实验仪器使用技术

植物生理实验使用的材料非常广泛，根据来源可划分为天然的植物材料（如植物幼苗、根、茎、叶、花等器官或组织等）和人工培养、选育的植物材料（如杂交种、诱导突变种、植物组织培养突变型细胞、愈伤组织、酵母等）两大类；按其水分状况、生理状态可划分为新鲜植物材料（如苹果、梨、桃果肉，蔬菜叶片，绿豆、豌豆芽下胚轴，麦芽、谷芽，鳞茎、花椰菜等）和干材料（小麦面粉，玉米粉，大豆粉，根、茎、叶干粉，干酵母等）两大类，因实验目的和条件不同，而加以选择。 植物材料 ...

植物根系是活跃的吸收器官和合成器官，根的生长情况和活力水平直接影响地上部的营养状况及产量水平。本实验练习测定根系活力的方法，为植物营养研究提供依据。 一、原理 氯化三苯基四氮唑（ TTC ）是标准氧化电位为 80 mV 的氧化还原色素，溶于水中成为无色溶液，但还原后即生成红色而不溶于水的三苯甲 （ TTF ），如下式： 生成的三苯甲 （ TTF ）比较稳定，不会被空气中的氧自动氧化，所以 TTC 被广泛用作酶试验的氢受体，植物根系中脱氢酶所引起的 TTC 还原，可因加入琥珀酸、延胡 ...

植物细胞的渗透势主要取决于液泡的溶质浓度，因此又称溶质势。渗透势与植物水分代谢、生长及抗逆性等有密切关系。已知在干旱、盐渍等条件下，一些植物常在细胞内主动积累溶质，以降低其渗透势，增加吸水能力，而在一定程度上维持细胞膨压，保障细胞的生长和气孔的开放，这种现象叫做渗透调节作用。渗透调节能力的大小可以用逆境条件下细胞的渗透势的降低值来表示，在水分生理与抗逆性生理研究中经常需要测定。 一、原理 将植物组织放入一系列不同浓度的 ...

1) wash 10' with tap H2O 2) wash w/ 4xV w/ 0.2% AgNO3 3) wash 2x w/ sterile H2O 4) wash 1x w/ 100mM NaCl 5) wash 3x w/ sterile H2O 6) plate w/ 100ug/ml chloramphenicol 100U/ml Nystatin OR 1) wash 1' 96% EtOH, aspirate 2) wash 10' 30% commercial bleach w/ 1ul/ml 20% TritonX100 3) wash 5x w/ sterile H2O 4) plate as above 上一篇：叶绿体色素的提取、分离和理化性质 下一篇：Pr ...

一、原理 叶绿体色素是植物吸收太阳光能进行光合作用的重要物质，主要由叶绿素 a 、叶绿素 b 、胡萝卜素和叶黄素组成。它们与类囊体膜相结合成为色素蛋白复合体。这两类色素都不溶于水，而溶于有机溶剂，故可用乙醇、丙酮等有机溶剂提取。提取液可用色谱分析的原理加以分离。因吸附剂对不同物质的吸附力不同，当用适当的溶剂推动时，混合物中各种成分在两相（固定相和流动相）间具有不同的分配系数，所以移动速度不同，经过一定时间后，可将各种色素分开。 当叶绿素 ...

1. Transform plants by infiltration, grow to seed (F1). Transgenes should be dominant (expressed constitutively), therefore phenotypes possible in these seeds although there are so relatively few transformants that they would be virtually impossible to see. 2. Plate F1 seeds on KAN and sel ...

Vapor-Phase Sterilization Useful when sterilizing several tubes of seeds. Use microfuge tubes marked with pencil instead of pen (pen will disappear). Poke a small hole into the top of the tube. Put seeds into tubes these tubes. Do not fill tube with seeds past the .1ml line on the tube. Place tubes closed (b ...

I. Prepare plant tissue samples: 1. Weigh ~0.5g fresh leaf tissue (3-5cm2), grind using liquid nitrogen. 2. Add ~0.4ml Plant Extraction Buffer, grind until slurry, transfer to 1.5ml tube.3. Spin down for 15 minutes at 4°C, transfer 0.2-0.3 ml supernatant to fresh tube. 4. Add an equal volume of 2X Laemmli's buff ...

1. Scoop dry soil into an autoclave bin. Fill until the soil is approximately one inch from the top after packing down with the scooper. 2. Cover the bins with aluminum foil and put a piece of autoclave tape on each one. Make sure the outsides of the bins are dirt free before putting in the autoclave. 3. Autoclave on grav ...

(Janine’s protocol)1. Add 200µl of acetone, vortex briefly. 2. Remove acetone and pipette into flat-bottom 96-well plate (polystyrene) 3. Centrifuge plate at 3000 x g for 3-5 minutes. 4. Invert plate carefully and blot dry with paper towels 5. Add 40µl of X-Gluc stain solution to each well (multi-pipett ...

PLANTING SEEDwith tweezers- recommended for pots: 1. Wet small tweezer tips with dH20. 2. Touch one tip to a seed. 3. Move seed to soil by touching tweezer tip to desired spot on soil. 4. Cover pots with humidomes. Let pots sit in 4°C room for 2 days and then move up to growth room. with 0.1% agar- recommended for flats 1. Weigh out s ...

0.01% boric acid 1 mM MgSO4 2mM CaCl2--this was used with media containing GABA. If no other nitrogen source is present, use 1mM CaCl2 and 1 mM CaNO3 18% sucrose pH 6.5 Air dry flowers for 1-2 hours (optional--this helps with release from anthers but is not essential), then dab anthers onto surface of a 25-30µL drop of ge ...

Complementation is performed to rescue the phenotype of a mutant plant with the cloned wildtype gene. This can prove that the gene cloned is in fact the gene that was mutated in the mutant plant. I. Cloning wildtype gene into the plant transformation vector: 1. Design primers for region to be amplified.** ...

Basta SelectionPlant 500 - 1000 plants per flat. (It's a good idea to plant two control pots, one with wild-type and another with a known Basta resistant strain). Apply BASTA solution after the cotyledons have emerged, but before the true leaves have extended. Apply the spray in the fume hood. Wait one day be ...

Fluorescein Diacetate (FDA)1. Place pollen on slide. 2. Incubate in humid chamber (wet Whatman in Tupperware) ~ 30 min. 3. Add drops of FDA (2mg/ml in acetone) to Pollen Growth Media (~/ml) until it becomes turbid. 4. Add 1 drop to pollen. Incubate 10’ 5. Add slide and view under FITC. Pollen Growth Media2X For 4 Liters: ...

Materials Gilson (or Warburg) Respirometer at 37° CMitochondria suspension from Exercise 8.4 Krebs Phosphate Ringers (KPR)10% (w/v) Glucose0.39% (w/v) Sodium azide18.4 mg% (w/v) Dinitrophenol (DNP)6.64% (w/v) Sodium Malonate10% (w/v) KOH Procedure When you enter the lab, check that the ref ...

Aniline Blue Staining of Pollen/Pollen Tubes花粉管染色Materials Fixative: 10% Acetic Acid in ethanol 1M NaOH 50 mM KPO4 buffer, pH 7.5: 4.17 mL 1M K2HPO4 0.83 mL 1M KH2PO4 0.01% aniline blue in 50 mM KPO4 buffer (dye) KPO4 buffer made with 50% glycerol (mounting media) Method 1.Submerge pistil tissue in about 250 ...

Materials 0.01 M Potassium Phosphate Buffer, pH 7.4Sat. K Fe(CN)0.6 M Sodium Malonate (Malonic acid, sodium salt)5 mM KCN0.02% (w/v) p-phenylenediamine oxalate (PPDO) Mitochondria suspension from Spectrophotometer and cuvettes Procedure Prepare a series of 5 tubes as follows: Substan ...

Materials Mitochondrial suspension from Exercise 8.4 Sodium dithionite0.2 M Sorenson Phosphate Buffer, pH 7.51% (w/v) Bovine Serum Albumin 0.005 M Potassium Cyanide0.00025 M Dichlorophenolindophenol0.6 M Sodium succinate, pH 7.50.6 M Sodium malonate, pH 7.50.033% (v/v) Phenazine M ...

Materials Chloroplast suspension from Exercise 8.1AcetoneSpectrophotometer and tubesClinical centrifuge and tubes Procedure Pipet 1.0 ml of a chloroplast suspension into a 15 ml centrifuge tube and add 8.0 ml of acetone. Mix thoroughly. Add 1.0 ml of distilled water, mix again and centr ...