Cloning and structural elucidation of a brassinosteroids biosynthetic gene (Atdwarf4) and genetic transformation of Indian mustard (Brassica juncea L.)

Phytohormones play critical roles in plant growth and development. Brassinosteroids (BRs) are essential group of phytohormones required for optimum growth of plants and their deficiency causes distinctive dwarf phenotypes in plants. Homeostasis of BRs in plants is maintained by DWARF4 enzyme that mediates multiple 22?-hydroxylation steps in brassinosteroid biosynthesis. Arabidopsis plants over-expressing DWARF4 show increase in inflorescence, number of branches and siliques; thereby increased number of seeds/plant. This suggests that engineering DWARF4 biosynthesis in Brassica plant can be strategized to enhance yield in mustard. In the present study (i) we cloned dwarf4 gene from Arabidopsis using gene specific PCR strategy, (ii) elucidated the three-dimensional structure of DWARF4 protein at molecular level where it revealed presence of four beta sheets and 20 alpha-helices, and (iii) transformed mustard cultivar Pusa Jaikisan with an objective to develop transgenic mustard with enhanced number of siliques. We obtained several putative transgenics with an average transformation efficiency of 3.3%. Molecular characterization with nptII specific primers confirmed presence of transgene in six putative transgenic plants.

Isolation and Characterization of Plant Growth-Promotion Diazotrophic Endophytic Bacteria Associated to Sugarcane (Saccharum officinarum L.) Grown in Paraíba, Brazil

Abstract: Sugarcane is an important Brazilian commodity, being usually cultivated in soils with low natural fertility. This study aimed to isolate diazotrophic endophytes from sugarcane tissues and evaluate the morphological and physiological characteristics of their colonies as well as their plant growth-promoting (PGP) traits in select diazotrophic endophytic bacteria. Fifty-six bacterial isolates were identified in the sugarcane tissues, and these isolates presented distinct morphological and physiological traits. A total of thirty-five bacterial isolates were biochemically evaluated. Overall, Bacillus was the dominant genus. Isolates of Methylobacterium spp. and Brevibacillus agri were present only in leaves, while Herbaspirillum seropedicae occurred only in stems. Except to IPA-CF45A, all isolates were nitrogenase positive. All endophytes exhibit production of indol 3-acetic acid. Over 50% of endophytes solubilize phosphate, release N-acyl homoserine lactones, and present the activity of 1-aminocyclopropane-1-carboxylic acid deaminase, catalase, lipase and protease. The network analysis showed that isolates belonged to Burkholderia, Herbaspirillum, and Methylobacterium interact with Bacillus. Bacterial endophytes exhibited distinct morphological, physiological, and PGP traits that are useful for sustainable agriculture, highlighting the isolates IPA-CC33, IPA-CF65, IPA-CC9 and IPA-CF27. Further studies on the effects of these diazotrophic endophytes and their potential for providing microbial inoculants for improving sugarcane fields will provide valuable information to maintain the sustainability and environment quality.

Research progress on mechanism of phytohormones in regulating flavonoid metabolism / 中国中药杂志

Phytohormones play an important role at all stages of plant growth, influencing plant growth and development and regulating plant secondary metabolism, such as the synthesis of flavone, flavonol, anthocyanin, and other flavonoids. Flavonoids, a group of important secondary metabolites ubiquitous in plants, have antioxidative, anti-microbial, and anti-inflammatory activities and thus have a wide range of potential applications in Chinese medicine and food nutrition. With the development of biotechnology, phytohormones' regulation on flavonoids has become a research focus in recent years. This study reviewed the research progress on the mechanism of common phytohormones, such as abscisic acid, gibberellin, methyl jasmonate, and salicylic acid, in regulating flavonoid metabolism, and discussed the molecular mechanism of the synthesis and accumulation of flavonoids, aiming at clarifying the key role of phytohormones in modulating flavonoid metabolism. The result is of guiding significance for improving the content of flavonoids in plants through rational use of phytohormones and of reference value for exploring the mechanism of hormones in regulating flavonoid metabolism.

In vitro multiplication of wild Manihot species with different naphthaleneacetic acid and benzylaminopurine concentrations

In vitro multiplication is an important tissue culture technique that is capable of efficiently producing seedlings at any scale. It is a propagation method based on the aseptic culture of small propagules in a suitable culture medium to enable plant regeneration. Multiplication experiments conducted in vitro to set protocols adapted to wild Manihot species have used modified mineral salts and MS vitamins as basic culture medium. Here, 25 treatments based on combinations of the regulators benzylaminopurine (BAP) and naphthaleneacetic acid (NAA) at 0, 0.025, 0.05, 0.075, and 0.1 mg L-1 were used for in vitro multiplication of three genotypes of wild Manihot species (M. violaceae Pohl Müll. Arg., M. pseudoglaziovii Pax & Hoff., and M. flabellifolia Pohl). Plant height and the number of 1 cm minicuttings, number of roots, shoots, green leaves and senescent leaves were recorded 120 days after explant inoculation. M. violaceae Pohl. Müll. Arg. and M. flabellifolia Pohl. presented favorable results with 0.05 and 0.025 mg L-1 NAA, respectively. Culture medium lacking NAA and BAP favored the in vitro growth of M. pseudoglaziovii Pax & Hoff.

Antagonic and plant growth-promoting effects of bacteria isolated from mine tailings at El Fraile, Mexico / Efecto antagónico y promotor del crecimiento vegetal de bacterias aisladas de losjales mineros de El Fraile, México

Abstract Mine tailings contain high concentrations of heavy metals such as As, Pb, Cu, Mn, andFe, which are detrimental to the health of humans and the environment. In tailings at the ElFraile mine in Guerrero, Mexico, some plant species are apparently tolerant of heavy metals andcan be found growing in the tailings. These plants could be associating with heavy metal-tolerantbacteria that promote plant growth and improve biomass production, and these bacteria couldbe a useful alternative for bacteria-assisted phytoremediation. The objective of this study wasto isolate bacteria detected in the mine tailings at El Fraile-Taxco, focusing on those in the soilfrom the rhizosphere, the inner tissue of the root, leachate, and water, which have the poten-tial to promote plant growth. The ability of the isolated bacteria to promote plant growth wasevaluated in vitro. Of the 151 morphotypes isolated, 51% fix nitrogen, 12% dissolve phosphates,and 12%, 39.7%, and 48.3% produce indole acetic acid, gibberellins, and siderophores, respec-tively. In addition, 66.7% were observed to produce lytic enzymes, such as proteases, celluloses,lipases, esterases, and amylases, which exhibited activity against Fusarium, Aspergillus, andColletotrichum. The use of 16S rRNA analysis led to the identification of the bacterial generaChryseobacterium, Bacillus, Pseudomonas, Mycobacterium, Staphylococcus, Curtobacterium,Enterobacter, Agrobacterium, Ochrobactrum, Serratia, Stenotrophomonas, and Acinetobac-ter. The bacteria isolated from the rhizosphere exhibited the greatest ability to fix nitrogenand produced indole acetic acid, gibberellins, siderophore, and lytic enzymes. In addition, theisolates collected from the soil samples demonstrated ability to solubilize phosphate.

Resumen Los jales mineros contienen una alta concentración de metales pesados como As, Pb, Cu, Mn y Fe. Estas altas concentraciones de metales son perjudiciales para la salud humana y el medio ambiente. En los jales mineros de El Fraile, México, es posible detectar especies de plantas tolerantes a los metales pesados; estas plantas podrían estar asociadas con bacterias capaces de promover su crecimiento, además de poseer actividad antagonista contra hongos. El objetivo de este estudio fue aislar de diferentes microambientes (suelo rizosférico, tejido de raíz, lixiviado y agua) del área del jale El Fraile bacterias con potencial de promover el crecimiento vegetal y actividad antagonista contra hongos fitopatógenos. Estudios in vitro demostraron que el 51% de los morfotipos aislados (151 en total) fijan nitrógeno y el 12% disuelven fosfatos. Asimismo, el 12, 39,7 y 48,3% producen ácido indolacético, giberelinas y sideróforos, respectivamente. Por otro lado, se observó que el 66,7% producía enzimas líticas como proteasas, celulasas, lipasas, esterasas y amilasas, además de exhibir actividad antagonista contra Fusarium, Aspergillus y Colletotrichum. Mediante análisis del gen 16S ARNr, se identificó a estas bacterias como pertenecientes a los géneros Chryseobacterium, Bacillus, Pseudomonas, Mycobacterium, Staphylococcus, Curtobacterium, Enterobacter, Agrobacterium, Ochrobac-trum, Serratia, Stenotrophomonas y Acinetobacter. Las bacterias de la rizosfera exhibieron la mayor capacidad para fijar nitrógeno y produjeron ácido indolacético, giberelinas, sideróforos y enzimas líticas. Además, se detectó que las cepas aisladas de suelo rizosférico eran las que tenían la capacidad de solubilizar fosfatos.

Principales reguladores hormonales y sus interacciones en el crecimiento vegetal / Main hormonal regulators and their interactions in plant growth

Resumen Una hormona vegetal o fitohormona es un compuesto producido internamente por una planta, que ejerce su función en muy bajas concentraciones y cuyo principal efecto se produce a nivel celular, cambiando los patrones de crecimiento de los vegetales y permitiendo su control. Los reguladores vegetales son compuestos sintetizados químicamente u obtenidos de otros organismos y son, en general, mucho más potentes que los análogos naturales. Es necesario tener en cuenta aspectos críticos como oportunidad de aplicación, dosis, sensibilidad de la variedad, condición de la planta, etc., ya que cada planta requerirá de unas condiciones específicas de crecimiento que pueden afectarse por la concentración de ellos en el medio. Los reguladores vegetales son productos sintéticos que se han convertido en las primeras herramientas capaces de controlar el crecimiento y actividad bioquímica de las plantas por lo que su uso ha aumentado en los últimos años. Esta revisión busca hacer una recopilación bibliográfica de los primeros acontecimientos de la aplicación de los reguladores de crecimiento vegetal. Se presentan las principales características fisiológicas que pueden desarrollar la aplicación de estos sobre el crecimiento vegetal a nivel celular y su repercusión a nivel fenotípico; además, se describen las principales fitohormonas más conocidas en la aplicación biotecnológica. Entre ellas se encuentran auxinas, giberelinas, citoquininas, ácido abscísico, ácido salicílico, poliaminas, jasmonatos y derivados, brasinoesteroides, etileno y estrigolactonas. Se detallan las principales funciones a nivel del metabolismo vegetal y sus posibles interacciones intra e intercelular.

Abstract A vegetal hormone or phytohormone is a compound produced inside by a plant, that work in low concentrations and whose mainly effect occurs at the cellular level, changing the patterns of grow in vegetal organism and allow their control. The plant grows regulators are synthetic chemical compounds that can be obtained by the organism different to the plants and are more stronger that natural analogues, is necessary have in consideration different aspects like application opportunity, concentration, sensibility, plant condition, it's because every plant requires specific conditions of grow that can be affected by phytohormonal concentration in the soil or medium. The vegetal regulators are synthetic products that it has been converted in the mainly tool available to control the growing and biochemical activity of the plant and for that reason their use are increased in the last years. This review makes a bibliography compilation of the first events in the beginning of vegetal plant regulators application and presents the mainly physiologic characteristics that can be developed by application of vegetal hormones and their action in cellular and phenotypic response, furthermore we explain some of the mechanisms of action of the mainly ten phytohormones more knowledge in the biotechnological application: auxins, gibberellins, cytokinins, abscisic acid, salicylic acid, polyamines, jasmonates and derived compounds, brassinosteroids, ethylene, and strigolactones. We describe their mainly functions to vegetal metabolism and their possible interactions intracellular and extracellular.

Towards the micropropagation of Euphorbia cyathophora Murray: a wild plant species with medicinal and ornamental potential / Micropropagação de Euphorbia cyathophora Murray: uma especie de planta selvagem com potencial medicinal e ornamental

ABSTRACT: Euphorbia cyathophora Murray is a plant species with medicinal and ornamental potential but whose in vitro propagation is unknown. Two experiments were performed to know 1) the effect of light (WL) at 25±2 °C and 20 μmol m-2 s-1 luminous intensity, or darkness at 20±2 °C, for the first 20 days and subsequent placement in light (DKL) as previously described, and 2) the best combination of plant growth regulators: benzyl amino purine (BAP), kynetin (KIN), 2-isopentenyl adenine (2iP) and thidiazuron (TDZ) in combination with naphthaleneacetic acid (NAA) and the effect of both light conditions. The DKL treatment did not promote the growth of leaves, cotyledons, or plantlet height; in contrast, WL produced a more developed root system with early appearance of tertiary roots. In regards to the interaction of plant regulators and light effect, only the treatment with 1 mg BAP L-1 and 0.1 mg NAA L-1 generated shoots in both light conditions (DKL and WL). Darkness (DKL) generated more indirect greatest morphogenic responses (callus formation).

RESUMO: Euphorbia cyathophora Murray é uma espécie vegetal com potencial medicinal e ornamental, mas cuja propagação in vitro é desconhecida. Dois experimentos foram realizados para conhecer 1) o efeito da luz (WL) a 25±2 °C e 20 μmol m-2 s-1 intensidade luminosa, ou escuridão a 20±2 °C, nos primeiros 20 dias e posterior colocação em luz (DKL) como descrito anteriormente, e 2) a melhor combinação de reguladores de crescimento vegetal: benzil amina purina (BAP), kynetina (KIN), 2-isopentenil adenina (2iP) e thidiazuron (TDZ) em combinação com ácido naftalenoacético (ANA) e o efeito de ambas as condições de luz. O tratamento com DKL não promoveu o crescimento de folhas, cotilédones ou altura das plântulas. Em contraste, a WL produziu um sistema radicular mais desenvolvido com o aparecimento precoce de raízes terciárias. Com relação à interação dos reguladores vegetais e efeito luminoso, apenas o tratamento com 1 mg de BAP L-1 e 0,1 mg de NAA L-1 gerou brotações em ambas as condições de luz (DKL e WL). Escuridão (DKL) gerou mais respostas morfogênicas mais diretas (formação de calo).

Coping with drought: stress and adaptive mechanisms, and management through cultural and molecular alternatives in cotton as vital constituents for plant stress resilience and fitness

Increased levels of greenhouse gases in the atmosphere and associated climatic variability is primarily responsible for inducing heat waves, flooding and drought stress. Among these, water scarcity is a major limitation to crop productivity. Water stress can severely reduce crop yield and both the severity and duration of the stress are critical. Water availability is a key driver for sustainable cotton production and its limitations can adversely affect physiological and biochemical processes of plants, leading towards lint yield reduction. Adaptation of crop husbandry techniques suitable for cotton crop requires a sound understanding of environmental factors, influencing cotton lint yield and fiber quality. Various defense mechanisms e.g. maintenance of membrane stability, carbon fixation rate, hormone regulation, generation of antioxidants and induction of stress proteins have been found play a vital role in plant survival under moisture stress. Plant molecular breeding plays a functional role to ascertain superior genes for important traits and can offer breeder ready markers for developing ideotypes. This review highlights drought-induced damage to cotton plants at structural, physiological and molecular levels. It also discusses the opportunities for increasing drought tolerance in cotton either through modern gene editing technology like clustered regularly interspaced short palindromic repeat (CRISPR/Cas9), zinc finger nuclease, molecular breeding as well as through crop management, such as use of appropriate fertilization, growth regulator application and soil amendments.

Molecular cloning and characterisation of scavenger receptor class B in pearl oyster Pinctada fuctada martensii

Background: Molluscs can accumulate carotenoids in their body tissues by predominantly feeding on aquatic plant sources. Carotenoid transport and absorption are determined by the regulation of various proteins such as Scavenger receptor class B(SR-BI). We report the identification and characterisation of pearl oyster Pinctada fuctada martensii SR-BI (PmSR-BI). The correlation between total carotenoid content (TCC) and gene expression was also estimated. Results: The full-length cDNA of PmSR-BI was 1828 bp, including an open-reading frame encoding of 1518 bp with a pI value of 5.83. PmSR-BI protein contains a hydrophobic CD36 domain and four centrally clustered cysteine residues for the arrangement of disulphide bridges. The deduced amino acid sequence had an identity of 30% to 60% with the SR-B of other organisms. Reverse transcription polymerase chain reaction analysis showed that mRNA transcripts were expressed in multiple tissues of adult pearl oyster. A higher expression of PmSR-BI gene was observed in the hepatopancreas than in the adductor muscle, gill and mantle. The TCC and gene expression of PmSR-BI were significantly correlated (P b 0.05), with a correlation coefficient of 0.978. Conclusions: The results suggested that PmSR-BI is involved in the absorption of carotenoids in the pearl oyster P. fuctada martensii.

Efficacy of a plant growth regulator in the culture of coffee / Eficácia de biorregulador na cultura do café

The purpose of this work was to evaluate the efficacy of a plant growth regulator applied at different growth stages in coffee. The experiment was conducted in the years 2010/2011 and 2011/2012 on the Docas 1 farm, using Mundo Novo IAC 379-19 coffee cultivar, with spacing 4 meter between rows and 0.8 meter between plants and 10 plants per plot. The experimental design was in blocks at random, with two factors and three repetitions. The first factor was the dose of the product (0.25 and 0.5 liter per hectare per application) and second factor was the time of applications according to the phenological stages (pre-flowering, post-flowering and pinhead). The control treatment (control) consisted of the absence of the application of the plant growth regulator. Biometric aspects (number of internodes and length of reproductive branches in centimeters, number of fruits on the fourth and fifth node on reproductive branches) and productivity were evaluated. Collected data were analyzed using Tukey test at 0.05 of significance. The application of a plant growth regulator at different growth stages of development of coffee leads to increased biometric variables in coffee: the number of internodes, the average number of fruits on the fourth and fifth node and the length of reproductive branches. The productivity of coffee can be increased with the use of plant growth regulators, particularly at the dose of 0.5 liter per hectare per application, regardless of the application period, promoting productivity up to 46.9%. It is not possible to state that the presence of substances similar to phytohormones in plant growth regulators is responsible for the increase in productivity without making isolate application of these substances.

O objetivo do trabalho foi avaliar a eficácia do biorregulador aplicado em diferentes estádios fenológicos na cultura do café. O experimento foi conduzido nos anos agrícola de 2010/2011 e 2011/2012 na fazenda Docas 1, utilizando café cultivar Mundo Novo variedade IAC 379-19, com o espaçamento de 4 m entre linhas e 0,8 m entre plantas e 10 plantas por parcela. O delineamento experimental utilizado foi o de blocos casualizados, com dois fatores e três repetições, sendo o primeiro fator dose do produto (0,25 L ha-1 e 0,5 L ha-1 em cada aplicação) e o segundo a época de aplicações de acordo com os estádios fenológicos (pré-florada, pós-florada e chumbinho) e o tratamento controle (testemunha) que constou-se da ausência de aplicação do biorregulador. Realizou-se avaliações dos aspectos biométricos (número de internódios e comprimento de ramos plagiotrópicos em centímetro, contagem dos frutos do 4º e 5º nó dos ramos plagiotrópicos) e a produtividade, e realizou-se o teste de Tukey a 0,05 de significância. A aplicação do biorregulador em diferentes estádios fenológicos de desenvolvimento do cafeeiro promove um aumento nas variáveis biométricas do cafeeiro, número de internódios, a média de frutos do quarto e quinto nó e o comprimento dos ramos plagiotrópicos. A produtividade do cafeeiro pode ser incrementada com o uso de biorreguladores, principalmente na dose de 0,5 L ha-1 por aplicação independente da época de aplicação, promovendo incrementos de até 46,9% na produtividade. Não é possível afirmar que a presença de substâncias semelhantes aos fitohormônios nos biorreguladores é responsável pelo incremento na produtividade, sem realizar a aplicação dessas isoladamente.

An Automated On-line Solid Phase Extraction-Liquid Chromatography-Tandem Mass Spectrometric Method for Six Endogenous Phytohormones Analysis in Rice / 分析化学

An automated on-line solid phase extraction-liquid chromatography-tandem mass spectrometry ( on-line SPE-LC-MS/MS ) method for the simultaneous quantification of six endogenous phytohormones including abscisic acid ( ABA ) , indole-3-acetic acid ( IAA ) , salicylic acid ( SA ) , jasmonic acid ( JA ) , indole-3-propionic acid ( IPA) and indole-3-butyric acid ( IBA) in rice tissues was described. Plant samples were extracted with methanol and on-line SPE procedure was performed with C18 SPE cartridges. Analytes were eluted to C18 analytical column with mobile phase and further analyzed by LC-MS/MS. The performance of the method was fully validated. Linearity showed good correlation (R2≥0. 99). Limits of detections (S/N=3) were in the range of 0. 1-0. 8 μg/kg. Recoveries varied between 71. 2% and 126%. The method was rapid and sensitive to determine multiple phytohormones in rice young panicles and the results were cross-validated with the off-line SPE-LC-MS/MS phytohormone analytical method. Finally, the method was applied to monitor the changes of ABA, IAA, SA and JA in wounded rice leaves. The trends were in accord with plant physiological processes.

Avaliação dos fitorreguladores auxina e giberelina na germinação e crescimento do arroz / Assessment of auxin and gibberellin on rice germination and growth / Evaluación de fitorreguladores auxina y giberelina en la germinación y crecimiento del arroz

O arroz (Oryza sativa L.) é uma planta da família das Poaceas (gramíneas), sendo um dos cereais mais cultivados no mundo. Os fitorreguladores como auxina e giberilina podem influenciar o crescimento e desenvolvimento das plantas como o arroz. O objetivo deste estudo foi avaliar a germinação e o crescimento do arroz sob diferentes doses de fitorreguladores (auxina e giberelina). Foram conduzidos dois experimentos, o primeiro para determinar a germinação e comprimento radicular das sementes de arroz. O segundo experimento foi conduzido para determinar a altura, massa seca da parte área (MSPA) e o conteúdo nitrogênio (N) da parte aérea (NPA). Nos dois experimentos foram testadas quatro doses de auxina + giberelina: 0 (testemunha); 200; 500 e 1000 mL ha­1. O delineamento experimental foi inteiramente casualizado com cinco repetições para cada tratamento nos dois experimentos. A germinação das sementes e o comprimento radicular foram aumentados com a aplicação de auxina e giberelina. A altura e o NPA foram aumentados significantemente com a aplicação de auxina e giberelina. Concluiu-se que a aplicação de auxina e giberelina na cultura do arroz aumentaram a germinação e o comprimento radicular das sementes. Além disso, aumentou a altura e o conteúdo de nitrogênio nas plantas de arroz.

Rice (Oryza sativa L.) is one of the most widely grown cereals in the world. Phytoregulators such as auxin and gibberellin can influence both growth and development of plants such as rice. The aim of this study was to assess the germination and growth of rice under different doses of phytoregulators (auxin and gibberellin). Two experiments were performed, one to determine the germination and root length of rice seeds, and the second one to evaluated height, shoot dry mass (SDM) and shoot nitrogen content (SNC). Both experiments tested four doses of auxin + gibberellin (0 (control), 200, 500 and 1000 mL ha-1). The experimental design was completely randomized with five repetitions for both experiments. The germination and root length of rice seedlings were increased by the application of auxin and gibberellin. Rice height and SNC were significantly increased with the application of auxin and gibberellin. Thus, it can be concluded that the application of auxin and gibberellin in rice increases germination, root length, plant height and nitrogen content.

El arroz (Oryza sativa L.) es una planta de la familia Poaceas (gramíneas), siendo uno de los cereales más cultivados en el mundo. Los fitorreguladores como auxina y giberelina pueden influir en el crecimiento y desarrollo de plantas, como el arroz. El objetivo de este estudio ha sido evaluar la germinación y el crecimiento del arroz bajo diferentes dosis de fitorreguladores (auxina y giberelina). Se realizaron dos experimentos, el primero para determinar la germinación y longitud de la raíz de las semillas de arroz. Se llevó a cabo un segundo experimento para determinar la altura, masa seca de la parte aérea (MSPA) y el contenido de nitrógeno (N) de la parte aérea (NPA). En ambos experimentos, se ensayaron cuatro dosis de auxina + giberelina: 0 (testigo); 200; 500 y 1000mL ha-1. El diseño experimental fue completamente al azar con cinco repeticiones para cada tratamiento en ambos experimentos. La germinación de las semillas y la longitud de la raíz se incrementaron mediante la aplicación de auxina y giberelina. La altura y el NPA se incrementaron de manera significativa mediante la aplicación de auxina y giberelina. Se concluye que la aplicación de auxina y giberelina en el cultivo de arroz incrementa la germinación y longitud de la raíz de las semillas. Además, incrementó la altura de plantas de arroz con nitrógeno.

A microdroplet cell culture based high frequency somatic embryogenesis system for pigeonpea, Cajanus cajan (L.) Millsp.

A protocol for high frequency production of somatic embryos was worked out in pigeonpea, Cajanus cajan (L.) Millsp. The protocol involved sequential employment of embryogenic callus cultures, low density cell suspension cultures and a novel microdroplet cell culture system. The microdroplet cell cultures involved culture of a single cell in 10 µl of Murashige and Skoog’s medium supplemented with phytohormones, growth factors and phospholipid precursors. By employing the microdroplet cell cultures, single cells in isolation were grown into cell clones which developed somatic embryos. Further, 2,4-dichlorophenoxyacetic acid, kinetin, polyethylene glycol, putrescine, spermine, spermidine, choline chloride, ethanolamine and LiCl were supplemented to the low density cell suspension cultures and microdroplet cell cultures to screen for their cell division and somatic embryogenesis activity. Incubation of callus or the inoculum employed for low density cell suspension cultures and microdroplet cell cultures with polyethylene glycol was found critical for induction of somatic embryogenesis. Somatic embryogenesis at a frequency of 1.19, 3.16 and 6.51 per 106 cells was achieved in the callus, low density cell suspension cultures and microdroplet cell cultures, respectively. Advantages of employing microdroplet cell cultures for high frequency production of somatic embryos and its application in genetic transformation protocols are discussed.

Coordinate Changes in Assimilatory Sulfate Reduction are Correlated to Salt Tolerance: Involvement of Phytohormones.

Salt stress affected areas are increasing significantly all over the world leading to the adverse effects on plant growth and development and productivity of agricultural crops. For better suitability of plants with the changing environment, plants show well-defined metabolic alterations primarily in response to nutrient availability in the environment as an adaptive response. Thus, the management of salt stress is a function of nutritional status of the plant. The uptake and assimilation of sulfur plays a pivotal role in development and metabolic processes of plants and is an integral part of several important compounds, such as vitamins, co-enzymes, phytohormones and reduced sulfur compounds that decipher growth and vigour of plants under optimal and stressful environments. Assimilatory sulfate reduction may induce salt tolerance by coordinating various physiological processes and molecular mechanisms which are likely to be induced by phytohormones. The present review provides an update on physiological and molecular approaches associated with salt stress and details out how sulfur assimilation and phytohormones induce salt tolerance.

Anti-HSV activity of Hypnea musciformis cultured with different phytohormones

Four extracts from the seaweed Hypnea musciformis (Wulfen in Jacq.) J.V. Lamour. (Rhodophyta), collected directly from its natural habitat or cultivated in the presence of phytohormones, were evaluated for their ability to inhibit the replication of acyclovir-resistant herpes simplex viruses types 1 (HSV-1) and 2 (HSV-2) strains. The main purpose was to determinate whether these growth conditions would affect the antiviral activity. Our results showed the possibility of improving the anti-HSV activity by using extracts from algae cultured in the presence of phytohormones.

Dynamic changes of endogenous phytohormones during after-ripening process of Panax notoginseng seeds / 中草药

Objective: The studies were carried out on the endogenous phytohormone changes of Panax notoginseng seeds during the after-ripening process in order to understand the machanism of endogenous phytohormones (GA, IAA, ZR, and ABA) in the process of embryo development and dormancy release. Methods: Dynamic changes of endogenous phytohormones of P. notoginseng seeds were measured by ELISA method through the entire after-ripening process. Results: The contents of endogenous GA, IAA, and ZR increased with the embryo development, while the ABA content decreased contrarily. Conclusion: The change of phytohormone balance is one of determining factors for relieving dormancy and promoting germination of P. notoginseng seeds.

In vitro shoot development in blepharis repnes.

An herb is a plant or plant part valued for its medicinal aromatic or savory qualities. In vitro shoot development of Blepharis repnes was achieved using thin sections of nodes and stems (1.0-1.5 mm) were cultured in MS medium containing varying concentrations of different combination of phytohormones Indole-3-butyric acid (IBA), Benzylaminopurine (BAP) & Gibberlic acid. The combination of BAP, IBA & Gibberellic acid at different concentrations showed the length of shoot development 0.6 -1.8cm with in three weeks in suitable laboratory condition.

In vitro Effects of Plant Extracts, and Phytohormones on Mycelial Growth of Anthracnose Fungi

Water extracts of six plants, such as Allium sativum, A. cepa, Zingiber officinale, Platycodon grandiflorum, Oenanthe javanica, and Capsella brusapastoris, were tested in vitro for inhibitory activity against mycelial growth of anthracnose fungi, Colletotrichum gloeosporioides, C. dematium, and C. coccodes. Among the plant extracts, an Allium sativum extract has good inhibitory effects in all the fungi. Four phytohormones namely, IAA (indole-3-acetic acid), NAA (a-Naphthyl acetic acid), 2,4-D (2,4-Dichloro phenoxy acetic acid) and BAP (Benzyl adenine purine) were used to find out the role over mycelial growth of these fungi. All the concentrations of BAP have good inhibitory effect against mycelial growth of these fungi than that of other tested plant hormones.

RNA polymerase activity in isolated nuclei of Nicotiana sanderae callus: Characteristics and modulation during differentiation.

Isolated nuclei from differentiating cultures of Nicotiana sanderae showed increased levels of RNA polymerase activity as compared to the nuclei from callus cultures. The RNA synthetic activity was dependent on nucleotide triphosphates and Mg2+ and was destroyed by RNase. Maximum activity was obtained in the presence of 50 mM (NH4)2 SO4 and α-amanitin inhibited 40% and 55% of the activity in the nuclei from callus and differentiating tissue respectively. The nuclei from differentiating tissue elicited a 3-fold increase in RNA polymerase I and a 4-fold augmentation in RNA polymerase II activities.

Effect of phytohormones on nuclear RNA synthesis in germinating seeds of Trigonella foenumgraeceum and its callus.

Treatment of Trigonella foenumgraeceum (fenugreek) seedlings with naphthalene acetic acid plus gibberellic acid enhanced the RNA synthesising capacity of nuclei isolated from the hypocotyl and cotyledonary regions. This increase was more pronounced in the nuclei from the hypocotyl region than from the cotyledonary region. In vitro addition of these phytohormones did not stimulate RNA synthesis by nuclei. The RNA synthesis by mitochondria was not affected by preincubating the seedlings with the hormones. The nuclei isolated from callus cultures of fenugreek hypocotyl treated with the hormone also showed increased RNA synthesis.