不同培养条件对拟南芥根细胞膜片钳记录的影响

本文以沙培养法、蛭石培养法、土培养法、水培养法和MS培养基等不同的方法培养拟南芥(Arabidopsis thaliana),分析了不同培养方法对根生长发育的影响,并分别分离根的原生质体.在膜片钳记录中对不同来源的根原生质体状态进行了比较.结果表明,土培养法分离的原生质体最适于膜片钳记录.     

一种改良的逼尿肌急性分离方法及离子通道电流检测

目的:建立一种稳定的适合膜片钳技术的逼尿肌细胞急性酶分离方法,为排尿相关障碍性疾病的研究提供必要的技术平台.方法:采用H型胶原酶和木瓜蛋白酶相混合的鸡尾酒酶液对新鲜离体的大鼠膀胱逼尿肌条在37℃条件下振荡消化,α-actin免疫荧光染色对分离并培养的原代细胞进行鉴定,在膜片钳工作台上分别对其进行L型钙电流和BKca钾电流的全细胞记录.结果:可获得大量的单个逼尿肌细胞.经过免疫荧光染色证实为平滑肌细胞.分离细胞活性良好,在膜片钳实验系统上可记录到多种通道电流.结论:建立了一种操作简单、成功率高、活性好的逼尿肌细胞急性酶分离方法并成功应用于膜片钳技术.     

影响决明无菌苗子叶原生质体分离和培养因素的研究

以决明(Cassia obtusi folia)无菌苗子叶为材料,对酶组合、无菌苗日龄,植物激素组合和培养方法对其原生质体的分离和培养的影响进行了研究。结果表明:用3％的纤维素酶和0.2％Pectinase Y-23的酶组合处理决明无菌苗子叶块8小时可以高效分离出有活力的原生质体;约14日龄的决明无菌苗子叶比较适合于原生质体的分离;适当浓度的2,4-D 有利于原生质体的分离。促进原生质体分裂的理想的植物激素组合为0.4 mg/L 2,4-D,1.0 mg/L NAA and 0.1 mg/L KT;漂浮培养法最有利于原生质体的分裂和发育。找出了适合于决明无菌苗子叶原生质体的分离和培养的酶组合、植物激索组合、有效培养方法和决明无菌苗子叶日龄。这为有效地从决明无菌苗子叶原生质体再生植株奠定了基础。     

恒流灌注分离心肌细胞及膜片钳记录钾电流方法学探讨

目的探讨膜片钳实验中豚鼠、大鼠心肌细胞急性分离过程中的观察指标和影响因素,并进行膜片钳记录。方法使用改进的Langendorff恒流灌注法分离心肌细胞和全细胞膜片钳记录钾通道电流。结果分离即刻,存活细胞可达80%以上,复钙后约40%～50%呈静息状态,余出现自发性收缩,存活细胞能够进行膜片钳封接,于室温KB液中可成活8 h以上。结论使用恒流灌注法,观测灌注压的变化,可以较客观的掌握酶解消化的程度,利于获取生理状态良好的心肌细胞。且细胞能够进行膜片钳实验。     

Na+和Ca2+对拟南芥根原生质体质膜内向K+通道电流的影响

以拟南芥(Arabidopsis thaliana Columbia)根为材料,利用膜片钳技术测定其根细胞原生质体质膜内向K^ 电流,并对Na^ 对其K^ 电流的影响进行了初步研究,发现Na^2 可明显抑制拟南芥根细胞原生质体的内向K^ 电流,外施Ca^2 可缓解Na^ 对内向K^ 电流的抑制．说明Ca^2 参与了质膜上K^ 通道对K^ ／Na^ 的选择性吸收的调节,从而使植物适应盐胁迫．     

坛紫菜原生质体的发育研究

用2％的海螺酶和1％的纤维素酶混合,将坛紫菜叶状体的4种不同细胞类型即:营养细胞、根丝细胞、精子囊和果孢子囊,分别解离成原生质体,并研究了这些不同部位和不同生长时期。细胞的生长、发育途径。由根丝细胞分离的原生质体能长成新的叶状体;由早期中部营养细胞分离的原生质体,有70％长成新的叶状体,其余的发育成精子囊和果孢子囊。再生叶状体在室内培养,能正常成熟。由精子囊和果孢子囊分离的原生质体,即精子细胞和果孢子细胞不能再生叶状体。前者形成新的精子囊放散精子,后者形成新的果孢子囊,放散果孢子发育成丝状体。晚期紫菜与早期紫菜比较,再生叶状体的数量显著减少,而发育成精子囊和果孢子囊的数量则大大增多。     

烟草幼嫩花粉原生质体分离与早期离体发育

建立了两种分离烟草（Ｎｉｃｏｔｉａｎａ ｔａｂａｃｕｍ ）幼嫩花粉原生质体的方法：一为通过花药漂浮培养释放出外壁裂开的花粉,后者转入酶液彻底脱去外壁,内壁降解而分离出原生质体（简称“花药预培养法”）;二为花粉经饥饿处理后,转入酶液释放原生质体（简称“花粉饥饿预处理法”）。对影响分离效果的主要因素作了研究。用花药预培养法分离的幼嫩花粉原生质体,在Ｋ３ 培养基中可再生细胞壁,启动１ 次细胞分裂,或萌发花粉管。表明具有孢子体发育与配子体发育的潜能。观察到花粉原生质体分裂成二细胞后,其中１ 个子细胞又长出花粉管的稀有现象,暗示其开始启动孢子体发育后又重新恢复配子体发育途径     

不同因素对金针菇原生质体制备和再生的影响

比较了不同浓度裂解酶及组合、渗透压稳定剂、酶解时间等因素对金针菇原生质体得率的影响以及不同渗透压稳定剂、培养基、接种方法等因素对金针菇原生质体再生的影响。试验结果表明:固体培养10d的金针菇菌丝,以0.5mol/L KCl作渗透压稳定剂,加入1%纤维素酶和1%溶菌酶在25℃下酶解1.5h,分离原生质体效果最佳,原生质体产量可达27.8×105个/ml以上;以0.5mol/L KCl作渗透压稳定剂,在25℃条件下,金针菇原生质体采用直接涂布法接种在RCM培养基上培养,再生率最高为0.5%。     

大豆原生质体培养再生植株

大豆原生质体培养诱导再生植株一直为国内外学者所关注。70年代初,Kao等、Miller等即由悬浮培养的大豆细胞分离原生质体,经培养获得了愈伤组织,但在以后的很多年中进展不大。近年来,虽然一些学者直接从大豆植株的不同器官或部位,如荚果组织、幼苗根、叶肉组织、子叶和下胚轴悬浮培养细胞游离和培     

一种适合膜片钳单通道记录的大鼠DRG神经元急性分离方法

目的：建立一种适合膜片钳单通道记录的脊髓背根神经节神经元急性分离方法。方法：用酶消化和机械分离相结合的方法急性分离大鼠DRG神经元。结果：用本方法分离的DRG细胞容易形成较高的封接电阻（〉5GΩ）,降低了噪音干扰,可记录到pA级的单通道电流。结论：本方法急性分离的DRG神经元适合单通道膜片钳实验研究。     

Regeneration of plants from mesophyll protoplasts of ground-cherry (Physalis minimal L.)

High yields of protoplasts were obtained by enzymatic digestion of leaf tissue derived from exenic shoot cultures of Physalis minima L. Isolated protoplasts underwent sustained mitotic divisions and produced calli. Shoot and root morphogenesis were achieved from these calli. The regenerated plants were established in soil.     

采伐干扰对华北落叶松细根生物量空间异质性的影响

以华北落叶松天然林为研究对象,选择采伐干扰林分(样地A)和未采伐干扰林分(样地B),利用根钻法分3层(0—10cm,10—20cm,20—30cm)获取各径级细根(≤1mm、1—2mm、2—5mm3级活细根,≤2mm死亡细根)生物量数据。采用地统计学变异函数和经典统计相结合的数据分析方法对采伐干扰造成的细根生物量空间异质性的变化进行定量研究。主要研究结果如下:采伐干扰林分样地A各经级细根生物量均值减少;同一土层相同径级细根生物量样地A与样地B相比差异显著(P<0.05);不同土层的细根生物量异质性具有显著差别(P<0.05)。0—10cm土层,未采伐干扰林分≤1mm细根生物量呈现较明显的空间自相关变异,采伐干扰林分则表现为随机性变异特征,采伐干扰导致≤1mm细根生物量空间分布特征更加复杂(分维数D=1.978);10—20cm土层,采伐干扰林分各径级细根生物量异质性程度明显降低,只有未采伐干扰林分的5.4%—88.9%。20—30cm土层,未采伐干扰林分≤1mm细根生物量在较小尺度范围(<2.9m)表现出明显的空间自相关变异(结构方差比86.1%),受采伐干扰林分各径级细根生物量异质性程度只有未采伐干扰林分的8.9%—45.9%,且呈现随机性变异。各径级细根生物量空间异质性的垂直分异均表现为随土层深度的增加异质性强度明显降低。     

三峡库区马尾松根系生物量的空间分布

以三峡库区主要植被马尾松人工林为研究对象,用内径为10 cm的根钻,分别在马尾松中龄林、近熟林和成熟林内,据树干0.5、1.0、1.5 m和2.0 m处设置取样点,各样点按0-10、10-20、20-30、30-40、40-60 cm将土壤分为5个垂直层次,对马尾松根系的空间分布格局进行调查。结果表明:(1)三峡库区马尾松总根系生物量(0-10 mm)为中龄林(4.72 t/hm²)显著高于成熟林(2.94 t/hm²)和近熟林(2.40 t/hm²)(P<0.05)。细根(0-2 mm)生物量随年龄增加而递减,差异不显著(P>0.05);(2)马尾松3个林龄中根系生物量表现出一定的水平分布特征,但具体趋势表现各异,细根生物量最大值均出现在距离样木1.0 m处;(3)细根主要分布在土壤上层,其中47.53%-71.73%的活细根集中在0-20 cm土壤深度内,且随土层的加深,其生物量明显减少。粗根(2-10 mm)则主要分布于20-60 cm土层范围内;(4)根系直径越小,受环境变化越明显。马尾松细根生物量分布主要受土壤深度的影响,树龄和不同水平距离对细根分布格局影响不显著(P>0.05),各因素对粗根生物量的影响均未达到显著水平(P>0.05)。     

Fertile plant regeneration from barley (Hordeum vulgare L.) protoplasts isolated from primary calluses

Barley (Hordeum vulgare L.) protoplasts were isolated from the immature embryo-derived primary calluses. These protoplasts were cultured with nurse cells, and they then divided to form colonies. After transfer of the colonies to regeneration medium, either green or albino shoots were regenerated from these colonies. A high agarose concentration (2.4% w/v) in the protoplast culture medium promoted protoplast division. The plantlets that developed strong root systems were transferred to the soil. These plants flowered and have set seeds.     

臭氧对无土栽培营养液的消毒作用研究

采用稀释平板法和MPN法,测定了臭氧对无土栽培营养液的消毒杀菌作用。结果表明,随着臭氧浓度(时间)的增加,对微生物杀灭率明显上升;连续处理20min,可杀灭90%以上的细菌、真菌和藻类;臭氧间歇曝气消毒营养液,有效控制微生物的滋生,对莴苣根系无不良影响,增加产量36.8%。     

KDC1, a novel carrot root hair K+ channel. Cloning, characterization, and expression in mammalian cells

Potassium is an essential nutrient which plays an important role in many aspects of plant growth and development. Plants have developed a number of highly specific mechanisms to take up potassium from the soil; these include the expression of K(+) transporters and potassium channels in root cells. Despite the fact that root epidermal and hair cells are in direct contact with the soil, the role of these tissues in K(+) uptake is not well understood. Here we report the molecular cloning and functional characterization of a novel potassium channel KDC1 which forms part of a new subfamily of plant K(in) channels. Kdc1 was isolated from carrot root RNA and in situ hybridization experiments show Kdc1 to be highly expressed in root hair cells. Expressing the KDC1 protein in Chinese hamster ovary cells identified it as a voltage and pH-dependent inwardly rectifying potassium channel. An electrophysiological analysis of carrot root hair protoplasts confirmed the biophysical properties of the Kdc1 gene product (KDC1) in the heterologous expression system. KDC1 thus represents a major K(+) uptake channel in carrot root hair cells.     

Prevention of aluminium toxicity with supplemental boron. II. Stimulation of root growth in an acidic, high-aluminium subsoil

Root growth inhibition is an early symptom of Al toxicity and B deficiency. Our hypothesis is that Al toxicity may induce B deficiency, and it was our objective to determine if incorporation of supplemental B would promote root penetration into an acidic, high-Al subsoil. Alfalfa (Medicago sativa L. cv. Hy-Phy) was grown in slanted tubes with a Plexiglas window along their length. The top half of the tub contained silt loam soil and the bottom half contained subsoil from the Bt1 horizon (26% Al saturation) of a Creldon silty clay loam. Both soils originally contained 0–9 kg B ha^?1. When root growth was measured in the bottom half of the high-A1 subsoil, all measurements—depth of rooting, total root growth, final root lengths and root dry weight—demonstrated greater root growth in treatments where additional B was incorporated into the high-Al subsoil. Results from this soil study extend those obtained in our hydroponic study in which supplemental B presented Al inhibition of root growth. Boron concentrations may need to he increased under acidic ‘high-Al’ soil conditions to promote root penetration into these soil zones, and this could be especially important during periods of drought stress.     

Morphology, RNase and transaminase of root protoplasts

RNase and transaminase activities were analysed for mechanicallyand enzymatically prepared protoplasts from Allium Cepa roots.The comparative analyses at three root regions show that theenzymes were less active in the protoplasts than in the cellsfrom which they had been obtained. The enzyme gradients (fromapex to base of the root: RNase increase and transaminase decrease)noted previously in the intact roots were found to be similarin the protoplasts, however to a lesser degree. On the otherhand, the relative activity of both tested enzymes was lowerin the enzymatically prepared protoplasts than in those obtainedby the mechanical technique. In connection with their physiologicalproperties, the respective effects of the mode of preparingthe protoplasts were discussed. (Received November 22, 1971; )     

轮叶党参营养成分的研究

本文研究了轮叶党参的茎叶、去皮根及根皮中的营养成分氨基酸、维生素、微量元素的含量。结果显示,这三种营养成分都在根皮中最多,茎叶和去皮根中的含量也都多于一般蔬菜中的含量,为轮叶党参的合理应用提供了科学依据。     

Abscisic Acid Response of Corn (<Emphasis Type="Italic">Zea mays</Emphasis> L.) Roots and Protoplasts to Lanthanum

Lanthanum ions antagonize calcium and are used as a Ca²⁺ channel blocker but their direct effects are unknown. We investigated lanthanum effects on endogenous abscisic acid (ABA) levels in protoplasts and intact primary roots of Zea mays L. Application of 1 mM La³⁺ reduced primary root elongation, caused swelling of root tips, and essentially doubled the ABA content in intact roots but decreased ABA in root protoplasts in a concentration-dependent manner. Osmotic stress increased ABA level in protoplasts more than in intact roots. Temporal ABA changes in response to La³⁺ treatment indicate that La³⁺ affects root growth at least partially via ABA pathway.