Adventitious root cultures of Castilleja tenuiflora Benth. as a source of phenylethanoid glycosides

Castilleja tenuiflora is a highly valued medicinal plant that grows in pine-oak woods in Mexico. In this study, we identified for the first time verbascoside and isoverbascoside as the major phenylethanoid glycosides (PhGs) in C. tenuiflora. These compounds have proven biological activities, including anti-inflammatory, antioxidant, and cytotoxic activities, which may be related to the traditional uses of this plant. We developed a reverse-phase high-performance liquid chromatography (RP-HPLC) procedure to analyze PhGs, and determined their concentrations in various different tissues of wild plants. Verbascoside accumulated mainly in roots and inflorescences (9.23 and 7.88 mg g⁻¹ dry biomass, respectively), while isoverbascoside accumulated mainly in the roots (7.13 mg g⁻¹ dry biomass). To provide an alternative source of material for production of bioactive compounds, we established in vitro adventitious root cultures in which roots were grown in B5 medium containing either 10 μM indole 3-acetic acid (IAA) or 10 μM α-naphthaleneacetic acid (NAA). The greatest dry biomass yield (30 g L⁻¹) was achieved at 30 days after transfer of roots into IAA-containing medium. The highest specific yields of PhGs were also obtained using this auxin; the maximum level of verbascoside was 14.62 mg g⁻¹ dry root biomass (438.6 mg L⁻¹) at 30 days after root transfer, and the maximum yield of isoverbascoside was 37.32 mg g⁻¹ dry root biomass (522.48 mg L⁻¹) at 23 days after root transfer. Adventitious root cultures of C. tenuiflora are a promising system for further studies on scale-up and phenylethanoid glycosides biosynthesis.     

Chitosan enhances withanolides production in adventitious root cultures of Withania somnifera (L.) Dunal

Calli were obtained from leaf, cotyledon and internode explants of in vitro-grown plants of Indian cultivar of Withania somnifera in MS medium supplemented with 2, 4-D (2.0 mg l⁻¹) and Kinetin (0.2 mg l⁻¹). The brown, semi-friable callus (500 mg FW) derived from leaf explants produced higher number of primary adventitious roots (9 roots/callus) in half strength MS medium fortified with IBA (0.5 mg l⁻¹) and NAA (0.1 mg l⁻¹). The primary adventitious roots with an inoculum mass of 15 g FW were cultured for 6 weeks in the same medium for secondary adventitious root proliferation. Elicitation of abiotic elicitor, aluminium chloride at 10 mg l⁻¹ at the end of 4 weeks culture with 4 h exposure time enhanced withanolides productivity. Under similar culture conditions, the biotic elicitor, chitosan at 100 mg l⁻¹ stimulated higher production of all withanolides when compared to aluminium chloride treatment. This is the first report on the use of callus-derived adventitious root culture for the enhanced production of withanolides upon chitosan elicitation.     

Synthesis of withanolide A depends on carbon source and medium pH in hairy root cultures of Withania somnifera

Withania somnifera (L.) Dunal. (Indian ginseng) is an important medicinal plant which yields pharmaceutically active compounds called withanolides. The present work deals with optimization of parameters of hairy root culture of W. somnifera for the production of biomass and withanolide A. We also investigated the effects of carbon source [sucrose, glucose, fructose, maltose, glucose + fructose (1:1), fructose + sucrose (1:1) and sucrose + glucose (1:1)], sucrose concentration (1%, 2%, 3%, 4%, 6% and 8%) and the initial medium pH (4.0, 4.5, 5.0, 5.5, 5.8, 6.0 and 6.5) on growth and production of withanolide A in hairy root cultures of W. somnifera. We found that biomass accumulation and production of withanolide A was highest when sucrose was used as the carbon source (11.92 g l⁻¹ DW and 11.96 mg g⁻¹ DW of withanolide A). Further 3% sucrose concentration was found to be optimal for biomass accumulation (11.92 g l⁻¹ DW) and 4% sucrose favoured the production of withanolide A (13.28 mg g⁻¹ DW) in the tested range of concentrations (1-8%). The biomass of hairy roots was optimal when the initial medium pH was 5.8 (12.1 g l⁻¹ DW) and the withanolide A production was highest in the medium pH set at 6.0 (13.84 mg g⁻¹ DW).     

Regulation of tissue differentiation by plant growth regulators on tTCLs of Panax ginseng adventitious roots

Differentiated tissue in Panax ginseng cultures was found to be very efficacious for saponin production. In order to increase the yield of saponins and preserve culture stability we were testing different plant growth regulators (PGR) and auxin/cytokinin combinations to regulate a level of tissue differentiation. For this purpose we used transverse thin cell layers (tTCLs) of adventitious roots of Panax ginseng. Adventitious roots were cultivated in Shenk and Hildebrand (SH) liquid medium supplemented with IBA (24.6 μM). Callus formation and root multiplication of adventitious root tTCLs was evaluated after 4 and following 12 weeks of cultivation, respectively, on SH basal medium containing various auxins (3 mg l⁻¹) or cytokinins (0.2 or 0.02 mg l⁻¹) or their combinations. We found that kinetin (Kin) in combination with auxin benzo[b]selenienyl acetic acid (BSAA), naphthalene acetic acid or indole-3-butric acidis the best for biomass production and following root multiplication. These combinations were tested in previously selected most suitable large-scale system—a temporary immersion system RITA. The best saponin production (15.94 ± 1.89 mg g⁻¹ dry weight) and growth value (5.62 ± 0.34) was reached on medium containing BSAA and Kin combination.     

Role of exogenous phytohormones on growth and plumbagin accumulation in Plumbago indica hairy roots and conservation of elite root clones via synthetic seeds

The present study describes the role of different exogenous hormones on morphology and plumbagin production in Plumbago indica hairy roots. It was also aimed to conserve elite root clones via synthetic seed technology. Insertion of rolB gene in transformed roots was confirmed by polymerase chain reaction followed by southern blot analysis. Hairy roots were treated with single or in combination of different phytohormones viz. IAA, IBA, 2, 4-D, NAA, BAP, GA₃ and ABA. Cultures incubated with GA₃ (0.5 mg l⁻¹) yielded highest root growth due to formation of profuse lateral branching while NAA (0.5 mg l⁻¹) treatment caused highest plumbagin accumulation. Cultures incubated with 2, 4-D exhibited highest inhibitory effect in terms of both root growth and plumbagin production. All phytohormones were found to be effective at lower concentration. In combinatorial study, GA₃ + NAA (0.5 mg l⁻¹, each) was found optimum for root biomass and plumbagin production at earlier stage of culture. Different combinations of auxins and BAP induced different morphologies ranging from reduction of lateral branching to rapid disorganization of root matrix. The combinations of ABA and selected auxins were not found promising at any of selected concentration. Based on the effect of exogenous hormones on hairy root culture, elite root clones were selected and encapsulated with sodium alginate matrix. Uniform shaped alginate coated synthetic seeds were conserved up to 6 months exhibited high regeneration potential without disturbing plumbagin content.     

Growth and accumulation of caffeic acid derivatives in Echinacea angustifolia DC. var. angustifolia grown in hydroponic culture

In separate experiments conducted in 2007 and 2008, growth and accumulation of selected caffeic acid derivatives (CADs; i.e., caftaric acid, chlorogenic acid, echinacoside, caffeic acid, cynarin, p-coumaric acid, ferulic acid and cichoric acid) were determined in Echinacea angustifolia DC. var. angustifolia seedlings grown in hydroponic culture (floating raft system) at a density of 122 plant m⁻² (at planting). Plants were harvested 11 (2007) or 16 (2008) weeks after transplanting (i.e., 15 or 20 weeks after sowing). In both years, plants grew vigorously and at harvest approximately half of the plants under observation had developed one to three inflorescences. In 2008, the root yield (2940 kg ha⁻¹) harvested in nearly eight months from two consecutive hydroponic cultures was within the yield reported in the literature for field cultivations lasting two to four years. None of the selected CADs was found in the leaves, while the inflorescences (stem and capitulum) contained only caftaric acid and echinacoside at concentrations higher than the detection limits (0.05 mg g⁻¹ dry weight). Echinacoside, cynarin and chlorogenic acid were found in root tissues at concentrations ranging from 0.36 to 5.25 mg g⁻¹ dry weight. In all plant samples, echinacoside, which is the marker compound for E. angustifolia material, did not reach the minimum quality standard (10 mg g⁻¹ dry weight) for the production of standardized extract. We concluded that short-cycle, high-density greenhouse hydroponic culture stimulates plant growth and root production in E. angustifolia, but it does not ensure sufficient CADs accumulation in dried roots.     

Large scale in vitro propagation of Stevia rebaudiana (bert) for commercial application: Pharmaceutically important and antidiabetic medicinal herb

Stevia rebaudiana is a valuable medicinal plant species and it is being used for the treatment of diabetes. Currently, there is a high demand for raw material of this medicinal herb due to ever increasing diabetes disorder among the population. In order to meet the increased demand an efficient in vitro propagation of S. rebaudiana was established. Nodal explants collected from the field were cultured on MS basal medium fortified with different concentrations of BAP (0.5-3.0 mg/l) and KIN (0.5-3.0 mg/l) individually for shoot bud induction. In vitro derived nodal buds were cultured on MS medium supplemented with different concentrations (0.5-3.0 mg/l) of BAP and KIN for multiple shoot bud regeneration. In the second experiment, in vitro derived buds were placed on MS medium supplemented with different concentrations of BAP (0.5-3.0 mg/l) in combination with 0.5 mg/l IAA or IBA or NAA for shoot bud multiplication. The highest frequency (94.50%) of multiple shoot regeneration with maximum number of shoots (15.69 shoots/explant) was noticed on MS medium supplemented with 1.0 mg/l BAP. For large scale plant production, in vitro derived nodal bud explants were cultured on MS medium fortified with 1.0 mg/l BAP, in which about 123 shoots/explant were obtained after three subcultures on the same media composition. Elongated shoots (>2 cm) dissected out from the in vitro proliferated shoot clumps were cultured on half-strength MS medium containing different concentrations of NAA (0.1-0.5 mg/l) and/or MS medium fortified with various concentrations (0.5-2.0 mg/l) of auxins (NAA, IAA and IBA) for root induction. Highest frequency of rooting (96%) was noticed on half-strength MS medium augmented with 0.4 mg/l NAA. The rooted plantlets were successfully transferred into plastic cups containing sand and soil in the ratio of 1:2 and subsequently established in the greenhouse. The present in vitro propagation protocol would facilitate an alternative method for rapid and large-scale production of this important antidiabetic medicinal plant.     

Year-Round Cultivation of Tetraselmis sp. for Essential Lipid Production in a Semi-Open Raceway System

There is increasing demand for essential fatty acids (EFAs) from non-fish sources such as microalgae, which are considered a renewable and sustainable biomass. The open raceway system (ORS) is an affordable system for microalgae biomass cultivation for industrial applications. However, seasonal variations in weather can affect biomass productivity and the quality of microalgal biomass. The aim of this study was to determine the feasibility of year-round Tetraselmis sp. cultivation in a semi-ORS in Korea for biomass and bioactive lipid production. To maximize biomass productivity of Tetraselmis sp., f medium was selected because it resulted in a significantly higher biomass productivity (1.64 ± 0.03 g/L) and lower omega-6/omega-3 ratio (0.52/1) under laboratory conditions than f/2 medium (0.70/1). Then, we used climatic data-based building information modeling technology to construct a pilot plant of six semi-ORSs for controlling culture conditions, each with a culture volume of 40,000 L. Over 1 year, there were no significant variations in monthly biomass productivity, fatty acid composition, or the omega-6/omega-3 ratio; however, the lipid content correlated significantly with photosynthetic photon flux density. During year-round cultivation from November 2014 to October 2017, areal productivity was gradually increased by increasing medium salinity and injecting CO₂ gas into the culture medium. Productivity peaked at 44.01 g/m²/d in October 2017. Throughout the trials, there were no significant differences in average lipid content, which was 14.88 ± 1.26%, 14.73 ± 2.44%, 12.81 ± 2.82%, and 13.63 ± 3.42% in 2014, 2015, 2016, and 2017, respectively. Our results demonstrated that high biomass productivity and constant lipid content can be sustainably maintained under Korean climate conditions.     

Improvement of growth and gymnemic acid production by altering the macro elements concentration and nitrogen source supply in cell suspension cultures of Gymnema sylvestre R. Br

Gymnema sylvestre is an important medicinal plant which bears bioactive compound namely gymnemic acids. The present work deals with optimization of cell suspension culture system of G. sylvestre for the production of biomass and gymnemic acid and we investigated effects of macro elements (NH₄NO₃, KNO₃, CaCl₂, MgSO₄ and KH₂PO₄ - 0.0, 0.5, 1.0, 1.5 and 2.0× strength) and nitrogen source [NH₄⁺/NO₃⁻ ratio of: 0.00/18.80, 7.19/18.80, 14.38/18.80, 21.57/18.80, 28.75/18.80, 14.38/0.00, 14.38/9.40, 14.38/18.80, 14.38/28.20 and 14.38/37.60 (mM)] of Murashige and Skoog medium on accumulation of biomass and gymnemic acid content. The highest accumulation of biomass (165.00 g l⁻¹ FW and 15.42 g l⁻¹ DW) was recorded in the medium with 0.5× concentration of NH₄NO₃ and the highest production of gymnemic acid content was recorded in the medium with 2.0× KH₂PO₄ (11.32 mg g⁻¹ DW). The NH₄⁺/NO₃⁻ ratio also influenced cell growth and gymnemic acid production; both parameters were greater when the NO₃⁻ concentration was higher than that of NH₄⁺. Maximum biomass growth (159.72 g l⁻¹ of FW and 14.95 g l⁻¹ of DW) was achieved at an NH₄⁺/NO₃⁻ ratio of 7.19/18.80, and gymnemic acid production was also greatest at the same concentration of NH₄⁺/NO₃⁻ ratio (11.35 mg g⁻¹ DW).     

Root growth dynamics and stomatal behaviour of rice (Oryza sativa L.) grown under aerobic and flooded conditions

Aerobic rice culture is a new technology designed to reduce water use, but the vulnerability of rice to aerobic condition has limited its development. The objective of this study was to characterize the root growth and stomatal behaviour of four rice cultivars grown in flooded and aerobic culture for 2 years. In aerobic culture, where the soil water potential at 20-cm depth averaged between −15 and −30 kPa, total root biomass was significantly lower than in flooded culture for the whole growth period, owing to a reduction in root biomass in the surface layer. Dry-matter partitioning to roots decreased, but the ratio of deep root biomass to total root biomass tended to be higher in aerobic culture than in flooded culture. The low root-to-shoot ratio and poor root growth in the surface layer in aerobic culture are attributable to the considerable reduction in adventitious root number. As a result, the varietal difference in total root biomass was due largely to individual root growth in aerobic culture. Stomatal closure was distinct at the vegetative stage in aerobic culture, even when the soil water potential was near field capacity, partly because of the poor rooting vigour. When the soil water potential at 20-cm depth was below −50 kPa, the stomatal behaviour reflected the root growth in the subsurface layer. These results suggest the role of vigorous root growth in soil water uptake and hence, in maintaining transpiration in aerobic rice culture.     

New approaches concerning the utilization of natural amendments in cadmium phytoremediation

Different vegetal biomass resources such as Picea abies bark, Castanea sativa chestnuts shell and Asclepias syriaca plant were considered to provide a source of natural bioactive compounds, which can be properly used in several directions such as: plant growth regulators and amendments in bioremediation. Having in mind a complex processing biomass technology, these raw materials are used in a first step to separate bioactive compounds by an aqueous extraction. The obtained extracts containing polyphenols were tested in germination tests to evaluate their influence on cadmium bioaccumulation in oat plant (Avena sativa). It was observed that the mentioned extracts modulated cadmium bioaccumulation, photosynthesizing pigments biosynthesis, plant growth and its development depending on metal ion concentration (5, 12.5, 25 mg/L), and extracts composition determined by vegetal raw material and their total polyphenols content (130, 122, 114 mg/L related to gallic acid). FTIR spectra and histo-anatomical cross section of the roots certify that P. abies bark and A. syriaca plant extracts amendments stimulated cadmium bioaccumulation, promoting the translocation of heavy metals to the aerial part of the plant, meanwhile chestnuts shell extract reduced the mobility of cadmium determining in situ inactivation of heavy metal ions.     

Plant regeneration from tuber discs of potato (Solanum tuberosum L.) using 6-benzylaminopurine (BAP)

Summary Shoots, roots and callus were formed from tuber discs of potato, cultivar Désirée, when grown in vitro on the basal medium of Murashige & Skoog (1962) (MS) supplemented with 2,4-D and/or BAP. Callus was formed in MS medium with 1 mg l⁻¹ BAP plus 0.5 mg l⁻¹ 2,4-D, callus and roots were formed in MS with 1 mg l⁻¹ BAP plus more than 0.5 mg l⁻¹ 2,4-D and shoots were formed directly on tuber discs cultured on MS medium with 1 mg l⁻¹ BAP without the addition of 2,4-D. Nodules produced at the explant surface after the 4th week increased in size following subculture onto the same medium (MS+BAP alone), and 2 to 6 shoots developed from each nodule. After 9 weeks total time in culture, these shoots were excised and transferred as cuttings to MS medium without growth regulators, after which roots developed and plantlets were formed. A histological study of the explants at the sites of nodule formation indicated that the shoots developed from meristematic zones initiated within small outgrowths of tissue similar to those occuring in adventive organogenesis but the presence of shoot and root meristems associated with the same axis suggests the formation of somatic embryos.     

In vitro propagation and regeneration of an industrial plant kenaf (Hibiscus cannabinus L.)

The present study report a protocol for the efficient in vitro propagation of kenaf (Hibiscus cannabinus L., an industrial crop having high cellulosic fiber content) on hormone free MS medium using the shoot apex and nodal explants. Shoot tips and nodes were isolated from 15 days old seedlings cultivated on MS medium. Different combinations and concentrations of auxin/cytokinin were used and added to the MS medium to assess the shoot and root induction of theses explants. Several subcultures were drived in order to enhance the multiplication rate. Healthy and well developed in vitro propagated shoots were transferred for acclimatization under greenhouse conditions in pots filled with different substrates (sand + compost or perlite). Our results showed that shoots could elongate and root within 4-6 weeks on MS basal medium without any callus formation. However, addition of growth regulators to the MS medium leaded to a decrease in shoot and root induction rates. Indeed, the highest shoot regeneration frequency (90.5%) was obtained on MS control medium. Elongated shoots were transferred onto the same hormone free MS medium using five subcultures where the multiplication rate reached the highest value (3.66) at the fifth and last step. The in vitro rooted plantlets were acclimatized in greenhouse and successfully transplanted to natural conditions with 70% survival.     

Effects of high salt stress on secondary metabolite production in the marine-derived fungus Spicaria elegans

To obtain structurally novel and bioactive natural compounds from marine-derived microorganisms, the effect of high salt stress on secondary metabolite production in the marine-derived fungal strain, Spicaria elegans KLA-03, was investigated. The organism, which was isolated from marine sediment, produced different secondary metabolites when cultured in 3% and 10% saline conditions. Four characteristic metabolites, only produced in the 10% salinity culture, were isolated, and their structures were identified as (2E,2'Z)-3,3'-(6,6'-dihydroxybiphenyl-3,3'-diyl)diacrylic acid (1), aspulvinone E (2), aspochalasin E (3) and trichodermamide B (6), according to their 1D and 2D NMR spectra. Compound 1 is a new compound. High salt stress may therefore be a promising means to induce the production of new and chlorinated compounds in halotolerant fungi. Compound 1 showed moderate antibacterial activity against Pseudomonas aeruginosa and Escherichia coli with minimum inhibitory concentration (MIC) values of 0.038 and 0.767 mM, respectively.     

甘蓝型油菜组培苗生根培养体系的优化

为解决油菜组织培养中组培苗生根细短、移栽不易成活的问题,以甘蓝型半冬性油菜转化pCAMBIA1301空载体的再生植株为材料,从生根时间、根长、根重、苗高、苗重、总重、生根数、生根率等方面,以MS和B5为基础培养基,分别使用琼脂糖、琼脂粉及植物凝胶为凝固剂时再生苗的生根效率,得到了两种基础培养基的最适凝固剂组合;在以上两种组合的培养基中分别加入不同浓度蔗糖,进一步比较生根情况和成本,认为含20 g/L蔗糖的B5琼脂粉培养基为最优配方。利用优化后的生根培养基配方,极大地促进油菜组培苗的生根长度和数目,提高后期移栽成活率。     

Field evaluation of upland rice lines selected for QTLs controlling root traits

A marker-assisted back-cross (MABC) programme was used to introgress four root quantitative trait loci (QTLs) from the tropical japonica rice variety Azucena into the Indian upland rice variety, Kalinga III. Previously we tested the products for root traits and reported that the introgressed QTL9 (on chromosome 9) significantly increased root length in the new genetic background. Here we describe field testing for agronomic traits in near-isogenic lines (NILs) that differ for introgressed QTLs. Four NILs were selected and characterised in replicated field experiments in eastern and western India over 3 years. They were tested by upland farmers in a target population of environments (TPE) in three states of eastern India, over 2 years. NILs out-performed Kalinga III for grain and straw yield and there was interaction between the genotypes and the environment (G × E). No effect was found for the root QTL9 on grain or straw yield, however, the presence of several introgressions significantly improved both traits. Some of this effect was due to introgression of Azucena alleles at non-target regions. Overall, the Azucena introgressions increased straw yield more than grain yield. While it has yet to be demonstrated whether this effect is due to improved root systems, this finding fits with the assumption that introgressed genes are involved in partitioning of biomass to the roots and stems, rather than to the grain. The NILs could replace Kalinga III for cultivation in medium upland environments in eastern India.     

A plant growth promoting rhizobacterium and temperature effects on performance of 18 clones of potato

A survey of genotypic responses to beneficial bacterium (Pseudomonas sp. strain PsJN) was conductedin vitro andex vitro, under two temperature conditions, using eighteen clones of potato of different heat stress tolerance: temperate adapted cultivars Kennebec and Russet Burbank; heat tolerant DTO-2, DTO-28, DTO-33, LT-1, LT-2, LT-5, LT-6, LT-7, LT-8, LT-9, Y84-02, NDD277-2, Désirée, and Maine-47; and heat sensitive abscissic acid (ABA)-deficient mutants 11401-01 and 9120-05. Nodal explants taken from 6-week-old bacterized and non-bacterized control plantlets were culturedin vitro on a hormone-free potato nodal cutting medium, and placed at either 20/15 C or 33/25 C day/night temperature, 12h photoperiod and 250 µE m^?2 s^?1 mixture of fluorescent and incandescent light, for six weeks. The tuberization response was studiedex vitro after two weeks acclimation of 2-week old plantlets at 33/25C. The acclimated plantlets were transplanted to 3L plastic nursery pots containing peat-based Pro-Mix growing medium and placed in growth chambers at either 20/15 or 33/25 C day/night temperature, 12 h photoperiod, 475 μE m^?2 s^?1 light and ≈80% RH, for 12 weeks. Compared to the non-bacterized controls, bacterization significantly increased stem length of 12, shoot biomass of 9, and root biomass of 2 clones at 20/15C; and stem length of 14, shoot biomass of 15, and root biomass of 13 clones at 33/25C. High temperature increased length of internodes and had either no effect or slightly decreased node number. Temperature increase had the most dramatic effect on root development. An average shoot to root ratio decreased from 3.7 at 20/15 C to 1.7 at 33/25 C for non-bacterized plantlets and, respectively, from 4.3 to 1.5 for bacterized. The beneficial effect of bacterization on root biomass was the most pronounced in LT-1 and Maine-47 at 20/15 C and LT-8, Maine 47, DTO-2, Kennebec, NDD277-2 and 11401-01 at 33/25C. The temperature elevation did not significantly affect root biomass of LT-6, DTO-28 and Désirée. Temperature stress caused severe reduction in tuber number and tuber fresh weight. ABA-deficient mutants did not produce any tubers and LT-8, LT-9, Y84-027 and DTO-28 tuberized very poorly at 33/25C. DTO-33, Désirée, LT-1 and Kennebec gave the highest number of tubers per pot and Kennebec, LT-1, Désirée and LT-7 the highest yields at this temperature. There was no significant effect of bacterization on tuberization at 20/15 C but at 33/25 C bacterization significantly enhanced tuber number and weight in LT-7 and reduced tuber weight in DTO-2. Although there was no clear link between thein vitro response of particular clones to bacterization and their heat stress tolerance, improvement ofex vitro performance of heat tolerant LT-7 indicates that rhizosphere bacteria may play a role in clonal adaptation of potato to heat stress.     

Optimization of ethanol production by Saccharomyces cerevisiae UFPEDA 1238 in simultaneous saccharification and fermentation of delignified sugarcane bagasse

Ethanol production by Saccharomyces cerevisiae UFPEDA1238 was performed in simultaneous saccharification and fermentation of delignified sugarcane bagasse. Temperature (32 °C, 37 °C), agitation (80; 100 rpm), enzymatic load (20 FPU/g cellulose and 10%, v/v β-glucosidase or 10 FPU/g cellulose and 5% β-glucosidase) and composition of culture medium were evaluated. Ethanol concentration, enzymatic convertibility of cellulose and volumetric productivity were higher than 25 g/L, 72% and 0.70 g/L h, respectively, after 30 h, when the culture medium 1 and 20 FPU/g cellulose/10%, v/v β-glucosidase or the culture medium 2 and 10 FPU/g cellulose/5% β-glucosidase were used in SSF at 37 °C and 80 rpm. In the SSF with culture medium 2 (supplemented with ammonium, phosphate, potassium and magnesium), 150 L ethanol/t bagasse was achieved, with minimum enzyme loading (10 FPU/g cellulose and 5%, v/v β-glucosidase) for 8%, w/v of solids, which is often an important requirement to provide cost-efficient second generation ethanol processes.     

Genotypic variation for root traits of maize (Zea mays L.) from the Purhepecha Plateau under contrasting phosphorus availability

Phosphorus (P) deficiency is a major constraint for maize production in many low-input agroecosystems. This study was conducted to evaluate genotypic variation in both root (root architecture and morphology, including root hairs) and plant growth traits associated with the adaptation of maize landraces to a P-deficient Andisol in two locations in the Central Mexican highlands. Two hundred and forty-two accessions from the Purhepecha Plateau, Michoacan were grown in Ponzomaran with low (23 kg P₂O₅ ha⁻¹) and high (97 kg P₂O₅ ha⁻¹) P fertilization under rain-fed field conditions, and subsequently a subset of 50 contrasting accessions were planted in the succeeding crop cycle in Bonilla. Accessions differed greatly in plant growth, root morphology and P efficiency defined as growth with suboptimal P availability. The accessions were divided into 3 categories of P efficiency using principal component and cluster analyses, and 4 categories according to the retained principal component and their relative weight for each genotype in combination with growth or yield potential. The distribution of accessions among three phosphorus efficiency classes was stable across locations. Phosphorus-efficient accessions had greater biomass, root to shoot ratio, nodal rooting, nodal root laterals, and nodal root hair density and length of nodal root main axis, and first-order laterals under P deficiency. Biomass allocation to roots, as quantified by the allometric partitioning coefficient (K) was not altered by P availability in the efficient accessions, but inefficient accessions had a lower K under low P conditions. Accessions with enhanced nodal rooting and laterals had greater growth under low P. Dense root hairs on nodal root main axes and first-order laterals conferred a marked benefit under low P, as evidenced by increased plant biomass. Late maturity improved growth and yield under low P. These results indicate that landraces of the Central Mexican highlands exhibit variation for several root traits that may be useful for genetic improvement of P efficiency in maize.     

Morphological traits and allocation patterns related to stress-tolerance and seed-yield in wild and domesticated evening primrose (Oenothera L. Onagraceae)

Wild evening primrose species (Oenothera spp.) native to Argentina, have been suggested as a new crop for irrigated valleys of semi-arid Patagonia. This paper describes patterns of biomass allocation, morphological traits related to stress-tolerance and seed-yield in four species of Oenothera grown in a common garden at three plant densities. Wild and domesticated species are compared. The effect of resource availability on those traits during three phenological stages (vegetative, reproductive and maturity) is described. Native species were characterized by traits related to stress-tolerance (high root allocation and low specific leaf area) during the vegetative stage. This suite of traits resulted in low biomass accumulation and low seed-yield. The domesticated O. biennis was characterized by a combination of traits related to stress-tolerance (low specific leaf area) and high productivity (high leaf allocation and leaf area ratio and low root allocation). Domesticated species accumulated more biomass than natives. Total biomass and total non-structural carbohydrates present in roots were positively correlated to seed-yield.Oenothera biennis showed the highest seed-yield, although this species showed yield instability in response to changes in the environmental quality. No changes in seed-yield in response to plant density were recorded for either O. lamarckiana or native species. Oenothera biennis showed an optimum density of 20 plants m⁻² and yielded 260 g m⁻², a seed-yield similar to that reported in other countries. Low seed-yield of native species is major drawback that must be overcome. Improving seed-yield in these species could be possible by selection oriented to increase total biomass. Since no detrimental effect of density was found in O. lamarckiana and natives, a higher plant density might increase yield production per unit area.