Study of germination efficiency and temperature/drowning resistance in some ornamental plants treated with ultra high dilute compounds

The ultra-high dilutions (UHDs) can be used for decreasing stress conditions causing by climate variations. The present research investigated the effects of ultra-highdilutions (UHDs) on some ornamental plants, germination, and hormonal variations.Methods: In order to study the effect of UHDs (Calendula officinalisCalen. andArnica montanaArn.) on the physiological, primary metabolite, and hormonal variations of theOryza sativa L. (rice), 104 experiments were designed and statistically analyzed using the Design Expert 7.0.1 software over the general factorial design methodology. Two qualitative factors, including the UHDs/placebo usage and the type of plant usage, and two quantitative factors, including temperature and irrigation, were studied. The validated analysis was subjected to more extended studies on the variations in physiological growth, carbohydrate, protein content, and levels of plant hormones, including gibberellic acids, indole acetic acid, abscisic acid, and salicylic acid.Results: The statistical analysis resulted in a prediction model which was more than 75% correlates with experimental results. The results showed that the UHDs increased the carbohydrate and protein content of seedlings. Also, compared to placebo, the levels of hormones GA3 and IAA in all samples increase, and the amount of GA4 decreases. The amount of ABA and SA hormones inS. officinalisincreased under UHDs treatment while decreasing in the other two samples.Conclusion: The use of UHDs leads to an increase in the production of carbohydrate and protein content. Moreover, it causes significant variations in the growth-inducing hormone and increases the tolerance of seeds under higher/lower temperatures and draught/drowning. The results of this study open up a window to reduce germination survival and increase their resistance to sudden climate change.

Effects of Exogenous Nitric Oxide on Growth and Alkaloid Accumulation of Catharanthus roseus Seedlings under Light-Shading Treatment / 中国药学杂志

OBJECTIVE: To explore the molecular mechanism of no regulating alkaloid accumulation under light- shading stress. METHODS: The seedlings of Catharanthus roseus (C. roseus) were used as material, and the leaves were sampled 7 d later. Leaves growth morphology were studied, the contents of alkaloids and hormones were determined by HPLC and the genes expression were analyzed by fluorescence quantitative RT-PCR. RESULTS: Under compound treatment (T3), the growth index of leaves from C. roseus significantly increased, and the content of hormones and alkaloids increased significantly. The expression of Myc1 and key genes (G10h, Tdc, Sgd and Str) in the pathway were significantly up-regulated. Correlation analysis showed that the contents of 6-BA and ABA were positively correlated with Myc1 expression (P<0.01); Myc1 expression was positively correlated with the expression of Tdc, Str, Sgd, and Dat (P<0.01); there was a significant positive correlation between the genes expression and alkaloid content (P<0.01). CONCLUSION: The exogenous NO donors (SNP) could act as a signaling molecule and regulate the increase of hormones content under light-shading stress, increase the high expression of transcription factors Myc1, activate the expression of key genes in the metabolic pathway of alkaloids, and increase the content of vinblastine and vincristine in C. roseus.

Selection of suitable reference genes for abiotic stress-responsive gene expression studies in peanut by real-time quantitative PCR

Background: Because of its strong specificity and high accuracy, real-time quantitative PCR (RT-qPCR) has been a widely used method to study the expression of genes responsive to stress. It is crucial to have a suitable set of reference genes to normalize target gene expression in peanut under different conditions using RT-qPCR. In this study, 11 candidate reference genes were selected and examined under abiotic stresses (drought, salt, heavy metal, and low temperature) and hormone (SA and ABA) conditions as well as across different organ types. Three statistical algorithms (geNorm, NormFinder and BestKeeper) were used to evaluate the expression stabilities of reference genes, and the comprehensive rankings of gene stability were generated. Results: The results indicated that ELF1B and YLS8 were the most stable reference genes under PEG-simulated drought treatment. For high-salt treatment using NaCl, YLS8 and GAPDH were the most stable genes. Under CdCl2 treatment, UBI1 and YLS8 were suitable as stable reference genes. UBI1, ADH3, and ACTIN11 were sufficient for gene expression normalization in low-temperature experiment. All the 11 candidate reference genes showed relatively high stability under hormone treatments. For organs subset, UBI1, GAPDH, and ELF1B showed the maximum stability. UBI1 and ADH3 were the top two genes that could be used reliably in all the stress conditions assessed. Furthermore, the necessity of the reference genes screened was further confirmed by the expression pattern of AnnAhs. Conclusions: The results perfect the selection of stable reference genes for future gene expression studies in peanut and provide a list of reference genes that may be used in the future.

Determination of Plant Hormones by Capillary Electrophoresis Based on Polymer Micelles / 分析化学

With the increasing number of food safety problems caused by the abusing and misusing of plant hormones, it will be increasingly important to strengthen the trace detection of plant hormones in foods. In this study, benzyl methacrylate_methyl methacrylate_based amphiphilic polymer micelles were prepared, and a method for trace determination of five plant hormones was established by using these micelles. This method was rapid, sensitive and reproducible. Through optimization, the best experimental conditions were obtained as follows: 2 g/L polymer micelles, 50 mmol/L NaOH_H3BO3 buffer (pH 9. 2), 15 kV running voltage. Based on this novel method, naphthalene acetic acid in the solution of rooting powder was determined.

Anatomical Changes during Rooting of Mangroves - Avicennia officinalis and Excoecaria agallocha.

Mangrove forests continue to disappear all over the world due to a number of reasons. This is the study made to screen the anatomical and biochemical changes during the rooting process. Two important mangrove species have been examined based on their salt relationship natures: Avicennia officinalis subsp. australasica (salt excreting plant) and Excoecaria agallocha L (salt accumulating plant). The plants were treated with different root promoting hormones like IBA, IAA and NAA at different concentrations for three minutes each. After 45 days of the growth period root growth and anatomical changes were observed. The roots originated from the deeper zone of the secondary xylem after several anatomical variations in the hormone treated plants. This is the first trial which may help to determine the mass propagation of these commercial and medicinally important mangroves.

Morphogenesis in vitro of chrysanthemum (Dendranthema grandiflora tzelev) from stem and pedicel explants / Morfogênese in vitro de crisântemo (Dendranthema grandiflora tzelev) a partir de explantes de caule e pedicelo

Visando a adequação de um protocolo de morfogênese in vitro para plantas de crisântemo (Dendranthema grandiflora Tzelev) analisou-se três fatores que afetam a frequência da regeneração. Foram avaliadas três concentrações de ácido indolil-acético (AIA) e 6- Benzilaminopurina (BAP), a fonte de explante (caule e pedicelo) e a orientação de inoculação do explante no meio de cultivo. Ao final de 15-30 dias de cultivo, observou-se que a fonte de explante, o tipo de explante e a interação explante e meio de cultivo afetaram significativamente a frequência de regeneração (FR) e o número de ramos por explantes (NRE) em crisântemo. Os explantes de pedicelo apresentaram maiores frequências de regeneração (15-20 dias) em meio de cultivo suplementado com 0,5 mg L-1 de BAP e 2,0 mg L-1 de AIA e os explantes caulinares (30 dias) em meio com 2,0 mg L-1 de BAP e 0,5 mg L-1 de AIA. Respostas diferenciais em relação à morfogênese foram observadas quando as duas posições do pedicelo foram inoculadas no meio de cultivo, a região seccionada em contato com o meio de cultivo suplementado com 0,5 mg L-1 de BAP e 2,0 mg L-1 de AIA proporcionou maior frequência de brotos. Foram obtidas plantas com flores 45-60 dias após a transferência para a casa de vegetação. As flores apresentavam os mesmos padrões e características das flores das plantas doadoras de explantes. Podese concluir que a o tipo de explante, sua orientação em relação ao meio de cultura bem como as combinações de reguladores de crescimento são fatores-chave envolvidos no sucesso da regeneração de plantas de crisântemo in vitro.

Aiming the optimization of a regeneration protocol for chrysanthemum plants (Dendranthema grandiflora), experiments of regeneration in vitro were accomplished, stem explants and pedicel inoculated in several shoots induction mediuns. The stem explants and of pedicels they were sectioned longitudinally and inoculated with the face opposed to the section in the medium of cultivation with the basic salts MS, with different concentrations and combinations of growth regulators BAP and AIA. At the end of 15-30 days of cultivation, significant differences were observed in the regeneration frequency, number of shoots for explante and in the length of the shoots among the appraised explantes. Explants of pedicel presented potential the best to the explants stem. The pedicel explants presented larger regeneration frequencies in medium of cultivation contends basic salts of MS, with 0,5 mg L-1 of BAP and 2,0 mg L-1 of AIA and the explantes of stem in medium with basic salts of MS + 2,0 mg L-1 of BAP and 0,5 mg L-1 of AIA. They were obtained plants with flowers 45-60 days after the transfer to the vegetation house. In function of the obtained results, it can be concluded that the choice of the explant type and the medium of cultivation are factor-key involved in the success of the regeneration of plants for posterior use in experiments of genetic transformation.

Effects of photoperiod, plant growth regulators and culture media on in vitro growth of seedlings of Cyrtochilum loxense (Lindl.) Kraenzl. an endemic and endangered orchid from Ecuador

Cyrtochilum loxense (Lindl.) Kraenzl. is an endemic and seriously endangered orchid species endemic in the Loja Province (Southern Ecuador). The main goals of this research were to analyze how culture media, plant growth regulators and photoperiod affect the growth of C. loxense. Eight month old plants (approximate 1 - 1.5 cm in height) obtained by in vitro germination, were cultivated on MS media or Knudson C; MS with three levels of naphthalene acetic acid (NAA) and 6-benzylaminopurine (BAP) (2/0.5; 1/0.5 y 0.5/ 0.5 mg-1L); and three photoperiodic regimes (24/0, 16/8, 8/16 h) on MS with and without plant growth regulators. No significant differences of shoot induction were observed on media with or without plant growth regulators, and all tested photoperiods. The highest growth (1.2 cm) was observed in plantlets cultivated on growth regulator-free media with a 16/8 photoperiod. Also the shoot and root formation was better in this species in absence of plant growth regulators. Probably this response is due to the endogenous hormone levels in the tissues or due to the kind and concentrations of PGRs used were too low to induce positive morphogenetic responses.

Cyrtochilum loxense (Lindl.) Kraenzl. es una orquídea endémica y críticamente amenazada, distribuida en la provincia de Loja (Sur del Ecuador). Los objetivos principales de esta investigación fueron analizar la influencia del medio de cultivo, reguladores de crecimiento vegetal y fotoperíodo en el crecimiento de C. loxense. Plantas de ocho meses (altura aproximada de 1 - 1.5 cm.) obtenidas por germinación in vitro, fueron cultivadas en medio MS y Knudson C; medio MS con tres niveles de ácido naftalen acético (ANA) y 6-bencil amino purina (BAP) (2/0.5; 1/0.5 y 0.5/ 0.5 mg-1L); y tres fotoperíodos (24/0, 16/8, 8/16 h) en MS con o sin reguladores de crecimiento vegetal (RCV). No se observaron diferencias significativas en la inducción de brotes en medio con o sin RCV, entre todos los fotoperíodos testados. El tamaño más grande (1.2 cm) se registró en plántulas cultivadas en medio libre de reguladores de crecimiento vegetal, incubadas en un fotoperíodo de 16/8. Además, la mayor brotación y enraizamiento se observó para esta especie en medio libre de reguladores de crecimiento vegetal. Probablemente esta respuesta es debida a los niveles endógenos de hormonas vegetales en el tejido o el nivel de RCV usado fue muy bajo para inducir alguna respuesta morfogénica.

Micropropagação e endorreduplicação em pitaya vermelha / Micropropagation and endoreduplication in red pitaya

A micropropagação facilita a obtenção de mudas de mudas de pitaya, Hylocereus undatus Haw; porém, pode propiciar alterações genéticas, como a polissomatia, endopoliploidia ou endorreduplicação. Neste trabalho, objetivou-se desenvolver protocolo para micropropagação e verificar a endorreduplicação em pitaya vermelha. Cladódios de pitaya foram micropropagados em meio de cultura MS, com 0,1 mg L-1 de ANA, acrescido de BAP e cinetina, nas dosagens 0, 1, 5 e 10 mg L-1, sendo o meio MS sem reguladores de crescimento utilizado como tratamento controle. O delineamento experimental utilizado foi inteiramente casualizado em fatorial 2x8 (ausência e presença de luz e 8 diferentes meios de cultura), totalizando 16 tratamentos, sendo 4 repetições de 3 explantes. O experimento foi conduzido na presença e ausência de luz em sala de crescimento a 25 ± 2ºC, irradiância média de 42 W m-2 e fotoperíodo de 16 horas, por 120 dias. A análise de citometria de fluxo foi utilizada em 3 explantes por tratamento. Amostras de cladódios, juntamente com o padrão interno (ervilha) foram triturados com 1 mL de tampão de extração LB01 e 25 mL de iodeto de propídeo, analisados em citômetro de fluxo. Os explantes submetidos à ausência de luz não responderam aos tratamentos. O meio de cultura que proporciona melhor crescimento e desenvolvimento in vitro é o MS acrescido de 0,1 mg L-1 de ANA e BAP ou cinetina nas concentrações de 1 ou 5 mg L-1. O meio de cultura que proporciona aos explantes maior variação na quantidade de DNA é MS com 10 mg L-1 de BAP e 0,1 mg L-1 de ANA. O fenômeno da endorreduplicação é observado em todos os tratamentos analisados.

Micropropagation makes the obtaining of seedlings of red pitaya easier, but, it can cause genetic mutations such as polysomaty, endopolyploidy or endoreduplication. In this work, it was aimed to develop a protocol for micropropagation and check the endoreduplication in red pitaya. Pitaya cladodes were micropropagated im MS culture medium with 0.1 mg L-1 of ANA added of BAP and kinetin at the dosages of 0, 1, 5 and 10 mg L-1, the MS medium without any growth regulators being used as the control treatment. The experimental design utilized was the completely randomized in factorial 2 x 8 (absence and presence of light and 8 different culture media), amounting to 16 treatments, namely, 4 replications of 3 explants. The experiment was conducted in the presence and absence of light in a growth room at 25 ± 2 ºC, average irradiance of 42W m-2 and photoperiod of 16 hours for 120 days. The flow cytometry analysis was utilized on three explants per treatment. Cladode samples, along with the internal pattern (pea) were ground with 1 ml of extraction buffer LB 01 and 25 mL of propide iodide, analyzed in flow cytometer. The explants submitted to the absence of light did not respond to the treatments. The culture medium which brings about the best in vitro growth and development is the MS added of 0.1 mg L-1 of ANA and BAP or kinetin at the concentrations of 1 or 0.5 mg L-1. The culture medium which provides to the explants greatest variation in the amount of DNA is MS with 10 mg L-1 of BAP and 0.1 mg L-1 of ANA. The phenomenon of endoreduplication is found in all the treatments investigated.

Ascorbic acid: an enigmatic molecule to developmental and environmental stress in plant.

Ascorbic acid (AA) is present in all eukaryotes including animals and plants and lack completely in prokaryotes with exception of cyanobactaria. Today, AA has gained significant place in plant science, mainly due to its properties (antioxidant and cellular reductant etc.), and multifunctional roles in plant growth, development, and regulation of large spectrum of plant defense mechanisms against environmental stresses. Some studies suggests that the endogenous AA has been implicated in the promotion of plant growth and development by involving in a complex array of phytohormonemediated signaling networks that ties together various developmental and environmental stresses. In the last few years, the role of AA in tolerance of plants to environmental stress has established much consideration. As it is evident from the present review, recent progress on AA potentiality in tolerance of plants to environmental stresses has been impressive. Indeed, AA also plays an important role in resistance to developmental programmes (stresses, e.g., bud induction, flowering and senescence) and environmental stresses (e.g., osmotic, temperature, pathogenesis and weedicides and herbicides).

Molecular battles between plant and pathogenic bacteria in the phyllosphere

The phyllosphere, i.e., the aerial parts of the plant, provides one of the most important niches for microbial colonization. This niche supports the survival and, often, proliferation of microbes such as fungi and bacteria with diverse lifestyles including epiphytes, saprophytes, and pathogens. Although most microbes may complete the life cycle on the leaf surface, pathogens must enter the leaf and multiply aggressively in the leaf interior. Natural surface openings, such as stomata, are important entry sites for bacteria. Stomata are known for their vital role in water transpiration and gas exchange between the plant and the environment that is essential for plant growth. Recent studies have shown that stomata can also play an active role in limiting bacterial invasion of both human and plant pathogenic bacteria as part of the plant innate immune system. As counter-defense, plant pathogens such as Pseudomonas syringae pv tomato (Pst) DC3000 use the virulence factor coronatine to suppress stomate-based defense. A novel and crucial early battleground in host-pathogen interaction in the phyllosphere has been discovered with broad implications in the study of bacterial pathogenesis, host immunity, and molecular ecology of bacterial diseases.

War of hormones over resource allocation to seeds: Strategies and counter–strategies of offspring and maternal parent.

It is suggested that maternal parent and offspring have conflicting interests over the extent of resource allocation to developing seeds. While maternal parent would be selected to allocate her resources optimally among her offspring, the latter would be selected to demand more. In animals, offspring are known to demand additional resources either visibly (through intense vocal calls) or subtly through the production of hormones. In plants though parent offspring conflict over resource allocation has been invoked, the mechanism through which the parent and offspring interact in regulating resource allocation into developing seeds is not yet clear. In this paper, we propose that the strategies and counter–strategies of the offspring and mother during the development of seeds might be manifested through the production of appropriate growth hormones. Accordingly, we predict (i) hormones that mobilize resources into seeds (e.g. auxins and gibberellic acid) shall be synthesized exclusively by the offspring tissue and (ii) hormones that inhibit resource flow in to seeds (e.g. abscisic acid) be produced exclusively by the maternal tissue. We show that these predictions are supported by existing literature on the temporal dynamics and source of production of growth hormones during seed development. Finally, we suggest that such analysis viewing the production of different hormones during early seed development, as strategies and counter–strategies of mother and offspring tissue, helps ofer a meaningful interpretation of the otherwise complex dynamics of hormone fluxes.