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In nature, plants are exposed to a dynamic light environment. Fluctuations in light decreased the photosynthetic light utilization efficiency (PLUE) of leaves, and much more severely in C4 species than in C3 species. However, little is known about the plasticity of PLUE under dynamic light in C4 species. Present study focused on the influence of planting density to the photosynthesis under dynamic light in maize (Zea mays L.), a most important C4 crop. In addition, the molecular mechanism behind photosynthetic adaptation to planting density were also explored by quantitative proteomics analysis. Results revealed that as planting density increases, maize leaves receive less light that fluctuates more. The maize planted at high density (HD) improved the PLUE under dynamic light, especially in the middle and later growth stages. Quantitative proteomics analysis showed that the transfer of nitrogen from Rubisco to RuBP regeneration and C4 pathway related enzymes contributes to the photosynthetic adaptation to lower and more fluctuating light environment in HD maize. This study provides potential ways to further improve the light energy utilization efficiency of maize in HD.
Asunto(s)
Luz , Zea mays , Zea mays/metabolismo , Fotosíntesis , Ribulosa-Bifosfato Carboxilasa/metabolismo , Hojas de la Planta/metabolismoRESUMEN
BACKGROUND: Agrobacterium rhizogenes-mediated (ARM) transformation is a highly efficient technique for generating composite plants composed of transgenic roots and wild-type shoot, providing a powerful tool for studying root biology. The ARM transformation has been established in many plant species, including soybean. However, traditional transformation of soybean, transformation efficiency is low. Additionally, the hairy roots were induced in a medium, and then the generated composite plants were transplanted into another medium for growth. This two-step operation is not only time-consuming, but aggravates contamination risk in the study of plant-microbe interactions. RESULTS: Here, we report a one-step ARM transformation method with higher transformation efficiency for generating composite soybean plants. Both the induction of hairy roots and continuous growth of the composite plants were conducted in a single growth medium. The primary root of a 7-day-old seedling was decapitated with a slanted cut, the residual hypocotyl (maintained 0.7-1 cm apical portion) was inoculated with A. rhizogenes harboring the gene construct of interest. Subsequently, the infected seedling was planted into a pot with wet sterile vermiculite. Almost 100% of the infected seedlings could produce transgenic positive roots 16 days post-inoculation in 7 tested genotypes. Importantly, the transgenic hairy roots in each composite plant are about three times more than those of the traditional ARM transformation, indicating that the one-step method is simpler in operation and higher efficiency in transformation. The reliability of the one-step method was verified by CRISPR/Cas9 system to knockout the soybean Rfg1, which restricts nodulation in Williams 82 (Nod-) by Sinorhizobium fredii USDA193. Furthermore, we applied this method to analyze the function of Arabidopsis YAO promoter in soybean. The activity of YAO promoter was detected in whole roots and stronger in the root tips. We also extended the protocol to tomato. CONCLUSIONS: We established a one-step ARM transformation method, which is more convenient in operation and higher efficiency (almost 100%) in transformation for generating composite soybean plants. This method has been validated in promoter functional analysis and rhizobia-legume interactions. We anticipate a broad application of this method to analyze root-related events in tomato and other plant species besides soybean.
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Agrobacterium/fisiología , Glycine max/genética , Raíces de Plantas/genética , Raíces de Plantas/microbiología , Plantas Modificadas Genéticamente , Rhizobium , Glycine max/microbiología , Transformación GenéticaRESUMEN
The detection of gas pollutants in atmosphere and indoor air is very important to human health and safety. Monoxide carbon (CO) is a common gas pollutant with high toxicity that mainly comes from the inadequacy oxidization of carbon such as oil, coal and petrol inadequacy combustion, auto-gas and some natural disasters whose limit value in air is lower than 6.0 mg·m-3 in the national standard. Due to its toxicity and uneasy detection, it is one of very dangerous component in the silent killer. Recently, several methods, including infra-red absorption, gas chromatography, potentiometry, Hg replacement, spectrophotometry, I2O5 and PdCl2 nake-eye, semiconductor sensor have been reportedly used for the detection of CO. To our best knowledge, there are no SPR absorption methods for CO, based on the NG SPR absorption. In this paper, the reaction between CO and HAuCl4 was studied with absorption spectrophotometry and transmission electron microscopy (TEM)while a simple and rapid SPR absorption method was developed for the determination of trace CO. In pH 7.2 phosphate buffer solutions, monoxide carbon reduced HAuCl4 to form nanogold (NG) particles with the size of about 45 nm that exhibited surface plasmon resonance (SPR) absorption peak at 540 nm and three energy spectral peaks at 1.70 keV, 2.20 and 9.70 keV for gold element. The analytical conditions were examined, and a pH 7.2 phosphate buffer solution with a concentration of 40 mmoL·L-1 PO3-4, a concentration of 40.0 µg· mL-1 HAuCl4 and a reaction time of 5 min was selected for use. Under the selected conditions, the SPR absorption peak value was linear to CO concentration in the range of 0.2ï½8.75 µg· mL-1, with a detection limit of 0.1 µg· mL-1 CO. According to the procedure, the influence of coexistent substances on the determination of 1.0 mg·L-1 CO was tested, with a relative error of ±5%. Results indicated that 200 times SO2-3, PO3-4, SO2-4, CO2-3 and NO-3, 100 times Zn2+, K+, BrO-3, Na2S, ethanol, methanol, 80 times Ni2+, Cr3+, Co2+, Ca2+, Mg2+, Fe3+, glucose, Pb2+, Al3+, SeO2-3, Na2S2O3, formaldehyde, 50 times Mn2+ do not interfere with the determination. It showed that this SPR method had good selectivity. The CO content in air samples was determined with the SPR method, with a relative standard deviation (RSD) of 1.8%ï½4.2%, the SPR method results were agreement with that of the gas chromatography (GC).
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To study the genetic characteristics and function of swine leukocyte antigen (SLA) class I from the Hebao pig, a rare inbreed in China, a pair of primers was designed to amplify the SLA-2 gene (SLA-2-HB) and then the genetic characteristics of the gene were analyzed. The 3D homology modeling was used to analyze the structure and function of SLA-2-HB proteins. After cloning, sequencing and computer analysis, four SLA-2-HB alleles were found, all of 1119 bp. Sites 3-1097 were an open reading frame encoding 364 amino acids with two sets of intra-chain disulfide bonds comprising four cysteines situated in sites 125, 188, 227 and 283. By alignment of SLA-2-HB sequences with other SLA-2 alleles in the IPD database, 11 key variable amino acid sites were found in the extracellular domain of the SLA-2-HB alleles at sites 23(F), 24(I), 43(A), 44(K), 50(Q), 73(N), 95(I), 114(R), 155(G), 156(E) and 216(S), which could be used to differentiate other SLA-2 alleles. The 3D homology modeling demonstrated that the eight of 11 key variable amino acid sites were all in antigenic binding groove of SLA-2-HB proteins. The amino acid identities between SLA-2-HB and other SLA-2, SLA-1 and SLA-3 alleles were 86.2-97.0%, 85.0-93.9% and 83.3-88.6%, respectively. The phylogenetic tree of SLA-2-HB showed that it was relatively independent of the other SLA-2 genes. Furthermore, the SLA-2-HB alleles were similar to HLA-B15 and HLA-A2 functional domains and preserved some functional sites of HLA-A2. It was concluded that SLA-2-HB are novel alleles of SLA-2 and that the Hebao pig might have evolved independently in China.
Asunto(s)
Antígenos de Histocompatibilidad Clase II/genética , Antígenos de Histocompatibilidad Clase II/inmunología , Alelos , Animales , China , Cartilla de ADN/genética , Disulfuros/metabolismo , Femenino , Variación Genética , Antígenos de Histocompatibilidad Clase I , Antígenos de Histocompatibilidad Clase II/química , Endogamia , Masculino , Modelos Moleculares , Filogenia , Reacción en Cadena de la Polimerasa , Conformación Proteica , Análisis de Secuencia de ADN , Homología de Secuencia de Aminoácido , PorcinosRESUMEN
In this paper, by introducing a combined method of sign function, homogeneous domination and adding a power integrator, and overcoming several troublesome obstacles in the design and analysis, the problem of state feedback control for a class of nonlinear high-order feedforward systems with the nonlinearity's order being relaxed to an interval rather than a fixed point is solved.
RESUMEN
The mechanisms of the palladium-catalyzed hydrothiolation of alkynes with thiols were investigated using density functional theory at the B3LYP/6-31G(d, p) (SDD for Pd) level. Solvent effects on these reactions were explored using the polarizable continuum model (PCM) for the solvent tetrahydrofuran (THF). Markovnikov-type vinyl sulfides or cis-configured anti-Markovnikov-type products were formed by three possible pathways. Our calculation results suggested the following: (1) the first step of the cycle is a proton-transfer process from thiols onto the palladium atom to form a palladium-thiolate intermediate. The palladium-thiolate species is attacked on alkynes to obtain an elimination product, liberating the catalyst. (2) The higher activation energies for the alkyne into the palladium-thiolate bond indicate that this step is the rate-determining step. The Markovnikov-type vinyl sulfide product is favored. However, for the aromatic alkyne, the cis-configured anti-Markovnikov-type product is favored. (3) The activation energy would reduce when thiols are substituted with an aromatic group. Our calculated results are consistent with the experimental observations of Frech and colleagues for the palladium-catalyzed hydrothiolation of alkynes to thiols.