scholarly journals Liquid Culture Systems Affect Morphological and Biochemical Parameters during Rosa canina Plantlets In Vitro Production

2018 ◽  
Vol 46 (1) ◽  
pp. 58-64 ◽  
Author(s):  
Małgorzata MALIK ◽  
Marzena WARCHOŁ ◽  
Bożena PAWŁOWSKA
Keyword(s):  
Solid Medium ◽  
Soluble Sugars ◽  
Dry Mass ◽  
Shoot Tips ◽  
Stem Explants ◽  
Biomass Growth ◽  
Mass Content ◽  
Temporary Immersion System ◽  
Culture Systems

Wild roses are an important group of plants used as decorative shrubs and cut flowers. They are also a row material for pharmaceutical, cosmetic and food industries. For rose in vitro propagation, solid media are commonly used. Up till now there is a few reports confirming the beneficial effect of liquid media on Rosa shoot growth and multiplication. The aim of the study was to investigate different culture systems, temporary immersion system (TIS) (immersion frequencies of 15 min every 6, 8 and 12 h), rotary shaker (RS) and stationary liquid (SL) for propagation of R. canina and compare with solid medium culture. Shoot tips and stem explants were grown on basic Murashige and Skoog medium with 20 mg dm-3 Fe EDDHA, 1 µM BA, 1.5 µM GA3 and 3% sucrose for six weeks. Liquid cultures stimulated biomass growth. The highest biomass growth in RS cultures were observed however, RS reduced the shoot dry mass content. TIS cultures immersed every 6 and 8 h increased dry mass content. In TIS and on solid medium shoot multiplication was 1.5-2 times better than in other systems and stem explants were more efficient. Solid medium improved the content of chlorophyll a, b, a+b and carotenoids. Higher contents of photosynthetic pigments were determined in shoot tips than stem explants. TIS-derived plantlets accumulated the largest amount of phenolic compounds. As the frequency of immersion increases the concentration of these compounds were reduced. In turn, SL cultures favored the accumulation of soluble sugars.

scholarly journals Micropropagation of Inula germanica L. from the Seedlings Explants

2018 ◽  
Vol 46 (1) ◽  
pp. 52-57 ◽  
Author(s):  
Alina TREJGELL ◽  
Monika KAMIŃSKA ◽  
Karolina LISOWSKA ◽  
Andrzej TRETYN
Keyword(s):  
Solid Medium ◽  
Positive Impact ◽  
Shoot Tips ◽  
Fluorescent Light ◽  
First Year ◽  
Naphthaleneacetic Acid ◽  
Multiplication Rate ◽  
Indolebutyric Acid ◽  
Regenerated Plants

This is the first communication of micropropagation system for Inula germanica using seedling explants germinated in vitro. The development of this system gives the possibility of future reintroduction of I. germanica providing a way to stabilize or re-establish its population. Shoot tips and fragments of cotyledons, hypocotyls and roots were isolated from ten-day-old seedlings. Explants were put on MS medium containing 1.0 mg l-1 benzylaminopurine and 0.1 mg l-1 naphthaleneacetic acid and cultured under continuous white fluorescent light (45 μmol.m-2.s-1) at 26 ± 1 °C. The highest percentage of shoot organogenesis (83.3%) was recorded for hypocotyl, while the highest average number of shoots per explant (12.0) was recorded for shoot tips. In subsequent subcultures, multiplication rate decreased to 3.0-4.9 shoots per explant. Less than 19% shoots were able to root on the solid medium without auxins. The highest rooting efficiency (69.3%) was recorded for solid medium supplemented with indolebutyric acid, but growth of roots was inhibited. The percentage of rooted shoots (62.2%) and number of roots per shoot (2.4 per shoot) into the liquid medium were comparable to medium with 0.1 mg·l-1 indolebutyric acid. showing a positive impact on the process of acclimatization. The regenerated plants were able to flowering in the first year after acclimatization. Developed micropropagation system for I. germanica is efficient and can be a useful tool for the active protection of this species.

scholarly journals Mikropropagasi planlet tebu menggunakan sistem perendaman sesaat (SPS) Micropropagation of sugarcane plantlets using temporary immersion system (TIS)

2016 ◽  
Vol 81 (1) ◽  
Author(s):  
Hayati MINARSIH ◽  
Imron RIYADI ◽  
. SUMARYONO ◽  
Asmini BUDIANI
Keyword(s):  
Solid Medium ◽  
Temporary Immersion System ◽  
Production Scale ◽  
Temporary Immersion ◽  
Biotechnology Research ◽  
Plantlet Production ◽  
Self Sufficiency ◽  
National Production ◽  
Planting Materials

bstractTo achieve Indonesian sugar self-sufficiency in2014, the national production needs to be escalatedthrough land extensification that requires a largenumbers of cane planting materials. This can be achievedby mass propagation of sugarcane through in vitroculture. Solid medium is commonly used for callusproliferation in sugarcane tissue culture. However, solidmedium is considered inefficient in terms of plantletproduction level, labour and space. The use of liquidmedium may solve the problem by allowing automationto increase plantlet production scale and uniformity.Temporary immersion system (TIS) is based on a shortperiodic immersion of explants in a liquid medium for aspecific frequency and duration. Research on in vitromass propagation of sugarcane using TIS was conductedat the Indonesian Biotechnology Research Institute forEstate Crops. Callus initiated from immature unfoldedleaves of PSJT 941 and PS 881 was cultured on liquidMS medium in TIS with different frequencies (12 and24 h) and durations (1 and 3 min) of immersion. Eachtreatment was replicated three times. The callus biomassof two elite cane varieties (PSJT 941 and PS 881)cultured in TIS for six weeks was higher (2 – 4 times fold)than that of on solid medium. The PSJT 941 varietyreached the highest calli biomass with immersion forthree min every 24 h. However, PS 881 variety reachedits highest biomass with immersion for one minute every24 h. The propagation of sugarcane using TIS culturewas proven to produce higher calli biomass up to fourfolds and to form more numbers and uniform shootscompared to the solid medium culture. The callus wassuccesfully regenerated to shoots and plantlets.AbstrakUntuk mencapai swasembada gula, perlu dilakukanpeningkatan produksi gula nasional melalui perluasanareal pertanaman tebu sehingga diperlukan bibit dalamjumlah besar. Hal tersebut dapat diatasi antara laindengan perbanyakan tebu melalui kultur in vitro. Peng-gunaan medium padat pada perbanyakan kalus tebumelalui kultur in vitro merupakan teknik yang umumdigunakan saat ini. Akan tetapi penggunaan mediumpadat dianggap kurang efisien dalam hal jumlah planletyang diproduksi, tenaga kerja dan ruang digunakan.Penggunaan medium cair dapat mengatasi kelemahantersebut dengan dimungkinkannya otomatisasi sehinggadapat meningkatkan skala produksi secara massal dankeseragaman planlet. Sistem perendaman sesaat (SPS)merupakan teknik kultur in vitro dalam medium cairmenggunakan bejana bersekat dimana kontak antaraeksplan dan medium terjadi hanya secara sesaat danperiodik. Penelitian perbanyakan massal bibit tebumelalui SPS dilakukan di Balai Penelitian BioteknologiPerkebunan Indonesia. Kalus diinisiasi dari daun meng-gulung varietas PSJT 941 dan PS 881 yang ditumbuhkanpada media MS cair dalam kultur SPS dengan frekuensiyang berbeda (12 dan 24 jam) dan lama perendaman (1dan 3 menit). Setiap perlakuan diulang tiga kali. Bobotbasah (biomassa) kalus dari dua varietas tebu (PSJT 941dan PS 881) yang ditumbuhkan dengan metode SPSsetelah enam minggu menunjukkan pening-katan yanglebih tinggi yaitu antara 2 - 4 kali lipat dibandingkandengan kontrol (media padat). Peningkatan biomassatertinggi pada varietas PSJT 941 diperoleh pada per-lakuan SPS dengan interval perendaman 24 jam dan lamaperendaman tiga menit. Sedangkan pada PS 881,peningkatan tertinggi biomassa diperoleh pada intervalperendaman 24 jam dan lama perendaman satu menit.Perbanyakan dengan metode SPS terbukti dapat mening-katkan biomassa kalus lebih dari empat kali lipat danpembentukan tunas yang lebih seragam dibandingkandengan pada media padat. Kalus yang dihasilkan dapatdiregenerasikan menjadi tunas dan planlet.

scholarly journals Monitoring the end of the in vitro phase of Anthurium andreanum Lindl. plantlets

2010 ◽  
Vol 22 (1) ◽  
pp. 61-68 ◽  
Author(s):  
Giulio Cesare Stancato ◽  
Maria Luiza Sant’Anna Tucci
Keyword(s):  
Culture Medium ◽  
Soluble Sugars ◽  
Dry Mass ◽  
Total Soluble Sugars ◽  
Gas Exchanges ◽  
Single Action ◽  
Fluorescent Lamps ◽  
Anthurium Andreanum

Estimulation of autotrophy in in vitro plantlets could be achieved through changes in the culture medium, or by changing the traditional hermetic caps by one that could allow gas exchanges between the culture and the environment. Besides that, the use of lamps with distinct emission spectrum irradiaction has propitiated successful results. This work was carried out aiming to evaluate the either the combined or the single action of some factors that can induce autotrophy on in vitro A. andraeanum cv. Eidibel plantlets. 3 sucrose concentrations were used: 0, 15 and 60 mM and for each one, to kinds of flasks according to the cap ventilation: under (0.038 L.h-1) and without ventilation. Flasks were kept under cold light fluorescent lamps or under gro-lux lamps. At the end of the experiment showing the highest shoot dry mass treatment was 60 mM, under ventilation and gro-lux, and the treatment which accumulate root dry mass to a lesser extent were 0 mM with ventilation and cold light and 15 mM without ventilation and cold light. In average, treatments with higher sucrose content in the culture medium, that is, 60 mM, under gro-lux lamps, presented the highest chlorophyll a, b and total contents, than those under cold lamp. Steps of carbohydrates metabolism could be associated with the total soluble sugars (sucrose and reducing sugars) levels, highlighting the steps where nutrient requirements were higher, showing the role of the plantlets sink.

scholarly journals Regeneration of plants from alginate-encapsulated shoots of Rhododendron dalhousiae Hook. F.

2011 ◽  
Vol 3 (1) ◽  
pp. 29-33 ◽  
Author(s):  
K. K. Singh ◽  
Bhusan Gurung
Keyword(s):  
Root Formation ◽  
Solid Medium ◽  
Shoot Proliferation ◽  
Shoot Tips ◽  
Nodal Segments ◽  
Synthetic Seed ◽  
Root Induction ◽  
Ms Medium ◽  
Direct Sowing

A method has been developed for plant regeneration from alginate-encapsulated nodal segments of Rhododendron dalhousiae. Shoot tips collected from in vitro proliferated shoots were used for synthetic seed production. For encapsulation, nodal segments were mixed with MS medium supplemented with 3% sodium alginate and incubated with calcium chloride (60 mM). The maximum frequency (69%) of conversion of encapsulated shoot tips into plantlets was achieved on MS medium containing 25 μM 2-isopentenyladenine (2iP) along with additive such as, 100 mg L-l polyvinyl pyrrolidone (PVP), 100 mg L -l ascorbic acid, 10 mg L-l citric acid. The presence of 2iP (25 μM) with IAA (0.6 μM) improved re-generation. Amongst the two gelling agents used higher shoot proliferation as well as better growth were observed in cultures grown on Agar in comparison to Phytagel medium. Encapsulated nodal segments stored at 4°C for 25 days also showed successful conversion, followed by development into complete plantlets when returned to regeneration medium. Liquid medium was superior over solid medium for root formation and growth. IBA (1.0 μM) was more effective than other auxins for root induction. Plantlets with developed shoot and roots were hardened off to survive ex vitro conditions and successfully established in greenhouse. Possibility of direct sowing of synthetic seeds in the soil was also examined.

scholarly journals In Vitro Regeneration of Capparis spinosa L. by Using a Temporary Immersion System

Plants ◽  
2019 ◽  
Vol 8 (6) ◽  
pp. 177 ◽  
Author(s):  
Valeria Gianguzzi ◽  
Paolo Inglese ◽  
Ettore Barone ◽  
Francesco Sottile
Keyword(s):  
Liquid Culture ◽  
Solid Medium ◽  
In Vitro Regeneration ◽  
Production Costs ◽  
Gelling Agent ◽  
Temporary Immersion System ◽  
Relative Growth ◽  
Temporary Immersion ◽  
Capparis Spinosa

Three caper (Capparis spinosa L.) biotypes grown on the Sicilian island of Salina (38°33′49″ N) were micropropagated to evaluate two different in vitro culture systems: one using the traditional solid medium, and the other based on liquid culture in a PlantForm bioreactor. PlantForm is a temporary immersion system (TIS), a new propagation method in which the shoots undergo temporary immersion in a liquid medium in order to avoid the accumulation of gas through forced ventilation. This study proposes a protocol to improve the efficiency of in vitro propagation of caper plants, while also reducing production costs, because of the elimination of the gelling agent, and manual labor, requiring limited subcultures and posing minimal contamination risks. Our results show that the caper shoots propagated in bioreactors demonstrated good adaptability and better growth rates than those grown in the conventional system. Statistically significant differences were observed between plants grown in the PlantForm liquid culture and those grown in solid medium regarding the number and length of shoots, which were further promoted by the addition of plant growth regulators (PGRs). The relative growth and real proliferation rate of the caper explants were higher when using meta-Topolin than when using 6-benzylaminopurine as a PGR. Overall, the TIS improved in vitro caper culture by promoting the proliferation, length, and vigor of the shoots.

scholarly journals The influence of different wavelengths of LED light on the production of glucosinolates and phenolic compounds and the antioxidant potential in in vitro cultures of Nasturtium officinale (watercress)

2021 ◽  
Author(s):  
Marta Klimek-Szczykutowicz ◽  
Barbara Prokopiuk ◽  
Kinga Dziurka ◽  
Bożena Pawłowska ◽  
Halina Ekiert ◽  
...  
Keyword(s):  
Phenolic Compounds ◽  
Blue Light ◽  
Photosynthetic Pigments ◽  
Soluble Sugars ◽  
Antioxidant Potential ◽  
Fluorescent Light ◽  
Biomass Growth ◽  
Led Light ◽  
Nasturtium Officinale

AbstractCultures of Nasturtium officinale were cultivated in vitro under illumination with different wavelengths of light-emitting diode (LED) light (white LED light—WLED, blue light—B, red light—R, 70% red and 30% blue light—RB, 50% green, 35% red and 15% blue light—RBG, 50% yellow, 35% red and 15% blue light—RBY, 50% far red, 35% red and 15% blue light—RBfR, 50% UV, 35% red and 15% blue light—RBUV, darkness—D), and under white fluorescent light (WF)—control conditions. The study investigated the influence of the applied lighting conditions on biomass growth and the production of glucosinolates, phenolic compounds, as well as photosynthetic pigments, and soluble sugars. The study showed a significant beneficial effect of the RBG light on biomass growth (Gi = 11.81 after 20 days) and the production of glucosinolates. The total glucosinolate content under these conditions increased 5.8 and 1.4 times in comparison with the WF light and D condition, respectively, reaching 237.92 mg 100 g−1 DW. The production of phenolic compounds, sugars, and photosynthetic pigments was comparable to the production under the control conditions. The antioxidant potential of extracts from the cultivated biomass was assessed by the CUPRAC, DPPH, and FRAP assays. Extracts obtained from the biomass of cultures grown under the RBG light had an antioxidant potential similar to that of the control cultures. This is the first report providing evidence of the stimulating effect of light quality on the biomass yield and production of glucosinolates by N. officinale microshoot cultures in vitro.

scholarly journals Mikropropagasi planlet tebu menggunakan sistem perendaman sesaat (SPS) Micropropagation of sugarcane plantlets using temporary immersion system (TIS)

2016 ◽  
Vol 81 (1) ◽  
Author(s):  
Hayati MINARSIH ◽  
Imron RIYADI ◽  
. SUMARYONO ◽  
Asmini BUDIANI
Keyword(s):  
Solid Medium ◽  
Temporary Immersion System ◽  
Production Scale ◽  
Temporary Immersion ◽  
Biotechnology Research ◽  
Plantlet Production ◽  
Self Sufficiency ◽  
National Production ◽  
Planting Materials

bstractTo achieve Indonesian sugar self-sufficiency in2014, the national production needs to be escalatedthrough land extensification that requires a largenumbers of cane planting materials. This can be achievedby mass propagation of sugarcane through in vitroculture. Solid medium is commonly used for callusproliferation in sugarcane tissue culture. However, solidmedium is considered inefficient in terms of plantletproduction level, labour and space. The use of liquidmedium may solve the problem by allowing automationto increase plantlet production scale and uniformity.Temporary immersion system (TIS) is based on a shortperiodic immersion of explants in a liquid medium for aspecific frequency and duration. Research on in vitromass propagation of sugarcane using TIS was conductedat the Indonesian Biotechnology Research Institute forEstate Crops. Callus initiated from immature unfoldedleaves of PSJT 941 and PS 881 was cultured on liquidMS medium in TIS with different frequencies (12 and24 h) and durations (1 and 3 min) of immersion. Eachtreatment was replicated three times. The callus biomassof two elite cane varieties (PSJT 941 and PS 881)cultured in TIS for six weeks was higher (2 – 4 times fold)than that of on solid medium. The PSJT 941 varietyreached the highest calli biomass with immersion forthree min every 24 h. However, PS 881 variety reachedits highest biomass with immersion for one minute every24 h. The propagation of sugarcane using TIS culturewas proven to produce higher calli biomass up to fourfolds and to form more numbers and uniform shootscompared to the solid medium culture. The callus wassuccesfully regenerated to shoots and plantlets.AbstrakUntuk mencapai swasembada gula, perlu dilakukanpeningkatan produksi gula nasional melalui perluasanareal pertanaman tebu sehingga diperlukan bibit dalamjumlah besar. Hal tersebut dapat diatasi antara laindengan perbanyakan tebu melalui kultur in vitro. Peng-gunaan medium padat pada perbanyakan kalus tebumelalui kultur in vitro merupakan teknik yang umumdigunakan saat ini. Akan tetapi penggunaan mediumpadat dianggap kurang efisien dalam hal jumlah planletyang diproduksi, tenaga kerja dan ruang digunakan.Penggunaan medium cair dapat mengatasi kelemahantersebut dengan dimungkinkannya otomatisasi sehinggadapat meningkatkan skala produksi secara massal dankeseragaman planlet. Sistem perendaman sesaat (SPS)merupakan teknik kultur in vitro dalam medium cairmenggunakan bejana bersekat dimana kontak antaraeksplan dan medium terjadi hanya secara sesaat danperiodik. Penelitian perbanyakan massal bibit tebumelalui SPS dilakukan di Balai Penelitian BioteknologiPerkebunan Indonesia. Kalus diinisiasi dari daun meng-gulung varietas PSJT 941 dan PS 881 yang ditumbuhkanpada media MS cair dalam kultur SPS dengan frekuensiyang berbeda (12 dan 24 jam) dan lama perendaman (1dan 3 menit). Setiap perlakuan diulang tiga kali. Bobotbasah (biomassa) kalus dari dua varietas tebu (PSJT 941dan PS 881) yang ditumbuhkan dengan metode SPSsetelah enam minggu menunjukkan pening-katan yanglebih tinggi yaitu antara 2 - 4 kali lipat dibandingkandengan kontrol (media padat). Peningkatan biomassatertinggi pada varietas PSJT 941 diperoleh pada per-lakuan SPS dengan interval perendaman 24 jam dan lamaperendaman tiga menit. Sedangkan pada PS 881,peningkatan tertinggi biomassa diperoleh pada intervalperendaman 24 jam dan lama perendaman satu menit.Perbanyakan dengan metode SPS terbukti dapat mening-katkan biomassa kalus lebih dari empat kali lipat danpembentukan tunas yang lebih seragam dibandingkandengan pada media padat. Kalus yang dihasilkan dapatdiregenerasikan menjadi tunas dan planlet.

scholarly journals Reproductive outcomes predicted by phase imaging with computational specificity of spermatozoon ultrastructure

2020 ◽  
Vol 117 (31) ◽  
pp. 18302-18309 ◽  
Author(s):  
Mikhail E. Kandel ◽  
Marcello Rubessa ◽  
Yuchen R. He ◽  
Sierra Schreiber ◽  
Sasha Meyers ◽  
...  
Keyword(s):  
Cell Viability ◽  
Contrast Agents ◽  
Assisted Reproductive Technologies ◽  
Reproductive Technologies ◽  
Dry Mass ◽  
Sperm Cells ◽  
Phase Imaging ◽  
Label Free ◽  
Mass Content

The ability to evaluate sperm at the microscopic level, at high-throughput, would be useful for assisted reproductive technologies (ARTs), as it can allow specific selection of sperm cells for in vitro fertilization (IVF). The tradeoff between intrinsic imaging and external contrast agents is particularly acute in reproductive medicine. The use of fluorescence labels has enabled new cell-sorting strategies and given new insights into developmental biology. Nevertheless, using extrinsic contrast agents is often too invasive for routine clinical operation. Raising questions about cell viability, especially for single-cell selection, clinicians prefer intrinsic contrast in the form of phase-contrast, differential-interference contrast, or Hoffman modulation contrast. While such instruments are nondestructive, the resulting image suffers from a lack of specificity. In this work, we provide a template to circumvent the tradeoff between cell viability and specificity by combining high-sensitivity phase imaging with deep learning. In order to introduce specificity to label-free images, we trained a deep-convolutional neural network to perform semantic segmentation on quantitative phase maps. This approach, a form of phase imaging with computational specificity (PICS), allowed us to efficiently analyze thousands of sperm cells and identify correlations between dry-mass content and artificial-reproduction outcomes. Specifically, we found that the dry-mass content ratios between the head, midpiece, and tail of the cells can predict the percentages of success for zygote cleavage and embryo blastocyst formation.

scholarly journals In Vitro Rooting Response of Yellow-Flowered Magnolia in Relation to the Phenolic Acids Content

Agronomy ◽  
2020 ◽  
Vol 10 (12) ◽  
pp. 1880
Author(s):  
Agnieszka Wojtania ◽  
Michał Dziurka ◽  
Edyta Skrzypek
Keyword(s):  
Chlorogenic Acid ◽  
Phenolic Acids ◽  
Late Phase ◽  
Soluble Sugars ◽  
Dry Mass ◽  
In Vitro Rooting ◽  
Total Phenolic ◽  
Naphthaleneacetic Acid ◽  
Indolebutyric Acid

The aim of this study was to analyze the profile of endogenous phenolic acids in yellow-flowered magnolias and to evaluate the effects of endogenous and exogenous phenolic acids on the in vitro rooting of three magnolia cultivars (‘Butterflies’, ‘Yellow Bird’, and ‘Elizabeth’). It has been shown that magnolia cultivars are phenolic acid-rich plants. Of the 16 phenolic acids tested, all were detected in each magnolia cultivar. The most abundant was gallic acid (max. 34,946 ng·mg−1 dry mass), followed by chlorogenic acid, ferulic acid, and caffeic acid. The amount of individual phenolic acids differed between the cultivars and media. The total phenolic production was enhanced by auxin, the main factor promoting rooting in magnolia in vitro. It has been found that the difficult-to-root ‘Butterflies’ and ‘Yellow Bird’ rooted better when they were grown on medium containing a mixture of auxins—3-indolebutyric acid (IBA) and 1-naphthaleneacetic acid (NAA)—as compared to IBA alone. The highest rooting frequency was observed for ‘Elizabeth’ (95.8%), followed by ‘Butterflies’ (46.1%) and ‘Yellow Bird’ (21.4%). In the case of ‘Yellow Bird’, the auxin treatment enhanced the leaf yellowing. The present work indicates a clear relationship between the overaccumulation of chlorogenic acid and coumaric acid in the late phase of rooting in vitro and the low rooting responses of magnolia ‘Butterflies’ and ‘Yellow Bird’. On the other hand, ‘Elizabeth’ produced more soluble sugars by 29.2% than easy-to-root ones. The biochemical status of the plantlets can influence their further ex vitro establishment, which was the highest for ‘Elizabeth’ (97.5%), followed by ‘Butterflies’ (49.9%) and ‘Yellow Bird’ (24.6%).

scholarly journals Manipulation of Low Temperature and Light Quality for Storage of Broccoli in Vitro

HortScience ◽  
1997 ◽  
Vol 32 (3) ◽  
pp. 471A-471
Author(s):  
Sandra B. Wilson ◽  
Keiko Iwabuchi ◽  
Nihal C. Rajapakse ◽  
Roy E. Young
Keyword(s):  
Blue Light ◽  
Carbon Balance ◽  
Light Quality ◽  
Storage Systems ◽  
Stem Elongation ◽  
Soluble Sugars ◽  
Dry Mass ◽  
Photon Flux ◽  
Zero Carbon

Storage systems for tissue-cultured plants offer versatility in managing labor to meet market availability. Storage systems that minimize growth and yet sustain photosynthetic and regrowth potential require temperature, light quality, and light intensity to be manipulated for plantlet quality during and after storage. Broccoli (Brassica oleracea L. Botrytis Group `Green Duke') plantlets were cultured photoautotrophically (without sugar) or photomixotrophically (with sugar) on cellulose plugs in liquid medium in vitro for 3 weeks at 23°C and 150 μmol·m–2·s–1 photosynthetic photon flux (PPF). To determine the conditions that yield a zero carbon balance, plantlets were subsequently stored for 3 days under different temperatures (1°C, 5°C, 10°C, 15°C), different light intensities (1.6 PPF, 4.1 PPF, 8.6 PPF), and different light spectra (white, blue, red). Plantlets stored under 5 PPF and 5°C maintained a zero carbon balance. Subsequently, plantlets were stored for 4, 8, or 12 weeks at 5°C under darkness or 5 PPF of white, red or blue light. Stem elongation was observed for plantlets stored under blue light. Plantlets stored under red light were characterized by increased chlorophyll, increased specific leaf mass (leaf dry mass per unit leaf area, SLM), increased starch in leaf tissue, and increased total soluble sugars in leaf and stem tissue. Plantlets grown with sucrose were characterized by increased dry mass, regardless of light treatment. After 8 weeks, plantlets grown with or without sucrose and stored in darkness did not survive acclimatization to greenhouse.

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