荣昌猪、长白猪先天免疫候选基因筛选

旨在探究影响荣昌猪和长白猪机体先天免疫功能的候选基因.以新生荣昌猪、长白猪胸腺为研究对象,利用RNA-seq分析荣昌猪、长白猪胸腺组织中的差异表达基因,同时对差异表达基因进行GO和KEGG富集分析,并通过qRT-PCR验证RNA-seq结果的准确性.结果表明:在新生荣昌猪和长白猪胸腺中,差异倍数在2倍以上的基因共421个,其中,荣昌猪相对长白猪上调的有231个,下调的有190个.GO富集分析发现,这些差异表达基因主要富集在清道夫受体活性、G蛋白偶联受体结合、T细胞活化、细胞分泌调节、胞吐作用调节、SNARE复合物组装调节等生物学过程.KEGG通路分析发现,这些差异表达基因主要参与细胞因子-细胞因子受体互作、cAMP信号、代谢(磷脂酶D、半胱氨酸和蛋氨酸)及一些疾病(非洲锥形病、癌症)相关信号通路.6个差异表达基因的RT-qPCR结果表明,此次RNA-seq的结果准确可靠.综合差异表达基因、GO和KEGG信号通路结果发现,TMSB15A和STAB2、SAA1和HP分别是荣昌猪和长白猪胸腺先天免疫的候选基因.     

甘蓝型油菜BnMAPK1超量表达及中油821的转录差异表达分析

甘蓝型油菜(Brassica napus L.)是我国重要的油料作物之一,具有较高的产量及营养价值。挖掘油菜广谱抗逆基因并深入探究其分子机制,对提高油菜抗逆性且保证产量及品质具有重要意义。MAPK级联(Mitogen-activated protein kinase cascades)是多种信号跨膜传递的交汇点,参与生长发育、生物及非生物胁迫应答等生物学过程。为了解甘蓝型油菜BnMAPK1基因的生物学功能,本研究以BnMAPK1超量表达(OE)和中油821(CK)油菜为材料进行数字基因表达谱分析。结果显示,与CK相比, 650个基因在OE油菜中差异表达;其中上调表达基因243个,下调表达基因407个。GO注释共富集到生物学过程118条途径,其中18条与光合作用相关(包含77个差异表达基因);分子功能7条途径,包括草酸氧化酶活性、锰离子结合、氧化还原酶活性等;细胞组分38条途径,包括叶绿体、质体、类囊体等。KEGG分析结果表明, BnMAPK1参与调控油菜光合作用、碳代谢、次生代谢物生物合成等14条过程。此外,利用实时荧光定量PCR验证8个候选基因在OE和CK油菜中的表达,其表达变化与测...     

假禾谷镰孢侵染小麦后3种植物激素相关基因的差异表达分析

假禾谷镰孢(Fusarium pseudograminearum)是我国近年新发现的小麦病原真菌,本研究目的是揭示植物激素信号传导途径中相关基因表达对假禾谷镰孢侵染的响应。利用假禾谷镰孢野生菌株WZ2-8A侵染小麦品种周麦24,对接种后5 d和15 d样品进行转录组测序,分析差异表达基因,并对候选基因进行q RT-PCR验证。假禾谷镰孢侵染后,小麦幼苗生长受到明显抑制,根长、株高、根重和地上部分鲜重均显著降低。转录组分析结果表明,植物激素信号传导途径中有29个基因差异表达,涉及到生长素、细胞分裂素和脱落酸3种植物激素。在接种后5 d有11个差异表达基因,其中2个上调,9个下调;在接种后15 d共有25个差异表达基因,其中8个上调,17个下调。在生长素信号传导途径中,生长素输入载体AUX1差异表达,影响生长素的极性运输,从而影响小麦根部的细胞伸长。在细胞分裂素信号传导途径中,起正调控作用的B-ARR上调表达,推测其促进细胞分裂素的信号传导,从而抑制细胞分裂,与生长素协同作用,造成小麦的长势减弱。在脱落酸途径中,脱落酸受体PYR/PYL下调表达;起到负调控作用的PP2C相关基因均上调表达。脱落酸使植物对真菌和细菌的抗性起到负调控作用,其信号传导途径与茉莉酸/乙烯途径相互拮抗,脱落酸信号传导途径的阻遏可能会使茉莉酸/乙烯途径信号通路打开。q RT-PCR结果基本能够和转录组测序结果相拟合,说明在假禾谷镰孢侵染胁迫下,脱落酸的信号传导被抑制可能是中抗品种周麦24对假禾谷镰孢产生一定的抗性的生理基础。     

高温胁迫下甜玉米雌穗发育基因差异表达谱分析

利用数字化基因表达谱技术, 对高温胁迫下优良甜玉米杂交种粤甜13雌穗发育相关基因的表达谱进行分析。结果表明, 在高温胁迫和适温下差异表达基因达949个, 其中有上调表达基因705个, 下调表达基因244个, 上调表达10倍以上的基因108个, 下调表达10倍以上的基因40个。对差异表达基因功能注释分析表明, 它们主要集中在细胞内组分和膜上, 主要具催化活性、结合活性、水解酶活性、氧化还原酶活性等, 参与代谢、细胞结构与功能、胁迫应答、物质运输和生物调节等生物学过程, 推测对雌穗发育过程中籽粒和果穗的形成具有重要作用。这些基因中有一半以上的基因功能未知, 下一步将对其开展克隆和功能方面的分析。     

马铃薯PYL基因家族的全基因组鉴定及表达分析

作为关键信号分子,脱落酸(ABA)通过其核心信号通路PYLs-PP2Cs-SnRK2s广泛调控植物的生长发育和逆境响应过程,PYLs蛋白作为ABA信号传导的核心组件,在ABA信号传导中发挥着不可替代的作用。为探究PYLs (PYR/PYL/RCARs)基因在马铃薯中的进化以及表达模式,本研究从马铃薯全基因组‘DM-v 6.1’共鉴定到17个StPYLs基因,并对其分布、蛋白理化性质、系统进化、基因结构特征以及基因表达模式进行了分析。结果表明,17个StPYLs基因不均匀分布在8条染色体上,其氨基酸大小在163～231aa之间,等电点在4.5～8.6之间,相对分子量在18.71～25.29 kD之间。根据基因结构和蛋白的系统发育特征, StPYL家族成员共分为3个亚组, motif 1存在于本家族所有基因中,说明它在StPYLs的进化过程中较为保守。基因表达模式分析表明,StPYL家族成员具有明显的组织表达特异性,且除StPYL1在外源激素(BAP、ABA和IAA)和非生物胁迫(高温、盐和干旱)下均上调表达外,其余基因存在功能分化,不同胁迫下的表达模式各异。本研究结果为进一步阐明StPY...     

植物耐受高温胁迫的分子机制研究进展

随着全球的温度持续上升,高温胁迫已经成为影响植物生长发育的重要因素之一,高温对水稻等农作物产量造成的损失对人类经济收益的影响尤为重要。为了了解植物应对高温胁迫的分子机制,本综述归纳了高温胁迫对植物形态、生理生化、光合作用等方面产生的不利影响并总结了信号传导途径、转录因子的调节、抗高温胁迫相关基因的表达这3种植物应对高温胁迫的分子机制。根据以上内容,本文建议利用生物信息学、基因工程、细胞生物学、分子生物学等手段继续深入探索植物耐受高温胁迫的分子机制,并对该领域未来的研究方向提出了展望。     

番茄系统创伤信号因子研究进展

当植物受到机械损伤或害虫伤害时,植物体会在受伤部位产生创伤信号分子启动防御基因的系统表达.许多研究表明系统素(或其前体原系统素)在受伤部位激活茉莉酸的合成,使之达到系统反应的水平,应对外来伤害;茉莉酸或其衍生物是重要的系统伤信号分子,它诱导伤防御基因的系统表达.自从系统素和茉莉酸被认为是伤诱导防御基因表达的信号分子后,番茄成为研究植物系统伤信号很好的模式植物.本文介绍了番茄的茉莉酸合成突变体、茉莉酸感知突变体和系统素功能突变体,及它们之间相互作用,并推测了茉莉酸和系统素在系统创伤信号调节中的作用.     

拟南芥PUB10和PUB11基因抵抗丁香假单胞杆菌番茄致病变种DC3000株的功能分析

丁香假单胞杆菌Pseudomonas syringae pv tomato(Pst)DC3000是研究植物免疫机制的常用病原菌。为分析拟南芥E3泛素连接酶PUB10、PUB11在免疫中的功能,本研究构建了pub10/pub11双突变体。在分析拟南芥PUB10、PUB11序列相似性和进化关系的基础上,比较了pub10/pub11双突变体和Col-0对Pst DC3000的敏感性、MAPK磷酸化水平、ROS产生的影响,并检测JA信号响应途径和免疫防御相关的部分代表性基因的表达量。结果表明PUB10、PUB11氨基酸序列高度保守,pub10/pub11双突变体对Pst DC3000的侵染更敏感,侵染后叶片病原菌数目明显增多,MAPK磷酸化水平迅速下降,ROS释放量降低,JA信号响应基因(LOX3,JR2)、免疫防御负调控基因(NIMIN1,WRKY38,WRKY62,RIN4)表达量上升显著高于Col-0,而免疫防御正调控基因(AZI1,EDS1,PAD4,EDS5,FMO1,NPR1)表达量显著低于Col-0。由此推断,AtPUB10、AtPUB11在防御Pst DC3000中起正调控作用,并且存在功能冗余。研究结果为进一步探索PUB10、PUB11调控植物防御的分子作用机制和分子育种提供了依据。     

马铃薯PIF家族成员鉴定及其对高温胁迫的响应分析

植物光敏色素作用因子(phytochrome interacting factors,PIFs)属于碱性-螺旋-环-螺旋(basic helix-loop-helix,bHLH)转录因子家族,通过将光和温度等外部环境信号与植物体内源信号途径相整合,进而形成复杂的信号转导网络来精密调控植物的生长发育进程。目前,关于马铃薯PIF家族基因的研究较少,鉴定和分析StPIF家族成员有助于进一步提高马铃薯的产量和品质。本研究运用生物信息学方法,以拟南芥PIF家族成员蛋白序列作为源序列,通过在马铃薯基因组数据库中进行BlastP分析鉴定出7个StPIFs家族成员,并对其进行系统进化、染色体分布、复制事件、蛋白理化性质、基因结构、Motif预测、启动子顺式作用元件、基因表达模式以及对高温胁迫的响应分析。结果显示,StPIF家族所有成员均含有Motif 1 (bHLH结构域)、Motif 2 (APB结构域)基序;在StPIF基因的启动子区域预测到多个参与光响应、激素、干旱、低温、昼夜节律以及防御和应激反应调控元件;基因表达模式和现蕾期高温胁迫响应分析表明,家族成员具有明显的组织表达特异性,基因存在功能...     

基于高通量测序技术鉴定松辽黑猪与长白猪背最长肌差异基因

旨在利用高通量测序技术对松辽黑猪和长白猪背最长肌组织样本进行mRNA测序和差异分析,筛选影响猪肌肉生长、肉质和脂肪沉积的关键基因,从而为猪肉品质研究提供新的参考信息。采集6头松辽黑猪和6头长白猪的背最长肌组织样本,提取其RNA,采用Illumina HiSeq 2500高通量测序技术对mRNA进行测序,并对获得的reads进行比对、注释和差异表达分析,用NOISeq筛选出差异表达基因并进行相关生物学功能富集分析。结果表明,从2个猪种间共筛选出了664个差异表达基因,其中,364个基因在松辽黑猪中高表达,300个基因在长白猪中高表达。通过对差异表达基因进行生物学功能分析,筛选出LPIN1、FADS1、FADS2、PLIN2、PPARGC1A、PRKAG2和ACSL1等基因参与脂类代谢和肌肉发育相关的调控,相关通路为脂肪酸代谢、PPAR信号通路、AMPK信号通路、胰岛素信号通路和脂肪细胞因子信号通路等。     

Location of a high-lysine gene and the DDT-resistance gene on barley chromosome 7

Summary The high-lysine gene in Risø mutant 1508 conditions an increased lysine content in the endosperm via a changed protein composition, a decreased seed size, and several other characters of the seed. The designation lys3a, lys3b, and lys3c, is proposed for the allelic high-lysine genes in three Risø mutants, nos 1508, 18, and 19. Linkage studies with translocations locate the lys3 locus in the centromere region of chromosome 7. A linkage study involving the loci lys3 and ddt (resistance to DDT) together with the marker loci fs (fragile stem), s (short rachilla hairs), and r (smooth awn) show that the order of the five loci on chromosome 7 from the long to the short chromosome arm is r, s, fs, lys3, ddt. The distance from locus r to locus ddt is about 100 centimorgans.     

Biological Activity and Quantification of Suspected Allelochemicals from Alfalfa Plant Parts

Autotoxicity restricts reseeding of alfalfa (Medicago sativa L.) after alfalfa until autotoxic chemical(s) breaks down or is dispersed into external environments. A series of aqueous extracts from leaves, stems, roots and seeds of alfalfa ‘Vernal’ were bioassayed against alfalfa seedlings of the same cultivar to determine their autotoxicity. The highest inhibition was found in the extracts from the leaves. Extracts at 40 g dry tissue l^?1 from alfalfa leaves were 15.4, 17.5 and 28.7 times more toxic to alfalfa root growth than were those from roots, stems and seeds, respectively. A high‐performance liquid chromatography (HPLC) analysis with nine standard compounds showed that the concentrations and compositions of allelopathic compounds depended on the plant parts. In leaf extracts that showed the most inhibitory effect on root growth, the highest amounts of allelochemicals were detected. Among nine phenolic compounds assayed for their phytotoxicity on root growth of alfalfa, coumarin, trans‐cinnamic acid and o‐coumaric acid at 10^?3 m were most inhibitory. The type and amount of causative allelochemicals found in alfalfa plant parts were highly correlated with the results of the bioassay, indicating that the autotoxic effects of alfalfa plant parts significantly differed.     

Simultaneous Determination of Four Phenolic Acids in Radix Salviae Miltiorrhizae Formula Granules by QAMS

[Objectives]This study aimed to establish a QAMS(quantitative analysis of multi-components by single-marker)method for simultaneous determination of four phenol...     

Changes in Seed Yield and Quality with Maturity in Onion (Allium cepa L., cv. 'Early Cream Gold')

Development of onion (Allium cepa L., cv. ‘Early Cream Gold’) seed under cool climate conditions in Tasmania, Australia occurred over a longer duration than previously reported, but similar patterns of change in yield components were recorded. In contrast to previous studies, umbel moisture content declined from 85 to 67 % over 57 days while seed moisture content decreased from 85 to 31 %. Seed yield continued to increase over the duration of crop development, with increasing seed weight compensating for seed loss resulting from capsule dehiscence in the later stages of maturation. Germination percentage was high and did not vary significantly from 53 to 77 days after full bloom (DAF), but mean germination time declined and uniformity of germination increased significantly over the same time period. The percentage abnormal seedlings declined with later harvest date, resulting in highest seed quality at 77 DAF. The results of this study suggest that the decision to harvest cool climate onion seed crops before capsule dehiscence will result in a loss of potential seed yield and quality.     

Water Extraction Process of Chinese Herbal Compound Man Gan Ning

[Objectives]To optimize the water extraction process of Chinese Herbal Compound Man Gan Ning and establish a method for its extraction and content determination...     

Present status and future strategy in breeding faba beans (Vicia Faba L.) for resistance to biotic and abiotic stresses

Progress is being made, mainly by ICARDA but also elsewhere, in breeding for resistance to Botrytis, AScochyta, Uromyces, and Orobanche; and some lines have resistance to more than one pathogen. The strategy is to extend multiple resistance but also to seek new and durable forms of resistance. Internationally coordinated programs are needed to maintain the momentum of this work.Tolerance of abiotic stresses leads to types suited to dry or cold environments rather than broad adaptability, but in this cross-pollinated species, the more hybrid vigor expressed by a cultivar, the more it is likely to tolerate various stresses.     

Pollen and pollination experiments. III. The viability of apple and pear pollen as affected by irradiation and storage

Summary Apple and pear pollen was irradiated with doses of 0, 50, 100, 250 and 500 krad (gamma rays) and stored at 4°C and 0–10% r.h. From the in-vitro germination percentages an average LD 50 dose of about 220 krad was estimated. For both irradiated and untreated pollen a close and corresponding lineair relationship existed between germination percentage and pollen tube growth.Irradiated pollen was much more sensitive to dry storage conditions than untreated pollen, resulting in less germination and more bursting. Apparently, irradiation caused the pollen cell membrane to lose its flexibility faster than normal. Rehydration of dry-stored, irradiated pollen in water-saturated air restored germination percentages up to their initial levels. The importance of this procedure in germination trials is stressed.     

Optimization of Extraction Technology and Determination of Content of Total Phenols of Leaves in Acanthopanax giraldii Harms

[Objectives] To determine the optimum extraction technology for total phenols of leaves in Acanthopanax giraldii Harms.[Methods]The single factor test and ortho...     

Cytoplasmic male sterility,resistance to gall mite and mildew,and season of leafing out in black currants

Summary Cytoplasmic male sterility (cms) is described in the F₁ hybrids Ribes × carrierei (R. glutinosum albidum × R. nigrum) and R. sanguineum × R. nigrum. In backcrosses to R. nigrum, progenies with R. glutinosum cytoplasm were either all male sterile, or segregated for full male fertility (F) and complete (S) and partial (I) male sterility. Ratios of F:I+S suggested that two linked genes controlled cms, F plants being dominant for one (Rf ₁) and recessive for the other (Rf ₂).Segregation for cms in relation to three linded genes, Ce (resistance to the gall mite, Cecidophyopsis ribes), Sph ₃(resistance to American gooseberry mildew, Sphaerotheca mors-uvae) and Lf ₁(one of two dominant additive genes controlling early season leafing out) indicated that Rf ₁and Rf ₂were in this linkage group. The gene order and approximate crossover values appeared to be: % MathType!MTEF!2!1!+-% feaafiart1ev1aaatCvAUfeBSjuyZL2yd9gzLbvyNv2CaerbuLwBLn% hiov2DGi1BTfMBaeXafv3ySLgzGmvETj2BSbqef0uAJj3BZ9Mz0bYu% H52CGmvzYLMzaerbd9wDYLwzYbItLDharqqr1ngBPrgifHhDYfgasa% acOqpw0xe9v8qqaqFD0xXdHaVhbbf9v8qqaqFr0xc9pk0xbba9q8Wq% Ffea0-yr0RYxir-Jbba9q8aq0-yq-He9q8qqQ8frFve9Fve9Ff0dme% aabaqaciGacaGaamqadaabaeaafaaakeaacaWGdbGaamyzamaamaaa% baGaaiiiaiaacccacaGGWaGaaiOlaiaacgdacaGG0aGaaiiiaiaacc% caaaGaaiiiaiaacccacaGGGaGaamOuaiaadAgaliaaigdakmaamaaa% baGaaiiiaiaacccacaGGGaGaaiiiaiaaccdacaGGUaGaaiOmaiaacs% dacaGGGaGaaiiiaiaacccacaGGGaGaaiiiaaaacaWGsbGaamOzaSGa% aGOmaOWaaWaaaeaacaGGGaGaaiiiaiaacccacaGGGaGaaiiiaiaacc% cacaGGGaGaaiiiaiaacccaaaGaamitaiaadAgaliaaigdakmaamaaa% baGaaiiiaiaacccacaGGGaGaaiiiaiaacccacaGGGaGaaiiiaiaacc% cacaGGGaGaaiiiaiaacccacaGGGaaaaiaadofacaWGWbGaamiAaSGa% aG4maaaa!6E4D!\[Ce\underline { 0.14 } Rf1\underline { 0.24 } Rf2\underline { } Lf1\underline { } Sph3\]. Crossover values of 0.36 for Ce-Lf ₁, and 0.15 for Lf ₁-Sph ₃were estimated from the relative mean differences in season of leafing out between seedlings dominant and recessive for Ce and Sph ₃.It is suggested that competitive disadvantage of lf ₁-carrying gametes and/or zygotes at low temperatures may be implicated in the almost invariable deficit of plants dominant for the closely linked mildew resistance allele Sph ₃. Poor performance of lf ₁- (and possibly lf ₂-) carrying gametes and young zygotes during periods of low temperature at flowering might also account for the liability of some late season cultivars and selections to premature fruit drop (running off).     

Screening techniques and sources of resistance to parasitic angiosperms

Parasitic angiosperms cause great losses in many important crops under different climatic conditions and soil types. The most widespread and important parasitic angiosperms belong to the genera Orobanche, Striga, and Cuscuta. The most important economical hosts belong to the Poaceae, Asteraceae, Solanaceae, Cucurbitaceae, and Fabaceae. Although some resistant cultivars have been identified in several crops, great gaps exist in our knowledge of the parasites and the genetic basis of the resistance, as well as the availability of in vitro screening techniques. Screening techniques are based on reactions of the host root or foliage. In vitro or greenhouse screening methods based on the reaction of root and/or foliar tissues are usually superior to field screenings and can be used with many species. To utilize them in plant breeding, it is necessary to demonstrate a strong correlation between in vitro and field data. The correlation should be calculated for every environment in which selection is practiced. Using biochemical analysis as a screening technique has had limited success. The reason seems to be the complex host-parasite interactions which lead to germination, rhizotropism, infection, and growth of the parasite. Germination results from chemicals produced by the host. Resistance is only available in a small group of crops. Resistance has been found in cultivated, primitive and wild forms, depending on the specific host-parasite system. An additional problem is the existence of pathotypes in the parasites. Inheritance of host resistance is usually polygenic and its transfer is slow and tedious. Molecular techniques have yet to be used to locate resistance to parasitic angiosperms. While intensifying the search for genes that control resistance to specific parasitic angiosperms, the best strategy to screen for resistance is to improve the already existing in vitro or greenhouse screening techniques.