Effects of graywater on the growth and survival of ornamental plants in nature-based systems.

The current paper investigates the development of two ornamental plants, canna lily (Canna x generalis) and giant horsetail (Equisetum giganteum), at both bench and pilot scale. Combinations of gravel-filled mesocosm, planted and unplanted (control), irrigated with light greywater (GWL) or tap water (WT), were used. Both species were able to grow under the tested conditions with no indication of toxicity that could affect the development. Irrigation with GWL, resulted in higher evapotranspiration (2.2 mm-2.8 mm) in canna lily than giant horsetail (1.7 mm-2.3 mm) in mesocosm system. When the plants were mature and the season was more humid and warmer, canna lily and giant horsetail irrigated with GWL evapotranspirated 69.23% and 30.77%, respectively as compared to the unplanted GWL-irrigated-mesocosm. Principal components and cluster analysis identified similarity between evapotranspiration (ET) and the characteristics of the plants. Both species can thus be used in constructed wetlands taking into consideration elements such as the space available, level of water and solar incidence so as to allow the full development of the plants. The roots of giant horsetail require high water availability. Low solar incidence is indicated for giant horsetail, and the opposite for canna lily, if flowering is desired.

Intensified nitrogen removal of constructed wetland by novel integration of high rate algal pond biotechnology.

High rate algal pond (HRAP) was combined with constructed wetland (CW) to intensify nitrogen removal through optimizing nitrification and denitrification. Nitrification and denitrification process mainly depends on the oxygen content and carbon source level in CWs. Algal biomass was enriched in HRAP, and dissolved oxygen (DO) concentration was increased via photosynthesis. Algal debris increased COD as degradable bioresource. The results showed that HRAP-CW hybrid systems effectively promoted the nitrogen removal performance due to rich DO and COD. The extension of hydraulic retention time in HRAP significantly improved NH4-N and TN removals by 10.9% and 11.1% in hybrid systems, respectively. The highest NH4-N and TN removals in hybrid systems respectively reached 67.2% and 63.5%, which were significantly higher than those in single CW. The study suggested that the hybrid system had the application potentials in nitrogen removal from wastewater.

Organogenesis and Ultrastructural Features of In Vitro Grown Canna indica L.

An efficient protocol for micropropagation of Canna indica L., an economically and pharmaceutically important plant, was standardized using rhizome explants, excised from two-month-old aseptic seedlings. Complete plant regeneration was induced on MS medium supplemented with 3.0 mg/L BAP plus 1.5 mg/L NAA, which produced the highest number of shoots (73.3 ± 0.5%) and roots (86.7 ± 0.4%) after 2 weeks. Furthermore, the optimum media for multiple shoots regeneration were recorded on MS enriched with 7.0 mg/L BAP (33.0 ± 0.5%). Plantlets obtained were transplanted to pots after two months and acclimatized in the greenhouse, with 75% survival. In addition, ultrastructural studies showed that rhizomes of in vitro grown specimens were underdeveloped compared to the in vivo specimens, possibly due to the presence of wide spaces. Meanwhile, the leaves of in vivo specimens had more open stomata compared to in vitro specimens, yet their paracytic stomata structures were similar. Hence, there were no abnormalities or major differences between in vitro regenerants and mother plants.

Identification and Expression Analyses of miRNAs from Two Contrasting Flower Color Cultivars of Canna by Deep Sequencing.

miRNAs are endogenous small RNA (sRNA) that play critical roles in plant development processes. Canna is an ornamental plant belonging to family Cannaceae. Here, we report for the first time the identification and differential expression of miRNAs in two contrasting flower color cultivars of Canna, Tropical sunrise and Red president. A total of 313 known miRNAs belonging to 78 miRNA families were identified from both the cultivars. Thirty one miRNAs (17 miRNA families) were specific to Tropical sunrise and 43 miRNAs (10 miRNA families) were specific to Red president. Thirty two and 18 putative new miRNAs were identified from Tropical sunrise and Red president, respectively. One hundred and nine miRNAs were differentially expressed in the two cultivars targeting 1343 genes. Among these, 16 miRNAs families targeting 60 genes were involved in flower development related traits and five miRNA families targeting five genes were involved in phenyl propanoid and pigment metabolic processes. We further validated the expression analysis of a few miRNA and their target genes by qRT-PCR. Transcription factors were the major miRNA targets identified. Target validation of a few randomly selected miRNAs by RLM-RACE was performed but was successful with only miR162. These findings will help in understanding flower development processes, particularly the color development in Canna.

The Potential of Canna lily for Wastewater Treatment Under Indian Conditions.

Low cost treatment of polluted wastewater has become a serious challenge in most of the urban areas of developing countries. The present study was undertaken to investigate the potential of Canna lily towards removal of carbon, nitrogen, and phosphorus from wastewater under sub-tropical conditions. A constructed wetland (CW) cell supporting vegetative layer of Canna lily was used to treat wastewater having high strength of CNP. Removal of biological oxygen demand (BOD3) and chemical oxygen demand (COD) varied between 69.8-96.4% and 63.6-99.1%, respectively. C. lily could efficiently remove carbon from a difficult to degrade wastewater at COD:BOD ratio of 24.4. Simultaneous reduction in TKN and nitrate pointed to good nitrification rates, and efficient plant assimilation as the dominant nutrient removal mechanism in the present study. Suitable Indian agro-climatic conditions favored plant growth and no evident stress over the Canna plant was observed. High removal rate of 809.8 mg/m(2)-day for TKN, 15.0 mg/m(2)-day for nitrate, and 164.2 mg/m(2)-day for phosphate suggests for a possible use of Canna-based CW for wastewater treatment for small, rural, and remote Indian communities.

Effect of Different Amendments on Growing of Canna indica L. Inoculated with AMF on Mining Substrate.

Canna indica L. (CiL) was used here in phytoremediation of mining soils. Our work evaluated the effect of AMF (i) on the growth and (ii) on the uptake of heavy metals (HM). The tests were conducted in the greenhouse on mining substrates collected from the Kettara mine (Morocco). The mine soil was amended by different proportions of agricultural soil and compost and then inoculated with two isolates of AMF (IN1) and (IN2) of different origins. After six months of culture, the results show that on mining soils (100%) only AMF (IN2) was able to colonize the roots of CiL with a frequency of 40±7% and an intensity of 6.5±1.5%. Also, the lowest values of shoot and root dry biomass are obtained on these mining soils with respectively 0.30 g and 0.27 g. In contrast, the accumulation of HM was higher and reached more than 50% of that contained in the mining soils, the highest values with 138 mg kg(-1) Cu2+, Zn2+ 270 mg kg(-1) and 1.38 mg kg(-1) Cd was recorded. These results indicate that the colonization of CiL roots by AMF (IN2) could significantly improve its potential to be used in phytoremediation of polluted soil.

Molecular evolution and patterns of duplication in the SEP/AGL6-like lineage of the Zingiberales: a proposed mechanism for floral diversification.

The diversity of floral forms in the plant order Zingiberales has evolved through alterations in floral organ morphology. One striking alteration is the shift from fertile, filamentous stamens to sterile, laminar (petaloid) organs in the stamen whorls, attributed to specific pollination syndromes. Here, we examine the role of the SEPALLATA (SEP) genes, known to be important in regulatory networks underlying floral development and organ identity, in the evolution of development of the diverse floral organs phenotypes in the Zingiberales. Phylogenetic analyses show that the SEP-like genes have undergone several duplication events giving rise to multiple copies. Selection tests on the SEP-like genes indicate that the two copies of SEP3 have mostly evolved under balancing selection, probably due to strong functional restrictions as a result of their critical role in floral organ specification. In contrast, the two LOFSEP copies have undergone differential positive selection, indicating neofunctionalization. Reverse transcriptase-polymerase chain reaction, gene expression from RNA-seq data, and in situ hybridization analyses show that the recovered genes have differential expression patterns across the various whorls and organ types found in the Zingiberales. Our data also suggest that AGL6, sister to the SEP-like genes, may play an important role in stamen morphology in the Zingiberales. Thus, the SEP-like genes are likely to be involved in some of the unique morphogenetic patterns of floral organ development found among this diverse order of tropical monocots. This work contributes to a growing body of knowledge focused on understanding the role of gene duplications and the evolution of entire gene networks in the evolution of flower development.

Phytoremediation of BTEX contaminated soil by Canna×generalis.

Bioaccumulation experiments showed that the canna (Canna×generalis) could accumulate BTEX (benzene, toluene, ethylbenzene and xylenes) from root zone and rhizome zone soil and translocate these compounds to the shoot. A comparison among these compounds showed that the sequences for accumulation in the root, rhizome and shoot were strongly related to their physicochemical properties (i.e. K(ow) values and molecular weight). For removal efficiency, the canna could remove about 80% of BTEX in the root zone and rhizome zone soil in 21 days. In addition, the removal efficiency in BTEX contaminated soil with 40% water content was a little higher than that found with 20% soil water content. This result indicated that the soil water content should also be considered when phytoremediation is employed.

Structure of the herb stratum under different light regimes in the Submontane Atlantic Rain Forest.

This study aimed to characterize the structure of the herb stratum in relation to light availability in the Submontane Atlantic Rain Forest at the Carlos Botelho State Park, SP, Brazil. Fortyone 10 x10 m plots were established under the closed canopy (18 plots), small and medium canopy gaps (11) and large canopy gaps dominated by Guadua tagoara (Ness) Kunth (12). Inside each plot, the line intercept method was applied to assess soil coverage as an estimate of density of herb stratum vegetation. Hemispherical photographs were taken at the centre of the plots to evaluate the annual light regime. Overall, Calathea communis Wanderley and S. Vieira had the greater mean coverage, followed by woody seedlings, ground ferns and other herbs (mainly, Araceae, Acanthaceae, Amaranthaceae and Cyperaceae). There were strong correlations among several groups of the herb stratum, such as the negative correlations between woody seedlings with the coverage of C. communis and with rocks. The analysis of the hemispherical photographs confirmed the difference among environments that led to significant differences in the soil coverage of the herb stratum vegetation but woody seedlings. For instance, C. communis showed great coverage in large gaps while ferns were more abundant in small and medium gaps and in the understorey. Other herbs, in turn, demonstrated bigger soil coverage in small and medium gaps. Although this study represents a rough assessment of the structure and composition of the herb stratum, the results found here illustrated the evident relation between herb species density and the environmental variation promoted by changes on canopy structure and topography.

Structure of the herb stratum under different light regimes in the Submontane Atlantic Rain Forest / Estrutura do estrato herbáceo sob diferentes regimes de luz na Floresta Pluvial Atlântica Submontana

This study aimed to characterize the structure of the herb stratum in relation to light availability in the Submontane Atlantic Rain Forest at the Carlos Botelho State Park, SP, Brazil. Fortyone 10 x10 m plots were established under the closed canopy (18 plots), small and medium canopy gaps (11) and large canopy gaps dominated by Guadua tagoara (Ness) Kunth (12). Inside each plot, the line intercept method was applied to assess soil coverage as an estimate of density of herb stratum vegetation. Hemispherical photographs were taken at the centre of the plots to evaluate the annual light regime. Overall, Calathea communis Wanderley and S. Vieira had the greater mean coverage, followed by woody seedlings, ground ferns and other herbs (mainly, Araceae, Acanthaceae, Amaranthaceae and Cyperaceae). There were strong correlations among several groups of the herb stratum, such as the negative correlations between woody seedlings with the coverage of C. communis and with rocks. The analysis of the hemispherical photographs confirmed the difference among environments that led to significant differences in the soil coverage of the herb stratum vegetation but woody seedlings. For instance, C. communis showed great coverage in large gaps while ferns were more abundant in small and medium gaps and in the understorey. Other herbs, in turn, demonstrated bigger soil coverage in small and medium gaps. Although this study represents a rough assessment of the structure and composition of the herb stratum, the results found here illustrated the evident relation between herb species density and the environmental variation promoted by changes on canopy structure and topography.

O objetivo deste estudo foi caracterizar a estrutura do estrato herbáceo em relação à disponibilidade de luz na Floresta Pluvial Atlântica Submontana do Parque Estadual Carlos Botelho, SP, Brasil. Para tanto, foram instaladas 41 parcelas de 10 x 10 m em ambientes sob o dossel fechado (18 parcelas), em clareiras pequenas e médias (11), e em clareiras grandes com dominância de Guadua tagoara (Ness) Kunth (12). Em cada parcela a percentagem de cobertura de solo, avaliada através do método de interceptação em linha, foi usada como estimativa da densidade da vegetação do estrato herbáceo. Fotografias hemisféricas foram tomadas ao centro de cada uma das parcelas para avaliar o regime anual de luz nos ambientes. No geral, a maior média obtida foi para Calathea communis Wanderley e S. Vieira, seguida por plântulas de regenerantes lenhosos, pteridófitas terrestres e outras ervas (principalmente Araceae, Acanthaceae, Amaranthaceae e Cyperaceae). Houve ainda fortes correlações entre vários grupos do estrato herbáceo, como as correlações negativas entre plântulas de regenerantes com a cobertura de C. communis e de rochas. A análise das fotografias hemisféricas confirmou a existência de ambientes com diferentes regimes de luz, que promoveram diferenças significativas na cobertura do solo de todos os grupos do estrato herbáceo, exceto para plântulas de regenerantes. Por exemplo, C. communis apresentou grande cobertura nas grandes clareiras, enquanto que as pteridófitas terrestres foram mais abundantes no subbosque e nas clareiras pequenas e médias. O grupo Outras Ervas, por sua vez, apresentou maiores coberturas nas clareiras pequenas e médias. Apesar de representar uma análise grosseira da estrutura e composição, os resultados encontrados aqui ilustraram uma evidente relação entre a densidade de formas de vida herbácea e as variações ambientais promovidas por mudanças na estrutura do dossel da floresta e na topografia.

Use of dewatered municipal sludge on Canna growth in pot experiments with a barren clay soil.

The present study evaluated the possibility of using the dewatered municipal sludge for non-agricultural purposes. The sludge was amended with soil and was applied at 0, 165, 330, 495 and 660 t/ha to promote the growth of Canna. The results showed that the Canna growth pattern exhibited a pronounced positive growth response in the range of 165-495 t/ha, and the Canna could not survive at an amendment rate of 660 t/ha. The analysis of chlorophyll fluorescence parameters showed that sludge did no harm to Canna, while under the conditions of barren soil alone, the plants were put into nutrients stress conditions. Due to the application of sludge, the concentration of heavy metals (Cu, Zn, Cr, Cd, Pb and Ni) in soil increased. However, by planting of Canna, contents of Cd, Ni and Zn showed trends of decline; Cd and Ni have shown a significant decline in concentration, while Zn had only limit response. As a result, dewatered sludge might be used to amend the barren soil and Canna could be used for phytoremediation of sludge.

Constructed wetland systems vegetated with different plants applied to the treatment of tannery wastewater.

Wastewaters from leather processing are very complex and lead to water pollution if discharged untreated, especially due to its high organic loading. In this study the survival of different plant species in subsurface horizontal flow constructed wetlands receiving tannery wastewater was investigated. Five pilot units were vegetated with Canna indica, Typha latifolia, Phragmites australis, Stenotaphrum secundatum and Iris pseudacorus, and a sixth unit was left as an unvegetated control. The treatment performance of the systems under two different hydraulic loading rates, 3 and 6 cmd(-1), was assessed. COD was reduced by 41-73% for an inlet organic loading varying between 332 and 1602 kgha(-1)d(-1) and BOD(5) was reduced by 41-58% for an inlet organic loading varying between 218 and 780 kgha(-1)d(-1). Nutrient removal occurred to lower extents. Phragmites australis and Typha latifolia were the only plants that were able to establish successfully. Despite the high removal of organic content from the influent wastewater, during 17 months of operation, no significant differences in performance were observed between units.

Inhibition of beta-amylase activity, starch degradation and sucrose formation by indole-3-acetic acid during banana ripening.

In order to observe the effect of indole-3-acetic acid (IAA) on carbohydrate metabolism, unripe banana (Musa acuminata AAA, cv. Nanicão) slices were infiltrated with the hormone and left to ripen under controlled conditions. The climacteric respiration burst was reduced by the action of IAA, and starch degradation and sucrose formation were delayed. Sucrose synthase (SuSy; EC 2.4.1.13) and sucrose-phosphate synthase (SPS; EC 2.4.1.14) activities and transcript levels were not affected, indicating that prevention of sucrose accumulation was not related to sucrose-metabolizing enzymes. Impairment of sucrose synthesis could be a consequence of lack of substrate, since starch degradation was inhibited. The increase in activity and transcript level of beta-amylase was delayed, indicating that this enzyme could be important in starch-to-sucrose metabolism in bananas and that it might be, at least partially, controlled at the transcriptional level. This is the first report showing that IAA can delay starch degradation, possibly affecting the activity of hydrolytic enzymes such as beta-amylase (EC 3.2.1.2).

Quarantine security of bananas at harvest maturity against Mediterranean and Oriental fruit flies (Diptera: Tephritidae) in Hawaii.

Culled bananas (dwarf 'Brazilian', 'Grand Nain', 'Valery', and 'Williams') sampled from packing houses on the islands of Hawaii, Kauai, Maui, Molokai, and Oahu identified specific "faults" that were at risk from oriental fruit fly, Bactrocera dorsalis (Hendel), infestation. Faults at risk included bunches with precociously ripened bananas, or bananas with tip rot, fused fingers, or damage that compromised skin integrity to permit fruit fly oviposition into fruit flesh. No Mediterranean fruit fly, Ceratitis capitata (Wiedemann), or melon fly, B. cucurbitae (Coquillett), infestations were found in culled banana samples. Field infestation tests indicated that mature green bananas were not susceptible to fruit fly infestation for up to 1 wk past the scheduled harvest date when attached to the plant or within 24 h after harvest. Recommendations for exporting mature green bananas from Hawaii without risk of fruit fly infestation are provided. The research reported herein resulted in a USDA-APHIS protocol for exporting mature green bananas from Hawaii.

Purification and characterization of cytosolic pyruvate kinase from banana fruit.

Cytosolic pyruvate kinase (PK(c)) from ripened banana (Musa cavendishii L.) fruits has been purified 543-fold to electrophoretic homogeneity and a final specific activity of 59.7 micromol of pyruvate produced/min per mg of protein. SDS/PAGE and gel-filtration FPLC of the final preparation indicated that this enzyme exists as a 240 kDa homotetramer composed of subunits of 57 kDa. Although the enzyme displayed a pH optimum of 6.9, optimal efficiency in substrate utilization [in terms of V(max)/K(m) for phosphoenolpyruvate (PEP) or ADP] was equivalent at pH 6.9 and 7.5. PK(c) activity was absolutely dependent upon the presence of a bivalent and a univalent cation, with Mg(2+) and K(+) respectively fulfilling this requirement. Hyperbolic saturation kinetics were observed for the binding of PEP, ADP, Mg(2+) and K(+) (K(m) values of 0.098, 0.12, 0.27 and 0.91 mM respectively). Although the enzyme utilized UDP, IDP, GDP and CDP as alternative nucleotides, ADP was the preferred substrate. L-Glutamate and MgATP were the most effective inhibitors, whereas L-aspartate functioned as an activator by reversing the inhibition of PK(c) by L-glutamate. The allosteric features of banana PK(c) are compared with those of banana PEP carboxylase [Law and Plaxton (1995) Biochem. J. 307, 807-816]. A model is presented which highlights the roles of cytosolic pH, MgATP, L-glutamate and L-aspartate in the co-ordinate control of the PEP branchpoint in ripening bananas.

Purification, characterization and structural analysis of an abundant beta-1,3-glucanase from banana fruit.

An abundant, catalytically active beta-1,3-endoglucanase (EC 3.2.1. 39) has been isolated from the pulp of ripe bananas. Biochemical analysis of the purified protein, molecular modelling, and molecular cloning of the corresponding gene indicate that this banana enzyme closely resembles previously characterized plant beta-glucanases with respect to its amino-acid sequence, structure and biological activity. The results described in this paper demonstrate both the occurrence of an abundant active beta-1,3-endoglucanases in fruits and also readdress the question of the possible involvement of these enzymes in the ripening and/or softening process.

A UV-light activated cinnamic acid isomer regulates plant growth and gravitropism via an ethylene receptor-independent pathway.

Naturally occurring cinnamic acids (CA) exist in both trans- and cis-isoforms. UV-light irradiation of trans-CA is able to produce cis-CA. cis-CA was found to possess auxin-like activity before. In contrast, the vapor of cis-CA induced an epinastic response in tomato plants just as ethylene does. Given the existence of a double bond in and the gaseous nature of cis-CA, we suspected that cis-CA might also function as an ethylene-like compound. To distinguish between these possibilities, we selected an ethylene perception-deficient tomato plant, Never-ripe (Nr), and an ethylene biosynthesis-deficient tomato plant, A11. Not only did the vapor of cis-CA fail to trigger A11 tomato fruit ripening but it also delayed the ripening of banana fruit. Moreover, the vapor of cis-CA induced epinasty and the 'triple response' in both the wild type and Nr tomato plants, indicating that the vapor of cis-CA does not act via an ethylene receptor-dependent pathway. Furthermore, the vapor of cis-CA inhibited the negative gravitropic response of stems of both etiolated Nr seedlings and young plants, whereas ethylene had little effect on the negative gravitropism of the Nr plants. These results support the conclusion that the action sites of the vapor of cis-CA and ethylene are fundamentally different.

Gene expression in the pulp of ripening bananas. Two-dimensional sodium dodecyl sulfate-polyacrylamide gel electrophoresis of in vitro translation products and cDNA cloning of 25 different ripening-related mRNAs.

mRNA was extracted from the pulp and peel of preclimacteric (d 0) bananas (Musa AAA group, cv Grand Nain) and those exposed to ethylene gas for 24 h and stored in air alone for a further 1 (d 2) and 4 d (d 5). Two-dimensional sodium dodecyl sulfate-polyacrylamide gel electrophoresis of in vitro translation products from the pulp and peel of these fruits revealed significant up-regulation of numerous transcripts during ripening. The majority of the changes were initiated by d 2, with the level of these messages increasing during the remainder of the ripening period. Pulp tissue from d 2 was used for the construction of a cDNA library. This library was differentially screened for ripening-related clones using cDNA from d-0 and d-2 pulp by a novel microtiter plate method. In the primary screen 250 up- and down-regulated clones were isolated. Of these, 59 differentially expressed clones were obtained from the secondary screen. All of these cDNAs were partially sequenced and grouped into families after database searches. Twenty-five nonredundant groups of pulp clones were identified. These encoded enzymes were involved in ethylene biosynthesis, respiration, starch metabolism, cell wall degradation, and several other key metabolic events. We describe the analysis of these clones and their possible involvement in ripening.

Differential gene expression in ripening banana fruit.

During banana (Musa acuminata L.) fruit ripening ethylene production triggers a developmental cascade that is accompanied by a massive conversion of starch to sugars, an associated burst of respiratory activity, and an increase in protein synthesis. Differential screening of cDNA libraries representing banana pulp at ripening stages 1 and 3 has led to the isolation of 11 nonredundant groups of differentially expressed mRNAs. Identification of these transcripts by partial sequence analysis indicates that two of the mRNAs encode proteins involved in carbohydrate metabolism, whereas others encode proteins thought to be associated with pathogenesis, senescence, or stress responses in plants. Their relative abundance in the pulp and tissue-specific distribution in greenhouse-grown banana plants were determined by northern-blot analyses. The relative abundance of transcripts encoding starch synthase, granule-bound starch synthase, chitinase, lectin, and a type-2 metallothionein decreased in pulp during ripening. Transcripts encoding endochitinase, beta-1,3-glucanase, a thaumatin-like protein, ascorbate peroxidase, metallothionein, and a putative senescence-related protein increased early in ripening. The elucidation of the molecular events associated with banana ripening will facilitate a better understanding and control of these processes, and will allow us to attain our long-term goal of producing candidate oral vaccines in transgenic banana plants.

[Optimal high-yield agronomic measures for Curcuma longa L].

Regression method of orthogonal conic substitution with factors was employed to build up a tuber yield simulation model. Three main measures (sowing time, plant population, application rate of fertilizer) affecting the tuber yield were analyzed by the mathematical model of three unknown second order orthogonal rotative regression. The optimal agronomic measures were obtained. The results indicate that sowing time and plant population play a important role in raising the tuber yield. Sowing time is clearly interrelated with plant population, and likewise, plant population application rate of fertilizer.