干旱对上部烟叶细胞结构和化学成分的影响

研究了烟草在土壤含水量为１２％、１６％、２０％的条件下叶片栅栏组织细胞的结构和烟昔日 化学成分的变化规律。结果表明：干旱处理的烟草,叶片栅栏组织细胞内淀粉粒变小,数量变少。叶片中还原性糖、可溶性总糖和Ｋ２Ｏ含量增加,淀粉和烟碱含量减少。干旱程度越深,其变化趋势越明显。     

大豆-根瘤菌共生体系光合与固氮关系研究

水培大豆和田间生长的大豆,接种根瘤菌 Rhizobium B16-11C 后植株全氮含量、叶片叶绿素含量和净光合速率及种子产量都明显增加。比较 Clark 大豆的结瘤品系和不结瘤品系获类似结果。摘除根瘤后3天内叶片净光合速率无明显变化。大豆植株遮阴、去叶或切掉地上部导致根瘤活性明显下降。但去豆荚不能提高根瘤固氮的比活性。根瘤活性的日变化不能用根瘤蔗糖、淀粉含量或周围温度的变化来解释,其控制因子尚待深入研究。     

锌胁迫对不知火杂柑和椪柑叶片光合特性及超微结构的影响

采用砂基培养法,运用CI－340便携式光合测定仪、H-7650透射电子显微镜研究了不同锌浓度如0mg／L（锌缺乏）、0．05mg／L（对照）和0．5mg／L（锌过量）处理下不知火杂柑（简称不知火）和槿柑叶片叶绿体色素含量、净光合速率及超微结构的变化。结果表明：（1）锌缺乏处理的不知火及锌过量处理的槛柑叶片叶绿素（a＋b）和类胡萝卜素含量、锌缺乏与锌过量处理的两者叶片净光合速率均相对低于对照。（2）锌缺乏处理下,两者叶片叶绿体出现变形或空室化,其中碰柑叶绿体内淀粉粒和质体小球增多,细胞核及线粒体正常,而不知火淀粉粒少,细胞核中出现许多黑色小颗粒物质。锌过量处理下,两者叶片叶绿体出现空室化或叶绿体膜模糊,线粒体解体,其中槿柑叶绿体内质体小球增多,淀粉粒少;而不知火淀粉粒明显增多、增大,基粒片层膨胀、松弛。可见,锌胁迫对两者叶片叶绿体色素、净光合速率及超微结构有着明显的影响,其影响程度因品种而异。     

菊花黄绿叶突变体不同类型叶片的叶绿素含量和结构特征比较

以菊花黄绿叶突变体-NAu04-1-31为试验材料,测定了黄叶、黄绿叶和绿叶3种不同类型叶片的叶绿素含量,并观察比较了叶片的显微与超微解剖结构.叶绿素含量测定表明:黄叶、黄绿叶的叶绿素含量显著低于绿叶,而黄叶叶绿素a与叶绿素b的比值大于绿叶.叶绿体显微与超微结构观察发现:黄叶细胞内叶绿体形状不规1则,缺乏正常的叶绿体膜结构,无类囊体,无淀粉粒,嗜锇颗粒较多;黄绿叶叶片栅栏组织绿色部分与绿叶的栅栏组织相似,黄色部分与黄叶的栅栏组织棚似,黄色部分的海绵组织中有类似于绿色叶片的海绵组织结构,而绿色部分含有类似于黄叶的海绵组织的结构特征.绿叶细胞内叶绿体较多,形状规则,基粒片层清晰,其内淀粉粒多而大,嗜锇颗粒较少.     

短期夜间低温后番茄光合作用及叶绿体超微结构的恢复效应

为明确短期夜间低温后番茄光合作用在常温下的恢复效应,研究了番茄幼苗经夜间15 ℃（对照）、12 ℃、9 ℃和6 ℃处理7 d后植株光合作用和叶绿体超微结构的变化.结果表明：短期夜间低温处理有利于恢复期番茄叶片的光合作用;12 ℃和9 ℃处理叶片叶绿素含量均可恢复至对照水平,6 ℃处理叶绿素含量则不可恢复;夜间低温处理并未严重抑制叶片气孔的形成,相反刺激了恢复期叶片气孔的形成与发育;6 ℃处理后叶绿体基粒片层排列较松散,片层数减少.恢复7 d时,夜间处理温度越低,叶绿体中淀粉粒越小、数量越少,基粒片层排列越整齐致密.另外,短期夜间低温处理促进了番茄植株叶片在恢复期的光合产物运输.     

玉米种子萌发过程幼叶细胞中淀粉粒的积累观察

研究玉米萌发初期幼叶的发育。在幼叶不同的发育时期 ,分别用 PAS反应 ,考马氏蓝处理不同叶片 ,结果发现 :叶片细胞内的叶绿体在叶片即将抽出时才形成 ;从浸种萌动到叶片进行光合作用前 ,植株的营养供给 ,主要靠叶片自身淀粉粒的积聚提供 ;在幼叶抽出以前 ,胚芽鞘的薄壁细胞中布满淀粉粒 ,随着叶片的发育 ,这些淀粉粒逐渐减少 ;而幼叶中的淀粉粒的变化情况正好相反 :在种子萌发初期 ,幼叶细胞内只有少量的淀粉粒 ,以后淀粉粒的积累逐渐增多 ;在这个阶段无蛋白质的积聚。幼叶中维管束的发生是先中间后两边 ,维管束中的韧皮部先形成 ,木质部后发生。     

Rpfan37在刺槐共生结瘤过程中的功能探究

目的:研究刺槐中与磷脂酰肌醇转运蛋白有较高同源性的基因Rpfan37的功能,为探究相关基因参与豆科植物与根瘤菌共生结瘤过程提供新的思路。方法:通过前期研究,建立豆科植物刺槐与共生根瘤菌互作的抑制差减杂交反交文库,筛选疑似与共生结瘤相关的基因。利用PCR技术快速克隆经实时荧光定量PCR技术(qRT-PCR)分析基因在不同接菌时间及不同植物组织的表达。构建RNA干扰(RNAi)重组载体,转农杆菌介导转化植物根部,接种根瘤菌后验证该基因在刺槐共生结瘤过程的功能。结果:基因表达分析显示,在接菌与未接菌的刺槐根中,处理后第15天,Rpfan37表达均显著上调,但接菌与未接菌处理对该基因表达无显著影响;在成熟的根瘤中,该基因仅为低水平表达。RNAi转化植株的鲜重、株高、根长及结瘤数较对照组显著降低。在显微镜下观察到RNAi植株根毛发育异常;与对照相比,RNAi转化植株形成的根毛卷曲、根毛侵染线及根瘤原基数目均显著降低。根瘤石蜡切片结果显示RNAi植株根瘤中的侵染细胞与对照相比明显减少,分析豆血红蛋白表达发现,RNAi植株中根瘤发育成熟过程明显受阻。结论:在豆科植物刺槐中发现的相关基因Rpfan37能够参与刺槐共生结瘤过程,为研究磷脂酰肌醇转运蛋白在共生结瘤过程中的作用提供了新的理论基础。     

小麦黄化突变体叶绿体超微结构研究

利用透射电镜对小麦自然黄化突变体及其突变亲本(西农1718)叶片细胞叶绿体的数目、形态及超微结构进行比较分析。结果发现:(1)3种不同黄化程度突变体的叶绿体分布、数目、形状及大小与突变亲本无明显差异;(2)突变体叶绿素含量为野生型58%的黄绿植株与其突变亲本叶绿体超微结构无明显差异,基质类囊体与基粒类囊体高度分化,基粒数目以及基粒片层数目较多;(3)突变体金黄和绿黄植株的叶绿素含量分别为野生型的17%、24%,其叶绿体超微结构与突变亲本明显不同,突变体的叶绿体发育存在明显缺陷,其中突变体金黄植株的叶绿体内无基粒、基质片层清晰可见,有淀粉粒,嗜锇颗粒较多,而突变体绿黄植株的叶绿体内有基粒,但明显少于突变亲本,且基粒片层较少,基质类囊体较发达。结果表明该黄化突变体叶绿体超微结构的改变,是由于叶绿素含量降低造成,推测,该黄化突变是由于叶绿素合成受阻导致的。     

施用硒肥对圆叶决明生长、酶活性及其叶肉细胞超显微结构的影响

通过盆栽试验 ,研究在红壤中施用不同浓度的硒肥对豆科牧草圆叶决明植株生长、叶片硝酸还原酶活性、根瘤固氮酶活性及其叶肉细胞超显微结构的影响。试验结果表明 ,施 0 .5～ 2 mg Se/ kg土不同用量的硒肥处理的圆叶决明植株株高、分枝数和植株干重 ,比不施硒肥的对照处理分别提高 11.71%～ 18.97%、0 .5 5 %～ 2 2 .6 1%和 5 2 .9%～ 14 4 .9% ;植株的硒含量随施硒浓度的提高而明显增加 ,不施硒 (CK)处理的植株含硒量仅为 0 .0 2 5 mg/ kg,当施硒肥量为 2 mg Se/ kg土 (S2 )时 ,植株的硒含量达到0 .2 2 2 mg/ kg,比对照增加了 7.88倍 ;施硒量为 1mg Se/ kg土 (S1)处理圆叶决明的叶片硝酸还原酶活性最大 ,比对照提高2 81.5 % ,比施 2 m g Se/ kg土的增加了 2 0 6 .1% ;根瘤固氮酶活性以施用 2 mg Se/ kg土的处理为最大 ,比对照增加了 4 9.4 1%。植株叶肉细胞超显微结构变化的观察结果表明 :施不同用量硒肥处理均能比对照处理增加植株叶绿体的数量 ,其中以1.5 mg Se/ kg土的施用量处理效果最好 ,而 1.5 m g Se/ kg土和 2 mg Se/ kg土处理对提高叶绿体的基粒数量和片层密度效果较佳 ,但对稳定叶绿体的双层膜结构效果则不明显 ;而施用 1.5 m g Se/ kg土和 1mg Se/ kg土处理则显示了减少线粒体的数量 ,降低线     

高CO2浓度对大平不同叶位叶片叶绿体淀粉粒积累的效应

对高ＣＯ２浓度下生长的大豆不同叶位的叶片进行电镜观察,揭示出大豆不同叶位叶片的叶绿体对倍增的ＣＯ２浓度反应不一。其显的超微结构差异特征是：１．叶位居中的叶片叶绿体积累的淀粉粒不仅很大,而且最多,有的叶绿体中的分粒可达２０个,几种充满着叶绿体的基质空间。２．下位叶叶绿体的淀粉粒积累较多,通常为２￣５个;３．上位叶叶绿体所含淀粉粒既小又少,虽然有的叶绿体中也积累有３￣４个淀粉粒,但大多叶绿体中所含淀     

Control of starch granule numbers in Arabidopsis chloroplasts

The aim of this work was to investigate starch granule numbers in Arabidopsis (Arabidopsis thaliana) leaves. Lack of quantitative information on the extent of genetic, temporal, developmental, and environmental variation in granule numbers is an important limitation in understanding control of starch degradation and the mechanism of granule initiation. Two methods were developed for reliable estimation of numbers of granules per chloroplast. First, direct measurements were made on large series of consecutive sections of mesophyll tissue obtained by focused ion beam-scanning electron microscopy. Second, average numbers were calculated from the starch contents of leaves and chloroplasts and estimates of granule mass based on granule dimensions. Examination of wild-type plants and accumulation and regulation of chloroplast (arc) mutants with few, large chloroplasts provided the following new insights. There is wide variation in chloroplast volumes in cells of wild-type leaves. Granule numbers per chloroplast are correlated with chloroplast volume, i.e. large chloroplasts have more granules than small chloroplasts. Mature leaves of wild-type plants and arc mutants have approximately the same number of granules per unit volume of stroma, regardless of the size and number of chloroplasts per cell. Granule numbers per unit volume of stroma are also relatively constant in immature leaves but are greater than in mature leaves. Granule initiation occurs as chloroplasts divide in immature leaves, but relatively little initiation occurs in mature leaves. Changes in leaf starch content over the diurnal cycle are largely brought about by changes in the volume of a fixed number of granules.     

墨兰幼叶和成熟叶不同部位叶绿体超微结构和光合作用

墨兰试管苗植株成熟叶片叶绿体基粒较发达,类囊本膜垛叠较紧密。幼叶叶绿体中少有亲锇颗粒,成熟叶的叶绿体中往往既有亲锇颗粒又有淀粉粒。幼叶中基粒数目比成熟叶的少,叶绿体也比成熟叶的小。幼叶的光合放氧速率比成熟叶的低。幼叶中叶尖部叶绿体最大而叶基部最小,但叶尖部的光合放氧速率比叶基部小。成熟叶中叶绿体大小及光合放氧速率区别不明显。通过对各部位叶绿素含量的测定发现,叶绿素含量与光合放氧速率之间没有正相关性     

杨树叶薄层培养中不定芽形态发生的细胞组织学研究

将杂种杨树(Populus nigra var.betulifolia×P.trichocarpe)NE299叶主脉用振动切片机横切成400μm或800μm的薄切片,培养在附加0.2mg/L BA和0.01mg/L NAA的木本植物培养基上。培养后,位于主脉维管束两侧中上部的维管束鞘薄壁细胞首先启动分裂。几乎同时,与其邻接的一些栅栏组织细胞也分裂,并很快形成胚性分生细胞团。主脉的愈伤组织主要由维管束鞘薄壁细胞,以及与其邻接的一些栅栏组织细胞和韧皮部的薄壁细胞分裂而来。不定芽通常发生在愈伤组织的周边区,也可以起源于维管组织结节(vascular nodules)周围的形成层状细胞。侧脉的维管束鞘细胞分裂活动很强,可不经愈伤组织直接长成不定芽。杨树叶主脉处的维管束鞘薄壁细胞在与叶肉组织相邻接的细胞中,通常含有少量较小的叶绿体,而位于背腹面的细胞中含有贮藏的淀粉。对形态发生的特定部位及其细胞进行了讨论。     

Altered cytokinin metabolism affects cytokinin, auxin, and abscisic acid contents in leaves and chloroplasts, and chloroplast ultrastructure in transgenic tobacco

Cytokinins (CKs) are involved in the regulation of plant development including plastid differentiation and function. Partial location of CK biosynthetic pathways in plastids suggests the importance of CKs for chloroplast development. The impact of genetically modified CK metabolism on endogenous CK, indole-3-acetic acid, and abscisic acid contents in leaves and isolated intact chloroplasts of Nicotiana tabacum was determined by liquid chromatography/mass spectrometry and two-dimensional high-performance liquid chromatography, and alterations in chloroplast ultrastructure by electron microscopy. Ectopic expression of Sho, a gene encoding a Petunia hybrida isopentenyltransferase, was employed to raise CK levels. The increase in CK levels was lower in chloroplasts than in leaves. CK levels were reduced in leaves of tobacco harbouring a CK oxidase/dehydrogenase gene, AtCKX3. The total CK content also decreased in chloroplasts, but CK phosphate levels were higher than in the wild type. In a transformant overexpressing a maize beta-glucosidase gene, Zm-p60.1, naturally targeted to plastids, a decrease of CK-O-glucosides in chloroplasts was found. In leaves, the changes were not significant. CK-O-glucosides accumulated to very high levels in leaves, but not in chloroplasts, of plants overexpressing a ZOG1 gene, encoding trans-zeatin-O-glucosyltransferase from Phaseolus lunatus. Manipulation of the CK content affected levels of indole-3-acetic and abscisic acid. Chloroplasts of plants constitutively overexpressing Sho displayed ultrastructural alterations including the occasional occurrence of crystalloids and an increased number of plastoglobuli. The other transformants did not exhibit any major differences in chloroplast ultrastructure. The results suggest that plant hormone compartmentation plays an important role in hormone homeostasis and that chloroplasts are rather independent organelles with respect to regulation of CK metabolism.     

Epidermal focussing and effects on light utilization in Oxalis acetosella

lmage analysis was used to quantify the lateral heterogeneity of the radiation field within the palisade of Oxialis acetosella L. leaves. Oxalis acetosella epidermal cells focus light up to four times incident irradiance, resulting in regions of high and low internal fluenec rate within the palisade. Chlorophyll fluorescence from leaves irradiated with directional light was found to originate primarily from palisade cell chloroplasts located within focal zones. When the internal radiation field was made more homogeneous by using diffuse light or by coating the leaf with a layer of mineral oil to eliminate epidermal focussing, the characteristics of the chlorophyll fluorescence signal were altered: non-photochemical quenching (qN) increased, while relaxation of qN was slowed. This indicates that upper palisade chloroplasts may fine-tune their light utilization to the intra leaf light microenvironment.     

Starch Accumulation and Photosynthetic Activity in Primary Leaves of Bean (Phaseolus vulgaris L.)

The effects of starch accumulation on photosynthesis and chloroplastultrastructure were studied in primary leaves of bean (Phaseolusvulgaris L. cv. Bulgarian). De-topping the shoot above the primaryleaf node, caused over an 8-day period, a considerable increasein the photosynthetic activity of the primary leaves, despitethe fact that a large quantity of starch had accumulated intheir chloroplasts. The accumulation of starch was greater inthe chloroplasts of spongy cells in comparison with that ofthe palisade cells. Initiation of starch grains was observedmainly in the peripheral part of the chloroplast, distant fromthe cell wall. As a result, most of the starch was accumulatedclose to the inner part of the cell, leaving a considerablemass of the chloroplast near the cell wall free of starch. Theaccumulation of starch was accompanied by the destruction, deformationand disorientation of grana and thylakoids. It is concludedthat the accumulation of starch is not inevitably a limitingfactor in photosynthesis and the results cast doubt on the hypothesisthat starch accumulation or dissipation is the main factor involvedin the regulation of photosynthesis. Phaseolus vulgaris L, bean, photosynthesis, starch accumulation, chloroplast ultrastructure     

Effect of water stress on cotton leaves : I. An electron microscopic stereological study of the palisade cells

Palisade cells from fully expanded leaves from irrigated and nonirrigated, field grown cotton (Gossypium hirsutum L. cv. Paymaster 266) were subjected to a microscopic examination to evaluate the effect of water stress on subcellular structures. The water potential difference between the two treatments was 13 bars at the time of sampling. The dimensions of the palisade cells and their density per unit leaf area were determined by light microscopy. Palisade cells from stressed plants had the same diameter, but were taller than their counterparts in irrigated plants. The density of the palisade cells was the same in both treatments as was the fractional volume of the intercellular space. It was concluded that the reduced leaf area observed in the stressed plants resulted primarily from a mitotic sensitivity to water stress. Further, expansion of palisade cells was not inhibited by the stress imposed in this study.

Morphometric analysis of electron micrographs was used to evaluate the subcellular structure of palisade cells from nonstressed and stressed plants. The fractional volumes of cell walls, total cytoplasm, chloroplasts, starch granules, intrachloroplast bodies, mitochondria, peroxisomes, and central vacuoles were determined. The surface densities of grana and stroma lamellae, outer chloroplast membranes, mitochondrial cristae, endoplasmic reticulum and Golgi cisternae were also measured. The number of chloroplasts, mitochondria, and peroxisomes were determined. These data were expressed as actual volumes, areas, and numbers per palisade cell for each treatment. Palisade cells from stressed plants had thinner cell walls, larger central vacuoles and approximately the same amount of cytoplasm compared to cells from nonstressed plants. Within the cytoplasm, stressed plants had more but smaller chloroplasts with increased grana and stroma lamellae surfaces, larger mithchondria with reduced cristae surfaces, smaller peroxisomes and reduced membrane surfaces of endoplasmic reticulum and Golgi cisternae.

     

CHLOROPLAST SPECIALIZATION IN DICOTYLEDONS POSSESSING THE C4-DICARBOXYLIC ACID PATHWAY OF PHOTOSYNTHETIC CO2 FIXATION

The C₄-dicarboxylic-acid pathway of photosynthetic CO₂ fixation found in tropical grasses has recently been demonstrated in members of the Amaranthaceae and Chenopodiaceae. In the tropical grasses this CO₂-fixation pathway is correlated with specialized leaf anatomy and chloroplast structure. This investigation was undertaken to determine if leaf cells of some representatives of these other families had structural features similar to those of tropical grasses. The leaf anatomy of Amaranthus edulis and a variety of Atriplex species is very similar and it resembles that of grasses such as sugar cane. Prominent bundle sheaths are surrounded by a layer of palisade cells. Bundle-sheath cells of Am. edulis have large chloroplasts containing much starch, but the chloroplasts have grana. The palisade cells have much smaller chloroplasts containing very little starch. The bundle-sheath cell chloroplasts of At. lentiformis have grana, their profiles tend to be ovoid, and they contain abundant starch grains. The palisade cell chloroplasts contain little starch and their profiles are discoid. The bundle-sheath cells of both species contain mitochondria which are much larger than those in the palisade cells. The chloroplasts in both types of cells in both species have a highly developed peripheral reticulum. This reticulum is composed of anastomosing tubules which are contiguous with the inner plastid membrane. The leaf anatomy and cell ultrastructure of these dicots are similar to those of the tropical grasses possessing this new photosynthetic carbon-fixation pathway. These morphological features are interpreted as adaptations for the rapid transport of precursors and end products of photosynthesis. A hypothesis is presented stating that the unique morphological and biochemical characters of these plants represent adaptations for efficient and rapid carbon fixation in environments where water stress frequently limits photosynthesis.