RNA Extraction from Herba Violae Roots with Low-temperature Sectioning Method

[ Objective ] This study aimed to investigate the optimal method for extracting RNA from roots of medicinal plant herba violae by comparing the effects of liquid nitrogen grinding method and low-temperature sectioning method on RNA extraction. [ Method] Roots of herba violae were respectively crushed by using liquid nitrogen grinding method and low-temperature sectioning method to extract RNA. The extraction effects of these two methods were compared based on detec- tion of RNA concentration, purity and integrity and amplification of GAPDH gene by RT-PCR. [Result] The concentration of RNA extracted by liquid nitrogen grinding method and low-temperature sectioning method was 1.21 and 3.57 p^g/~, respectively. Both RNA extracted by these two methods showed two distinct bands after agarose gel electrophoresis. The ratio of brightness of the 28S rRNA to the 18S rRNA bands was greater than 1. PCR amplification showed that the length of GAPDH gene was about 230 bp, which was consistent with the expected result. [ Conclusion ] The experimental results indicated that using low-tempera ture sectioning method to crush the roots of herba violae can meet the needs of most molecular biological experiments including gene cloning and expression analysis, which is an effective and simple method for extracting RNA from plant roots.

Identification and functional analysis of miRNAs in developing kernels of a viviparous mutant in maize

Given the important roles of miRNAs in post-transcriptional gene regulation, identification of differentially expressed miRNAs will facilitate the elucidation of molecular mechanisms underlying kernel development. In this study, we constructed a small RNA library to comprehensively represent the full complement of individual small RNAs and to characterize miRNA expression profiles in pooled ears of maize(Zea mays L.) at 10, 15,20, 22, 25 and 30 days after pollination(DAP). At least 21 miRNAs were differentially expressed. The differential expression of three of these miRNAs, i.e., miR528a, miR167a and miR160b, at each stage was verified by qRT-PCR. The results indicated that these miRNAs might be involved in kernel development. In addition, the predicted functions of target genes indicated that most of the target genes are involved in signal transduction and cell communication pathways, particularly the auxin signaling pathway. The expression of candidate germination-associated miRNAs was analyzed by hybridization to a maize genome microarray, and revealed differential expression of genes involved in plant hormone signaling pathways. This finding suggests that phytohormones play a critical role in the development of maize kernels. We found that in combination with other miRNAs, miR528a regulated a putative laccase, a Ring-H2 zinc finger protein and a MADS box-like protein, whereas miR167a and miR160b regulated multiple target genes,including ARF(auxin response factor), a member of the B3 transcription factor family. All three miRNAs are important for ear germination, development and physiology. The small RNA transcriptomes and mRNA obtained in this study will help us gain a betterunderstanding of the expression and function of small RNAs in the development of maize kernel.

Exceptionally Preserved Conodont Natural Assemblages from the Middle Triassic Luoping Biota,Yunnan Province,China:Implications for Architecture of Conodont Feeding Apparatus

Articulated natural assemblages contain direct evidence of the element numbers,morphologies,positions and structures for reconstructing the feeding apparatuses of conodont animals,but these kind of materials are very rare in fossil records.Here we report ten new conodont natural assemblages from Member II of the Guanling Formation in Luoping County,eastern Yunnan Province,southwestern China.These assemblages were obtained from the fossil-bearing layers of the Luoping Biota,dated to the early Middle Triassic Anisian stage(Pelsonian substage).These fossiliferous laminated limestones mainly consist of calcites and dolomites,with platy clay minerals and pyrites as subordinate components,indicating that the conodont assemblages were preserved in a lower energy and anoxic sedimentary environment.The new natural assemblages preserve the primary collapse orientations of the Nicoraella feeding apparatus from the Luoping Biota,showing the relative original positions of the S,M and P elements,that could be further used to refine the architecture of this apparatus in space variable M elements and the position unclear P1 elements.Integrating previously reported three-dimensional and bilaterally symmetrical fused clusters,we statistically analyze the size of the positional homogenous elements within different sized materials,in particular S4,S3,or S2 elements,indicates that elements ontogenic changes do not affect the apparatus architecture.Architecture of Nicoraella apparatus remains stable in the process of ontogeny,so our reconstructed model is credible.It is the first time that ample types of materials have been used to restore a conodont apparatus within one genus,namely natural assemblages(articulated and disarticulated),fused clusters(articulated,disarticulated,compressed,and relatively three-dimensional),and abundant discrete elements,respectively.These materials together record the most complete information on architecture in the gondolelloid apparatuses,thus enabling us to reconstruct a reliable fifteen-element apparatus and propose it as a standard template for gondolelloid apparatus reconstruction.

山地模式下剪力墙结构基础设计问题探讨

在山地建筑设计过程中,基础设计是结构设计的重点与难点。山地条件下剪力墙结构相对于框架结构的基础设计则显得更加复杂,该文结合工程实例,就山地条件下剪力墙结构基础设计中的相关问题进行分析,根据分析结果提出相应处理方案以供参考。

Permian integrative stratigraphy,biotas,paleogeographical and paleoclimatic evolution of the Qinghai-Tibetan Plateau and its surrounding areas

The Permian Period was a critical time interval during which various blocks of the Qinghai-Tibetan Plateau have experienced profound and complex paleogeographical changes.The supercontinent Pangea was formed to its maximum during this interval,hampering a global east-to-west trending equatorial warm ocean current.Meanwhile,a semi-closed Tethys Ocean warm pool formed an eastward-opening oceanic embayment of Pangea,and became an engine fostering the evolutions of organisms and environmental changes during the Paleozoic-Mesozoic transition.Stratigraphy and preserved fossil groups have proved extremely useful in understanding such changes and the evolutionary histories of the Qinghai-Tibetan Plateau.Widely distributed Permian deposits and fossils from various blocks of the Qinghai-Tibetan Plateau exhibited varied characteristics,reflecting these blocks’different paleolatitude settings and drifting histories.The Himalaya Tethys Zone south to the Yarlung Zangbo suture zone,located in the northern Gondwanan margin,yields fossil assemblages characterized by cold-water organisms throughout the Permian,and was affliated to those of the Gondwanaland.Most of the exotic limestone blocks within the Yarlung Zangbo suture zone are Guadalupian(Middle Permian)to Early Triassic in age.These exotic limestone blocks bear fossil assemblages that have transitional affinities between the warm Tethys and cold Gondwanan regions,suggesting that they most probably represent seamount deposits in the Neo-Tethys Ocean.During the Asselian to Sakmarian(Cisuralian,also Early Permian),the Cimmerian microcontinents in the northern part of Gondwana preserved glacio-marine deposits of Asselian to Sakmarian,and contained typical Gondwana-type cold-water faunas.By the middle Cisuralian(~290-280 Ma),the Cimmerian microcontinents rifted off from the Gondwanaland,and drifted northward allometrically due to the active magmatism of the Panjal Traps in the northern margin of the Indian Plate.Two slices of microcontinents are discerned as a result of such allometic drifting.The northern Cimmerian microcontinent slice,consisting of South Qiangtang,Baoshan,and Sibuma blocks,drifted relatively quickly,and preserved widespread carbonate deposits and warm-water faunas since Artinskian.By contrast,the southern Cimmerian microcontinent slice,consisting of Lhasa,Tengchong,and Irrawaddy blocks,drifted relatively slowly,and were characterized by widespread carbonate deposits containing warm-water faunas of late Kungurian to Lopingian(Late Permian).As such,these blocks rifted off from the northern Gondwanan margin since at least the Kungurian.Thus,it can be inferred that these blocks were incorperated into the low latitude,warm-water regions later than the northern Cimmerian slice.Such discrepancies in depositional sequences and paleobiogeography imply that the rifting of Cimmerian microcontinents resulted in the formation of both Meso-Tethys and Neo-Tethys oceans during the Cisuralian.By contrast,the North Qiangtang block,because of its further northern paleogeographical position,contains warm-water faunas throughout the whole Permian Period that are affiliated well with the faunas from the South China,Simao,and Indochina blocks.Together,these blocks belonged to the members of the northern Paleo-Tethys Ocean.Thus,an archipelagic paleogeographical framework divided by Paleo-,Meso-,and Neo-Tethys oceans was formed,fostering a global biodiversity centre within the Tethys warm pool.Since most of the allochthonous blocks assembling the Qinghai-Tibetan Plateau were situated in the middle to high latitude regions during the Permian,they preserved most sensitive paleoclimate records of the Late Paleozoic Ice Age(LPIA),the Artinskian global warming event,and the rapid warming event at the end-Permian.Therefore,sedimentological and paleontological records of these blocks are the unique window through which we can understand global evolutions of tectonic movement and paleoclimate,and their impacts on spatiotemporal distributions of comtemporaneous biotas.

Paleontology Knowledge Graph for Data-Driven Discovery

A knowledge graph(KG)is a knowledge base that integrates and represents data based on a graph-structured data model or topology.Geoscientists have made efforts to construct geosciencerelated KGs to overcome semantic heterogeneity and facilitate knowledge representation,data integration,and text analysis.However,there is currently no comprehensive paleontology KG or data-driven discovery based on it.In this study,we constructed a two-layer model to represent the ordinal hierarchical structure of the paleontology KG following a top-down construction process.An ontology containing 19365 concepts has been defined up to 2023.On this basis,we derived the synonymy list based on the paleontology KG and designed corresponding online functions in the OneStratigraphy database to showcase the use of the KG in paleontological research.

Characteristics of Paleontological Communities in Surface Sediments of the Southern South China Sea and Their Paleoclimatic and Paleoenvironmental Significance

Understanding the spatial and temporal distribution of different paleontological communities in the southern South China Sea(SCS)is fundamental to explore its paleoclimatic and paleoenvironmental changes.In this study,foraminifera,pollen and diatoms from 100 surface sediment samples covering 40000 km^(2) of sea floor in the southern SCS were comprehensively investigated in terms of their assemblage and distribution.The results showed the existence of abundant foraminifera and pollen in most of the samples,although diatom communities were relatively scarce.Foraminifera were dominated by G.sacculifer,G.menardii,G.ruber,while diatoms were characterized by T.simonsenii,T.nitzschioides and T.longissima,indicating a typical tropical marine environment.The pollen assemblages showed a better reflection of montane rainforest,tropical rainforest and mangrove.The spatial differences among foraminifera assemblages may indicate the effect of water depth and a warmer environment in the southeast part of the study area,while the spatial patterns of diatom and pollen assemblages imply the influence of coastal current.Our study also noted that the compositions of paleontological communities in the SCS can vary significantly in a short distance,and synthesized studies on multiple biological groups are needed to reconstruct the Quaternary climate and the oceanographic environment.

Palaeoecological Analysis of Trace Fossil Sinusichnus sinuosus from the Middle Triassic Guanling Formationin Southwestern China

The Luoping Biota discovered from the early Middle Triassic （Anisian） Guanling Formation of southwestern China represents a fully recovered shallow marine ecosystem, marking the end point of Early Triassic biotic recovery following the end-Permian mass extinction. Contemporaneously preserved are prolific trace fossils, which offer good opportunities to understand the palaeoecology of marine invertebrates from a fully recovered shallow marine ecosystem. Here we present a newly discovered sinuous branching burrow from the fossil-bearing unit in Member II of the Guanling Formation. Several features, including the horizontal regular sinuous nature, the branching pattern, typical H-junction, and the small wavelength/amplitude ratio of these sinuous structures within the burrow systems justify assignment of these traces as Sinusichnus sinuosus, a trace possibly produced by decapod crustaceans. Close association ofS. sinuosus with Rhizocorallium commune suggests a deposit-feeding strategy of these trace makers. The newly reported Anisian material from the Guanling Formation in Luoping represents first report of Sinusichnus from South China. The global record of Sinusichnus occurrence suggests that these burrows might have an older history than Early Middle Triassic.

Microbially Induced Carbonate Precipitation in a Middle Triassic Microbial Mat Deposit from Southwestern China:New Implications for the Formational Process of Micrite

Lime mud(i.e.,micrite)is a major component of carbonate deposits.Various mechanisms(biotic versus abiotic)have been proposed for the formation of lime mud in Earth’s history.However,the detailed role that microbes play in the nucleation and subsequent precipitation of micrites remains to be resolved.Herein we undertook a detailed geobiological characterization of laminated lime mudstone from the Middle Triassic Guanling Formation in Yunnan Province,southwestern China.Morphological features,together with previous geobiological investigations,suggest that the laminated lime mudstones represent the former presence of microbial mats.These lime mudstones consist mainly of calcite,dolomite and quartz,with clay minerals and pyrites as subordinate components.In particular,micro-analysis shows copious nano-globules(65–878 nm)and capsule-shaped nano-rods in laminations.These low-Mg calcite nano-globule aggregates are closely associated with mucilaginous biofilms resembling extracellular polymeric substances(EPS).Nano-sized globules coalesce to form semi-euhedral micrite crystals.We suggest that a decaying hydrolytic destruction of the EPS by microbial communities within microbial mat leads to the precipitation of the nano-globules by enhancing alkalinity in local micro-environment.As an intermediate,these nano-globules further aggregate to form micrite crystals possibly through a dissolution-reprecipitation process.

Great Paleozoic-Mesozoic Biotic Turnings and Paleontological Education in China: A Tribute to the Achievements of Professor Zunyi Yang

Professor Zunyi Yang is a pioneer paleontologist who established the earliest Paleontological education and research in China, and has contributed his lifetime to promotion of Chinese paleontological education and researches as well as the studies on the Permian-Triassic （P-Tr） mass extinction and its possible causes. Yang has studied six fossil clades and trace fossils, together with his colleagues, he has established 6 new species of cephalopods, 1 new genus and 15 new species of gastropods, 8 new genera and 31 new species of bivalves, 17 new genera and 66 new species of brachiopods, 1 new genus and 4 new species of ophiuroids, 2 new genera and 7 new species of triopsids （Crustacea）, and 3 new ichnogenera and 7 new ichnospeeies of trace fossils. Yang led the 2nd IGCP working on the P-Tr mass extinction in the world. His group＇s excellent works on basic stratigraphy and paleontology enable the GSSP of P-~r boundary （PTB） to be ratified in China. Yang＇s earlier works on three-episode extinction pattern and volcanism-causing extinction hypothesis are also highlighted here to show how their first-hand data and initiative hypothesis have influenced the current and ongoing debates on the P-Tr crisis and possible causation. Yang school＇s extinction pattern is reviewed here, and their 2nd phase of extinction is marked by a dramatic loss in biodiversity, pointing to a widely accepted mass extinction. The 3rd extinction is characterized by ecological collapse of ecosystem structures and disappearance of the PTB microbialite ecosystem, while the 1st extinction （also prelude extinction） is indicated by the collapses of deep-water and reef ecosystems. Updated studies show that the volcanic ashes near the PTB originated from silicic, subduction-related igneous activity with little or no basaltic input. This subduction zone activity is related to closure of the Paleo-Tethys Ocean, and the intensity and frequency of the volcanic activity appear to increase near the P-Tr extinction interval. Hg anomalies （Hg/TOC ratios and Hg isotopes） were also detected from the P-Tr extinction interval, and they are interpreted as the results of enhanced volcanic-generated atmospheric mercury, which was injected by the violate eruption of the Siberian traps. Thus, the peak felsic volcanism is coeval with violate eruption of Siberian traps, and the coupled relationship between both types of volcanisms and biotic extinction suggests a causal relationship.