Sink trap: duckweed and dye attractant reduce mosquito populations

Duckweeds, such as Lemna minor Linnaeus (Alismatales: Lemnaceae), are common in aquatic habitats and have been suggested to reduce larval mosquito survivorship via mechanical and chemical effects. Furthermore, pond dyes are used increasingly in aquatic habitats to enhance their aesthetics, although they have been shown to attract mosquito oviposition. The present study examined the coupled effects of L. minor and black pond dye on the oviposition selectivity of Culex pipiens Linnaeus (Diptera: Culicidae) mosquitoes in a series of laboratory choice tests. Subsequently, using outdoor mesocosms, the combined influence of duckweed and pond dye on mosquito abundances in aquatic habitats was quantified. Mosquitoes were strongly attracted to duckweed, and oviposited significantly greater numbers of egg rafts in duckweed-treated water compared with untreated controls, even when the duckweed was ground. The presence of pond dye interacted with the duckweed and further enhanced positive selectivity towards duckweed-treated water. The presence of duckweed caused significant and sustained reductions in larval mosquito numbers, whereas the relative effects of dye were not evident. The use of floating aquatic plants such as duckweed, combined with dye, may help reduce mosquito populations via the establishment of population sinks, characterized by high rates of oviposition coupled with high levels of larval mortality.     

DNA barcoding as a tool for early warning and monitoring alien duckweeds (Lemna sp.pl.): the case of Central Italy

Aquatic habitats are vulnerable to the invasion of alien species, so early warning protocols are necessary for eradication. The presence in Italy of two alien duckweeds in freshwaters has been documented: Lemna minuta, that showed high invasivity, and L. valdiviana, still confined to south Lazio. These two species may be mistaken for each other and for the domestic L. minor and L. gibba due to morphological variation. Here, we assess the applicability of DNA barcoding as a complement to morphological analysis for monitoring the spread of alien Lemna. We chose two chloroplast genome sequences for their ability to discriminate all Lemna species: the 5’ intron of the trnK gene and the matK gene. Among 48 samples of Lemna collected at 20 sites in Central Italy, 20 were identified as L. minor, 19 as L. minuta, five as L. trisulca and four as L. gibba. L. minuta was present at most sampling sites; in particular, at six locations of Lake Trasimeno, eight L. minuta samples were found. We demonstrate that DNA sequence analyses with cost-effective barcoding techniques can effectively support expert efforts in species determination for an early alert system of invasive Lemna species.     

Short-term chromium(VI) stress induces different photosynthetic responses in two duckweed species, <Emphasis Type="Italic">Lemna gibba</Emphasis> L. and <Emphasis Type="Italic">Lemna mino</Emphasis>r L.

Physiological responses of two duckweed species, Lemna gibba and Lemna minor, to hexavalent chromium [Cr(VI)] were studied in axenic cultures using short-term (48 h) treatments by K₂Cr₂O₇ (0–200 μM). Chlorophyll (Chl) fluorescence parameters and photosynthetic pigment composition of plants were screened to determine the effects of Cr(VI) exposures. The two duckweed species exhibited different sensitivity in the applied Cr(VI) concentration range. Chl fluorescence parameters of dark-adapted and light-adapted plants and electron transport inducibility were more sensitive to Cr(VI) in L. minor than in L. gibba. We also found fundamental differences in quantum yield of regulated, Y(NPQ), and nonregulated, Y(NO), non-photochemical quenching between the two species. As Cr(VI) concentration increased in the growth medium, L. minor responded with considerable increase of Y(NPQ) with a parallel significant increase of Y(NO). By contrast, in L. gibba only 200 μM Cr(VI) in the growth medium resulted in elevation of Y(NPQ) while Y(NO) remained more or less constant within the regarding Cr(VI) concentration range during 48 h. Photosynthetic pigment content did not change considerably during the short-term Cr(VI) treatment but decrease of Chl a/b and increase of Car/Chl ratios were observed in good accordance with the changes in Chl fluorescence parameters. The data suggest that various duckweed species respond with different sensitivity to the same ambient concentrations of Cr(VI) in the growth medium, and presumably to other environmental stresses too, which may have an influence on their competitive relations when heavy metal pollution occurs in aquatic ecosystem.     

Preference,performance, and impact of the water-lily aphid on multiple species of duckweed

1. The role of herbivores in driving the structure of freshwater macrophyte communities remains poorly understood in comparison with terrestrial ecosystems. For instance, although duckweed (subfamily Lemnoideae) are globally distributed, can be locally highly abundant and ecologically dominant, and are of growing economic importance, their interactions with herbivores remain understudied.
2. To address how herbivores may impact duckweed species composition, we here experimentally quantify the preference and performance of a common duckweed herbivore, the water-lily aphid (Rhopalosiphum nymphaeae) on four widespread duckweed species.
3. Our two-way choice experiments reveal that aphids display a preference for Spirodela polyrhiza > Landoltia punctata = Lemna minor > > Wolffia brasiliensis. These results are rarely influenced by natal host species.
4. By evaluating the growth of aphid populations on each duckweed species, we find that preference may be adaptive in certain ecological conditions.
5. Quantifying the population growth rate of duckweed growing in the presence and absence of aphids revealed differential tolerance of herbivory across duckweed species.
6. This study shows that aphids, through preferential feeding and significant differential effects on duckweed growth, can have a significant impact on duckweed population dynamics and potentially community composition.

     

气候变化情景下西藏入侵植物印加孔雀草的潜在分布预测

入侵植物通常由于具有较强的适应性而能够快速繁殖扩散,影响本土物种的生长繁殖,进而威胁到当地生态安全、景观格局和农业生产等。西藏生态环境非常脆弱,一旦发生恶性物种大面积入侵,生态后果不堪设想。为了探究入侵植物印加孔雀草(Tagetes minuta L.)对西藏东南生态安全的影响趋势,基于野外实地调查数据,采用最大熵(MaxEnt)模型,应用R语言平台对模型和数据进行优化筛选,探讨影响其地理分布的主要环境因子,并模拟预测了当代及2种气候变化情景(RCP 4.5、RCP 8.5)下,其在西藏的潜在适生区分布情况。结果表明：(1)训练数据集和测试数据集的受试者工作特征曲线下的面积(AUC) 均为0.997,模拟效果较好;底层土壤酸碱度、最暖季降水量、土壤有效含水量、最暖月最高温度为影响印加孔雀草分布的主导环境因子,贡献率总和超过90%。(2)加查县、朗县是印加孔雀草分布密集区域,米林县、林芝市区、察隅县、墨脱县等地为入侵高风险地区。(3)中短期(2050年)内印加孔雀草适生面积增加明显,2070年时面积则会减少;印加孔雀草适生区在藏东南地区进一步向东北区域扩张,分布质心由当前的墨脱县域向波密县域转移。总体而言,印加孔雀草分布受土壤环境、温度和降水影响较大,气候变化将使其向西藏东部、南部扩张。研究结果对于西藏自治区制定植物入侵防控管理办法具有重要参考价值。     

Genetic transformation of duckweed Lemna gibba and Lemna minor

Summary We developed efficient genetic transformation protocols for two species of duckweed, Lemna gibba (G3) and Lemna minor (8627 and 8744), using Agrobacterium-mediated gene transfer. Partially differentiated nodules were co-cultivated with Agrobacterium tumefaciens harboring a binary vector containing β-glucuronidase and nptII expression cassettes. Transformed cells were selected and allowed to grow into nodules in the presence of kanamycin. Transgenic duckweed fronds were regenerated from selected nodules. We demonstrated that transgenic duckweed could be regenerated within 3 mo. after Agrobacterium-mediated transformation of nodules. Furthermore, we developed a method for transforming L. minor 8627 in 6 wk. These transformation protocols will facilitate genetic engineering of duckweed, ideal plants for bioremediation and large-scale industrial production of biomass and recombinant proteins.     

Non-native fish species in Neotropical freshwaters: how did they arrive,and where did they come from?

The invasion of aquatic ecosystems by introduced invasive alien species (IAS) has become a worldwide phenomenon, and often leads to competitive interactions with native species. At high-nutrient levels, native species mostly are outcompeted by the introduced species. We performed an outdoor competition experiment between IAS free-floating Lemna minuta and native Lemna minor in a eutrophicated pond to examine whether the invasive species is the better competitor. We additionally performed an indoor experiment resembling mesotrophic phosphorus (P) conditions to investigate both species’ competitiveness in low P availability and compared with previous experiments at high-nutrient levels. Our results showed that in field conditions, the alien L. minuta was the better competitor. In the mesotrophic indoor condition, however, the native L. minor was the better competitor. Both species produced longer roots in the indoor experiment compared to field conditions. The species’ relative growth rates were also lower in the indoor experiment. A P reduction to mesotrophic condition in the water column thus might reduce invasive L. minuta growth and competitive performance. Additionally, introduction and recovery of L. minor could reduce L. minuta cover, but only following P reduction. Field experiments in mesotrophic ponds are needed to confirm these indoor findings.     

Diatom ecology in the phyllosphere of the common duckweed (Lemna minor L.)

Epiphyton associated with thick, floating mats of the common duckweed (Lemna minor L.) was studied at four sites in western Canada between 1985 and 1988. Maximum epiphyton abundance generally occurred in spring as biomass of the duckweed mat was increasing. Epiphytic biomass was low during summer and increased at some sites in autumn with mat decomposition. The community was composed mostly of diatoms and, during summer, photosynthetic bacteria. Species richness of the diatom flora was low, suggesting that duckweed mats are environments to which few species are adapted. Photosynthesis - irradiance curves indicated that duckweed epiphyton was not adapted to low light levels that occurred in the mat (< 1 % of ambient), suggesting they may survive via other means of nutrition. The mat phyllosphere was also characterized by wide spatial and temporal variation in temperature, and sharp vertical profiles of dissolved oxygen and nutrients.     

印加孔雀草入侵生态风险评估及其管理措施

【目的】印加孔雀草为菊科一年生草本植物,原产南美洲,现已在我国多个地区发现,局部呈现生态危害。分析印加孔雀草生态适应性特点,可以有针对地提出其风险指标体系和判断标准,确定其危险级别。【方法】对印加孔雀草的分布、危害情况及生物学特性进行调查,定性分析其生态适应性特点,定量分析评估其生态风险,确定其危险级别。【结果】根据风险指标体系准则层的赋值,其中入侵性R_1为83,适应性R_2为88,扩散性R_3为65,危害性R_4为84,计算得出印加孔雀草风险值(R)=8061,属于具有危害性和侵入性的物种。【结论】印加孔雀草为具有危害性和侵入性的物种,应严格禁止引入。由于该植物已经传入我国,应采取必要的应急措施,开展检疫、监测和灭除,防止其传入其他还未发生的地区。     

Frond transformation system mediated by <Emphasis Type="Italic">Agrobacterium tumefaciens</Emphasis> for <Emphasis Type="Italic">Lemna minor</Emphasis>

The Lemnaceae, known as duckweed, the smallest flowering aquatic plant, shows promise as a plant bioreactor. For applying this potential plant bioreactor, establishing a stable and efficient genetic transformation system is necessary. The currently favored callus-based method for duckweed transformation is time consuming and genotype limited, as it requires callus culture and regeneration, which is inapplicable to many elite duckweed strains suitable for bioreactor exploitation. In this study, we attempted to establish a simple frond transformation system mediated by Agrobacterium tumefaciens for Lemna minor, one of the most widespread duckweed species in the world. To evaluate the feasibility of the new transformation system, the gene CYP710A11 was overexpressed to improve the yield of stigmasterol, which has multiple medicinal purposes. Three L. minor strains, ZH0055, D0158 and M0165, were transformed by both a conventional callus transformation system (CTS) and the simple frond transformation system (FTS). GUS staining, PCR, quantitative PCR and stigmasterol content detection showed that FTS can produce stable transgenic lines as well as CTS. Moreover, compared to CTS, FTS can avoid the genotype constraints of callus induction, thus saving at least half of the required processing time (CTS took 8–9 months while FTS took approximately 3 months in this study). Therefore, this transformation system is feasible in producing stable transgenic lines for a wide range of L. minor genotypes.     

Lemna minor tolerance to metal‐working fluid residues: implications for rhizoremediation

For the first time in the literature, duckweed (Lemna minor) tolerance (alone or in combination with a consortium of bacteria) to spent metal‐working fluid (MWF) was assessed, together with its capacity to reduce the chemical oxygen demand (COD) of this residue. In a preliminary study, L. minor response to pre‐treated MWF residue (ptMWF) and vacuum‐distilled MWF water (MWFw) was tested. Plants were able to grow in both residues at different COD levels tested (up to 2300 mg·l^?1), showing few toxicity symptoms (mainly growth inhibition). Plant response to MWFw was more regular and dose responsive than when exposed to ptMWF. Moreover, COD reduction was less significant in ptMWF. Thus, based on these preliminary results, a second study was conducted using MWFw to test the effectiveness of inoculation with a bacterial consortium isolated from a membrane bioreactor fed with the same residue. After 5 days of exposure, COD in solutions containing inoculated plants was significantly lower than in non‐inoculated ones. Moreover, inoculation reduced β+γ‐tocopherol levels in MWFw‐exposed plants, suggesting pollutant imposed stress was reduced. We therefore conclude from that L. minor is highly tolerant to spent MWF residues and that this species can be very useful, together with the appropriate bacterial consortium, in reducing COD of this residue under local legislation limits and thus minimise its potential environmental impact. Interestingly, the lipophilic antioxidant tocopherol (especially the sum of β+γ isomers) proved to be an effective plant biomarker of pollution.     

The role of the hemiparasitic annual Rhinanthus minor in determining grassland community structure

Summary Rhinanthus minor (yellow-rattle) is a widespread hemiparasitic plant of grassland habitats throughout Great Britain. It is usually considered to be indicative of species-rich grassland, but in a survey of 14 habitats throughout Britain it was found that R. minor at the time of flowering normally occupied relatively low-diversity patches within areas of high diversity as determined by the number of species, Simpson's Index and the Shannon-Wiener Index. Following the death of adult plants of R. minor in the summer it was shown that the pattern of species diversity changed such that by the time R. minor germinated in the following spring the differences between the areas containing and not containing R. minor were much less distinct. A perturbation experiment in which R. minor was removed from four sites indicated that the effect of the removal of R. minor on the development of community structure over the next year was to increase species diversity on three of the sites and decrease it on the fourth. Those species which responded to the removal of R. minor by an increase in abundance were shown to be preferred hosts. All three lines of evidence point to the fact R. minor has a significant effect on the species diversity of the communities in which it grows by selectively parasitizing components of the flora and modifying the competitive relationships between plants. However, as the communities generally responded to the removal of R. minor by an increase in diversity and as the general survey indicated that R. minor is generally associated with areas of low diversity it would appear that the plants which are selectively parasitized are generally not the competitive dominants in the community.     

Bioremoval of Aqueous Lead Using Lemna Minor

The aquatic plant Lemna minor (duckweed) was examined for its ability to sorb soluble lead from water under laboratory conditions. The use of biomass as a sorption medium provides a simple alternative separation technology. Laboratory-scale (700-mL) batch reactors containing soil-based sediments were used to expose L. minor to concentrations of 0.0, 5.0, and 10.0 mg/L of lead for 7 d. Overall removal amounts of 95% were observed, with 85% removal occurring within the first day. A sorption process model was developed based on a mass balance and a power law rate of sorption equation.     

入侵植物印加孔雀草在不同生境的种群构件生物量及其分配特征

印加孔雀草是中国西藏新近入侵种,其危害已初见端倪,但我国鲜有对其入侵机理的研究。为探究印加孔雀草在异质环境下的种群构件生物量及其分配特征,进一步深入了解其生存策略和易入侵生境。该研究在菜园、果园、路边、荒地和河滩等五种典型入侵生境内对其花果期的种群构件生物量进行了测定和分析,计算了表型可塑性指标值。结果表明:(1)印加孔雀草种群各构件生物量的基本规律为茎>花果>叶>根,在路边最大,菜园最小,两地间各值均存在显著差异(P<0.05)。(2)印加孔雀草各构件生物量的总变异系数(CV)和可塑性指标(PI)的均值分别为46.93%和61.44%。(3)印加孔雀草繁殖与营养构件生物量比的关系为荒地>路边>果园>河滩>菜园,根冠比表现为菜园>荒地>河滩>路边>果园。(4)印加孔雀草各构件之间、各构件与总生物量间都存在显著的正相关关系,体现了其整体协调的生存策略。上述结果表明,印加孔雀草能通过各构件生物量的调整来适应异质生境,具有较高的可塑性;高繁殖输出和对异质环境的适应性可能是其成功入侵的重要原因。     

Evaluation of heat shock protein 70 as a biomarker of environmental stress in Fucus serratus and Lemna minor

Heat shock proteins (Hsps) are known to be induced in response to short-term stress. The present study aimed to evaluate the potential of Hsp70 as a biomarker of stress produced by increased temperature, osmotic pressure, and exposure to cadmium and sodium chloride in marine macroalgae and fresh water plant species. An indirect competitive enzyme-linked immunosorbent assay (IC-ELISA) was developed with a working range of 0.025–10 μg?ml^?1 using a monoclonal antibody raised against purified Hsp70 of Phaseolus aureus (mung bean). Fucus serratus (toothed wrack), Chondrus crispus (Stackhouse or Carrageen moss), Ulva lactuca (sea lettuce) and Lemna minor (common duckweed) sample extracts were stressed for up to 24 h and then tested in the IC-ELISA. The presence of Hsp70 and cross-reactivity of the monoclonal antibody was confirmed by Western blot. The heat shock response was confirmed in each species using a 2-h 42°C treatment. Following heat shock, Hsp70 concentrations increased to a peak at 2 h (F. serratus) or 4 h (L. minor), after which concentrations decreased. Osmotic and cadmium stresses also resulted in elevated Hsp70 concentrations in samples of F. serratus and L. minor when compared with unstressed controls. In both, osmotic and metal stress, the production of Hsp70 increased to a maximum and subsequently decreased as the stressor levels increased. Results suggest that Hsp70 IC-ELISA could potentially be applied to the detection of stress in these aquatic species, although it would probably be most effective when used in conjunction with other measurements to provide a stressor-specific biomarker profile or fingerprint.     

Effects of co-inoculation of two different plant growth-promoting bacteria on duckweed

Aseptic Lemna minor was soaked for 4 h in pond water where wild L. minor was naturally flourishing. Seven of the eight surface-colonizing bacterial strains were found capable of promoting the growth of L. minor. This high appearance of plant growth-promoting bacteria (PGPB) suggests that association of environmental bacteria is generally beneficial rather than harmful for host plants. One of the PGPB, Pseudomonas sp. Ps6, enhanced the growth of L. minor by 2–2.5-fold in 10 days. This activity was higher than that previously reported for Acinetobacter calcoaceticus P23, which enhanced growth of L. minor by 1.5–2-fold. Ps6 mostly adhered to and colonized the root rather than the frond, a leaf-like structure of duckweed where P23 preferentially adheres. It was expected that these two strains can share niches, coexist, and enhance the growth of duckweed additively upon co-inoculation. However, contrary to expectation, the growth of L. minor was enhanced by only 2.3-fold by co-inoculation of these two bacteria. P23 lowered the initial adhesion of Ps6 cells by 98.2% on the fronds and by 79.5% on the roots. However, initial adhesion of P23 cells to the roots increased dramatically, by 47.2-fold, following co-inoculation with Ps6. However, the number of P23 cells decreased dramatically to 0.7% on the root and to 3.6% on the frond after 10 days, whereas Ps6 cells increased by 12.5-fold on the frond and kept 69% on the root, thereby eventually restoring the population on the plant surfaces. Because duckweed is the fastest growing vascular plant and it is easy to grow an aseptic and axenic plant, the duckweed/bacteria co-culture system will be a model platform for studying multiple interactions among host plants and the associated bacteria.     

Arsenic Uptake by Lemna minor in Hydroponic System

Arsenic is hazardous and causes several ill effects on human beings. Phytoremediation is the use of aquatic plants for the removal of toxic pollutants from external media. In the present research work, the removal efficiency as well as the arsenic uptake capacity of duckweed Lemna minor has been studied. Arsenic concentration in water samples and plant biomass were determined by AAS. The relative growth factor of Lemna minor was determined. The duckweed had potential to remove as well as uptake arsenic from the aqueous medium. Maximum removal of more than 70% arsenic was achieved at initial concentration of 0.5 mg/l arsenic on 15th day of experimental period of 22 days. Removal percentage was found to decrease with the increase in initial concentration. From BCF value, Lemna minor was found to be a hyperaccumulator of arsenic at initial concentration of 0.5 mg/L, such that accumulation decreased with increase in initial arsenic concentration.     

Phytoremediation applications in natural condition and in mesocosm: The uptake of cadmium by Lemna minuta Kunth,a non-native species in Italian watercourses

Metal pollution in water and soil is an environmental and public health issue. Cadmium (Cd) is included in the list of priority hazardous substances in the European Water Framework Directive. Phytoremediation system is a cost-effective, plant-based approach that takes advantage of the ability of plants to concentrate elements and compounds from the environment and to metabolize various molecules in their tissues. We studied the presence and the importance of an invasive species, such as Lemna minuta, in the environment and the effects of Cd pollution on this species. Growth, removal, and tolerance were evaluated for different Cd concentrations and different times of plant exposure. Overall, the results show that L. minuta has a good capacity of growth, metal bioconcentration, and tolerance up to 3 days of exposure at 0.5 and 1.5 mg L^?1 of Cd. In particular, L. minuta was able to accumulate Cd up to 3771 mg kg^?1 on dry mass basis. We can conclude that L. minuta possesses a great capability of Cd absorption and accumulation, thus supporting a potential use of this species in designing a metal bioremediation system in phytoremediation field.     

Low levels of intraspecific genetic variation at a rapidly evolving chloroplast dna locus in North American duckweeds (Lemnaceae)

Although most previous studies on chloroplast (cp) DNA variation in plants have concentrated on systematics and evolution above the species level, intraspecific variation in cpDNA is common and has provided useful insights into population-level evolutionary processes. Polymerase chain reaction methods were used to examine restriction site and sequence variation in the chloroplast rpLI6 gene within and among populations of duckweed species (Spirodela and Lemna) from the southern and eastern United States. To our knowledge, the rpL16 region has not previously been used to investigate cpDNA variation in nature. While considerable restriction site and sequence variation were detected among species, no variation was found within populations of either of the two species (S. punctata and L. minor) selected for sequence analysis, and S. punctata showed no interpopulational variation. Two cpDNA haplotypes were identified in L. minor, with one haplotype restricted to three sites in Louisiana and the other found in all other populations sampled. This paucity of variation cannot be readily explained as the result of a low mutation rate. In general, group II introns appear to be subject to very little functional constraint, and extensive sequence differences have been found between species in the chloroplast rpL16 intron in particular. However, factors such as historical range expansions and contractions, founding effects, fluctuations in local population size, and natural selection may play a role in reducing cpDNA sequence variability in these species.     

Real-time in vivo visualization of oxidative stress in duckweed (Lemna minor L.)

An experimental set-up which enabled non-invasive, real-time reactive oxygen species (ROS) visualization on a whole plant level was constructed. In the test organism, Lemna minor L. (common duckweed), apoplastic and symplastic oxidative stress was evaluated by exposure to menadione (50 μM), menadione (50 μM) + ascorbate (100 μM) or neither for control. Menadione (50 μM) caused a statistically significant increase in H₂DCFDA fluorescence in the apoplast after 60 minutes of exposure. The addition of ascorbate (100 μM) in the test medium significantly decreased apoplastic oxidative stress. 50 μM menadione caused an increase in symplastic H₂DCFDA fluorescence in 57% of fronds. The exposure of L. minor plants to both menadione and ascorbate decreased the rate of fluorescence intensity accumulation in the symplast to control levels. The method has proven to be quick and straightforward and could be applied to a range of chemicals in various physiological and toxicological plant studies. The advantages of the set-up and different possible artefacts are discussed.