植物细胞壁伸展测定仪在蚕豆扩张蛋白特性研究中的应用

扩张蛋白(expansin)在细胞扩张和果实成熟中起着极为重要的作用.植物细胞壁伸展测定仪是研究扩张蛋白必不可少的仪器.为此以电涡流传感器为核心部件装配了一种具有结构简单、操作方便和测量准确等优点的新型测定仪,并利用该仪器研究了蚕豆(Vicia faba)扩张蛋白的特性.结果表明蚕豆根、茎、上胚轴和成熟叶片中均存在扩张蛋白,而且叶片和幼根的扩张蛋白活性最强;免疫印迹证实在蚕豆根、茎、上胚轴和成熟叶片中确实存在扩张蛋白.以上结果说明本仪器灵敏且可靠,用此仪器首次发现在成熟叶片中存在扩张蛋白.     

藤茶香树脂醇合成酶基因编码区克隆及表达分析

为研究藤茶(Ampelopsis grossedentata)三萜类化合物的生物合成,采用RT-PCR方法,从藤茶叶片cDNA中扩增得到香树脂醇合成酶(amyrin synthase)基因,命名为AgAS。该基因编码区长2 250bp,编码750个氨基酸,与葡萄的同源蛋白亲缘关系最近。亚细胞定位和Southern blot结果表明,AgAS编码蛋白主要定位于细胞核与细胞膜,在藤茶基因组中存在2个拷贝。实时荧光定量PCR结果显示,AgAS基因在紫芽藤茶的根、茎和叶均有表达,其中叶片中表达最高,茎次之,根中表达量最少,且随着叶片成熟程度的增加,表达量呈先升后降趋势;而在绿芽藤茶中,AgAS基因在叶片中的表达量随着成熟程度的增加呈上升趋势,根与茎中表达量相对较少。     

油橄榄下胚轴诱导完整植株

油橄榄离体茎培养出根已有报道,但无诱导完整植株的试验结果。我们以油橄榄下胚轴、子叶、胚根、嫩茎、茎尖和叶片等作为外植体进行了试验,除从茎尖诱导发生小植株外(待发表),在下胚轴上也已诱导出完整植株。从甘蓝下胚轴诱导发生小植株已有报道,本文就油橄榄下胚轴诱导完整植株作简要报道。油橄榄(Olea europaea L.)种子经消毒处理后,通过无菌培养得到无菌苗。在幼苗下胚轴伸长到2—3厘米时,在无菌条件下切除茎尖、子叶     

双子叶植物出土幼苗根茎转变区维管组织发育动态

关于根茎初生维管系统之间的连接以及与子叶的关系,在文献中已有广泛的论述,有过各种不同的解释。大部分早期关于根茎转变区的文献研究的是初生组织已完成发育的幼苗。这些研究者认为转变区域是根和茎这两种轴器官之间维管组织发生转变、相互连接的区域。但由于茎中的初生维管组织可以认为是叶迹及叶迹的延伸的综合,转变区域应被看作是轴维管系统与叶迹维管系统之间的连接。因此,转变区的研究必须说明根维管系统与最早的真叶叶迹之间的关系。通过对北乌头和大豆胚胎及幼苗维管组织的解剖学研究,本工作显示在出土萌发的双子叶植物中,初生维管组织在根-下胚轴-子叶中形成一连续系统,并完成根与子叶叶迹之间的维管组织过渡转变。而上胚轴中的维管组织是位于根-下胚轴-子叶上方独立形成的第二维管系统。上胚轴中维管组织的分化起始于第一真叶叶迹基部,向上分化进入叶片,向下进入胚轴并在子叶节下方与根-下胚轴-子叶维管系统相连接。真叶叶迹的木质部与下胚轴中靠近韧皮部的后生木质部或次生木质部连接。根与上胚轴之间不存在维管组织的过渡、转变,而只是在同样发育方向的组织中有一种直接的简单的连接．     

钴在蚕豆中的积累分布及其对叶片光合作用和抗氧化酶活性的影响

以6叶期蚕豆(Vicia faba L.)幼苗为材料,用不同浓度钴(0、20、40、80、120mg·kg-1)土培处理14d,研究不同浓度钴处理对蚕豆体内钴的富集和分布特征,以及叶片光合色素含量、叶绿素荧光参数、光合作用参数、生长指标和POD、SOD、CAT活性的影响。结果显示:(1)蚕豆的根、茎、叶均能吸收一定量的钴,但不同器官间积累量有显著差异,并表现为根叶茎,且在不同钴浓度处理下根吸收量也存在显著差异。(2)不同浓度钴处理下,蚕豆叶片叶绿素(a、b)含量、气孔导度(Gs)、最大光化学效率(Fv/Fm)、PSⅡ潜在活性(Fv/F0)等参数与对照组相比表现出0~40mg·kg-1浓度下促进,40~120mg·kg-1浓度下抑制,而净光合速率(Pn)、胞间CO2浓度(Ci)以及光系统Ⅱ最小荧光(F0)、最大荧光(Fm)表现为在0~80mg·kg-1浓度下促进,80~120mg·kg-1浓度下抑制,蒸腾速率(Tr)和类胡萝卜素含量则在各浓度下一直表现为促进,并且以上光合作用相关参数与对照组相比都在40mg·kg-1浓度下促进作用达到最大。(3)蚕豆幼苗的根长、株高、生物量以及抗氧化酶POD、SOD和CAT活性均随着钴浓度增加表现出先升高后降低的趋势。研究表明,蚕豆的根、茎、叶均能积累钴,且吸收量有随土壤钴浓度增加而增加的趋势,但以根部的富集能力最强、积累量最大;低浓度钴可以诱导蚕豆中抗氧化酶活性增强,促进其叶片光合色素含量、光合作用效率提高和植株生长发育,而高浓度钴则表现出相反趋势,抑制蚕豆的光合效率和生长发育。     

刺槐幼苗根茎过渡区初生维管组织的观察

对不同苗龄(1-12d)的刺槐(Robinia pseudoacacia L.)幼苗进行观察,比较了其初生维管组织的发育变化。结果表明:根中的初生维管组织借助下胚轴过渡到子叶中,而茎中的初生维管组织与下胚轴中的维管组织仅是简单的连接。这与Esau等认为幼苗的外部形态可以分为根-下胚轴-子叶和上胚轴苗两个系统的观点是一致的。本研究亦证实了可以将刺槐幼苗的外部形态描述为这两个系统,二者的维管组织只是简单的连接,不存在过渡。     

中学生物实验材料准备月历提纲(华东地区)

(一)植物月份准备材料8月 洋葱鳞茎。番茄、苹果或沙果、西瓜等果实。菜豆、玉米、小麦、棉花、苹果、柿、水稻、高粱的种子。小麦粉。单子叶植物和双子叶植物的根及变态根。水培荞麦和番茄。培养金鱼藻和黑藻。采集和制作葫芦科、菊科植物标本9月 萝卜种子。准备南瓜茎、玉米茎(浸制)。采集和制作各种花、果实和种子标本10月 胡萝卜的圆锥根、马铃薯的块茎。1一3年生杨树、极、木横枝茎。蚕豆叶片。准备水稻、凤仙花的茎。采集丁香和杨树的茎芽。种小麦和蚕豆11月 莲的根状茎,葡萄茎的扦插。果树(苹果、梨、桃)芽接材料2月整理装订植物学十…     

油菜子叶、下胚轴和根愈伤组织的诱导和植株再生

我们从1978年开始,用油菜幼苗的子叶、下胚轴和根进行组织培养,探索在油菜无性系快速繁殖中的应用。油菜组织培养方面的资料,目前仅见有茎段培养、花药培养,花器培养和叶肉原生质体培养。由花器各部分诱导形成的植株有早花现象,由茎段再生的植株,从茎段外植体至再生植株成熟只要90天。用油菜子叶、下胚轴和根诱导产生愈伤组织并从子叶和下胚轴愈伤     

广东连州自然分布的3种植物体内及根际环境中稳定性碳和氮同位素的分析

以广东连州自然分布的3种国家重点保护野生植物南方红豆杉〔Taxus chinensis var.mairei(Lemée et Lévl.)Cheng et L.K.Fu〕、半枫荷(Semiliquidambar cathayensis Chang)和金荞麦〔Fagopyrum dibotrys(D.Don)Hara〕为研究对象,分析了根、茎和叶片及根际土壤和岩石的C、N含量和C:N比以及δ13 C和δ15 N值的差异;在此基础上,通过δ13 C和δ15 N值的散点图比较了3种植物生态位的差异.结果表明:在同种植物中,根、茎和叶片的C和N含量及C:N比总体高于根际土壤和岩石,其中,叶片中C和N含量均最高,茎的C:N比最高;而根际土壤和岩石的δ13 C和δ15 N值总体高于根、茎和叶片.在供试的3种植物间,根际土壤和岩石中C和N含量总体上无明显差异,但根、茎和叶片中C和N含量以及根、茎和叶片及根际土壤和岩石的C:N比、δ13 C和δ15 N值均有一定差异;其中,金荞麦根中C含量显著(P<0.05)低于南方红豆杉和半枫荷,其根、茎和叶片中N含量和δ15 N值均极显著(P<0.01)高于后二者,其根、茎和叶片的C:N比和δ13 C值均极显著低于后二者,其根际土壤和岩石的C:N比和δ13 C值总体上也低于后二者;南方红豆杉和半枫荷的叶片中C和N含量以及茎和叶片的δ13 C值、根际土壤和岩石的δ13 C和δ15 N值均存在显著差异,但二者的整体差异相对较小.从散点图上看,金荞麦的生态位远离南方红豆杉和半枫荷,而后二者的生态位有交集.综合分析结果显示:草本植物金荞麦与木本植物南方红豆杉和半枫荷的C和N含量以及δ13 C和δ15 N值的差异不仅与植物自身的生活型有关,而且与各自生境中的光照和土壤因子等相关.另外,供试3种植物的根、茎和叶片的δ13 C值变幅为-31.69‰~-26.46‰,符合C3植物的δ13 C值范畴.     

喜树毛状根的诱导及其喜树碱含量分析

研究了不同外植体类型(包括真叶、茎段、子叶及胚轴)、胚轴年龄和不同发根农杆菌菌株(包括A4、15834、R1601、C58C1)等因素对喜树毛状根诱导频率的影响,并用PCR对诱导出的毛状根进行了分子鉴定.结果表明:(1)最佳外植体为胚轴,5~10 d是胚轴最佳诱导年龄段,最佳诱导菌株为15834.(2)PCR鉴定结果表明,发根农杆菌的rolB基因已整合到喜树毛状根基因组中.(3)对不同菌株诱导的毛状根进行HPLC检测表明,C58C1菌株诱导的毛状根的喜树碱和羟基喜树碱含量最高,分别为1.219 mg/g和0.305 mg/g.研究结果为喜树碱的药源开发提供了一条新途径,并为进一步利用基因工程技术调控喜树碱的代谢合成奠定了基础.     

Expansins in growing tomato leaves

An expansin-like protein from growing tomato leaves was identified by its ability to restore the 'acid-growth' response to heat-inactivated tomato walls and by its similarity to expansins from cucumber hypocotyls. Native walls from growing tomato leaves exhibit an endogenous acid-induced extension (creep) that resembles in various biochemical characteristics the acid-growth activity of cucumber hypocotyls. For example, the acid-growth activity is lost when the walls of tomato leaves are briefly heated and is largely restored by addition of a crude protein extract from the walls of growing leaves. Wall proteins from growing leaves enhance the stress relaxation spectrum of tomato walls in a fashion characteristic of cucumber expansins. HPLC fractionation of the crude wall protein from tomato leaves yielded an active fraction containing a major 27 kDa protein that cross-reacts with an antibody raised against cucumber expansin. The results show that tomato leafwalls possess at least one expansin that is responsible for the acid-growth property of leaves and indicate that cell wall extension in leaves shares an underlying protein mechanism common to cell wall expansion in stems.     

Growth maintenance of the maize primary root at low water potentials involves increases in cell-wall extension properties, expansin activity, and wall susceptibility to expansins

Previous work on the growth biophysics of maize (Zea mays L.) primary roots suggested that cell walls in the apical 5 mm of the elongation zone increased their yielding ability as an adaptive response to low turgor and water potential (psi w). To test this hypothesis more directly, we measured the acid-induced extension of isolated walls from roots grown at high (-0.03 MPa) or low (-1.6 MPa) psi w using an extensometer. Acid-induced extension was greatly increased in the apical 5 mm and was largely eliminated in the 5- to 10-mm region of roots grown at low psi w. This pattern is consistent with the maintenance of elongation toward the apex and the shortening of the elongation zone in these roots. Wall proteins extracted from the elongation zone possessed expansin activity, which increased substantially in roots grown at low psi w. Western blots likewise indicated higher expansin abundance in the roots at low psi w. Additionally, the susceptibility of walls to expansin action was higher in the apical 5 mm of roots at low psi w than in roots at high psi w. The basal region of the elongation zone (5-10 mm) did not extend in response to expansins, indicating that loss of susceptibility to expansins was associated with growth cessation in this region. Our results indicate that both the increase in expansin activity and the increase in cell-wall susceptibility to expansins play a role in enhancing cell-wall yielding and, therefore, in maintaining elongation in the apical region of maize primary roots at low psi w.     

小麦胚芽鞘扩展蛋白特性及对水分胁迫的响应

扩展蛋白是植物细胞壁延伸过程中的关键调节因子,在植物的生长发育以及对逆境的响应过程中起着重要作用。本文选用小麦(HF 9703)胚芽鞘为材料,采用Hepes法和SDS法分别提取小麦胚芽鞘扩展蛋白,通过改良的植物组织伸长测定仪测定其活性,并利用扩展蛋白抗体进行免疫印迹以检测其丰度,主要研究了小麦胚芽鞘扩展蛋白的特性及对水分胁迫的响应。结果表明:Hepes法提取的扩展蛋白活性较高,而SDS法的提取效率高;离体小麦胚芽鞘扩展蛋白的活性具有pH依赖性,且随缓冲液的交替更换(pH 4.5:pH 6.8)而反复逆转;扩展蛋白主要定位于细胞壁中;小麦胚芽鞘扩展蛋白和黄瓜下胚轴扩展蛋白具有交叉重组活性,但这种活性具有种属特异性。水分胁迫诱导小麦胚芽鞘扩展蛋白的活性和丰度提高,扩展蛋白活性的提高在小麦对水分胁迫的抗性方面可能具有重要作用。     

Protein and Peroxidase Modulations in Sunflower Seedlings (Helianthus annuus L.) Treated with a Toxic Amount of Aluminium

The aim of this study is to investigate the effect of aluminium treatment on peroxidases activities and protein content in both soluble and cell-wall-bound fractions of sunflower leaves, stems and roots. Fourteen-day-old seedlings, grown in a nutrient solution, were exposed to a toxic amount of aluminium (500 μM AlNO₃) for 72 h. Under stress conditions, biomass production, root length and leaf expansion were significantly reduced. Also, our results showed modulations on soluble and ionically cell-wall-bound peroxidases activities. In soluble fraction, peroxidases activities were enhanced in all investigated organs. This stimulation was also observed in ionically cell-wall-bound fraction in leaves and stems. Roots showed a differential behaviour: peroxidase activity was severely reduced. Lignifying peroxidases activities assayed using coniferyl alcohol and H₂O₂ as substrates were also modulated. Significant stimulation was shown on soluble fraction in leaves, stems and roots. In ionically cell-wall-bound fraction lignifying peroxidases were enhanced only in stems but severely inhibited in roots. Also, aluminium toxicity caused significant increase on cell wall protein content in sunflower roots.     

Variations in the level of enzyme activity and immunolocalization of calcium-dependent protein kinases in the phloem of different cucumber organs

Calcium-dependent protein kinases (CDPKs) constitute a unique family of enzymes in plants that are characterized by a C-terminal calmodulin (CaM)-like domain. Through protein kinase assays, we have examined the levels of cucumber calcium-dependent kinase (CsCDPK) activity in various organs of cucumber seedlings and plants. The activity of CsCDPK was highest in cucumber plant leaves followed by seedling roots and hypocotyls; however, cucumber plant flowers, seedling cotyledons, and hooks had levels that were barely detectable. The CsCDPKs were immunolocalized using polyclonal antibodies that are highly specific against a part of the kinase domain of a calcium-dependent protein kinase (CsCDPKS) in the phloem sieve elements (SEs) in various organs of cucumber. In addition, this study indicates the presence of CsCDPKs in organelle-like bodies associated with the plasma membrane of sieve elements in mature stems and roots as well as in the storage bodies of immature seeds. These findings are discussed in terms of the likely roles played by CDPKs in the signal transduction pathways for Ca2+-regulated phloem transport of assimilates from leaves to various organs during growth and development of cucumber seedlings and plants.     

硫水平对玉米氮、硫代谢特性及根系活性的影响

通过Hoagland营养液水培玉米,定量研究了供S水平对玉米根系活性和植株S、N代谢特性的影响．当玉米缺S时。玉米根系活性和叶片硝酸还原酶(NR)活性显著降低,植株冠／根比增大;根系与地上部对S、N的吸收均减少。S在根系和新生叶中分配相对减少、在衰老叶和茎鞘中分配相对增多;N在根系中分配增多,新生叶和完全展开叶中分配相对减少;植株蛋白质、蛋白氮、蛋白硫、无机硫的含量均显著降低,非蛋白氮比率增多,无机硫占总S的比率变小,全N／全S比增大;蛋白质中的N／S比在地上部无显著变化。而在根系中显著增大;根系对S缺乏的反应更为敏感．研究表明。缺S植株蛋白质含量与NR活性之间呈极显著正相关;S的缺乏或过量均影响玉米的N、S代谢;以根系作为S素营养诊断器官。以N／S比或无机S／全S比作为诊断指标较灵敏。正常条件下其数值分别为10．7和0．302．     

Swollenin, a Trichoderma reesei protein with sequence similarity to the plant expansins, exhibits disruption activity on cellulosic materials.

Plant cell wall proteins called expansins are thought to disrupt hydrogen bonding between cell wall polysaccharides without hydrolyzing them. We describe here a novel gene with sequence similarity to plant expansins, isolated from the cellulolytic fungus Trichoderma reesei. The protein named swollenin has an N-terminal fungal type cellulose binding domain connected by a linker region to the expansin-like domain. The protein also contains regions similar to mammalian fibronectin type III repeats, found for the first time in a fungal protein. The swollenin gene is regulated in a largely similar manner as the T. reesei cellulase genes. The biological role of SWOI was studied by disrupting the swo1 gene from T. reesei. The disruption had no apparent effect on the growth rate on glucose or on different cellulosic carbon sources. Non-stringent Southern hybridization of Trichoderma genomic DNA with swo1 showed the presence of other swollenin-like genes, which could substitute for the loss of SWOI in the disruptant. The swollenin gene was expressed in yeast and Aspergillus niger var. awamori. Activity assays on cotton fibers and filter paper were performed with concentrated SWOI-containing yeast supernatant that disrupted the structure of the cotton fibers without detectable formation of reducing sugars. It also weakened filter paper as assayed by an extensometer. The SWOI protein was purified from A. niger var. awamori culture supernatant and used in an activity assay with Valonia cell walls. It disrupted the structure of the cell walls without producing detectable amounts of reducing sugars.     

Two enzymatic sources of nitric oxide in different organs of apple plant

Nitric oxide production, nitric oxide synthase (NOS) and mitochondrial nitrite-reducing activities in roots, leaves and stems of different developmental stages were investigated, using potted 3-year-old apple (Malus domestica Borkh.) trees. The arginine-dependent NOS activity is sensitive to NOS inhibitor L-NAME and aminoguanidine (AG), with L-NAME being more effective than AG. Endogenous NO production, NOS and mitochondrial nitrite-reducing activities are predominately presented in young leaves and especially in young white roots and young stems. Root and stem mitochondria can reduce nitrite to nitric oxide at the expense of NADH, however, this mitochondrial nitrite-reducing activity is absent in leaves.