高浓度氧或HCl处理条件下家蚕胚胎早期比较转录组分析

本研究以阐明阻断蚕卵滞育机制为目标,通过获得连续时间点的转录组数据,深入解析滞育过程的分子变化.采用高通量测序技术对高浓度氧、HC1处理的滞育蚕卵(以下简称"处理组")和未处理的对照组,在产后26 h、32 h和44h取样进行转录组测序,在产后26 h到32h阶段,处理组和对照组的共同差异基因(DE-Gs)233个,占...     

仁用杏果肉3个发育阶段的转录组分析

仁用杏果肉糖酸比低是造成其鲜食性差的主要原因。该研究以仁用杏‘山苦2号’开花后30、60和90d的果肉为材料,利用RNA-seq对仁用杏果肉进行转录组测序及差异表达基因的筛选,以了解可能与糖酸合成相关的基因及其表达模式,为进一步解析仁用杏果肉糖酸比低的分子调控模式与机制,深入研究仁用杏果肉糖酸代谢转化机制提供依据。结果显示:(1)在仁用杏果肉发育的3个阶段共获得差异表达的单基因簇(unigene)28 989条;根据单基因簇的功能注释,共发掘出19条单基因簇编码5个与仁用杏果肉中糖合成代谢相关的酶,54条单基因簇编码15个与仁用杏果肉中酸合成代谢相关的酶。(2)在所选3个发育阶段的仁用杏果肉中共发现9条与糖代谢相关的单基因簇和27条与酸代谢相关的单基因簇呈现差异表达。(3)编码与蔗糖合成积累相关的蔗糖磷酸酶和蔗糖合成酶的单基因簇主要呈下调表达,而与糖分解相关的果糖激酶和己糖激酶却呈上调表达。(4)与仁用杏果肉中主要有机酸组分苹果酸积累相关的苹果酸脱氢酶主要呈上调表达,而分解苹果酸的苹果酸酶呈下调表达。研究推测,仁用杏果肉在发育中糖积累受阻,而有机酸却得到大量的合成积累,这可能是造成仁用杏果肉糖酸比例小,鲜食口感差的主要内在因素之一。     

基于转录组测序的扎龙湿地野大麦耐盐性分析

通过转录组测序技术对扎龙湿地野大麦幼苗在200 mmol/L NaCl胁迫2 d后的叶片进行分析。对测序结果进行De novo拼接后,将差异表达基因在GO、KEGG数据库中进行比对注释。结果表明:NaCl胁迫2 d后,对照组与处理组分别获得Unigene序列61038个和46754个,检测到差异表达基因25465个,差异基因GO功能注释到3个大类的55个功能组。差异表达基因被注释到135个Pathway上,直观地显示出NaCl胁迫下野大麦幼苗体内发生调节及改变的代谢过程和信号通路,其中主要涉及光合作用、脯氨酸代谢、叶绿素代谢等途径。通过对野大麦幼苗叶片转录组分析,发掘相关耐盐基因,有助于培育野大麦及其近缘作物新品种,并对揭示植物耐盐性分子机制及相关代谢途径具有重要意义。     

基于转录组学和代谢组学联合分析草珊瑚萜类化合物生物合成的组织特异性分布

本研究旨在探究草珊瑚叶和根中萜类化合物的组织特异性分布差异,解析其药效品质差异形成的分子机制。采用液相色谱-质谱联用技术(liquid chromatography-mass spectrometry, LC-MS)和Illumina HiSeq^TM高通量测序技术获得草珊瑚[Sarcandra glabra (thunb) naka]叶和根的代谢组学和转录组学数据。代谢组学结果表明参与叶和根中萜类合成的差异代谢物有50个,包括法尼西基半胱氨酸、甘油醛-3-磷酸、甲羟戊酸-5-磷酸等。转录学结果表明差异代谢酶基因有57条,包括ACTC、HMGCR、MVK、DXS与KS等,并预测了MYB、C2H2、AP2/ERF-ERF等7个转录因子参与调控草珊瑚不同组织部位中萜类的合成和积累差异。实时荧光定量PCR (quantitative real-time polymerase chain reaction, qRT-PCR)结果显示,随机选取的8个参与萜类合成的酶基因在草珊瑚不同组织部位中的表达趋势与转录组学测序结果一致。本研究有助于阐明草珊瑚叶和根临床疗效差异形成的分子机制,同时为草珊瑚的资源开发利用奠定基础。     

枝干树皮宏基因组DNA的提取

旨在得到一种适用于提取多种枝干树皮的高质量DNA的方法。参考前人提取植物DNA的经验,综合了研磨时加PVP、缓冲液洗涤、SDS与CTAB结合使用、高浓度KAc低温冻融、高浓度Na Cl条件下异丙醇沉淀、DNA完全溶解后加微量RNase A处理等操作,所得5种基因组DNA浓度介于190-970 ng/μL,纯度都很高,皆能被限制性内切酶切割,都可用作模板扩增到细菌16S r RNA基因片段,以苹果树皮DNA为模板扩增到苹果BIP和PDI基因且苹果树皮基因组DNA经测序公司检测,质量满足高通量测序法研究微生物多样性对环境宏基因组的要求。     

空间分辨代谢组学进展和挑战

空间分辨代谢组学即整合质谱成像和代谢组学技术,对动/植物组织和细胞中内/外源性代谢物的种类、含量和差异性空间分布进行精准测定。质谱成像技术因其具有无标记、非特异、高灵敏度、高化学覆盖、元素/分子同时检测等优势,被广泛应用于动/植物组织中各类代谢物、多肽和蛋白的时空分布研究。首先介绍了代谢组学和质谱成像技术的研究现状,然后重点综述了空间分辨代谢组学在动物组织、植物组织和单细胞水平上的前沿应用。最后展望了空间分辨代谢组学技术的现有瓶颈和未来发展方向。空间分辨代谢组学是继代谢组学之后又一门新兴的分子成像组学技术,能够无标记、可视化检测动物组织中外源性药物的吸收、分布、代谢和排泄,以及植物组织中多种代谢产物的生物合成、转运途径和积累规律。该技术将推动靶向药物发现、病理机制解析和动植物生长发育密切关联的空间代谢网络调控等前沿应用研究。     

高通量转录组测序技术及其在鳞翅目昆虫上的应用

转录组是指组织或者细胞在某一特定状态下转录出来的所有RNA的集合。高通量第2代测序技术使转录组学的研究模式发生了巨大的改变,所衍生出的转录组测序迅速成为研究非模式生物的先进技术。转录组测序能够在整体水平上探究细胞内基因表达的种类和数量,揭示在特定条件下机体生理生化发生过程以及其中的分子机理。本文简要阐述了转录组测序技术的基本概念、技术流程与原理,详细介绍了转录组测序在解决鳞翅目昆虫的分类、毒理、发育、与寄主互作以及非编码RNA调控等问题上做出的贡献,并对该技术现存的困难进行了系统的阐述并对其未来的发展趋势作出了简要的预测与剖析。     

款冬花不同发育阶段的代谢组学和比较转录组学分析

以不同发育阶段款冬花（发育初期、中期、中后期、解封期及花朵期）为对象,基于核磁共振的代谢组学技术,分析其次生代谢物种类及含量的合成累积规律,并进行高通量转录组学测序,从差异表达的基因中寻找次生代谢物生物合成的关联酶基因。代谢组学分析发现,款冬花不同发育阶段的次生代谢物代谢组成明显不同,苯丙素类成分在发育初期至中后期含量较高,之后逐渐降低;黄酮类成分（芦丁、山奈酚）在发育的各个阶段含量均有波动,但总体变化不大。转录组学分析结果显示,与苯丙素类成分生物合成相关的酶基因（COMT、HCT）,随着花蕾的发育表达量逐渐降低;与黄酮类成分合成相关的酶基因（FLS、F3H、DFR）,其表达量在不同阶段变化不大。转录组测序中的相关酶基因表达量同次生代谢物成分的变化趋势基本一致。本文采用的代谢组学和转录组学结合的方式,对款冬花的次生代谢物累积规律进行分析,为今后款冬花次生代谢物的生物合成调控研究奠定了基础。     

多组学技术及其在生命科学研究中应用概述

随着新一代测序技术、高分辨质谱技术、多组学整合分析方法及数据库的发展,组学技术正从传统的单一组学向多组学技术发展。以多组学驱动的系统生物学研究将带来生命科学研究的新范式。本文简要概述了基因组学、表观基因组学、转录组学,蛋白质组学及代谢组学的进展,重点介绍多组学技术平台的组成和功能,多组学技术的应用现状及在合成生物学及生物医学等领域的应用前景。     

基于转录组分析鉴定苹果茉莉素响应基因

为阐明外源茉莉酸甲酯(MeJA)诱导的苹果(Malus domestica)抗病分子机制, 以生长30天的Gala组培苗为试材, 用100 μmol?L ^-1MeJA处理叶片12小时, 通过转录组测序, 结合生物信息学分析鉴定出苹果叶片中受MeJA诱导表达的基因。结果表明, 外源MeJA主要影响苹果叶片倍半萜类、三萜和类黄酮的生物合成, 以及芸薹素(BR)信号转导途径间接诱导的抗病性; 倍半萜类、三萜及类黄酮生物合成途径中的关键基因为MDP0000702120和MDP0000692178; MDP0000123379是联系芸薹素信号转导途径和植物-病原菌互作途径的关键调控基因。     

基于转录组分析鉴定苹果茉莉素响应基因

为阐明外源茉莉酸甲酯(MeJA)诱导的苹果(Malus domestica)抗病分子机制, 以生长30天的Gala组培苗为试材, 用100 μmol∙L ^-1MeJA处理叶片12小时, 通过转录组测序, 结合生物信息学分析鉴定出苹果叶片中受MeJA诱导表达的基因。结果表明, 外源MeJA主要影响苹果叶片倍半萜类、三萜和类黄酮的生物合成, 以及芸薹素(BR)信号转导途径间接诱导的抗病性; 倍半萜类、三萜及类黄酮生物合成途径中的关键基因为MDP0000702120和MDP0000692178; MDP0000123379是联系芸薹素信号转导途径和植物-病原菌互作途径的关键调控基因。     

The phenylpropanoid pathway affects apple fruit resistance to Botrytis cinerea

Apple fruits are rich in phenolic compounds that may enhance resistance to grey mould disease caused by Botrytis cinerea. Using Malus domestica Borkh. cultivars Fuji and Qinguan, we analysed the contents of total phenols, total flavonoids, eight individual phenolic compounds, H₂O₂ and O₂^.? as well as the activities of key enzymes in the phenylpropanoid pathway in the flesh of control and B. cinerea‐inoculated fruits. Chlorogenic acid contents increased for a short period in the less susceptible cultivar Qinguan fruits, but decreased in the disease‐susceptible Fuji fruits. Additionally, ferulic acid production was induced in both cultivars in response to B. cinerea. Furthermore, the activities of phenylalanine ammonia lyase, cinnamate 4‐hydroxylase, 4‐coumarate:coenzyme A ligase and cinnamyl alcohol dehydrogenase were differentially induced between the two apple cultivars. Remarkably, the contents of H₂O₂ and O₂^.? as well as the activities of enzymes in phenolic metabolism tested in this study were always higher in Qinguan fruits than in Fuji fruits. Our data imply that phenylpropanoid metabolism is closely associated with apple fruit resistance to grey mould disease. These findings may be useful for characterizing the mechanism(s) underlying plant resistance to B. cinerea, with potential implications for the screening of grey mould disease‐resistant apple varieties in breeding programmes.     

高氧对果蔬采后生理影响研究进展

高氧（21%-100%O2）贮藏是近几年发展起来的果蔬采后处理技术之一.概述了高氧对果蔬采后生理如呼吸作用、乙烯合成、组织褐变、活性氧代谢及抗氧化能力、风味等方面的影响,还介绍了高氧对病原微生物生长和采后果蔬腐烂控制的作用.     

Air pollution effects on structure,proteins and flavonoids in pollen grains of Thuja orientalis L. (Cupressaceae)

Increase in the levels of air pollution due to the increase in industrial and agricultural technology has prompted investigation of mechanisms that contribute to air pollution tolerance in plants. Pollen grains of Thuja orientalis were collected from controlled (less polluted) and polluted areas (mainly SO₂, NO₂, CO, HC and APM). Thuja pollen is considered inaperturate and granulate. The exine is shed during rehydration, leaving the male gametophyte naked. The pollen grains collected from polluted areas are smaller and more fragile compared to control ones. The exine splitting is faster and higher in polluted pollen grains. SDS–PAGE (sodium dodecyl sulphate-polyacrylamide gel electrophoresis) pattern do not show significant differences in polluted pollen than those in the control group. HPLC (high pressure liquid chromatography) analysis demonstrates that air pollution induces flavonoids accumulation to significantly higher levels in polluted pollen than in controlled ones. These observations suggest that plants try to respond suitably by adjusting their metabolism so that minimum damage is done due to air pollutants. Their protective responses may include an increase in antioxidant enzymes and metabolites and induction of protection-related secondary metabolite genes especially flavonoids.     

宁夏枸杞‘中科绿川1号’果实和叶片的主要成分分析

以国家林业局审定的宁夏枸杞新品种‘中科绿川1号’为实验材料,研究不同月份采集的枸杞叶片芦丁含量,以及果实总黄酮、总多糖、总类胡萝卜素、玉米黄素双棕榈酸酯的含量变化。结果显示,叶片芦丁含量在枸杞盛果期较低,抽条期较高;果实总黄酮、总类胡萝卜素、玉米黄素双棕榈酸酯的含量在头茬果中含量较高,盛果期果实含量较低;果实总多糖可能与果实量呈负相关;褐化果(俗称"油果")总黄酮含量与正常果差异不大,可用作提取枸杞黄酮类物质的原材料。研究结果可为枸杞果实分级、成分提取提供新思路。     

Some aspects of enzymatic browning in apples

The investigations outlined in ‘Browning reactions in fruits’, part of the special study of food science of the Nuffleld Advanced Chemistry course, are modified to make the material meaningful and relevant to pupils in the middle school. This article discusses a series of simple experiments on apple browning which were performed by third formers. A summary of the nature of the browning process and its control are included.     

Two differentially expressed MATE factor genes from apple complement the Arabidopsis transparent testa12 mutant

Proanthocyanidins (PAs) are a class of flavonoids with numerous functions in plant ecology and development, including protection against microbial infection, animal foraging and damage by UV light. PAs are also beneficial in the human diet and livestock farming, preventing diseases of the cardiovascular system and lowering the risk of cancer, asthma and diabetes. Apples (Malus x domestica Borkh.) are naturally rich in flavonoids, but the flavonoid content and composition varies significantly between cultivars. In this work, we applied knowledge from the model plant Arabidopsis thaliana, for which the main features of flavonoid biosynthesis have been elucidated, to investigate PA accumulation in apple. We identified functional homologues of the Multidrug And Toxic compound Extrusion (MATE) gene TRANSPARENT TESTA12 from A. thaliana using a comparative genomics approach. MdMATE1 and MdMATE2 were differentially expressed, and the function of the encoded proteins was verified by complementation of the respective A. thaliana mutant. In addition, MdMATE genes have a different gene structure in comparison to homologues from other species. Based on our findings, we propose that MdMATE1 and MdMATE2 are vacuolar flavonoid/H(+) -antiporters, active in PA accumulating cells of apple fruit. The identification of these flavonoid transporter genes expands our understanding of secondary metabolite biosynthesis and transport in apple, and is a prerequisite to improve the nutritional value of apples and apple-derived beverages.