The responsive characteristics of phytochrome genes to photoperiod, abiotic stresses and identification of their key natural variation sites in foxtail millet (Setaria italica L.) / 生物工程学报

The responsive patterns of phytochrome gene family members to photoperiod and abiotic stresses were comparatively analyzed and the favorable natural variation sites of these genes were identified. This would help understand the mechanism of phytochrome gene family in photoperiod-regulated growth and development and abiotic stress response. In addition, it may facilitate the molecular marker assisted selection of key traits in foxtail millet. In this study, we used RT-PCR to clone three phytochrome genes SiPHYA, SiPHYB and SiPHYC from ultra-late maturity millet landrace variety 'Maosu'. After primary bioinformatics analysis, we studied the photoperiod control mode and the characteristics of these genes in responding to five abiotic stresses including polyethylene glycol (PEG)-simulated drought, natural drought, abscisic acid (ABA), high temperature and NaCl by fluorescence quantitative PCR. Finally, we detected the mutation sites of the three genes among 160 foxtail millet materials and performed haplotype analysis to determine the genes' functional effect. We found that the cloned cDNA sequences of gene SiPHYA, SiPHYB and SiPHYC were 3 981, 3 953 and 3 764 bp respectively, which contained complete coding regions. Gene SiPHYB and SiPHYC showed closer evolutionary relationship. Photoperiod regulated all of the three genes, but showed more profound effects on diurnal expression pattern of SiPHYB, SiPHYC than that of SiPHYA. Under short-day, when near heading, the expression levels of SiPHYA and SiPHYB were significantly lower than that under long-day, indicating their roles in suppressing heading of foxtail millet under long-day. SiPHYB and SiPHYC were responsive to PEG-simulated drought, natural drought, ABA and high temperature stresses together. SiPHYA and SiPHYB responded differently to salt stress, whereas SiPHYC did not respond to salt stress. Re-sequencing of 160 foxtail millet materials revealed that SiPHYB was highly conservative. Two missense mutations of SiPHYA, such as single nucleotide polymorphism (SNP) 7 034 522C→T and SNP7 036 657G→C, led to delaying heading and increasing plant height. One missense mutation of SiPHYC, such as SNP5 414 823G→T, led to shortening heading under short-day and delaying heading under long-day, as well as increasing plant height and panicle length regardless of photo-thermal conditions. Photoperiod showed different regulatory effects on SiPHYA, SiPHYB and SiPHYC. SiPHYB and SiPHYC jointly responded to various abiotic stresses except for the salt stress. Compared with the reference genotype, mutation genotypes of SiPHYA and SiPHYC delayed heading and increased plant height and panicle length.

Germination and seed conservation of a pioneer species, Tecoma stans (Bignoniaceae), from tropical dry forest of Colombia / Germinación y conservación de semillas de la especie pionera, Tecoma stans (Bignoniaceae), del bosque seco tropical de Colombia

Abstract Seed germination and seed longevity under different environmental conditions are fundamental to understand the ecological dynamics of a species, since they are decisive for its success within the ecosystem. Taking this into account, seed germination and seed storage behavior of a pioneer species of tropical dry forest (Tecoma stans) were studied in the laboratory, to establish the effect of different environmental conditions on a local tree population. Two seed lots collected in July 2011, from Cali (Colombia), were evaluated under three alternating temperatures (20/30, 20/25, 25/30 ºC; 16/8 h) and four light qualities (12-hour white light photoperiod, darkness, and 15 minutes of red light -R and far red light -FR). Final germination was recorded for all treatments; for white light treatment the daily germination was recorded to calculate mean germination rate, mean germination time, and two synchronization indices. To assess the effect of light quality on physiological variables, a destructive germination test was carried out. For this test, another seed lot was evaluated under the same light conditions using an alternating temperature of 20/30 °C - 16/8 h, recording germination during six days for every treatment. In addition, seeds were stored at two different moisture contents (7.7, 4.1 %) and three storage temperatures (20, 5, -20 ºC), during two time periods (one and three months); a seed germination test was conducted for each treatment. Four replicates of 35 seeds per treatment were used for all experiments. Germination was high (GP > 90 %) with all alternating temperatures under white light, whereas under R, FR, and darkness germination was evenly successful at low temperatures, but at higher temperature, half of the seeds entered into secondary dormancy (GP= 45-65 %). However, mean germination rate and synchronization under R and FR decreased significantly in comparison to white light treatment and, consequently, mean germination time increased. Seed storage behavior of this species is orthodox due to the high germination (GP > 90 %) obtained under all treatments. In conclusion, T. stans seeds have a negative germination response at high incubation temperatures in the absence of white light, entering into a secondary dormancy. In contrast, an environment with a lower temperature and without white light delays the germination, but at the end seeds are able to reach the same germination values. This seed dependence on incident light in limiting conditions suggests a physiological mechanism on the seed tissues of this species, probably mediated by phytochromes. Finally, the orthodox seed storage behavior of T. stans is a reason to include this species in ex situ seed conservation programs for restoration and recovery of the tropical dry forest; however, long-term studies should be conducted in order to evaluate the maintenance of this characteristic throughout longer periods of time. Rev. Biol. Trop. 66(2): 918936. Epub 2018 June 01.

Resumen La germinación y la longevidad de las semillas de una especie bajo diferentes condiciones ambientales son fundamentales para las dinámicas ecológicas de una especie, debido a que son decisivas en el éxito de la misma en un ecosistema. Teniendo en cuenta esto, se estudió la germinación y el comportamiento en el almacenamiento de las semillas de una especie pionera de bosque seco tropical (Tecoma stans) a nivel de laboratorio, para establecer el efecto de diferentes condiciones ambientales en una población local de árboles. Dos lotes de semillas recolectados en julio 2011, de Cali (Colombia), se evaluaron a tres temperaturas alternadas (20 / 30, 20 / 25, 25 / 30 °C; 16 / 8 h) y cuatro calidades de luz (fotoperiodo de 12 horas de luz blanca, oscuridad, y 15 minutos de luz roja -R y roja lejana -RL). Se registró la germinación final para todos los tratamientos; para el tratamiento de luz blanca se registró la germinación diaria para calcular la tasa media de germinación, el tiempo medio de germinación y dos índices de sincronización. Para evaluar el efecto de la calidad de luz sobre las variables fisiológicas, se realizó una prueba de germinación destructiva. Para esta prueba, otro lote de semillas fue puesto a las mismas condiciones de luz usando una temperatura alternada de 20 / 30 °C - 16 / 8 h, registrando la germinación durante seis días para cada tratamiento. Además, se almacenaron semillas a dos contenidos de humedad (7.7, 4.1 %) y a tres temperaturas de almacenamiento (20, 5, -20 °C), durante dos periodos de tiempo (uno y tres meses); se realizó una prueba de germinación a cada tratamiento. Cuatro repeticiones de 35 semillas por cada tratamiento se usaron en cada experimento. La germinación fue alta (PG > 90 %) en todas las temperaturas alternadas con luz blanca, mientras que en los tratamientos de luz R, RL y en oscuridad, la germinación fue igualmente exitosa a bajas temperaturas, pero a temperaturas más altas la mitad de las semillas entraron en latencia secundaria (PG= 45-65 %). Sin embargo, la tasa media de germinación y la sincronización en R y RL disminuyeron significativamente en comparación con el tratamiento de luz blanca y consecuentemente el tiempo medio de germinación aumentó. El comportamiento de las semillas de T. stans en el almacenamiento es ortodoxo debido a la alta germinación obtenida (PG > 90 %) en todos los tratamientos. En conclusión, las semillas de T. stans tienen una respuesta germinativa negativa a temperaturas de incubación alta en ausencia de luz blanca, donde entran a latencia secundaria. En contraste, un ambiente con baja temperatura y sin luz blanca retrasa la germinación, pero al final las semillas son capaces de alcanzar los mismos valores de germinación. Esta dependencia de las semillas a la luz incidente en condiciones limitantes sugiere la presencia de un mecanismo fisiológico en los tejidos de esta especie, probablemente mediado por fitocromos. Finalmente, el comportamiento ortodoxo de las semillas de T. stans en el almacenamiento abre la posibilidad de incluirla en programas de conservación ex situ para la restauración y recuperación del bosque seco tropical; no obstante, se deben llevarse a cabo pruebas más largas para evaluar el mantenimiento de esta característica por periodos de tiempo más largos.

Photosynthate partitioning and morphoanatomical aspects of photomorphogenic mutants of tomato / Partição de fotoassimilados e aspectos morfoanatômicos em tomateiros mutantes fotomorfogenéticos

The aim of this study was to analyze photosynthate partitioning in tomato photomorphogenic mutants at the ends of the vegetative (40 days after emergence [DAE]) and reproductive (69 DAE) stages and to determine its interaction with morphoanatomical aspects. The mutants aurea (au), phytochrome-deficient, high pigment-1 (hp1), light-exaggerated response, were studied along with the non-mutant Micro-Tom (MT) cultivar. The plants were analyzed at 40 and 68 DAE to identify photosynthate source organs and tissues as well as the target organs of remobilized photosynthate during the reproductive stage. The plants were evaluated for their internal and external morphology as well as the percentage of dry mass of their organs. Photosynthate allocation in the hp1 mutant occurred primarily in the roots and leaves, and allocation in the au mutant occurred primarily in fruits. The au mutant showed a high capacity for photosynthate remobilization to fruit during the reproductive stage, and the predominant sources of these remobilized photosynthates were the leaf spongy parenchyma, the root vascular cylinder and the marrow stem.

O objetivo deste estudo foi analisar a partição de fotoassimilados em tomateiros mutantes fotomorfogenéticos ao final da fase vegetativa, aos 40 dias após a emergência (DAE), e ao final da fase reprodutiva, aos 69 DAE, e sua interação com aspectos morfoanatômicos. Foram estudados os mutantes aurea (au), deficiente em fitocromo, e hp1, o qual expressa resposta exagerada à luz, e o tomateiro selvagem cultivar Micro-Tom (MT). As plantas foram analisadas 40 dias após a emergência (DAE) e 68 DAE, tentando identificar os órgãos e tecidos dos fotoassimilados remobilizados e seus órgãos de destino durante o estádio reprodutivo. As plantas foram avaliadas quanto à sua morfologia interna e externa e percentagem de massa seca entre os órgãos. A alocação de fotoassimilados no mutante hp1 ocorreu prioritariamente em raízes e folhas comparativamente aos demais órgãos, e no mutante au ocorreu prioritariamente em frutos comparativamente aos demais órgãos. O mutante au deteve alta capacidade de remobilização de fotoassimilados durante sua fase reprodutiva para os frutos e os fotoassimilados remobilizados tiveram origem preponderante do parênquima lacunoso foliar, do cilindro vascular radicular e da medula caulinar.

Growth responses of Betula pendula ecotypes to red and far-red light

The effect of Red light (R), Far-red light (FR) and R/FR combinations on shoot growth of latitudinal ecotypes of B. pendula was studied using special diodes that emit monochromatic lights. When a 12 hrs PAR (110 µmol m-2 s-1) was extended with R, FR or R/FR ratios, lower intensities of monochromatic lights could not prevent growth cessation. At 25 µmol m-2 s-1, FR compared to R enhanced stem elongation in all ecotypes. This was due to the inhibitive effect of R on internode elongation. When day-length was extended by R/FR at various ratios, there was continuous shoot elongation, but was found to be declining with increasing ratios. The more the R, the shorter were the internodes of each plant. B. pendula ecotypes produced branches when PAR light during the day was extended by incandescent light, but did not do so when the light extensions were made by monochromatic R or FR or their combination. Branching increased with decreasing latitude of the ecotype.

Study on the reconstitution in vitro and photochemical activities of phytochrome from the Synechocystis sp. PCC6803 / 生物工程学报

Genomic DNA sequence analysis of phytochrome like photoreceptors in a number of bacteria revealed several open reading frames (ORFs) encoding proteins with amino acid sequences homologous to plant phytochromes. The phytochrome like photoreceptors, collectively called bacteriophytochromes, contain an N-terminal domain homologous to the chromophore-binding domain (CBD) of higher plants and a C-terminal domain of histidine kinase domain( HKD). Due to their simple structure, bacteriophytochromes broaden the view of phytochrome evolution and provide us with a simple model to investigate phytochrome-mediated light signal in higher plants. In this report, the bacteriophytochromes from Synechocystis sp. PCC6803 were investigated. The gene cph1 and its fragment cph1 (C-435) were isolated from the Synechocystis sp. PCC6803 genomic DNA by polymerase chain reaction(PCR) using specific primers. Then, the genes were cloned with the vector pBluescript, yielding plasmids pBlu-cphl and pBlu-cph1 ( C-435), before they are subcloned with the vector pET30, using the EcoRV and Xho I restriction sites. pBlu-cph1, pBlu-cph1 (N-435) were cleaved with Sma I and Xho I, and the released genes were ligated to the pET30a fragment. The E. coli [strain BL21 (DE3)] cells containing recombinant pET30a were grown in medium RB at 20 degrees C, and harvested 6 h later after induction with isopropyl thio-beta-D-galactoside (IPTG). Then, reconstitution systems were employed to study the characteristics of the genes. In the reconstitution system, autoassembly of aprotein of phytochrome with PCB was investigated. The chromophore addition was an autocatalytic process. Reconstitution products were red/infrared (R/FR) photochromic, which was similar to that of the phytoehrome in higher plants. How ever, the spectral change ratios (deltaAmax/deltaAmin) of the two fragments differed from each other. It was also shown that PCB was covalently bound to apo-protein via Zn2+ fluoresc ence SDS-PAGE. After irradiation by light of 700 nm, the maximum absorption spectrum o f holo-Cphl was 650nm. The absorption of it after denaturatior in the dark with ur ea in the presence of hydrochloric acid (pH = 2) was 660nm, which was similar with th at of cis-PCB. In addition, after irradiation by light of 650nm, the maximum absorption spectrum of holo-Cph1 was 700nm. The absorption of it after denaturation in the dark with urea in the presence of hydrochloric acid (pH = 2) was 600nm, which was similar with that of trans-PCB. The result showed that the photochromism of phytochrome resulted from the isomerizaation of chromophore (PCB in this report). The reconstitution of Cph1 (C-435) under the same condition supported the conclusion. Fluorescence emission spectrum of the products suggested that bacteriophytochrom e structure with cis-PCB was more stable than that with trans-PCB. The new reconstitution system in this report sets a base for the application of phytochrome as photochromic biomaterials in biosensors. In addition, phytochrome shows great potential in food, cosmetic and biological engineering, etc.

A search for homologues of plant photoreceptor genes and their signaling partners in the sugarcane expressed sequence tag (Sucest) database

A partir dos dados do projeto de sequenciamento de Ests da Cana de Açúcar (Sucest/FAPESP) e utilizando BLAST (tblastn) como ferramenta, foi realizada uma busca de genes homólogos aos elementos envolvidos nos processos de foto-recepçäo e já descritos para outras plantas, principalmente Arabidopsis. Foram obtidas altas identidades para os fitocromos A, B e C assim como para os críptocromos 1, 2 e a fototropina. Diversos elementos identificados como reguladores primários ou secundários na transduçäo de sinal de foto-receptores também foram identificados com baixos valores de E-value.

Light signal transduction mediated by phytochromes: preliminary studies and possible approaches.

Phytochromes mediate a variety of developmental and growth processes involved in the photomorphogenesis of plants. In this article, we review the current understanding of the structure and function of the photoreceptor, discuss some very preliminary results, and offer speculations and even conjectures that may elicit future studies into the molecular mechanisms of the phytochrome-mediated light signal transduction in plants.

Phytochemistry of the genus asparagus

A comprehensive review on the genus Asparagus which covers the period till 1994. The review also includes the significance of saponins as this genus is rich in saponins

Pharmacognostic studies of indigenous medicinal plants-I

The comparative cytomorphological studies of Conyza ambigua, Eclipta alba and Sonchus asper were carried out with fresh transverse section and with dried powder of their various parts under light microscope; while the irritant constituents were extracted successively with pet. ether [40-60], CHCI3 and MeOH. The CHCI3 and MeOH extracts of leaves, roots and flowers of all the species exhibited a prominent erythema on mice's skin. The CHCI3 and MeOH extracts of the flower of S. asper were more poten irritant than the other extracts of the same species and also the other species. The possible relationship of anatomical structures with phytochemistry and irritancy of these species have been discussed

NMR spectral data of triterpenoidal saponins isolated from Gualacum officinale linn [Zygophyllaceae]

The NMR spectral data of 24 naturally occurring triterpenoidal saponins isolated from Guaiacum officinale [Zygophyllaceae] and published uptill 1991 are reviewed. These saponins are arranged with respect to their aglycone moieties and then by their number of sugar units

Phytochemical studies on the species of genus abutilon

The medicinal properties attributed to various Abutilon species created wide interest in their phytochemistry. This article deals with a summarized account of various chemical constituents isolated so far from Abutilon species

Pharmacognostical and Phytochemical investigations on the leaves of mallotus philippinensis muell Arg

In the present work leaves of M. Philippinensis Muell. Arg. has been subjected to pharmacognostical and phytochemical investigations. The anatomical characteristics of the leaf showed the typical tissue layer as is found in dorsiventral leaf. The petroleum ether extract showed the presence of free sterols i.e. sitosterol and this was found to be 0.55%. The phenolic acid found was p-coumaric acid which was identified by paper chromatography, melting point and U.V. spectra. A phloroglucinol dervative rottlerin was isolated and characterized from the pericarp of Mallotus philippinensis

Comparative phytochemical investigation of certain Melaleuca species grown in Egypt

Comparative TLC, PC and GLC analysis of the extracts of five Melaleuca species grown in Egypt viz. M. armillaris Sol.ex Gaerther, M. bracteata. F. Mne, M. elliptica Smith, M.quinquenervia Cav, and M. styphelloides Smith [Fam. Myrtaceae], revealed similarities in their phytoconstituents. TLC and GLC analysis of the unsaponifiable matter indicated that cholesterol, beta-sitosterol, stigmasterol and oleanolic acid are present in all species except, stigmasterol which could not be detected in M. armillaris while beta-amyrin was detected in M. armillaris leaf and stem. GLC analysis of the fatty acid methyl esters revealed high percentage of saturated fatty acids with palmitic acid as the main constituent. Flavonoids were studied using paper chromatography, while saponins were studied by TLC and their percentages were determined by the haemolytic index