Using LED lighting in somatic embryogenesis and micropropagation of an elite sugarcane variety and its effect on redox metabolism during acclimatization

This study investigated the ability of a light emitting diode (LED) to induce somatic embryogenesis (SE), shoot multiplication, and rooting of sugarcane (RB98710). We also accessed the effects on acclimatization. MS medium was used for all stages and was supplemented with different concentrations of growth regulators according to the culture stage. The material was maintained in a growth room under fluorescent (FL) or LED (82?% red, 18?% blue) lighting after rooting plants were acclimatized. We conducted both biometric and biochemical analyses before and after acclimatization. The LED conditions favored the formation of callus; however, the FL was more efficient at plant regeneration. A histological analysis showed the formation of somatic embryos occurred through direct and indirect pathways. The plants obtained through SE and grown under LED had a higher multiplication rate over six subcultures. Shoots rooted in both light sources, but the number of shoots and the weight gain of the roots were higher under LED. The malondialdehyde (MDA) level did not differ among treatments. Our results indicate the SE induction phase should be conducted under FL and the remaining micropropagation process should be performed using LED. After acclimatization the plants grown under LED did not change the SOD and CAT activities during the first 5 days, which suggests there was no acclimatization impact. The H₂O₂ and MDA values observed do not suggest damage to membranes. There was better development, lower water loss, and higher survival rate in plants from in vitro culture under LED conditions when compared to FL.     

Experimentally comparing the attractiveness of domestic lights to insects: Do LEDs attract fewer insects than conventional light types?

LED lighting is predicted to constitute 70% of the outdoor and residential lighting markets by 2020. While the use of LEDs promotes energy and cost savings relative to traditional lighting technologies, little is known about the effects these broad‐spectrum “white” lights will have on wildlife, human health, animal welfare, and disease transmission. We conducted field experiments to compare the relative attractiveness of four commercially available “domestic” lights, one traditional (tungsten filament) and three modern (compact fluorescent, “cool‐white” LED and “warm‐white” LED), to aerial insects, particularly Diptera. We found that LEDs attracted significantly fewer insects than other light sources, but found no significant difference in attraction between the “cool‐” and “warm‐white” LEDs. Fewer flies were attracted to LEDs than alternate light sources, including fewer Culicoides midges (Diptera: Ceratopogonidae). Use of LEDs has the potential to mitigate disturbances to wildlife and occurrences of insect‐borne diseases relative to competing lighting technologies. However, we discuss the risks associated with broad‐spectrum lighting and net increases in lighting resulting from reduced costs of LED technology.     

Conserving energy at a cost to biodiversity? Impacts of LED lighting on bats

Artificial lighting is a key biodiversity threat and produces 1900 million tonnes of CO ₂ emissions globally, more than three times that produced by aviation. The need to meet climate change targets has led to a global increase in energy‐efficient light sources such as high‐brightness light‐emitting diodes (LEDs). Despite the energetic benefits of LEDs, their ecological impacts have not been tested. Using an experimental approach, we show that LED street lights caused a reduction in activity of slow‐flying bats ( Rhinolophus hipposideros and Myotis spp.). Both R. hipposideros and Myotis spp. activities were significantly reduced even during low light levels of 3.6 lux. There was no effect of LED lighting on the relatively fast‐flying Pipistrellus pipistrellus, Pipistrellus pygmaeus and Nyctalus/Eptesicus spp. We provide the first evidence of the effects of LED lights on bats. Despite having considerable energy‐saving benefits, LED lights can potentially fragment commuting routes for bats with associated negative conservation consequences. Our results add to the growing evidence of negative impacts of lighting on a wide range of taxa. We highlight the complexities involved in simultaneously meeting targets for reduction of greenhouse gas emissions and biodiversity loss. New lighting strategies should integrate climate change targets with the cultural, social and ecological impacts of emerging lighting technologies.     

LED light for in vitro and ex vitro efficient growth of economically important highbush blueberry (<Emphasis Type="Italic">Vaccinium corymbosum</Emphasis> L.)

Light-emitting diodes (LEDs) are useful for the growth of many plants, but not known for blueberry species. This study examined the effects of fluorescent lamps and 100 % red, 80 % red plus 20 % blue, 50 % red plus 50 % blue, and 100 % blue LEDs on the growth and development of highbush blueberry shoots under aseptic and non-aseptic conditions. Results revealed that monochromatic blue LEDs accumulated the highest contents of leaf chlorophylls. In contrast, monochromatic red LEDs inhibited chlorophyll accumulation, but produced the longest shoots and roots and provided high percentages of side shoot formation from ex vitro plants. Mixed LEDs, particularly 50 % red plus 50 % blue light, improved plant growth with respect to notably increased shoot and root biomass. Direct rooting of in vitro shoots under non-aseptic conditions was readily achieved using a commercial mixture of perlite and peat moss with high humidity controls. These findings obviously suggest the efficient use of LEDs to replace traditional fluorescent lamps in large-scale propagation of the highbush blueberry, and also pave the way for future studies on LEDs for standardizing micropropagation protocols to shrub crops and woody plants.     

不同光源对柚木组培苗生长发育的影响

该研究采用不同光源处理柚木组培继代苗,通过测定株高、叶宽、叶鲜重、全株干重、叶绿素含量和观察叶片上下表皮结构,研究不同光源对柚木组培苗生长发育的影响,并筛选适宜其快速生长发育的光环境。结果表明:不同光照下顶芽培养的植株高生长顺序为H_(FL)﹥H_(1RB)﹥H_S﹥H_(2RB)﹥H_(3RB)﹥H_D,茎段培养的腋芽高生长顺序为h_(FL)﹥h_S﹥h_D﹥h_(1RB)﹥h_(2RB)﹥h_(3RB)。LED红蓝组合灯(2RB、3RB)处理下植株最大叶宽显著大于荧光灯(FL)、全光谱灯(S)和无补光设备(D)处理,并随RB光强增加,最大叶宽及叶鲜重显著增加。全株干重以D和S光照处理显著低于其它光照处理。RB系列和S光照处理下的叶绿素总含量高于对照FL和D处理组。2RB和3RB光照处理下叶片的上表皮细胞不规则多角形,相互镶嵌排列,其它光照处理下上表皮细胞呈圆形。下表皮气孔数量在有补光设备处理(S、RB、FL)时是无补光设备处理(D)的1.5倍以上,2RB和3RB光照处理下气孔张开度大于对照FL和D处理组,且保卫细胞呈井型突起。参试光源中,柚木组培苗增殖阶段选择荧光灯或全光谱灯为好,高生长显著,腋芽分化和生长快,有利于提高增殖率;而RB红蓝组合灯(3RB、2RB)适合壮苗培养,叶大苗壮,全株生物量大,且叶表面结构发育成熟,有利于提高光合作用。     

Long‐term effects of artificial nighttime lighting and trophic complexity on plant biomass and foliar carbon and nitrogen in a grassland community

The introduction of artificial nighttime lighting due to human settlements and transport networks is increasingly altering the timing, intensity, and spectra of natural light regimes worldwide. Much of the research on the impacts of nighttime light pollution on organisms has focused on animal species. Little is known about the impacts of daylength extension due to outdoor lighting technologies on wild plant communities, despite the fact that plant growth and development are under photoperiodic control. In a five‐year field experiment, artificial ecosystems (“mesocosms”) of grassland communities both alone or in combination with invertebrate herbivores and predators were exposed to light treatments that simulated street lighting technologies (low‐pressure sodium, and light‐emitting diode [LED]‐based white lighting), at ground‐level illuminance. Most of the plant species in the mesocosms did not exhibit changes in biomass accumulation after 5 years of exposure to the light treatments. However, the white LED treatment had a significant negative effect on biomass production in the herbaceous species Lotus pedunculatus. Likewise, the interaction between the white LED treatment and the presence of herbivores significantly reduced the mean shoot/root ratio of the grass species Holcus lanatus. Artificial nighttime lighting had no effect on the foliar carbon or nitrogen in most of the grassland species. Nevertheless, the white LED treatment significantly increased the leaf nitrogen content in Lotus corniculatus in the presence of herbivores. Long‐term exposure to artificial light at night had no general effects on plant biomass responses in experimental grassland communities. However, species‐specific and negative effects of cool white LED lighting at ground‐level illuminance on biomass production and allocation in mixed plant communities are suggested by our findings. Further studies on the impacts of light pollution on biomass accumulation in plant communities are required as these effects could be mediated by different factors, including herbivory, competition, and soil nutrient availability.     

Growth of Tsuru-rindo (Tripterospermum japonicum) culturedin vitro under various sources of light-emitting diode (LED) irradiation

We studied the effects of light generated by LEDs on the growth of Tsururindo (Tripterospermum japonicum) shoots. Apical shoots (2–3 cm long) were cultured on MS basal media supplemented with 3% sucrose, and were maintained for four weeks under five different light qualities: F (fluorescent lamp), red LED (R), 70% red + 30% blue LED (R7B3), 50% red + 50% blue (R5B5), or blue LED (B). Rooting was promoted by red light (100%) but was inhibited by blue light. Plant growth, as defined by root number, fresh weight, and chlorophyll content, was generally healthier for cultures irradiated with mixed LEDs, and was the best under R7B3. Ventilation resulted in more rapid apical shoot growth and rooting compared with control plants, when both were treated with the R7B3 system. We demonstrated here that plant growth can be controlled by using LEDs to adjust for the most effective irradiation conditions, compared with the performance observed when conventional fluorescent lamps are utilized.     

Growth and photosynthesis of Chinese cabbage plants grown under light-emitting diode-based light source

We compared growth and the content of sugar, protein, and photosynthetic pigments, as well as chlorophyll fluorescence parameters in 15- and 27-day-old Chinese cabbage (Brassica chinensis L.) plants grown under a high-pressure sodium (HPS) lamps or a light source built on the basis of red (650 nm) and blue (470 nm) light-emitting diodes (LEDs) with a red to blue photon ratio of 7: 1. One group of plants was grown at a photosynthetic photon flux (PPF) level of 391 ± 24 μ mol/(m² s) (normal level); the other, at a PPF level of 107 ± 9 μ mol/(m² s) (low light). Plants of the third group were firstly grown at the low light and then (on the 12th day) transferred to the normal level. When grown at the normal PPF level, the plants grown under LEDs didn’t differ from plants grown under HPS lamps in shoot fresh weight, but they showed a lower root fresh and dry weights and the lower content of total sugar and sugar reserves in the leaves. No differences in the pigment content and photosystem II quantum yield were found; however, a higher Chl a/b ratio in plants grown under LEDs indicates a different proportion of functional complexes in thylakoid membranes. The response to low light conditions was mostly the same in plants grown under HPS lamps and LEDs; however, LED plants showed a lower growth rate and a higher nonphotochemical fluorescence quenching. In the case of the altered PPF level during growth, the plant photosynthetic apparatus adapted to new conditions of illumination within three days. Plants grown under HPS lamps at a constant normal PPF level and those transferred to the normal PPF level on the 12th day, on the 27th day didn’t differ in shoot fresh weight, but in plants grown under LEDs, the differences were considerable. Our results show that LED-based light sources can be used for plant growing. At the same time, some specific properties of plant photosynthesis and growth under these conditions of illumination were found.     

The effect of light quality on the growth and development of in vitro cultured Doritaenopsis plants

The influence of light quality on growth and development of in vitro grown Doritaenopsis hort. (Orchidaceae) plants was investigated. Growth parameters like leaf and root fresh/dry mass and leaf area were highest with plants grown under red plus blue light emitting diodes (LEDs). Leaf length was greater with the plants grown under red LED. Carbohydrate (starch, sucrose, glucose and fructose) and leaf pigment (chlorophylls and carotenoids) biosynthesis of the plants was significantly increased in plants grown under red plus blue LEDs compared to red or blue LED and fluorescent light treatments. This study suggests that the production of quality Doritaenopsis plants is possible by culturing the plants in vitro under a mixture of blue plus red light sources.     

Effects of Red and Blue LED Light on the Growth and Photosynthesis of Barley (Hordeum vulgare L.) Seedlings

According to the action spectrum of photosynthesis, photosynthetic efficiency is highest for red light. However, long-term growth with only red light leads to unfavorable changes in plant morphology, decrease in photosynthetic capacity and plant productivity. Detailed mechanisms behind these changes are still poorly understood. We studied the effects of narrow-band red (RL) and blue (BL) LED lighting on the morphology and photosynthesis of barley (Hordeum vulgare L.) seedlings at 9 days old, when energy for plant growth comes mostly from the endosperm, and light has a mainly morphogenic effect on plant growth. Plants grown with white fluorescent lamps (WL) were used as a control. At this developmental stage, light spectrum had small but significant effects on most morphometric parameters, which may become more prominent as the plant grows. These effects were more pronounced in RL-grown plants and were similar to the ‘shade-avoidance response’, which is unusual as in nature it occurs when the fraction of red light in the spectrum is low. RL-grown plants also had impaired photosynthetic photochemical efficiency (as assessed by PAM-fluorometry and leaf absorption). BL-grown plants had a stronger similarity to control plants in their morphology and photosynthetic characteristics than RL-grown plants; however, they had higher NPQ and different NPQ induction kinetics than WL- and RL-grown plants. Our results suggest that photoregulation of plant morphology and photosynthesis evolutionarily adapted to natural light is miscoordinated in narrow-band LED light. We discuss possible reasons for this miscoordination and for the formation of observed phenotypes on the level of photoreceptors.

     

Field performance of transgenic potato plants compared with controls regenerated from tuber discs and shoot cuttings

Summary The objective of this study was to separate and determine effects on the field performance of transgenic potatoes that originate from the tissue culture process of transformation and from the genes inserted. The constructs introduced contained the reporter gene for betaglucuronidase (GUS) under the control of the patatin promoter (four different constructs) and the neomycin phosphotransferase gene under the control of the nopaline synthase promoter. Both genes might be expected to have a neutral effect on plant phenotype. The field performance of transgenic plants (70 independent transformants) was compared with non-transgenic plants regenerated from tuber discs by adventitious shoot formation and from shoot cultures established from tuber nodal cuttings. Plants from all three treatments were grown in a field trial from previously field-grown tubers, and plant performance was measured in terms of plant height at flowering, weight of tubers, number of tubers, weight of large tubers and number of large tubers. There was evidence of somaclonal variation among the transgenic plants; mean values for all characters were significantly lower and variances generally higher than from plants derived from nodal shoot cultures. A similar change in means and variances was observed for the non-transgenic tuber-disc regenerants when compared with shoot culture plants. Plant height, tuber weight and tuber number were, however, significantly lower in transgenic plants than in tuber-disc regenerants, suggesting an effect on plant performance either of the tissue culture process used for transformation or of the genes inserted. There were significant differences between constructs for all five plant characters. The construct with the smallest segment of patatin promoter and the lowest level of tuber specificity for GUS expression had the lowest values for all five characters. It is proposed that the nature of GUS expression is influencing plant performance. There was no indication that the NPTII gene, used widely in plant transformation, has any substantial effect on plant performance in the field.     

Impact of light-emitting diode irradiation on photosynthesis,phytochemical composition and mineral element content of lettuce cv. Grizzly

Light-emitting diodes (LEDs) are a promising technology with a potential to improve the irradiance efficiency, light quality, and the light spectrum for increasing plant yield and quality. In this experiment, we investigated the impacts of various LED light qualities, including 100% red, 100% blue, 70% red + 30% blue, and 100% white, on the growth and photosynthesis, phytochemical contents, and mineral element concentrations in lettuce (Lactuca sativa L. cv. ‘Grizzly’) in comparison to normal greenhouse conditions. Photon flux of 300 µmol m^?2 s^?1 was provided for 14 h by 120 LEDs set on a 60 cm × 60 cm sheet of aluminum platform in the growth chambers, where plants were grown for 60 d. Fresh mass per plant was significantly higher when grown under 100% blue and 70% red + 30% blue LEDs compared to the other environments including greenhouse conditions. Phytochemical concentrations and a nutritive value of lettuce were also significantly affected by the light treatments. Chlorophyll and carotenoid concentrations increased in the plants grown under 70% red + 30% blue LEDs compared to those grown in the greenhouse. Vitamin C content was 2.25-fold higher in the plants grown under 100% blue LEDs compared to those grown in the greenhouse. Higher photosynthesis and maximal quantum yield of PSII photochemistry were also observed in the plants treated with LED lights. The application of LED light led to the elevated concentrations of macro-and micronutrients in lettuce possibly because of the direct effect of LED light and lower stress conditions in the growth chambers compared to the greenhouse. Although the mechanism of the changes in lettuce grown under LED is not well understood, the results of this study demonstrated that LED light could be used to enhance the growth and nutritional value of lettuce in indoor plant production facilities.     

二十世纪我国植物学家对植物组织培养的贡献

回顾了上一世纪我国植物组织培养的发展。 1934年以来 ,我国的植物组织培养研究一直与国际发展同步进行。我国学者在离体器官发生、茎尖培养、花药培养、子房培养、胚乳培养、原生质体培养和细胞大量培养等分支领域都取得重要进展。本文在引证我国研究者发表的植物组织培养论文的基础上 ,着重评述了那些被国际同行公认的研究成果。此外 ,还介绍了植物组织培养在我国农业和工业上应用的情况     

Role of AUX1 in the control of organ identity during in vitro organogenesis and in mediating tissue specific auxin and cytokinin interaction in <Emphasis Type="Italic">Arabidopsis</Emphasis>

Classic plant tissue culture experiments have shown that exposure of cell culture to a high auxin to cytokinin ratio promotes root formation and a low auxin to cytokinin ratio leads to shoot regeneration. It has been widely accepted that auxin and cytokinin play an antagonistic role in the control of organ identities during organogenesis in vitro. Since the auxin level is highly elevated in the shoot meristem tissues, it is unclear how a low auxin to cytokinin ratio promotes the regeneration of shoots. To identify genes mediating the cytokinin and auxin interaction during organogenesis in vitro, three allelic mutants that display root instead of shoot regeneration in response to a low auxin to cytokinin ratio are identified using a forward genetic approach in Arabidopsis. Molecular characterization shows that the mutations disrupt the AUX1 gene, which has been reported to regulate auxin influx in plants. Meanwhile, we find that cytokinin substantially stimulates auxin accumulation and redistribution in calli and some specific tissues of Arabidopsis seedlings. In the aux1 mutants, the cytokinin regulated auxin accumulation and redistribution is substantially reduced in both calli and specific tissues of young seedlings. Our results suggest that auxin elevation and other changes stimulated by cytokinin, instead of low auxin or exogenous auxin directly applied, is essential for shoot regeneration. Electronic supplementary material  The online version of this article (doi:) contains supplementary material, which is available to authorized users.     

二十世纪我国植物学家对植物组织培养的贡献

回顾了上一世纪我国植物组织培养的发展.1934年以来,我国的植物组织培养研究一直与国际发展同步进行.我国学者在离体器官发生、茎尖培养、花药培养、子房培养、胚乳培养、原生质体培养和细胞大量培养等分支领域都取得重要进展.本文在引证我国研究者发表的植物组织培养论文的基础上,着重评述了那些被国际同行公认的研究成果.此外,还介绍了植物组织培养在我国农业和工业上应用的情况.     

Cloning of medicinal plants through tissue culture--a review

In order to have standardized formulations, the chemical constituents from plants and their parts are required to be uniform both qualitatively and quantitatively. Furthermore, an ever increasing demand of uniform medicinal plants based medicines warrants their mass cloning through plant tissue culture strategy. A good number of medicinal plants have been reported to regenerate in vitro from their various parts, but a critical evaluation of such reports reveals that only a few complete medicinal plants have been regenerated and still fewer have actually been grown in soil, while their micropropagation on a mass scale has rarely been achieved, particularly in those medicinal plants where conventional propagation is inadequate, like, the mass clonal propagation of Dioscorea floribunda leading to its successful field trials. Such facts make it imperative to document the factual position of micropropagation of medicinal plants bringing out the advancements made along with the short falls, in this important area. The present review deals with the futuristic view on the said subject restricted to higher plants.     

Review: role of carbon sources for in vitro plant growth and development

In vitro plant cells, tissues and organ cultures are not fully autotrophic establishing a need for carbohydrates in culture media to maintain the osmotic potential, as well as to serve as energy and carbon sources for developmental processes including shoot proliferation, root induction as well as emission, embryogenesis and organogenesis, which are highly energy demanding developmental processes in plant biology. A variety of carbon sources (both reducing and non-reducing) are used in culture media depending upon genotypes and specific stages of growth. However, sucrose is most widely used as a major transport-sugar in the phloem sap of many plants. In micropropagation systems, morphogenetic potential of plant tissues can greatly be manipulated by varying type and concentration of carbon sources. The present article reviews the past and current findings on carbon sources and their sustainable utilization for in vitro plant tissue culture to achieve better growth rate and development.     

Developmental and hormonal regulation of direct shoot organogenesis and somatic embryogenesis in sugarcane (Saccharum spp. interspecific hybrids) leaf culture

Rapid and efficient in vitro regeneration methods that minimise somaclonal variation are critical for the genetic transformation and mass propagation of commercial varieties. Using a transverse thin cell layer culture system, we have identified some of the developmental and physiological constraints that limit high-frequency regeneration in sugarcane leaf tissue. Tissue polarity and consequently the orientation of the explant in culture, size and developmental phase of explant, and auxin concentration play a significant role in determining the organogenic potential of leaf tissue in culture. Both adventitious shoot production and somatic embryogenesis occurred on the proximal cut surface of the explant, and a regeneration gradient, decreasing gradually from the basal to the distal end, exists in the leaf roll. Importantly, auxin, when added to the culture medium, reduced this spatial developmental constraint, as well as the effect of genotype on plant regeneration. Transverse sections (1-2 mm thick) obtained from young leaf spindle rolls and orienting explants with its distal end facing the medium (directly in contact with medium) are critical for maximum regeneration. Shoot regeneration was observed as early as 3 weeks on MS medium supplemented with alpha-naphthalenencetic acid (NAA) and 6-benzyladenine, while somatic embryogenesis or both adventitious shoot organogenesis and somatic embryogenesis occurred on medium with NAA and chlorophenoxyacetic acid. Twenty shoots or more could be generated from a single transverse section explant. These shoots regenerated roots and successfully established after transplanted to pots. Large numbers of plantlets can be regenerated directly and rapidly using this system. SmartSett, the registered name for this process and the plants produced, will have significant practical applications for the mass propagation of new cultivars and in genetic modification programs. The SmartSett system has already been used commercially to produce substantial numbers of plants of orange rust-resistant and new cultivars in Australia.     

LED light mediates phenolic accumulation and enhances antioxidant activity in Melissa officinalis L. under drought stress condition

The popularity of lemon balm in conventional medicine is suggested by the existence of two groups of compounds, namely essential oil and phenylpropanoids pathway derivatives. One of the promising approaches to improve tolerance to drought stress induced oxidative damage in seedlings grown in greenhouses and plant growth chambers is replacing the traditional and high-cost light technologies by recently developed light emitting diodes (LED). In this experiment, we analyzed the role of various LED lights including red (R), blue (B), red (70%) + blue (30%) (RB), and white (W) as well as normal greenhouse light (as control) to stimulate defense mechanisms against drought stress in two genotypes of Melissa officinalis L. The present study demonstrates that pre-treatment with LEDs with high-intensity output for 4 weeks alleviated harmful effects of drought stress in the two genotypes. Under drought stress, RB LED pre-treated plantlets of the two genotypes exhibited the highest relative growth index of shoot and root and total phenolic and anthocyanin content compared to those irradiated with other LEDs and greenhouse lights. The highest amount of malondialdehyde level was detected in R LED exposed plants. In response to drought, LED-exposed plants especially RB light-irradiated plants of the two genotypes maintained significantly higher antioxidant and phenylalanine ammonia-lyase (PAL) enzyme activities and higher expression level of the PAL1 and 4CL-1 genes compared to those irradiated with greenhouse light. We concluded that RB LED light provides a better growth condition and resistance to drought stress for the two genotypes of lemon balm by the highest antioxidant activity and the least amount of damage to the cell membranes. Our data suggest that LED light pre-treatments as moderate stress activate antioxidant systems, enhance the scavenging of ROS and induce drought stress tolerance in the two genotypes of lemon balm plants.