扁桃愈伤组织诱导及分化的研究

以扁桃优良品种'Naporeil'的茎段、叶片和花药作为外殖体,分别对其进行愈伤组织诱导和分化研究,以筛选愈伤组织的最佳诱导增殖培养基、分化培养基和生根培养基.结果表明,该品种以茎段、花药作为外殖体最易诱导获得愈伤组织,叶片不适宜作为外殖体诱导愈伤组织;愈伤组织的最佳诱导增殖培养基均为B5+0.5 mg/L 2,4-D+0.2 mg/L 6-BA+1.0 mg/L NAA,愈伤组织诱导率为100%,增殖倍数最高可达7倍;茎段愈伤组织的分化培养基为MS+0.2 mg/L NAA+0.8 mg/L 6-BA+0.5 mg/L ZT,分化率为71%;花药愈伤组织未见分化.由茎段愈伤组织再分化获得的不定芽在1/2MS+0.5 mg/L IBA培养基上诱导生根,并给以黑暗预处理可使生根率达80%以上.     

红叶石楠离体植株再生频率的影响因素研究

以红叶石楠带芽茎段及叶片为外植体,分析激素和培养条件等因子对愈伤组织诱导及植株再生的影响。结果表明,MS+0.10mg/L 2,4-D+0.50mg/L NAA+0.50mg/L 6-BA+0.50mg/L KT为最佳愈伤组织诱导培养基,暗培养的愈伤组织诱导率高于光培养,其愈伤组织诱导率可达100%(带芽茎段)和98%(叶片)。MS+0.50mg/L IBA+2.00mg/L 6-BA+2.00mg/L KT为最佳分化增殖培养基,分化率91%以上,增殖倍数6.8以上,均达到最高。1/2MS+0.50mg/L IBA+0.01mg/L NAA为最佳生根培养基,生根率92%,生根量4.4根/株,均达到最高。     

玉竹的组织培养与快速繁殖

以玉竹[Polygonatum odoratum (Mill.) Druce]根状茎、叶片和茎段为外植体,于附加不同激素配比的MS培养基中诱导愈伤组织、不定芽和不定根,探讨增殖培养和植株再生的条件.结果表明,叶片和茎段外植体诱导愈伤组织和芽的分化率很低;而根状茎外植体易于培养,有较高的诱导率和增殖倍数,其愈伤组织、不定芽和不定根的诱导率分别可达87%、90%和99%以上.适宜根状茎外植体愈伤组织诱导的培养基为MS+1.0 mg/L 6-BA+0.5 mg/L NAA,有利于增殖和丛生芽分化的培养基为MS+2.0 mg/L 6-BA+0.5 mg/L IBA和MS+3.0 mg/L 6-BA+0.1 mg/L NAA,而1/2MS+3.0～5.0 mg/L NAA适宜诱导试管苗生根培养.试管苗的移栽成活率可达85%以上.     

匙羹藤愈伤组织培养研究

以匙羹藤组培苗为外植体,研究茎段、茎尖和叶片以及不同植物生长调节剂组合对愈伤组织形成的影响;采用4因素4水平正交设计,用SPSS软件其结果进行分析。结果显示:各种外植体诱导愈伤的能力不同,依次是茎段、叶片。叶片诱导出非胚性愈伤组织,茎段、茎尖利于诱导胚性愈伤组织和胚状体;愈伤诱导的最佳培养基为MS+2,4-D 2.0 mg/L+6-BA 0.5 mg/L,诱导率为84.5%,愈伤生长旺盛,分裂快,颜色为淡黄色。愈伤组织分化成苗效果最好的条件是MS+NAA 1.0 mg/L+IBA 0.1 mg/L,分化率达到90.6%,最佳的生根条件是1/2 MS+NAA1.6 mg/L,生根率为100%。这为研究匙羹藤的组织的脱分化、分化以及胚状体的形成机制提供了参考。     

栀子带芽茎段愈伤组织诱导分化及丛生芽增殖和生根研究

以栀子带芽茎段为试材,对其愈伤组织诱导分化及丛生芽增殖与生根进行初步研究。结果表明,栀子带芽茎段愈伤组织快速诱导和分化最佳培养基是MS+KT 1 mg/L+NAA 0.05 mg/L;栀子带芽茎段丛生芽增殖和生根最佳培养基是MS+KT 2.0 mg/L+NAA 0.05 mg/L。     

睫毛萼凤仙花的组织培养及其愈伤组织的超微观察

试验以睫毛萼凤仙花无菌苗的叶片、茎和根为外植体进行离体培养,并对两种具有不同再分化能力的愈伤组织进行电镜超微观察.结果表明:在培养基MS+NAA 0.5 mg/L+6-BA 0.5 mg/L上愈伤组织诱导率达最高,叶片愈伤组织的诱导率为100%,茎的愈伤组织诱导率为67%,而根却没有愈伤组织的发生;叶片愈伤组织在MS+...     

黄山药愈伤组织的诱导与分化

以黄山药的叶片、茎段和叶柄作外植体,以MS为基本培养基,试验了不同激素组合对愈伤组织诱导的影响,采用正交试验法研究了愈伤组织的分化效果。结果表明,以MS+2,4-D1~2mg/L+6-BA2mg/L培养基对叶片的愈伤组织诱导效果最好,接种12d后初见愈伤组织,20d后可形成大量的愈伤组织,而茎段和叶柄的诱导效果较差。分化试验结果表明,生根率最高可达85.3%,而出芽率最高仅达29.6%。     

金线莲外植体筛选及愈伤组织诱导研究

以金线莲茎段、叶片、茎片、不定芽为试材,分别在添加6-BA、ZT、NAA、KT 5个不同处理的MS及1/2MS培养基上培养,诱导愈伤组织。结果表明,以茎段、茎片、不定芽为外植体,在MS + 6-BA 2.0 mg/L + NAA0.5 mg/L、MS + NAA 2.0 mg/L + KT 0.1 mg/L和MS + 6-BA 2.0 mg/L + NAA 0.5 mg/L + ZT 0.2 mg/L培养基中培养,均能成功诱导愈伤组织。不定芽为诱导愈伤组织最佳外植体,最佳培养基为MS + 6-BA 2.0 mg/L + NAA 0.5 mg/L + KT 0.2 mg/L。     

罗汉果叶片离体再生快繁技术

以罗汉果(Ssiraiti grosvenori)叶片为外植体,探讨培养方式、激素组合对愈伤组织诱导和不定芽分化的影响。结果表明:罗汉果叶片在培养基MS+BA 1.0 mg/L+IBA 0.7 mg/L上培养4周愈伤组织的诱导率达90%以上;罗汉果愈伤组织在培养基MS+BA 1.0 mg/L+IBA0.2 mg/L+赛苯隆(TDZ)0.1 mg/L上不定芽的分化率可达70%,平均出芽指数3.7;罗汉果试管苗在培养基MS+BA 2.0 mg/L+IBA 0.2 mg/L上茎芽增殖比较稳定,在培养基MS+IBA0.1 mg/L上培养2周开始分化不定根,其生根率在90%以上。     

不同外植体来源和培养条件对拟似棉植株再生的影响

对拟似棉的不同器官(叶、叶柄、茎)在离体培养中的反应以及影响植株分化的各种因素(继代培养时间、激素、光、温度)进行了研究。结果表明:茎段作为外植体效果最佳,并且愈伤组织的诱导能力与茎段切口的面积成正比。外植体的方向性,也就是指茎表皮靠着培养基,切口面暴露于空气中为愈伤组织诱导的重要条件。愈伤组织诱导的最适培养基是MS+2mg/l IAA+1mg/l KT,最佳培养条件是强光(10,000勒克斯)、高温(29±1℃)。在MS+0.1mg/l NAA+3mg/l 2ip+0.5g/l LH分化培养基上,在4,000—5,000勒克斯的光强,28℃条件下,愈伤组织分化成小苗的频率可达20％。     

产藏红花素栀子愈伤组织的诱导和筛选

为解决藏红花资源问题,研究了栀子愈伤组织的诱导,并首次应用目视法和HPLC 方法对产藏红花素细胞系进行了筛选。结果表明栀子叶子愈伤组织优化的诱导条件为: 在添加1mg/L 2,4-D和 0.25 mg/L 6-BA的MS 培养基上, 25±1℃和31.74 mmol/m2s光照强度下每天光照16小时,培养15d后可获得100%的诱导率。种子愈伤组织也易诱导,在添加0.5mg/L 2,4-D 和 0.25 mg/L 6-BA的MS培养基上,25±1℃全时暗培养15d后可获得100%的诱导率。首次应用建立的目视法和HPLC 方法, 从90株细胞系中筛选出细胞系seed4,其藏红花素1 的含量是0.348mg/g,生长较快、不易褐化,兼顾了此次生代谢产物的含量和细胞的生长速度,为采用植物细胞工程法解决藏红花素资源短缺问题提供了种质资源。     

生姜离体叶片愈伤组织悬浮细胞系建立与植株再生

以‘莱芜大姜’为试材,研究了生姜离体叶片愈伤组织的诱导以及细胞悬浮系建立与植株再生。结果表明,以生姜试管苗叶片为外植体,接种到MS+1.0 mg/L 2,4-D+0.5 mg/L 6-BA+30 g/L蔗糖的培养基上,可有效诱导出生长迅速、质地疏松的愈伤组织。将获得的愈伤组织接种到MS+0.15 mg/L 2,4-D+6.0 mg/L 6-BA+30 g/L蔗糖的液体培养基上,25℃黑暗条件下震荡培养25-30 d,可建立分散性好、生长迅速的悬浮细胞系,细胞悬浮系培养的适宜参数为:初始接种量为1.0-1.5 g,继代培养的适宜间隔期为15 d,继代培养液体培养基更新比例为3/4。将悬浮细胞接种到固体培养基MS+0.2 mg/L NAA+10.0 mg/L 6-BA+30 g/L蔗糖上可获得再生植株。     

细毛樟愈伤组织培养

从富含香叶醇的细毛樟（ＣｉｎｎａｍｏｍｕｍｔｅｎｕｉｐｉｌｕｍＫｏｓｔｅｒｍ）叶片外植体诱导出愈伤组织。愈伤组织生长较合适的培养基为ｐＨ５．８附加ＩＡＡ２ｍｇ／Ｌ和ＫＴ０１ｍｇ／Ｌ的ＭＳ培养基,在２５℃下暗培养。愈伤组织无性系继代培养到第６代后芳香成分初步分析结果表明主成分仍为香叶醇。     

Isolation,Culture and Plant Regeneration of Protoplasts from an Embryogenic Callus Tissue of Gossypium hirsutum

Protoplasts were isolated and cultured from hypocotyl embryogenic callus tissue of Gossypium hirsutum L. cv. "Lumian 6". The highest yields of viable protoplasts were obtained from a vigorous embryogenic callus 7 to 9 d old subcultured on MS medium supplemented with 2 mg/L IAA and 1 mg/L KT using a solution of 1% cellulase Onozuka R-10, 1% pectinase, 0.7 mmol/L KH2PO4, 2.5 mmol/L Ca2+ , and 0.5 mol/L osmoticum (mannitol), at pH 5.8 and at a temperature of 30 ℃. After separation and purification (in 21% sucrose floatation medium), the protoplasts were laid up in a quiet liquid protoplast culture medium containing K3 salts, NT vitamins with 0.1 mg/L 2,4-D, 0.2 mg/L KT and 0.45 mol/L glucose for 10 to 15 min. The protoplasts were fractioned into an upper and a lower layer in the centrifugal tube. Most of the protoplasts in the lower layer were smaller, round and rich in cytoplasts in which contain many granular substances. When this kind of protoplasts were cultured in the thin liquid protoplast culture medium with a density of 1 x l0s to 5 x los protoplasts/mL, the division and the callus formation of the regenerated cells were easily observed. The first divisions occurred in 3 days and small cell clusters could be seen after 2 to 3 weeks in the culture. At this moment, the addition of the protoplast culture medium with decreased osmoticum once or twice is needed for the continuous protoplasts division to form calli. Regenerated calli, 3 to 5 mm in diameter, were transferred in succession on MS medium with 2 mg/L IAA and 1 mg/L KT for the initiation of embryogenesis. The embryoids germinated on the hormonefree MS medium and a number of plantlets were obtained. It seems that using vigorous embryogenic callus and decreasing osmoticum are the two critical factors for plant regeneration of cotton protoplasts.     

Mechanical stimulation-induced chilling tolerance in tobacco suspension cultured cells and its relation to proline

Mechanical stimulation (MS), widely existing but usually ignored in nature, is one of the major environmental stress factors. MS by increasing the rotational speed of shaker incubator could alleviate a decrease in vitality of tobacco (Nicotiana tabacum L.) suspension cultured cells and reduce the accumulation of MDA under chilling stress at 1°C, which in turn improved survival percentage under chilling stress and regrowth ability of tobacco suspension cells after chilling stress. In addition, MS could increase the activity of Δ¹-pyrroline-5-carboxylate synthetase (P5CS) and induce the accumulation of endogenous proline in tobacco cells; exogenously applied proline also could enhance its endogenous level under normal culture conditions and survival percent-age of the cells under chilling stress. These results suggest that MS could improve chilling tolerance of tobacco suspension cells and the acquisition of this chilling tolerance was related to proline.     

提取工艺和植物生长调节剂对罗布麻愈伤组织中黄酮含量的影响

以罗布麻愈伤组织粉末为材,在单因素实验的基础上,利用响应曲面法对罗布麻愈伤组织中黄酮的提取工艺进行优化。响应曲面分析结果表明,提取试剂和提取温度对提取的黄酮含量存在显著影响。通过响应曲面分析得到罗布麻愈伤组织中黄酮提取的最佳条件为:提取试剂为70%甲醇,物料比1∶40,提取时间为4 h,提取温度为70℃。培养并比较了30种不同植物生长调节剂浓度与配比诱导100 d生长的愈伤组织中黄酮的含量,结果得出MB+KT(1.0 mg/L)+NAA(0.2 mg/L)上培养约100 d的愈伤组织中黄酮含量最高,为73.90mg/g。测定愈伤组织培养30 d内黄酮的积累动态,探明从培养的愈伤组织提取黄酮的最佳时段。通过优化提取工艺和筛选最佳植物生长调节剂浓度与配比,运用组织培养技术提高了罗布麻愈伤组织中黄酮含量。     

野牛草幼穗愈伤组织的诱导及植株再生

以野牛草[Buchloe dactyloides(Nutt．)Engelm．]幼穗为外植体,建立了愈伤组织诱导、继代培养和植株再生体系。结果表明,雌穗比雄穗难以脱分化形成愈伤组织;小于8mm雄幼穗在2mg／L2,4-D培养基上的愈伤组织诱导率为80．0％～86．8％;添加10mg／L AgNO3对愈伤组织诱导率影响不明显,但可改善愈伤组织质量。2mg／L 2,4-D结合0．1mg／L 6-BA的培养基有利于愈伤组织的继代培养;继代超过3次、继代间隔超过3周,愈伤组织分化能力明显下降。雄穗愈伤组织在含1．0mg／L 6-BA培养基上,弱光条件下分化出芽的频率较高,达31．8％～35．0％;附加3％麦芽糖既可减轻褐化程度,又利于丛生芽的分化。分化苗在1/2MS 0．3mg／L IBA培养基上的生根率为62．5％。     

紫果猕猴桃幼胚愈伤组织诱导及植株再生

以紫果猕猴桃(Actinidia arguta var．purpurea)幼胚为外植体,诱导愈伤组织并进行植株再生。结果表明：不同的培养基和不同的培养条件对幼胚愈伤组织的诱导率及分化率不同;0．2mg／L ZT与0．5mg／L GA。配合使用有利于促进愈伤组织的诱导;7％蔗糖、600mg／L CH与400mg／L Gln都有利于促进愈伤组织的形成;在添加0．5mg／L 6-BA、0．05mg／L NAA与0．5mg／L GA3的MS培养基中植株的再生率达93．3％。     

Participation of H2O2 in Enhancement of Cold Chilling by Salicylic Acid in Banana Seedlings

The possible physiological mechanism of enhancement of cold tolerance by salicylic acid (SA) in banana seedlings (Musa acuminata cv. Williams 8188) was explored. Measurements of leakage electrolyte after 2 d of recovery at 30/22 ℃ (day/night) following 3 d of cold stress at 7 ℃ showed that pretreatment with hydroponic solution containing SA 0.3－0.9 mmol/L as foliar spray under normal growth conditions (30/22 ℃) could significantly enhance cold tolerance of banana plants. The highest enhancing effect of SA occurred at 0.5 mmol/L and it showed the lowest leakage rate of electrolyte or smaller leaf wilting area after 2 d of recovery at normal temperature from 3 d of 7 ℃ or 5 ℃ cold stress. Higher concentrations (≥2.5 mmol/L) of SA, however, caused more electrolyte leakage, indicating that they aggravated chilling damage. Enhanced cold tolerance by SA could be related to H2O2 metabolism. Compared with water-treated seedlings (control), SA 0.5 mmol/L treatment inhibited activities of catalase (CAT) and ascorbate peroxidase (APX), increased peroxidase (POX) activity, but did not affect the activity of superoxide dismutase (SOD) under normal growth conditions, and these changes might lead to an accumulation of H2O2, whereas SA pretreatment enhanced the activities of CAT and APX, and reduced the increase in productions of H2O2 and thiobarbituric acid-reaction substances (TBARS) during subsequent 7 ℃ cold stress and recovery periods. Exogenous H2O2 treatments (1.5－2.5 mmol/L) also increased cold tolerance of banana seedlings. Furthermore, pretreatment of banana seedlings with dimethylthiourea (a trap for H2O2) significantly inhibited cold tolerance induced by SA. These results suggested that endogenous H2O2 may be required for SA-enhanced cold tolerance. The significance of the interaction of SA, H2O2 and H2O2-metabolizing enzymes during cold stress has been discussed.     

茄子幼苗耐低温性生理机制研究

选择耐低温性不同的6份茄子高代自交系幼苗为材料,进行白天10℃/夜间7℃低温处理10d,调查其耐低温性、生长指标(株高、茎粗、鲜重、干重)、生理指标(POD、SOD、APX活性及脯氨酸含量),以探讨低温对茄子幼苗生长过程中的生长和生理指标的影响.结果显示:(1)各材料的上述性状在常温下均差异不显著,而在低温处理下差异极显著;与对照相比较,幼苗株高、茎粗、鲜重和干重下降,而POD、SOD、APX活性及脯氨酸含量上升.(2)各材料指标的升降幅度与其低温耐受性密切相关,耐低温性强的材料生长指标下降幅度较小,而生理指标提高幅度较大,耐低温性弱的材料则表现相反.(3)各指标间相比较,茄子耐低温性与株高、茎粗、干重、POD活性、SOD活性呈显著正相关,与其余性状的相关未达显著水平.研究表明,低温使茄子幼苗的生长受到一定程度的抑制,但耐低温材料能够通过调节自身的保护酶系统活性来减轻低温的伤害,维持植物体的正常生长及生理代谢功能.