Cd^2＋或Hg^2＋水污染对菱体细胞的细胞核及叶绿体超微结构的影响

用Ｃｄ２＋、Ｈｇ２＋两种重金属离子溶液培育菱（ＴｒａｐａｂｉｃｏｒｎｉｓＯｓｂｅｃｋ．）植物体后,观察体细胞的细胞核及叶绿体超微结构的变化。处理后第８天,菱浮水叶叶片和不定根细胞中细胞核的染色质与核质遭到破坏,不定根中细胞核的核仁消失。但在各处理浓度（１０μｍｏｌ／Ｌ～５０μｍｏｌ／ＬＣｄ２＋或Ｈｇ２＋）下,核膜均保持完整。叶片细胞的叶绿体基粒数目减少,基粒片层解体,叶绿体双层膜断裂,叶绿体中的质体球流入细胞基质中。两种结构的破坏程度随处理的离子浓度提高而增大。说明菱体细胞超微结构的变化观测可作为监测重金属污染的一种方法。     

汞,镉污染对黑藻叶细胞伤害的超微结构研究

黑藻（Ｈｙｄｒｉｌｌａ ｖｅｒｔｉｅｉｌｌａｎ（Ｌ．ｆ．）Ｒｏｙｌｅ）植株分别在Ｈｇ＾２＋、Ｃｄ＾２＋梯度浓度的污染水中培养,在培养的时间段中（Ｈｇ＾２＋为３ｘｄ,Ｇｄ＾２＋为６ｄ）,随着浓度的递增,叶片逐渐出现均匀退绿症状。电镜观察发现,叶细胞遭受Ｈｇ＾２＋、Ｃｄ＾２＋毒害初期,高尔基体消失,内质网膨胀后解体,叶绿体中的类囊体和线粒体中的脊突胀成呈囊泡状,核中染色质涕 集。随着叶细胞遭受毒害程度     

Pb~(+2)、Cd~(+2)、Hg~(+2)对蚕豆(Vicia faba L.)乳酸脱氢酶的影响

Ｐｂ＾＋２,Ｃｄ＾＋２,Ｈｇ＾＋２浓度分别小于５．００ｍｇ／ｋｇ,２．００ｍｇ／ｋｇ,０．５０ｍｇ／ｋｇ时,蚕豆（ＶｉｃｉａｆａｂａＬ．）乳酸脱氢酶（ＬＤＨ）的活性主于对照组,当处理剂量分别长高超过上述剂量时,ＬＤＨ活性显著地降低,分析不同污染处理条件下ＬＤＨ同工酶,发现其５种同工酶在不同金属以及同一金属的不同剂量处理条件下,有差异表达的特性,其中ＬＤＨ２在小剂量重金属作用下被诱导表达;ＬＤＨ４对     

Cr^6＋对莼菜冬芽叶片急性毒害与保护酶系活性变化关系的研究

研究了Ｃｒ＾６＋的急性毒害对莼菜冬芽叶片的伤害程度和可溶性蛋白质、ＳＯＤ、ＣＡＴ、ＰＯＤ活性和ＭＤＡ含量变化之间的关系。在以０．５ｍｍｏｌ／Ｌ￣２．０ｍｍｏｌ／Ｌ浓度的Ｃｒ＾６＋处理时,冬芽叶片受害的程序明显地与处理浓度和处理时间呈正相关;可溶性蛋白质只在处理４ｄ时出现较大变化,其含量随处理浓度的提高而急剧增加;随着处理浓度的增加,ＳＯＤ、ＣＡＴ和ＰＯＤ的活性峰出现的时间不断后推,并且ＰＯＤ的活性     

Cd^2＋污染对莼菜叶片形态学伤害反应的研究

重点研究了Ｃｄ＾２＋胁迫下莼菜叶片的受害症状及叶肉细胞超微结构的变化。当Ｃｄ＾２＋浓度大于２μｇ·ｇ＾－１时,莼菜叶面将在短期内出现不同程度的受害症状。莼菜叶片遭受Ｃｄ＾２＋毒害初期,叶肉细胞质体中的少数基粒会发生膨胀或解体,线粒体的脊出现瓦解,部分核仁裂解成数块大小不等的颗粒。随着叶肉细胞遭受毒害程度的加深,质体中较多的类囊体便出现解体,多种膜性结构如：质体、线粒体的外包膜以及核膜、液泡膜等均遭     

不同浓度Hg^2＋对睡莲的毒害影响研究

主要研究了Ｈｇ＾２＋对睡莲的外部形态及叶绿素含量、过氧化物酶活性、硝酸还原酶活性。可溶性蛋白含量等生理指标的影响。结论是：Ｈｇ＾２＋对其外部形态的毒害程度与处理浓度和时间成正相关。１－５ｍｍｏｌ／Ｌ处理使叶绿素含量、硝酸还原酶活性上升,８－１０ｍｍｏｌ／Ｌ处理则下降;１－８ｍｍｏｌ／Ｌ处理过氧化物酶活性随浓度增加而上升,１０ｍｍｏｌ／Ｌ处理则下降,但仍高于对照;可溶性蛋白含量随处理浓度增加呈下降趋     

TFP刺激裂殖酵母细胞钙离子内流的质膜效应

ＴＦＰ（１０－１００μｍｏｌ／Ｌ）可引起裂殖酵母（Ｓｃｈｉｚｏｓａｃｃｈａｒｏｍｙｃｅｓ ｐｏｍｂｅ）细胞Ｃａ＾２＋内流,ＴＦＰ浓度不同,促进Ｃａ＾２＋内流程度也不一样,５０μｍｏｌ／Ｌ ＴＦＰ的促进作用最大。并且ＴＦＰ浓度越大,Ｃａ＾２＋内流出现峰值也越早,１０、２０、５０、１００μｍｏｌ／Ｌ ＴＦＰ处理后,胞内总钙出现峰值时间分别为４５、４５、３０、１５分钟。胞外Ｈ＾＋浓度也会对ＴＦＰ引起的     

香艳梨离体培养研究

本试验以香艳梨的茎尖,不带芽茎段,带芽茎段,叶片,叶柄为试材,以１／２ＭＳ为基本培养基,分别附加０．５～２．０ｍｇ／Ｌ的６－卡基氨基嘌呤（６－ＢＡ）和０．１ｍｇ／Ｌ的萘乙酸（ＮＡＡ）、吲哚乙酸（ＩＡＡ）,２,４－二氯苯氧乙酸（２,４－Ｄ）进行离体培养研究。结果表明,试材用０．１％ＨｇＣｌ２灭菌５～６分钟为宜;在７月份接种感染率低,愈伤组织形成较快;茎尖,不带芽茎段,带芽茎段,叶片均可作为离体培养的外植体,叶片的脱分化,分化的效果尤为明显,叶柄不宜作外植体;１／２ＭＳ＋１．５ｍｇ·Ｌ－１６－ＢＡ＋０．１ｍｇ·Ｌ－１ＮＡＡ有利于脱分化;１／２ＭＳ＋１．０ｍｇ·Ｌ－１６－ＢＡ有利于分化;瓶外生根优于瓶内生根。本试验可为香艳梨的工厂化育苗提供参考。     

马铃薯离体再生及人工种子研究

马铃薯３个品种虎头,克４和Ｆａｖｏｒｉｔａ的茎叶外植体在ＭＳ＋１ｍｇ／ＬＮＡＡ＋１ｍｇ／ＬＢＡ培养基上形成愈伤组织。在ＭＳ＋０２ｍｇ／ＬＮＡＡ＋１ｍｇ／ＬＢＡ培养基上,愈伤组织分化产生不定芽。在ＭＳ＋００５ｍｇ／ＬＮＡＡ培养基上,不定芽生根形成再生完整植株。０２～０３ｃｍ大小的不定芽反复继代可持续增殖。有３个以上叶片的不定芽在ＭＳ＋５ｍｇ／ＬＢＡ＋００５ｍｇ／ＬＩＢＡ培养基上和黑暗条件下,在侧芽或顶芽部位形成微型薯。用４％海藻酸钠和２％氯化钙溶液包裹０２～０３ｃｍ大小的不定芽或直径为０２～０３ｃｍ的微型薯制成微芽人工种子和微薯人工种子。在４℃下贮存２个月后,微芽人工种子和微薯人工种子在有菌腐殖土壤中播种２１ｄ的萌发率分别是１５７％和９６２％。     

介质Ca^2＋和La^3＋对酿酒酵母生长的影响

采用正交实验研究了外加Ｃａ＾２＋和Ｌａ＾３＋对酿酒酵母生长的影响。结果表明：外加Ｃａ＾２＋和Ｌａ＾３＋对酿酒酵母的生长均有显的影响,都呈现出低浓度对正效应和高浓度时负效应,当Ｃａ＾２＋浓度为１ｍｍｏｌ／Ｌ及Ｌａ＾３＋浓度为１５μｍｏｌ／Ｌ时酿酒酵母生长最好。     

Ultrastructural Study of Proplastid Ontogeny in Tobacco Mesophyll Cells in Vitro

Since the discovery of plastid DNA the continuity of plastids has well been established. It is known that in plant cultures a form of plastid can differentiate into others. However, only a little has been made in studing chloroplast dedifferentiation in vitro. In the work present here, we reported on ultrastructural changes of chloroplasts dedifferentiation and the proplastid origin in the mesophyll cells of cultured tobacco leaf explant. Fully expanded leaves of haploid tobacco (cv. Ge Xin No. 1) were cut into pieces of 5–6 mm width. These were inoculated on MS medium supplemented with 1 mg/L 2,4-D and 1 mg/l kinetin. The cultures were maintained at (30±2) ℃ and illuminatied by a bank of fluorescent lamps. For electronmicroseopic investigation, after 0, 1, 2, 3, 6 days of culture small leaf fragments were cut off along the cut edges of the explants. The samples were fixed and processed in the manner as described earlier. The sections were examined with a Hitachi HU-11A or a JEM-100CX electronmicroscope. Electronmicroscopic observation shows that the uncultured mesophyll cells are highly vacuolete, with a thin peripheral layer of cytoplasm in which a nucleus and some chloroplasts and other organelles are found in it. But these cells do not contain proplastids (Fig. l). In the explants cultured for 1 day there are no obviously changes in mesophyll cells, except a few cytoplasmic strands extend from periphery to central vacuole. At 2 days of culture quite obvious changes can be detected. A increase in the amount of cytoplasm becomes apparent and transvacuolar cytoplasmic strands grow up. Following cytoplasmic growth, the nucleus and chloroplasts move away from the peripheral cytoplasm and enter the central vacuolate zone (Fig. 2). At this stage some of mesophyll cells have completed the first cell division. After 3 days of culture numerous mesophyll cells have undergone several divisions and formed multicellular masses. In those subdivided cells a more important change of the chloroplasts is the occurrence of protrusions which we call proplastid buds. This phenomenon has also been named as chloroplast budding. According to observations on a large amount of sections chloroplast budding is a common phenomenon in the dedifferentiating mesophyll cells of tobacco leaf explants. Fig ure 3 exhibits a typical profile of a chloroplast with a proplastid bud. The proplastid buds observed are generally long-oval in shape and 1.0–2.5 μm long and about 0.5–0.7 μm thick. These dimensions agree with those of proplastids in meristematie cells. Inside of proplastids ribosomes and electron opaque areas containing DNA fibrils can be seen (Fig. 3). Near the proplastid buds proplastids can often be found (Fig.5). According to above observations we can conclude that the proplastids in dedifferentiating mesophyll cells originate from the proplastid buds by chloroplast budding. The newly formed proplastids usually surround the nucleus and sometimes undergo equal division to increase their number (Figs.5, 6). There are no inner membranes in the newly formed proplastids except vesicles connected with inner membrane of the envelope (Fig.7). While the proplastids are continuously produced, the chloroplasts themselves are filled with starch and gradually turned to large amyloplasts (Fig.5). On the other hand, a few of chloroplasts can divide into equal parts following the chloroplast budding (Fig.4). Israel and Steward (1967) suggested that when cultured carrot cells developed into plantlets the chloroplasts turned into leucoplastids, chromoplastids or proplastids. However, they did not describe how chloroplast became a proplastid. Several investigators reported that the chloroplasts in the dedifferentiating cells gradually lost their grana and intergranal lamellae and then became eueoplasts or proplastids. But according to our observation in tobacco explants, the initiation of proplastids is due to unequal division of chloroplasts, i.e. “budding fission” as described by Malzan and Miihlethaler in Splachnum ampullaceum. Since the proplastid is an organelle characteristic of meristematie cells, the ontogeny of proplastids and its control mechanism should be very important in studing cell dedifferentiation.     

枸杞组织培养中过氧化物酶和可溶性蛋白质的变化

枸杞(Lycium barbarum L.)叶组织切块接种在附加6-BA 0.5毫克/升和NAA 0.5毫克/升的MS固体培养基上可大量诱导产生愈伤组??织,培养至30天分化出芽,到50天长成小苗。过氧化物酶活性和可溶性蛋白质的含量分别在大量分生细胞团形成和芽形成时出现两个峰值,酸性蛋白质的种类也在这两个时期迅速增加。过氧化物酶同工酶阳极端出现的四条酶带中,A_2、A_3较稳定,A_1、A_4则在不同发育时期呈现规律性的变化。     

5-Aminolevulinic Acid Ameliorates the Growth, Photosynthetic Gas Exchange Capacity, and Ultrastructural Changes Under Cadmium Stress in Brassica napus L.

Heavy-metal toxicity in soil is one of the major constraints for oilseed rape (Brassica napus L.) production. One of the best ways to overcome this constraint is the use of growth regulators to induce plant tolerance. Response to cadmium (Cd) toxicity in combination with a growth regulator, 5-aminolevulinic acid (ALA), was investigated in oilseed rape grown hydroponically in greenhouse conditions under three levels of Cd (0, 100, and 500 μM) and three levels of foliar application of ALA (0, 12.5, and 25 mg l^?1). Cd decreased plant growth and the chlorophyll concentration in leaves. Foliar application of ALA improved plant growth and increased the chlorophyll concentration in the leaves of Cd-stressed plants. Significant reductions in photosynthetic parameters were observed by the addition of Cd alone. Application of ALA improved the net photosynthetic and gas exchange capacity of plants under Cd stress. ALA also reduced the Cd content in shoots and roots, which was elevated by high concentrations of Cd. The microscopic studies of leaf mesophyll cells under different Cd and ALA concentrations showed that foliar application of ALA significantly ameliorated the Cd effect and improved the structure of leaf mesophyll cells. However, the higher Cd concentration (500 μM) could totally damage leaf structure, and at this level the nucleus and intercellular spaces were not established as well; the cell membrane and cell wall were fused to each other. Chloroplasts were totally damaged and contained starch grains. However, foliar application of ALA improved cell structure under Cd stress and the visible cell structure had a nucleus, cell wall, and cell membrane. These results suggest that under 15-day Cd-induced stress, application of ALA helped improve plant growth, chlorophyll content, photosynthetic gas exchange capacity, and ultrastructural changes in leaf mesophyll cells of the rape plant.     

Morphological and physiological modifications of Cistus salvifolius L. winter leaves in response to the rise in winter temperatures

The global climate is predicted to change in the next century; for the Mediterranean Basin, an increase in air temperature more than 4°C and a higher frequency of extreme climatic events such as drought and heat waves are expected. In this work, the response of Cistus salvifolius L. to the rise in winter temperature has been studied. Plants acclimated to winter conditions [outdoor (OUT)] were moved into a greenhouse [indoor (IND)] at higher temperature and eco-physiological behaviour was analysed on leaves after 15 days from plant transferring (IND_15d) and on leaves developed IND. IND leaves were characterized by reduced thickness, higher specific leaf area, higher CO₂ mesophyll conductance and photosynthetic rate, and lower respiratory rate than leaves grown OUT upon current winter conditions. In IND_15d leaves, no improvement of photochemical activity was found. When IND leaves were subjected to a rapid increase in air temperature, the CO₂ fixation was not limited indicating a high thermotolerance of photosynthetic machinery. The results for IND leaves indicate the occurrence of a strategy that merging changes in leaf structure as well as in photosynthetic regulation allow C. salvifolius to maintain an elevated carbon gain in response to temperature increase.     

甜瓜子叶离体培养直接再生不定芽的形态学和解剖学观察

对甜瓜品种“西莫洛托”子叶离体培养直接再生不定芽过程进行了形态学和解剖学观察,结果表明：以子叶远轴面接触培养基培养能直接再生不定芽,以子叶近轴面接触培养基培养只能得到无序组织块;不定芽再生过程中,培养4d时,子叶变绿,但还不见有细胞分裂,约6d时,在子叶外植体的形态学下端切口处的近轴面到有局部的表皮细胞和亚表皮细胞分裂活跃,初步形成了拟分生组织,7－9d时,这些拟分生组织形成了肉眼可见的小突起,10－14d时,这些突起变得狭长,发育成幼叶或叶状体,它们在外植体上成族存在,但此时还没有典型的“芽”结构出现,15－20d时,成族突起形成幼叶丛,一些幼叶和叶状体的近轴面的基部出现叶腋分生组织,这些将外植体转入伸长培养基,3－5d后,幼叶充分展开,成丛叶状,多个叶腋分生组织同时发育成芽,再过2－7d,相继有不定芽从丛叶外植体上伸长,将伸长的不定芽切下,可促使外 植体上的其它不定芽伸长。     

Responses of Photosynthetic Electron Transport in Stomatal Guard Cells and Mesophyll Cells in Intact Leaves to Light, CO2, and Humidity

High-resolution images of the chlorophyll fluorescence parameter Fq'/Fm' from attached leaves of commelina (Commelina communis) and tradescantia (Tradescantia albiflora) were used to compare the responses of photosynthetic electron transport in stomatal guard cell chloroplasts and underlying mesophyll cells to key environmental variables. Fq'/Fm' estimates the quantum efficiency of photosystem II photochemistry and provides a relative measure of the quantum efficiency of non-cyclic photosynthetic electron transport. Over a range of light intensities, values of Fq'/Fm' were 20% to 30% lower in guard cell chloroplasts than in mesophyll cells, and there was a close linear relationship between the values for the two cell types. The responses of Fq'/Fm' of guard and mesophyll cells to changes of CO2 and O2 concentration were very similar. There were similar reductions of Fq'/Fm' of guard and mesophyll cells over a wide range of CO2 concentrations when the ambient oxygen concentration was decreased from 21% to 2%, suggesting that both cell types have similar proportions of photosynthetic electron transport used by Rubisco activity. When stomata closed after a pulse of dry air, Fq'/Fm' of both guard cell and mesophyll showed the same response; with a marked decline when ambient CO2 was low, but no change when ambient CO2 was high. This indicates that photosynthetic electron transport in guard cell chloroplasts responds to internal, not ambient, CO2 concentration.     

Hg2+和Cd2+胁迫对满江红生理和细胞超微结构的影响

研究了在梯度浓度Hg2+和Cd2+胁迫下,满江红(Azolla imbricata (Roxb.) Nakai)的叶绿素含量、叶绿素a/b比值、光合放氧速率、呼吸速率、抗氧化酶系(超氧化物歧化酶(SOD)、过氧化氢酶(CAT)、过氧化物酶(POD))和细胞超微结构受Hg2+和Cd2+的毒害影响.结果显示:随着胁迫程度的增大,叶绿素含量、叶绿素a/b比值、光合放氧速率明显下降,呼吸速率均在2 mg/L浓度下达到峰值,尔后下降; SOD、CAT、POD的活性均出现不同程度的应激性升高(除POD在Cd2+处理时下降),尔后下降.电镜观察发现,随着污染物浓度的增加和胁迫时间的延长,叶绿体出现膨大、破损和解体;线粒体嵴突膨胀和线粒体变形及空泡化;核染色质凝集,核仁消失,核膜破裂.实验结果表明: Hg2+和Cd2+污染不仅损害植物的生理活性,而且也破坏细胞的超微结构,最终导致植物死亡;随着Hg2+和Cd2+胁迫的增大,细胞超微结构的损伤程度和植物的生理变化是同步的;植物受毒害的程度表现出明显的剂量效应关系;在同一处理时间和浓度下,Cd2+对满江红的毒性大于Hg2+.Hg2+对满江红的致死浓度为3.5～4.0 mg/L,Cd2+为3.0～3.5 mg/L.对满江红鱼腥藻(Anabaena azollae Strasburger)细胞的超微结构变化观察表明,满江红鱼腥藻对Hg2+和Cd2+的耐受性明显高于满江红.     

Hg^2＋和Cd^2＋胁迫对满江红生理和细胞超微结构的影响

研究了在梯度浓度Hg^2 和Cd^2 胁迫下,满江红（Azolla imbricata(Roxb.)Nakai)的叶绿素含量,叶绿素a/b比值,光合放氧速率,呼吸速率,抗氧化酶系（超氧化物歧化酶（SOD）,过氧化氢酶（CAT）,过氧化物酶（POD）和细胞超微结构受He^2 和Cd^2 的毒害影响。结果显示：随着胁迫程度的增大,叶绿素含量,叶绿素a/b比值,光合放氧速率明显下降,呼吸速率均在2mg/L浓度下达到峰值,尔后下降;SOD,CAT,POD的活性均出现不同程度的应激性升高（除POD在Cd^2 处理时下降）,尔后下降,电镜观察发现,随着污染物浓度的增加和胁迫时间的延长,叶绿体出现膨大,破损和解体;线粒体嵴突膨胀和线粒体变形及空泡化;核染色质凝集,核仁消失。核膜破裂,实验结果表明：Hg^2 和Cd^2 污染不仅损害植物的生理活性,而且也破坏细胞的超微结构,最终导致植物死亡,随着Hg^2 和Cd^2 为3.0-3.5mg/L。对满江红鱼腥藻（Anabaena azollae Strasburger)细胞的超微结构变化观察表明,满江红鱼腥藻对Hg^2 和Cd^2 的耐受性明显高于满江红。     

Effect of ABA on the in Vitro Induction of Floral Buds of Dendrobium candidu Wall.ex Lindl.

An in vitro flowering system of Dendrobium candidurn Wall. ex Lindl., a wild species of orchid, was established. Callus was induced from seeds and protocorms formed on MS agar medium supplemented with 0.3 mg/L NAA under diffused light. Floral buds were induced when protocorms were cultured on MS agar medium supplemented with 2 mg/L 6-BA and 0.5 mg/L NAA, the frequency of floral bud induction being 27.0%. When protocorms were precultured on MS medium supplemented with 0. 5 mg/L ABA for 15 days and then transfered onto MS medium supplemented with 2 mg/L 6-BA, the frequency of floral bud induction increased greatly reach 84.0 % during a period of 5 months. Meanwhile the number of branches and floral buds also increased and in some instances inflorescence appeared. Never the less, no floral bud was formed if protocorms continued to be cultured on the ABA-containing MS medium.     

Establishment of an Experimental System for the Study of Tracheary Element Differentiation from Single Cells Isolated from the Mesophyll of Zinnia elegans

Single cells were isolated mechanically from the mesophyll of adult plants and of seedlings of Zinnia elegans L. cv. Canary bird. When single cells isolated from the first leaves of seedlings were cultured in a liquid medium in the dark with rotation, they differentiated to tracheary elements with a reasonable degree of synchrony in the 24-hour period between days 2 and 3 after culture. The proportion of tracheary elements as a percentage of total cells reached nearly 30% 3 days after culture. Factors favoring cytodifferentiation were certain optimum levels of both α-naphthalene-acetic acid (0.1 milligram per liter) and benzyladenine (1 milligram per liter), a low concentration of ammonium chloride (0 to 1 millimolar), and an initial cell population density in the range 0.4 to 3.8 × 10⁵ cells/ml. It was possible to follow analytically the sequence of cytodifferentiation in individual cells in this system.