Search

Understanding the essential processes of heredity, mechanisms of metabolism, and evolutionary traits of this ginkgo (Ginkgo biloba L.) in terms of molecular biology is necessary. The extraction of high-quality and high-integrity RNA from ginkgo is one of the basic techniques of molecular biology. Therefore, this study aimed to compare the efficacy of the six commonly used RNA extraction methods for detection. TRIzol, modified TRIzol, cetyltrimethylammonium bromide (CTAB), modified CTAB, sodium dodecyl sulfate (SDS), and RNeasy Plant Mini Kit (Qiagen) methods, were evaluated for their capacity to extract high-quality and high-quantity RNA from the embryo and endosperm of ginkgo. The advantages and disadvantages of each method were analyzed to evaluate the isolation process. The optimized modified CTAB method for embryo prevented the precipitation of RNA for a long period and reduced complex separation and precipitation processes. In addition, the modified TRIzol method is more effective in endosperm. Obvious differences in RNA quality and quantity were observed between the embryo and endosperm when the same method is used. Modified TRIzol, modified CTAB, SDS and RNeasy Plant Mini Kit (Qiagen) methods were applied to RNA extraction from other different ginkgo tissues. The modified CTAB is efficient and recommended for the isolation of good-quality total RNA from all tissues with reduced chemical use, costs, and RNA loss. The results of quantitative real-time PCR confirmed that modified CTAB methods could obtain showing low Ct values and high amplification efficiency and enabled the successful isolation of RNA from all tissues.

The oleaginous alga Botryococcus braunii (Chlorophyta) is known to accumulate hydrocarbons up to 80% of cell dry weight. Races A, B, and L of B. braunii species accumulate alkadiene/triene, botryococcene, and lycopadiene, respectively. Transcriptome analyses in race A and race B have identified various transcripts encoding enzymes involved in the biosynthesis of alkadiene/triene and botryococcene. However, the transcriptome of B. braunii race L has not been analyzed. In this study, we report the de novo assembly of the transcriptome of B. braunii AC768 race L. AC768 culture in 2×BB medium reaches a maximum density (cell dry weight) of 5.5 g L−1, of which, 6% is lycopadiene. Transcriptional profiles of AC768 show that ESTs involved in energy metabolisms are most abundantly expressed, suggesting an energy requirement for lipid accumulation. Transcriptomic analysis indicates the presence of a putative lycopaoctaene synthase that synthesizes the precursor of lycopadiene in AC768. Comparative analysis between races reveals that botryococcene synthase and lycopaoctaene synthase are specific to races B and L of B. braunii, respectively. Taken together, our results indicate that hydrocarbon chemicals accumulated in different races of B. braunii are determined by their transcriptional profiles.

Magnolia sieboldii K. Koch seed is characterized with having deep dormancy. The inner molecular regulation mechanism has not been investigated because of the absence of a protocol for total RNA (ribonucleic acid) extraction. The extraction of high-quality RNA is important and can be a limiting factor in plant molecular biology experiments. Sufficient high-quality total RNA is required to elucidate the molecular regulation mechanism of germination. However, M. sieboldii seeds with large amounts of secondary metabolites also contain recalcitrant tissues for RNA isolation. We found two simple and low-cost RNA extraction methods for M. sieboldii seeds by evaluating and selecting eight types of methods and further optimizing these methods. The two methods were not only suitable for extracting M. sieboldii seed RNA but also applicable to RNAs from several other tissues. Total RNA extracted through these approaches was applicable for general molecular biology experiments such as qRT-PCR (quantitative real-time polymerase chain reactions). The protocols also meet the strict harsh requirements for transcriptome sequencing and small RNA sequencing. This study provides a powerful approach for future studies at the transcription level.

To elucidate the action mechanism of Xingnaojing Injection (醒脑静注射液, XNJI) for sepsis, and to target screen the potential bioactive ingredients. An integrated protocol that combines in silico target screen (molecular docking) and database mapping was employed to find the potential inhibitors from XNJI for the sepsis-related targets and to establish the compound–target (C-T) interaction network. The XNJI's bioactive components database was investigated and the sepsis-associated targets were comprehensively constructed; the 3D structure of adenosine receptor A2a and 5-lipoxygenase proteins were established and evaluated with homology modeling method; system network pharmacology for sepsis treatment was studied between the bioactive ingredients and the sepsis targets using computational biology methods to distinguish inhibitors from non inhibitors for the selected sepsis-related targets and C-T network construction. Multiple bioactive compounds in the XNJI were found to interact with multiple sepsis targets. The 32 bioactive ingredients were generated from XNJI in pharmacological system, and 21 potential targets were predicted to the sepsis disease; the biological activities for some potential inhibitors had been experimentally confirmed, highlighting the reliability of in silico target screen. Further integrated C-T network showed that these bioactive components together probably display synergistic action for sepsis treatment. The uncovered mechanism may offer a superior insight for understanding the theory of the Chinese herbal medicine for combating sepsis. Moreover, the potential inhibitors for the sepsis-related targets may provide a good source to find new lead compounds against sepsis disease.

Four novel polymers, poly(3,6-9-decyl-carbazole-alt-1,3-benzene) (PB13CZ), poly(3,6-9-decyl-carbazole-alt-bis(4-phenyl) (phenyl) phosphine oxide) (PTPPO38CZ), poly(3,6-9-decyl-carbazole-alt-2,4-phenyl(diphenyl) phosphine oxide) (PTPPO13CZ) and poly(3,6-9-decyl-carbazole-alt-bis(3-phenyl) (phenyl) phosphine oxide) (PTTPO27CZ) were synthesized, and their thermal, photophysical properties and device applications were further investigated to correlate the chemical structures with the photoelectric performance of bipolar host materials for phosphorescent organic light emitting diodes. All of them show high thermal stability as revealed by their high glass transition temperatures and thermal decomposition temperatures at 5% weight loss. These polymers have wide band gaps and relatively high triplet energy levels. As a result, the spin coating method was used to prepare the green phosphorescent organic light emitting diodes with polymers PTPPO38CZ, PTPPO13CZ and PTTPO27CZ as the typical host materials. The green device of polymer PTPPO38CZ as host material shows electroluminescent performance with maximum current efficiency of 2.16 cd·A−1, maximum external quantum efficiency of 0.7%, maximum brightness of 1475 cd·m−2 and reduced efficiency roll-off of 7.14% at 600 cd·m−2, which are much better than those of the same devices hosted by polymers PTTPO27CZ and PTPPO13CZ.

This is the first reported proteomic analysis to study the dormancy breaking of Magnolia sieboldii seeds. Our results provide a fundamental reference for further studies on the regulation of protein expression during seed germination. Magnolia sieboldii K. Koch is an ornamental tree. The deep dormancy of its seeds hinders its cultivation for economic purposes. The biochemical basis of the regulation of seed germination remains poorly understood. The present study aimed to identify differentially expressed proteins in germinated seeds of M. sieboldii using polyethylene glycol fractionation. In total, 59 differentially expressed protein spots from two-dimensional gel maps were detected, 33 of which were identified by mass spectrometry. They were assigned to eight functional classes on the basis of their putative biological functions: photosynthesis (3 %), chaperonin/heat shock protein (9 %), protein and amino acid synthesis (9 %), stress/defense (18 %), cytoskeleton structure (3 %), metabolism (18 %), hormone and polyamine (9 %) and storage proteins (31 %). Among the other functions, the effects of plant hormones on seed germination may be one of the most important functions in plant growth. Gibberellins and ethylene positively regulate seed germination. The activities of several hormone-associated proteins possibly influencing seed germination were increased. The characterization of these proteins will be of great help in identifying the molecular mechanism underlying seed germination.

The existence and release of fluid inclusions in bornite and its associated minerals, namely, quartz and calcite were investigated and confirmed. The structures, forms, and phases of these large quantities of fluid inclusions were also studied. A mass of fluid inclusions with various sizes, distributions, shapes, and phases exist in bornite and its associated minerals. Their sizes vary from a few micrometers to tens of micrometers, and the forms appear as negative crystals, or elongated, elliptical, and irregular. At room temperature, fluid inclusions were mainly characterized as gas-liquid twophase. However, small amounts of fluid inclusions with pure gas phase and pure liquid single-phase were also observed in quartz and calcite. These fluid inclusions initially broke during the ore crushing and grinding process and then released into the flotation pulp in the flotation process. The quantitative analysis of fluid inclusions in the solution and the comparisons of mineral dissolution show that the amount of copper and iron released by fluid inclusions in the bornite sample is higher than the amount dissolved by the mineral; fluid inclusions in the associated gangue minerals, quartz, and calcite also make contribution.

In this study, we analyzed 12 species in the subgenus Potamogeton (8 tetraploids and 2 diploids, 2 putative tetraploid hybrids) at the chromosomal level, including counting the chromosome number and physically mapping the rDNA. The extent of variation in the chromosome number and rDNA loci was determined in the Potamogeton species on both the inter- and intra-specific level. Moreover, one of the hybrid sets (P. perfoliatus ♂ × P. wrightii ♀) was picked for performing artificial pollination for producing F1 generation adults and for sequential morphological character analysis and FISH detection. After comparing the parental species and natural hybrids from three different geographic locations, the intraspecific variations of rDNA loci were revealed in sampled plants; in addition, comparisons between artificial F1 and natural hybrids (P. × intortusifolius) showed a rapid change of 45S rDNA loci in response to the interspecific hybridization. We also compared our results concerning rDNA patterns with phylogenetic documents to derive complementary clues to karoytype evolution and interspecific relationships. Based on frequent hybridizations and active clonal reproduction with weak genetic selection, the rDNA chromosomal repatterning, such as the gain or loss of rDNA loci together with rDNA movements, might be one trend of chromosome evolution in this genus.

Interspecific hybridization played an important role in speciation and evolution of angiosperms. Although the widespread occurrence of natural hybrids in the genus Potamogeton has been studied intensively, few successful experimental hybridization studies have been reported in this genus. In the present study, critical experimental hybridization was conducted using Potamogeton x intortusifolius, a natural hybrid widely distributed in China, and its parents (P. perfoliatus and P. wrightii). The absence of prezygotic barriers between P. wrightii and P. perfoliatus was observed, which contributes to the frequent hybridization of these species in nature. The pollen tube growth rates of P. perfoliatus were much faster than those of P. wrightii in the style of that species. However, the conspecific pollen tubes were competitively advantageous in P. perfoliatus styles. The interspecific hybridization could be applied bidirectionally, and 28 F1 hybrid individuals were successfully obtained from P. wrightii x P. perfoliatus, despite the low germination possibility of the hybrid seeds. Both the artificial and natural hybrids exhibited intermediate morphological characters but presented much lower fertility. The sterility of P. x intortusifolius is mainly due to its low female fitness. However, the offspring from P. wrightii x P. x intortusifolius indicated the potential for backcrossing in nature. This is the most successful attempt at artificial hybridization in this genus so far. The possible route for restoration of fertility and the fitness of the hybrids are also discussed in this paper.

Pollen longevity in seven Potamogeton species representing different pollination systems (anemophily, epihydrophily and hydroautogamy) was assessed both under aerial condition and in contact with water to investigate how water impacts the sexual reproduction in these aquatic taxa. Stainability of pollen with MTT was considered as an indicator of pollen viability. The half-life of pollen longevity was calculated using exponential decay regression. Overall, pollen viability decreased relatively rapidly with time. Pollen grains of obligate anemophilic species had lower initial viability and shorter half-lives than those of facultative anemophilic species. Pollen in these latter species may take more time to reach the stigma. The pollen of Potamogeton may be categorized as partially hydrated pollen owing to its generally spherical shape and lack of furrows, rapid loss of viability, and fast pollen tube initiation. The half-life is positively correlated with pollen size. Smaller-sized grains are at greater risk of desiccation than larger grains. In contrast with the situation observed in most terrestrial angiosperms, contact with water increases pollen longevity in Potamogeton species. In our present study the half-lives of pollen longevity of Potamogeton species in which the pollen had come into contact with water (mean of 10.65 h) were markedly higher than those under aerial conditions (mean of 5.79 h, t = 2.622, P = 0.039). The results of our study contradict a widely held belief that water is detrimental to pollen viability in angiosperms and furthermore indicate that close proximity to water results in selection for wettability. The transition to a hydrated status together with its morphology, make Potamogeton pollen more adapted to the aquatic environment and thus serves to ensure reproductive process. Results of our present study may have direct implications for understanding the evolution of the sexual reproductive system in aquatic angiosperms.