4mg/lBAP/NAA combinations were studied. Each group consist of seven replicates and group A without Growth Regulators (GR) serves as control. The results at 4 Weeks after Inoculation (WAI) showed that effects of the growth regulators were significant on shoot length and number of adventitious shoots while number of roots and leaves were closely related. Treatment E produced highest number of adventitious shoots (3.6) which was higher than 0.9 shoots from treatment G and closely related to others. Maximum number of leaves (16.6) was produced by treatment F followed by E (15.7) while the least (12) was obtained in treatment A. The highest number of roots (4.9) was obtained from treatments B, followed by E (4.3) with the lowest being recorded in C (2.43). Liquid MS medium supplemented with 0.3/0.2mg/lBAP/NAA shows some promise for plantlets generation for the purpose of multiple shoot production of N. diderrichii in TIBs.

Influence of silicon and in vitro culture systems on the micropropagation and acclimatization of “Dwarf Cavendish” banana

In vitro culture systems based on liquid culture media are considered to be more effective than semisolid culture medium systems. Liquid culture media systems provide better nutrient availability for plant tissues, easier culture handling, and the potential for scaling up and automation. However, in vitro liquid culture requires more careful handling due to the potential for contamination and the possibility of negative effects, such as hyperhydricity or vitrification, that hinder the growth and development of the plant material. Temporary immersion bioreactors have emerged as a workable alternative for capturing the benefits of liquid media, though semisolid systems are still traditional. Many studies have shown that silicon (Si) is a beneficial plant nutrient. Silicon might have a positive effect in both semisolid and liquid in vitro systems. The objective of this study was to evaluate the effect of silicon on the micropropagation and acclimatization of banana plants cultivated in vitro by comparing liquid temporary immersion bioreactor technology and semisolid traditional culture systems. Different silicon concentrations (0 and 1 mL L-1) and culture systems (liquid temporary immersion bioreactor and semisolid traditional culture) were evaluated over a 36-day period. The growth characteristics plant size, fresh and dry weight, and number and length of leaves and roots were evaluated. After the 36-day in vitro growth period, plants were transferred to a greenhouse for acclimatization and were evaluated after 30 days for the same growth characteristics used in the in vitro studies. The temporary immersion bioreactor system resulted in greater growth of banana plants compared to the traditional semisolid system. Temporary immersion bioreactors also showed a positive interaction with Si and resulted in higher values for all growth characteristics in the acclimatization phase.

In vitro propagation of three Iranian apricot cultivars

In vitro propagation is a useful method for clonal propagation of apricot; however, it is a highly genotype-dependent procedure. Eight different experiments were conducted to optimize and establish an efficient in vitro propagation protocol for three Iranian apricot cultivars, including Ordubad, Shams, and Qaysi. Sterilization, in vitro establishment, proliferation, root induction, and acclimatization steps were assessed. The fungal and bacterial infections were significantly decreased when nanosilver (2.5%) was applied in sterilization of initial explants. The highest number of active buds was obtained from summer-season collected lateral bud explants. The establishment responses of apricot cultivars to investigated basal culture media were different and the best results for Ordubad, Shams, and Qaysi cultivars were obtained from Driver and Kuniyuki (DKW), Woody Plant Medium (WPM), and modified Quoirin and Lepoivre (QL) basal media, respectively. The highest number of induced lateral shoots in Ordubad cultivar (2.33) was obtained from basal QL medium supplemented with BAP (4.44 μmol L−1), GA3 (1.44 μmol L−1), and IBA (0.098 μmol L−1). In Shams and Qaysi cultivars, the highest numbers of induced lateral shoots were obtained from WPM basal medium. The best combinations of BAP and GA3 hormones to increase the number of induced lateral shoots in three Iranian apricot cultivars were 11.56 μmol L−1 GA3 and 4.44 μmol L−1 BAP. In comparison with routine solid culture system, a 10 min h−1 temporary immersion bioreactor system led to the significant increase of the number of induced lateral shoots, with higher shoot quality, in all investigated cultivars. The half strength QL medium supplemented with 19.68 μmol L−1 of IBA resulted in the highest rooting percentage in all investigated apricot cultivars. The results of the present study would be applicable for cost-effective large-scale clonal propagation of other valuable cultivars of apricot.