AtWRKY40参与拟南芥干旱胁迫响应过程

以拟南芥(Arabidopsis thaliana)野生型、AtWRKY40缺失突变体和过表达株系为材料,研究AtWRKY40在植物干旱胁迫响应过程中的作用及其生理和分子机制。结果显示,AtWRKY40受干旱胁迫诱导;AtWRKY40缺失导致干旱胁迫下种子萌发率降低,叶片失水加剧,而AtWRKY40过表达植株呈现出相反的表征;干旱胁迫下,At WRKY40缺失突变体植株叶片过氧化氢(H_2O_2)、超氧阴离子(O_2~-·)及丙二醛(MDA)含量显著高于野生型及其过表达株系,超氧化物歧化酶(SOD)和过氧化氢酶(CAT)活性、脯氨酸(Pro)和可溶性糖含量以及相关基因AtCu/ZnSOD、AtCAT1、AtP5C1S、AtG6PD5和AtBAM4表达量显著低于野生型,同时AtWRKY40过表达株系的渗透物质含量和保护酶活性及其基因表达量则高于野生型。由此说明,AtWRKY40通过调节植株抗氧化能力及渗透调节能力参与拟南芥干旱胁迫响应过程。     

拟南芥黑芥子酶TGG1对抗旱性的作用

为阐明拟南芥中黑芥子酶TGG1对抗旱性的影响,构建了35S启动子驱动的TGG1过表达载体,并将其转入拟南芥获得了转基因植株。以野生型和过量表达TGG1的转基因植株为材料,进行干旱胁迫实验,结果显示,在甘露醇模拟的干旱胁迫下35S∶TGG1种子平均发芽率显著高于野生型,平均相对电导率则显著低于野生型;自然干旱胁迫下,35S∶TGG1的相对失水速率显著慢于野生型,而干旱复水后的平均存活率则显著高于野生型。对气孔的观察结果表明,过表达TGG1的转基因植株气孔对ABA处理具有更高的敏感性,气孔关闭程度显著高于野生型植株。以上研究结果表明,过量表达TGG1基因可显著提高拟南芥的抗旱能力,而且其抗性机制很可能与气孔在逆境下的关闭程度有关。     

油菜素内酯合成酶基因DAS5促进杨树生长及提高抗旱性的作用

以河北杨(Populus hopeiensis)为材料, 研究拟南芥(Arabidopsis thaliana)油菜素内酯(BR)生物合成酶基因DAS5对其生长表型、生物量及抗旱性的影响。结果表明: (1) 转DAS5基因河北杨植株的根长、地径、叶柄及叶片长度均显著大于野生型植株, 且地上、地下部分干重及根冠比显著高于野生型, 其拥有发达的根系; (2) 在干旱胁迫下, 转DAS5基因河北杨植株失水褪绿速度较野生型植株缓慢, 在复水后转基因植株能够较早较好地恢复活力, 萌发较多的新幼芽且长势良好; (3)控水期间, 转基因河北杨的相对生长率显著高于野生型, 且随着干旱胁迫程度的加剧, 其可溶性糖含量、游离脯氨酸含量、过氧化氢酶(CAT)活性、超氧化物歧化酶(SOD)活性均显著高于野生型。实验结果表明, 与野生型相比, 转基因植株具有较高的生长量与较强的抗干旱胁迫能力, 说明来自拟南芥的BR生物合成酶基因DAS5可以显著增加河北杨的生长量并在抵御干旱胁迫机制中发挥重要作用。     

转拟南芥P5CS1基因增强羽衣甘蓝的耐旱性

为提高羽衣甘蓝的耐旱性,本文将拟南芥Δ1-吡咯啉-5-羧酸合成酶（P5CS1）基因经农杆菌介导转入羽衣甘蓝植株中,检测转基因株系与野生型植株在干旱胁迫下P5CS1 mRNA表达量、幼苗脯氨酸含量、株系根系性状、整株干重、鲜重和整株存活率。结果表明,在15%PEG6000渗透胁迫下,转基因植株的P5CS1基因mRNA表达量明显增加,转基因植株脯氨酸含量是野生型的2.4倍;主根长、最长侧根长、侧根数目、整株干重和鲜重均高于野生型,干重/鲜重则低于野生型,转基因植株的平均存活率为78%,极显著高于野生型。数据显示,AtP5CS1基因在羽衣甘蓝中的表达明显改善了转基因植株的耐旱性。     

高粱ATP合成酶E亚基基因(SbATPase-E)的克隆及其抗逆功能研究

高粱是一种抗旱性较强的禾谷类作物。本研究在高粱中克隆到一个全长为693 bp的编码ATP合成酶E亚基的基因(SbATPase-E)。在高粱幼苗期,SbATPase-E基因受Na Cl和脱落酸(ABA)处理诱导上调表达。该基因在拟南芥中过量表达可提高转基因植株的耐旱性和耐盐性,在逆境胁迫条件下转基因拟南芥植株较野生型植株根系发达,可能是转基因植株耐旱性和耐盐性提高的主要原因。在干旱胁迫条件下,转基因植株中DREB2A、P5CS1、RD29A、RAB18和ABI1基因的表达量相对于野生型植株中的表达量提高更为显著;在高盐处理条件下,转基因植株中SOS1和SOS2基因的表达量也较野生型植株中的表达量明显提高。这些抗逆相关基因的上调表达可能是转基因植株抗逆性提高的主要分子机制。     

过量表达谷胱甘肽转移酶基因对转基因拟南芥抗旱能力的影响

盐地碱蓬谷胱甘肽转移酶基因（OST）在拟南芥中过量表达后,在干旱胁迫下,转基因拟南芥植株的干重比野生型植株高,其总谷胱甘肽含量和谷胱甘肽库的氧化水平都比野生型植株的高,而丙二醛含量则比野生型的低。这些显示转基因拟南芥的抗干旱胁迫能力有所增强。     

羊草LcPIP基因遗传转化露地菊及其抗盐性鉴定

PIP是重要的水孔蛋白之一,与植物的抗逆性有关。本文用叶盘法将羊草LcPIP遗传转化露地菊火焰,通过常规PCR和GUS染色方法鉴定转基因株系,Real-time PCR检测盐胁迫下转基因和野生植株中CmPIP1和CmPIP2的相对表达量,并测定SOD、POD活性和MDA含量。结果显示,在转LcPIP基因的露地菊植株叶片中,CmPIP1和CmPIP2基因表达量均上升,是野生型植株的2倍;根部的CmPIP1与CmPIP2基因的表达量均上升,CmPIP1上升程度高于CmPIP2。在200 mmol·L~(-1) NaCl的胁迫下,野生型植株中CmPIP1基因表达量明显下降,而转基因植株中CmPIP1基因表达量在12~48 h出现明显的上升;在盐胁迫12 h后,转基因植株中CmPIP2基因表达量上升程度明显高于野生型植株。盐胁迫12 h后,转基因植株的SOD活性提高更为明显。在盐胁迫前期(0~24 h)转基因植株MDA含量增加幅度低于野生型植株;在盐胁迫后期(48~72 h)转基因露地菊POD活性出现明显上升,而野生型露地菊呈下降趋势。     

蒙古冰草MwMYB4基因功能鉴定

MwMYB4基因是从蒙古冰草中克隆得到的MYB类转录因子家族成员之一。该研究以转MwMYB4基因的拟南芥后代为材料,通过在干旱和低温胁迫下对转基因植株进行表型分析、理化指标测试和分子鉴定,分析并验证MwMYB4基因的功能。结果显示:(1)蒙古冰草MwMYB4基因已成功整合到转基因拟南芥T_1代的基因组中并实现转录水平的表达。(2)转基因拟南芥T_2代植株在干旱胁迫条件下,转基因植株叶片枯黄程度较轻,相对电导率较野生型变化幅度低,脯氨酸含量明显高于野生型对照,且MwMYB4基因的表达量随干旱胁迫时间延长而增加。(3)在低温胁迫条件下,转基因拟南芥叶片的枯白程度明显低于野生型,且MwMYB4基因的表达量随低温胁迫时间增加而增加。研究表明,过量表达蒙古冰草MwMYB4基因能够提高转基因拟南芥对干旱和低温的耐受性,该基因可能在干旱胁迫和低温胁迫调控机制中发挥调控作用,可作为改良农作物和其他牧草抗旱、抗寒性的重要候选基因。     

小麦E3泛素连接酶基因TaAIRP2-1B克隆及功能分析

干旱等非生物胁迫严重影响农作物生产。本研究克隆了小麦(Triticum aestivum L.)TaAIRP2-1B基因,探讨其对非生物胁迫的响应机制,为促进小麦抗旱性的遗传改良提供基因资源。组织特异性表达模式分析显示,TaAIRP2-1B基因在小麦抽穗期的各个组织中均有表达,在茎组织中的表达水平较高,而根系中的表达水平较低。非生物胁迫表达模式分析显示,Ta AIRP2-1B受ABA、PEG及冷胁迫诱导表达。过表达TaAIRP2-1B拟南芥在0.4μmol/L的ABA处理条件下,种子发芽率显著低于野生型,表明TaAIRP2-1B提高了拟南芥种子萌发期对ABA的敏感性。ABA处理抑制转基因和野生型拟南芥幼苗的根系生长,但转基因拟南芥受抑制程度显著高于野生型,表明TaAIRP2-1B提高了拟南芥幼苗对ABA的敏感性。转基因结果表明超表达TaAIRP2-1B增强了拟南芥的抗旱性,并且转基因株系的保水率显著高于野生型。总之,本研究发现小麦基因Ta AIRP2-1B参与了植物对非生物胁迫的应答,可能是通过ABA途径正向调控植物的抗旱性。     

AtNHX5基因过量表达对拟南芥耐盐性的影响

为了研究AtNHX5基因在植物耐盐中的作用,构建了植物过量表达载体pROKⅡ-AtNHX5,并转化拟南芥。结果显示:(1)RT-PCR检测表明,转基因拟南芥中AtNHX5基因的表达大幅提高。(2)对转基因纯合株系进行耐盐性分析显示,AtNHX5过量表达提高了植株在种子萌发和苗期的耐盐性。(3)转基因植株在盐处理下的干重、鲜重以及地上部分Na+、K+含量均高于野生型对照。在200mmol/L NaCl处理下,以转基因株系a1-4为例,其地上部分单株鲜重、单株干重、K+含量分别是野生型的1.27、1.54、1.16倍,较野生型显著升高。研究表明,过量表达AtNHX5基因促进了盐胁迫下转基因植株对K+的吸收,转基因拟南芥的耐盐性明显提高。     

六种藓类植物茎的比较解剖学研究

通过对6种藓类植物,即褶叶青藓(Brachythecium salebrosum(Web.et Mohr.)B.S.G.)、湿地匐灯藓(Plagiomnium acutum(Lindb.)Kop.)、侧枝匐灯藓(Plagiomnium maximoviczii(Lindb.)Kop.)、大凤尾藓(Fissidensnobilis Griff.)、大羽藓(Thuidium cymbifolium(Doz.et Molk.)B.S.G.)和大灰藓(Hypnum plumaeforme Wils.)嫩茎和老茎的石蜡切片和显微观察发现,同一藓类植株的嫩茎和老茎,茎结构稳定,不同种藓类植物茎横切面具有不同特征.植物体茎横切面形状、表层细胞的层数、细胞大小和细胞壁厚薄、皮层细胞大小和形状、中轴的有无以及比例等特征可以作为藓类植物的分科分类依据之一.     

Cause of Senescence of Nine Sorts of Flowers

The levels of endogenous phytohormones and respiratory rate in nine sorts of flowers such as Cymbidium faberi Rolfe, Nopalxochia ackermannii Kunth and others were investigated both at full bloom and senescence and meanwhile the effect of exogenous phytohormones on prolonging the blossoms and promoting ethylene production were tested. There is a high content of endogenous ethylene in all the long-lived flowere, about 3–16 folds higer than the short-lived ones. There is a high level of ABA at full blooming flowers of short-lived flowers, in which there is no or only some cytokinins in it, but the ratio of CTK (6BA+zeatin)/ABA is smaller(l.7). The endogenous ABA reached a much higher level at senescence in all nine sorts of flowers, so it is reasonable to consider that it is ABA which plays an important role of regulation in controlling flower's senescence. There is a much higher level of GA3 and zeatin in the long-lived flowers which is not demonstrated in the shortlived ones. The respiratory rate is one of the factors controtling the longevity of flowers, but it does not play a decided role. Application of 6BA and zeatin prolongs distinctly orchid’s longevity, however exogenous IAA through the promotive action on ethylene production, evidently extends the longevity of the flowers of the Nopalxochia ackermannii Kunth.     

龙胆科药用植物化学成分的研究现状

龙胆科植物在我国的分布范围很广,且多数为药用植物,其多数种属的药用植物,至今其化学成分尚未被系统研究。综述了目前龙胆科药用植物的化学成分的研究现状及一般提取方法,对近年来发现的环烯醚萜及裂环烯醚萜类化合物进行了总结,为本科药用植物的更深入研究提供了参考。     

真菌类遗传学分析的知识结构教学

本文以认知结构理论为指导,讨论了真菌类遗传分析与高等动植物遗传分析的内在联系,认为利用这种内在联系进行教学可收到好的效果并说明了作者的具体教学过程。 Abstract:In the paper, the relationship between genetic analysis of Fungi and genetic analysis of high animal and plant was discussed.A good results were obtained when we adopted this method in the teaching.     

The effects of glutamine of the maintenance of embryogenic cultures of Cryptomeria japonica

Summary Embryogenic tissues of sugi (Cryptomeria japonica) were induced on a modified Campbell and Durzan (CD) medium containing 1 μM 2,4-dichlorophenoxyacetic acid (2,4-D) and 600 mg l⁻¹ glutamine, and subcultured in the medium of the same composition for over 1 yr. This resulted in a mixed culture of embryogenic and non-embryogenic cells. When embryogenic cells were isolated and cultured independently, their capacity to form embryogenic aggregates was lost. Thus, the non-embryogenic cells present within a mixed culture system were essential to the formation of embryogenic aggregates. When embryogenic tissues were isolated and cultured independently on a high glutamine-containing (2400 mg l⁻¹) medium, dry weights and endogenous levels of glutamine increased, and the tissue could generate a large number of embryogenic aggregates. Amino acid analysis of embryogenic and non-embryogenic cells from the maintenance culture indicated a higher level of glutamine was present in the latter. The high endogenous level of glutamine in the non-embryogenic portion of mixed cell masses may be the supplier of glutamine for maintaining the embryogenic property of the tissues.     

Specificity of action of piperidylcholine derivatives as substrates of cholinesterases of various origin

The review deals with study of enzymologic properties of a novel highly specific acetylcholinesterase substrate, N-(β-acetoxyethyl) piperidinium iodomethylate (“piperidylcholine”), and its 30 derivatives that were tested as effectors of cholinesterases of mammals and various species of Pacific squids. It was proven for the first time that responsible for specificity of action was structure of cyclic ammonium grouping of the alcohol part of molecule of the ester substrate. Analysis of specificity is performed based on enzymatic hydrolysis parameters—activity of catalytic center of cholinesterases and bimolecular constant of the reaction rate that are determined at optimal and low substrate concentrations. Among the specially synthesized group of thioester compounds there is revealed one more highly specific acetylcholinesterase substrate—N-(β-acetoxyethyl) piperidinium.     

医疗机构合理用药评价模型的构建研究

目的 针对医疗机构的合理用药水平进行评价研究。方法 根据医疗机构合理用药的具体要求,构建医疗机构合理用药评价指标体系,采用基于模糊群决策的方法和多指标评价分析法构建医疗机构合理用药评价模型。结果 构建了基于模糊群决策的医疗机构合理用药评价模型,并通过实例分析证明了评价模型的可行性。结论 建立的基于模糊群决策的医疗机构合理用药评价模型能够对医疗机构的合理用药水平进行科学评价,为提高医疗机构合理用药水平奠定基础。     

Photoregulation of seed germination of wild-type and of an aurea-mutant of tomato

Seed germination of an aurea mutant of tomato ( Lycopersicon esculentum Mill.) is promoted by continuous irradiation with red, far-red or long-wavelength far-red (758 nm) light as well as by cyclic irradiations (5 min red or 5 min far-red/25 min darkness). Far-red light applied immediately after each red does not change the germination behaviour. Seed germination of the isogenic wild-type, cv. UC-105, is promoted by continuous and cyclic red light while it is inhibited by continuous and cyclic far-red light and by continious 758 nm irradiation. Far-red irradiation reverses almost completely the promoting effect of red light. The promoting effect (in the aurea mutant) and the inhibitory effect (in the wild-type) of continuous far-red light do not show photon fluence rate dependency above 20 nmol m⁻² s⁻¹. It is concluded that phytochrome controls tomato seed germination throgh low energy responses in both the wild type and the au mutant. The promoting effect of continuous and cyclic far-red light in the au mutant can be attributed to a greater sensitivity to P_fr.