不同分化状态的白百利烟草愈伤组织在生长过程中核酸蛋白质的变化

本文报道了在正常分化芽和根、诱导芽或恨定向发生的白百利烟草(Nico-tiana tabacum Baibaili)愈伤组织在生长过程中DNA、RNA和蛋白质变化的结果。MS+0.2mg/1NAA+0.2mg/1 KT诱导白百利烟草愈伤组织正常分化出芽和根,MS+0.05mg/1NAA+2mg/1KT诱导愈防组织定向地芽发生,MS+0.5mg/1NAA+0.05mg/1KT诱导愈伤组织定向地根发生。在定向诱导芽或根发生愈伤组织里的RNA和蛋白质合成的第一个高峰出现,比正常发生芽和根的愈伤组织里DNA、RNA和蛋白质的第一个高峰迟5天,在芽发生的愈伤组织里DNA峰出现也迟5天,在根发生的愈伤组织里DNA蜂,则相同于正常分化的愈伤组织DNA峰出现。外源的植物生长物质诱导器官定向发生的作用表现在RNA水平上。在三种分化状态的愈伤组织里,蛋白质组成在第8天表现出明显的差异。41KD和46KD蛋白质在器官的定向发生中可能起着相当重要的作用。     

尾巨桉不同类型愈伤组织抗性相关酶活性差异与不定芽分化关系研究

本实验以尾巨桉无性系32-29无菌苗的茎段为外植体,应用不同植物生物调节剂处理,诱导愈伤组织形成,分别测量4种类型愈伤组织的蛋白质含量及过氧化物酶(POD)、多酚氧化酶(PPO)、超氧化物歧化酶(SOD)等保护酶的指标活性,比较不同愈伤组织的酶活性强弱与不定芽分化的关系。结果表明:不同处理间愈伤组织诱导率差异达到极显著水平,且不同愈伤组织蛋白质含量及3种酶活力差异明显,POD、SOD活力高的愈伤组织不定芽诱导率也高,而PPO含量高的愈伤组织诱芽率低。     

尾叶桉茎段再生系统的建立

本文以尾叶桉(Eucalypt urophylla)U6的无菌苗茎段为材料,选用MS培养基为基本培养基,研究不同IAA浓度对尾叶桉茎段愈伤组织诱导,不同IAA、6-BA浓度对尾叶桉茎段愈伤组织诱导芽、生根的影响。结果表明,当外植体的接种数一样,IAA的浓度为20.0mg/L时,愈伤组织诱导率达到最高,为95%,且此浓度下,出芽率、生根率最高,分别达43.3%和100%,而添加0.5mg/L6-BA则会抑制芽的再生、不能产生根。因此最适合诱导尾叶桉愈伤组织、再生芽和再生根的培养体系是MS+20.0mg/L IAA+30g/L蔗糖+8g/L琼脂。该再生系统的建立将为转基因技术改良桉树性状提供前提条件。     

大叶补血草LgNHX1基因对烟草的遗传转化及耐盐性分析

目的:通过农杆菌介导法将大叶补血草液泡膜Na+/H-逆向转运蛋白基因LgNHX1转化烟草,并对转化烟草的耐盐性进行分析.方法:采用叶盘法进行遗传转化,对得到的转化植株经PCR和RT-PCR鉴定后,用不同浓度的NaCl溶液胁迫,分别测定了它们的丙二醛含量、相对电导率及根和叶中的K+和Na+含量.结果:在200～300mmol/L NaCl胁迫下,转基因植株叶片的丙二醛含量分别为2.85nmol/g FW、3.50nmol/g FW,对照为3.25nmol/g FW、4.39nmol/g FW;转基因植株叶片的相对电导率分别为42.22%、74.10%,对照为53.2%、84.33%;在300mmol/L NaCl胁迫下,转基因植株根和叶中的K+/Na+比分别为5.80和3.10,对照为4.32和2.35.结论:在NaCl胁迫下,转基因烟草的耐盐性有了一定程度的提高.     

禾谷镰刀菌粗毒素对不同抗性小麦品种成熟胚脱分化与再分化的影响

以5个赤霉病抗性水平不同的小麦品种为材料,研究了禾谷镰刀菌粗毒素对小麦成熟胚脱分化与再分化的影响.结果表明,小麦成熟胚愈伤组织的诱导率和分化率在品种间、毒素浓度间、毒素浓度×品种间差异极显著.低浓度（5 g/L）禾谷镰刀菌粗毒素能够促进抗性较强品种的出愈速度,高浓度禾谷镰刀菌粗毒素（10 g/L和15 g/L）对成熟胚愈伤组织的出愈速度起到抑制作用,随着浓度提高,抑制作用增强.成熟胚愈伤组织的诱导率和分化率随着禾谷镰刀菌粗毒素浓度升高而下降,品种的抗性越差下降幅度越大.10 g/L毒素浓度,5个品种成熟胚愈伤组织的SOD活性均比对照（0 g/L）增加;20 g/L毒素浓度,5个品种成熟胚愈伤组织的SOD活性均有所下降,抗性品种苏麦3号和郑9023的SOD活性仍高于对照,其它3个感性品种低于对照,SOD活性与小麦品种的赤霉病抗性水平存在一定关系.研究结果为小麦赤霉病抗性育种提供了一定的技术支撑.     

电晕电场对沙生植物愈伤组织诱导及分化的影响

用不同电场剂量处理花棒和杨柴的下胚轴后进行愈伤组织诱导及分化培养,以探讨电晕电场对其愈伤组织诱导与分化的影响.结果表明,电场处理可以显著提高愈伤组织诱导率和分化率,并有效降低愈伤组织的褐化率.花棒在电场强度为5 kV/m下处理60 min时,出愈率可达90.5%,分化率可达88.7%;杨柴在电场强度为3 kV/m下处理100 min时,出愈率可达95.8%,分化率可达78.9%.     

文心兰原球茎形态发生与可溶性物质及抗氧化酶的关系

以文心兰切花品种'南茜'无菌苗为材料,取其茎尖通过组织培养诱导形成原球茎和幼苗,观察并分析了原球茎各形态发生阶段的特征及其可溶性糖和蛋白质含量、抗氧化酶(POD、CAT和SOD)活性以及相关同功酶(POD、EST和SOD)的变化.结果显示:(1)文心兰原球茎形态发生可分为外植体期、外植体膨大期、愈伤组织期、原球茎形成期、原球茎成熟期、叶鞘伸展期、顶端腋芽发育期及幼苗期8个阶段.(2)可溶性糖和蛋白质含量均在叶鞘伸展期出现最大峰值;POD活性在外植体膨大期、CAT和SOD活性在愈伤组织期分别出现最大峰值.SOD同工酶的2条酶带在愈伤组织期到幼苗期交替出现;EST同工酶在原球茎形成期有2条特异酶带.研究表明,可溶性糖和蛋白质的含量以及POD、CAT、SOD活性的特异变化与文心兰茎尖脱分化及原球茎再分化的实现密切相关,不同类型的同工酶在原球茎同一发生阶段表现出较大差异,EST同工酶的2条特异酶带可作为原球茎形成的标志.     

低温与植物生长调节剂预处理对小麦成熟胚培养特性的影响

为提高小麦成熟胚愈伤组织的分化率,以3个黄淮麦区冬小麦品种(系)‘小偃22’、‘西农1013’和‘千斤早’为材料,研究了4℃低温和3种植物生长调节剂(TDZ、2,4-D、多效唑)预处理小麦种子对小麦成熟胚愈伤组织分化特性的影响。结果表明:4℃低温和植物生长调节剂预处理对小麦成熟胚出愈率无显著影响,但能够显著促进愈伤组织的分化,并表现出处理及基因型间差异。‘小偃22’、‘西农1013’、‘千斤早’感应低温预处理的最好时间节点分别为6h、12h、12h,其成熟胚愈伤组织的分化率比对照分别显著提高16.2%、14.2%、12.8%;3个材料分别在10mg/L、5 mg/L、20 mg/L TDZ预处理下成熟胚愈伤组织分化率最高,比对照分别显著提高39.1%、29.7%、16.7%;3个材料的最佳2,4-D预处理浓度均为10mg/L,其愈伤组织分化率分别比对照显著提高17.3%、11.2%、25.2%;3个材料的成熟胚愈伤组织分化率分别在10mg/L、20mg/L、20mg/L多效唑预处理时最高,分别比对照显著提高6.2%、11.6%、7.2%。研究表明,低温和植物生长调节剂预处理均可有效提高小麦成熟胚愈伤组织的分化率,但促进植物生长的调节剂(TDZ和2,4-D)对小麦成熟胚愈伤组织再分化的效应远大于抑制植物生长的调节剂(多效唑)。     

棉花大田植株叶柄组织培养体系的建立

以早熟棉花品种'中棉所24'为材料,通过对大田棉花植株叶柄灭菌条件、叶柄愈伤组织诱导与分化、胚状体的萌发及植株再生所需的培养基等进行研究,以建立棉花大田植株叶柄组织培养体系.结果表明:棉花植株叶柄经0.1%HgCl2灭菌4～5 min后,横切成0.8 cm左右的小段,在MSB+0.05 mg·L-1IAA+0.05 mg·L-1KT+0.05 mg·L-12,4-D+25 g·L-1葡萄糖+2 g·L-1Gel(pH 5.8)培养基上培养,易获得分化的愈伤组织;愈伤组织在添加有KT/IAA(MSB+0.08 mg·L-1IAA+0.08 mg·L-1KT+25 g·L-1葡萄糖+2 g·L-1Gel,pH6.5)或ZT/IBA(MSB+0.1 mg·L-1ZT+0.1 mg·L-1IBA+25 g·L-1葡萄糖+2 g·L-1Gel,pH 6.5)的分化培养基中,均可以获得胚性愈伤组织.胚性愈伤组织在MSB+0.05 mg·L-16-BA+0.05 mg·L-1IAA+25 g·L-1蔗糖+2 g·L-1Gel(pH 6.5)培养基上,形成胚状体、生长发育成再生植株.     

云南重楼的组织培养与植株再生

1植物名称云南重楼[Paris polyphylla Smith vaY.yunnanensis(Franch.)Hand.-Mazz.]. 2材料类别芽. 3培养条件愈伤组织诱导培养基:(1)MS 6-BA 2 mg·L-1(单位下同) NAA 0.1;愈伤组织增殖和分化培养基:(2)MS 6-BA 2 NAA 0.5 KT 0.5;生根培养基:(3)I/2MS NAA 0.5 IAA 0.5.上述培养基中均附加3%蔗糖和0.7%琼脂,pH 5.8,在121℃下高压灭菌20 min.培养温度控制在(20±2)℃,光照时间8 h·d-1,光照强度20～30μmol·m-2·s-1.     

静电场对苜蓿愈伤组织抗寒能力的影响

用正负高压静电场 (HVEF)处理苜蓿叶片来源的愈伤组织后 ,在4℃下暗培养。我们发现静电场处理可不同程度地提高愈伤组织的淀粉酶 ,过氧化物酶 ,和超氧化物岐化酶的活性和可溶性蛋白的含量 ,促进愈伤组织对冷害的防御能力。在冷处理过程中 ,细胞膜中饱和脂肪酸 (棕榈梭和硬脂酸 )和不饱和脂肪梭含量先升高然后降低 ,而高压静电场处理组的不饱和脂肪酸 (油酸、亚油酸、α -亚麻酸、β-亚麻酸 )含量高于对照组 ,脂肪酸不饱和指数 (UIFA)也高于对照组 ,这说明静电场处理可促进愈伤组织细胞抗寒能力提高。此外 ,丙二醛 (MDA)含量在静电场处理组低于对照组的测定结果也验证了前面的结果。     

Effect of heavy metals on the growth of tissue cultures (II)

The effect of toxic concentrations of three heavy metal compounds on the growth of the secondary callus tissue of Nicotiana tabacum L. and Ruta graveolens L. was studied. The metal compounds examined were ZnSO4, NiSO4, CuSO4. The metal compounds used were placed in Murashige, Skoog (1962) and White (1943) culture medium at 10(-6) and 10(-4) M concentration, respectively, before autoclaving. The culture media containing macro- and microelements and vitamins were completed with carbon source and regulators (IAA, GA, kinetin for Nicotiana and IAA, 2, 4-D for Ruta). The cultures were kept for 4 weeks at 25 (+2) degrees C under 16/8 n light/dark conditions. The value of pH was 5.6 before the autoclave treatment. The increase in fresh weight of the secondary callus tissue was inhibited by the metal compounds applied with both plant species (to 75-87% by zinc, 7-97% by nickel, 5-98% by copper with tobacco; to 47-69% by zinc, 5-88% by nickel, 57-90% by copper with rue). The cell number and dry weight per g of callus tissue partly increased, partly decreased compared to the control in response to the heavy metal treatment. The growth values obtained with various concentrations of the heavy metals were different in the two plant species due to differences in metabolism and organization potential between them.     

烟草花柄愈伤组织花芽分化的能力(简报)

来源于开花植株的外植体(如花柄、花序轴等)具有在离体培养条件下直接分化花芽的能力,这一现象已在数十种植物的组织培养中得到证实。但是,这种成花能力能否保留在由这些外植体形成的愈伤组织之中?已有报道在风信子、布罗瓦利亚花、石龙芮、大蒜、矮通泉草等值物的愈伤组织中得到无     

Effect of Host Plants and Inoculum Forms on the Inoculum Potential of Arbuscular Mycor rhizal Fungi

The effects of inoculum forms (single-spore, multi-spores, or colonized root pieces) and host plants (Nicotiana tabacum L., Sorghum sudanense(Piper) Stapf, and Trifolium repens L. ) on the development and inoculum potential (IP) of the arbuscular mycorrhizal fungi (AMF): Glomus macrocarpum Tul & Tul, Glomus mosseae (Nicol & Gerd. ) Gerdemann & Trappe, Glomus versiforme (Karsten) Berch, and Sclerocystis sinuosa Gerdemann & Bakhi cultured in pots were investigated. The lag phase of treatment with 50 spores or 0.5 g (fresh weight) of colonized root pieces was 4 weeks, much shorter than that of the treatment with 1 spore (8 weeks); the value of IP(VIP) and percentage of root colonization(PRC) of the former were greater than those of the latter. Only on the early stages of colonization was there difference between the 50 spores and the 0.5 g (fresh weight) of colonized root piece inoculation treatments. The IP per plant inoculated with 0. 5 g (fresh weight) of colonized mot pieces of AMF was greater than that of the other two treatments except G. vers/forme on Nicotiana tabacum, while the PRC of the plants inoculated with 50 spores and 0. 5 g (fresh weight) of colonized root pieces of AMF was higher than that of the 1 spore inoculation after 10 weeks. The VIP of AMF on Trifolium repens was significantly higher than that on the other two hosts. The VIP of G. mosseae, G. versiforme, and S. sinuosa was respectively greater than that of G. macrocarpum. This suggested that different species of AMF produced different VIP of the inoculum. Nicotiaha tabacum was much better than the other host plants which used to be inoculated with single spore, and to produce inocula of AMF.     

De novo root formation in thin cell layers of tobacco: changes in free and bound polyamines

Thin cell layers excised from tobacco ( Nicotiana tabacum L. cv. Samsun) stem internodes, with an appropriate exogenous hormonal balance, were able to form a greater number of roots, and in a larger percentage of the explants (93%) than when they were excised from pedicels (40%). The developmental sequence of root formation and explant growth were followed by histological analysis. Free and bound [trichloroacetic acid (TCA)-soluble and -insoluble] putrescine and spermidine increased in the explants, particularly when root meristemoids appeared. These meristemoids originated in the superficial (day 6 in culture) or deep (days 10–11) layers and inside the newly formed callus (day 25). At those times, TCA-soluble and, to a lesser extent, TCA-insoluble bound putrescine predominated over the other polyamines. Spermine was always present in trace amounts. Polyamines decreased again when root and callus formation was completed (day 30). The involvement of these three classes of polyamines (free, TCA-soluble and -insoluble) in morphogenic processes is discussed.     

Ethylene production in habituated and auxin-requiring tobacco callus cultures. Does ethylene play a role in the habituation?

Ethylene production of habituated and auxin-requiring tobacco ( Nicotiana tabacum L. cv. Xanthi) callus cultures were compared. More ethylene was produced by auxinrequiring i.e. auxin-heterotrophic cultures than by habituated ones. Treatment with 2,4-dichlorophenoxyacetic acid increased the ethylene evolution of habituated cultures over the range 10⁻⁷ to 10⁻⁴ M , which suggests that the higher ethylene production of auxin-dependent callus is caused by the 2,4-D in the medium. The IAA levels depended on the age of both types of callus cultures.     

高压静电场对银杏愈伤组织生长的影响

银杏愈伤组织经１ ．０ｋＶ／ ｃｍ 强度的高压静电处理后, 愈伤组织生长速率提高, 超氧物歧化酶（ＳＯＤ） 活性和蛋白质及糖含量增加,过氧化物酶（ＰＯＤ） 和吲哚乙酸（ＩＡＡ） 氧化酶活性下降。实验表明,正高压静电场促进银杏愈伤组织细胞的生长,与ＨＶＥＦ 影响体内ＳＯＤ、ＰＯＤ 和ＩＡＡ 氧化酶活性以及蛋白质和糖含量的变化相关