尾叶桉的不同类型愈伤组织芽分化与某些相关生理生化指标的关系

切取8d苗龄的尾叶桉幼苗下胚轴,培养7d后愈伤组织的诱导率达100%。8周后,愈伤组织分化成褐色、白色和浅绿色3种类型。其中只有浅绿色愈伤组织能分化出不定芽,分化率达57%。浅绿色愈伤组织的过氧化物酶（POD）、超氧化物歧化酶（SOD）、过氧化氢酶（CAT）和多酚氧化酶（PPO）的活性最高。除SOD外,浅绿色愈伤组织的POD、CAT和PPO活性与其他两类愈伤组织相比具有显著性差异（P〈0.05）。H2O2和丙二醛含量则是褐色愈伤组织的最高。     

银杏愈伤组织生长、褐化与黄酮积累研究

旨在建立稳定的银杏愈伤组织继代体系,筛选细胞活性强且黄酮产量高的细胞培养条件,为工业化生产提供一定参考。以MS培养基为基本培养基,在40 d的继代周期内设计正交实验研究外植体月份、外源激素与抗褐化剂交互作用下的银杏愈伤组织生长、褐化与黄酮积累,并对比了4、5、6月中旬银杏叶片黄酮含量。结果表明,银杏愈伤组织继代中的最佳生长组合为:4月叶片诱导的愈伤组织+3.0 mg/L NAA+0.5 mg/L KT+5 mg/L VC;最佳抗褐化组合为:4月叶片诱导的愈伤组织+1.0 mg/L NAA+1.0mg/L KT+10 mg/L VC;银杏叶黄酮含量从4-6月呈上升趋势,且不同月份叶片诱导的愈伤组织黄酮含量与之呈正相关,以6月叶片诱导的愈伤组织+2.0 mg/L NAA+1.0 mg/L KT+5 mg/L VC为最佳黄酮积累条件;结合愈伤组织干重,总黄酮实际产量最高组合为:5月叶片诱导的愈伤组织+2.0 mg/L NAA+0.5 mg/L KT+5 mg/L VC。银杏愈伤组织的适宜继代周期为24-32 d,生长量较大且褐化率低。     

高压静电场（HVEF）对苜蓿叶片愈伤组织诱导的影响

用一定强度的负高压静电场处理苜蓿叶片外值体,可显著促进外植体愈伤组织的发生和形成。处理组叶片愈伤化进程比对照组提前二天,愈伤组织诱导率平均提高了３６％,愈伤组织鲜重增长率提高了３３％,而且形成的愈伤组织质量较高。生理生化分析显示：静电场处理使愈伤组织诱导过程中培养物的可溶性蛋白质含量较对照高,且峰值来临较早;过氧化物酶（ＰＯＤ）,淀粉酶（Ａｍｙｌａｓｅ简称ＡＭＬ）,超氧物歧化酶（ＳＯＤ）活性和峰值也较对照组有不同程度的提高和提前,与观察结果一致。本文对其作用机理做了初步探讨。     

月季愈伤组织的诱导及植株再生

以可在黑龙江地区露地越冬的5个现代月季（Rosa chinensis）品种为实验材料,分别以其无菌苗的叶片和茎段为外植体,研究了愈伤组织诱导及植株再生方法。实验结果表明：5个寒地月季品种的叶片和茎段均可诱导出愈伤组织,2,4-D诱导愈伤组织的效果较好,高浓度的细胞分裂素不适合用于月季叶片和茎段愈伤组织的诱导;TDZ在月季愈伤组织分化培养过程中具有重要作用,光照培养可促进月季愈伤组织的分化,愈伤组织的分化能力随着继代次数的增加呈下降趋势。该实验成功地从2004-8和2004-9（2个月季品种）愈伤组织中诱导出再生植株,其愈伤组织的分化率分别为45%和38%。     

不同树龄白桦的不同器官及其组培苗诱导的愈伤组织中白桦酯醇和齐墩果酸的分布和含量变化

检测一年生、四年生、六年生、八年生及四十年生白桦树各器官以及白桦组培苗诱导的愈伤组织和悬浮培养细胞中齐墩果酸和白桦酯醇的分布和含量。结果表明,不同树龄白桦的叶、茎外皮和根皮中都能检测出齐墩果酸和白桦酯醇,但主要集中在茎外皮,叶片和根皮中含量极低,齐墩果酸和白桦酯醇含量分别以四年生和八年生白桦茎外皮中最高。由白桦组培苗茎段诱导的愈伤组织及其悬浮培养细胞和组培苗根诱导的愈伤组织中都能测出白桦酯醇,其含量以茎段诱导的悬浮培养细胞中为最高。组培苗、茎段诱导的愈伤组织及其悬浮培养细胞、叶片和根诱导愈伤组织中均可以检测出齐墩果酸,其含量以悬浮培养细胞中的为最高。     

枇杷叶片胚性愈伤组织诱导与内源激素含量的关系

为揭示胚性愈伤组织发生过程中内源激素的变化规律,本研究以枇杷叶片为实验材料,通过诱导胚性愈伤组织获得体细胞胚,采用高效液相色谱法测定枇杷叶片及愈伤组织的赤霉素（GA₃）、生长素（IAA）、脱落酸（ABA）和玉米素（ZT）4种内源激素的含量,探讨胚性愈伤组织发育过程中4种内源激素的动态变化。结果显示,不同成熟度叶片内IAA/ZT比值对胚性愈伤组织的发生有正效应,而GA₃/IAA的比值具有负效应。胚性愈伤组织的发生需要较低含量的GA₃及高含量的IAA和ABA,IAA/ZT比值高有利于胚性愈伤组织形成,培养后期及时添加一定量的外源激素有利于胚性保持。本研究可为离体培养时选择外植体、添加外源激素及控制继代时间提供理论指导,并为快速获得枇杷胚性材料、开展基因转化研究奠定基础。     

暴马丁香成熟胚愈伤组织和体胚诱导及其生理状态解析

以暴马丁香成熟胚为外植体进行愈伤组织和体胚发生诱导,通过调节诱导培养基中植物生长调节剂的种类和浓度,分析其对愈伤组织诱导和体胚诱导的影响,同时对培养过程中的外植体进行形态发生观察和生理状态分析。结果表明:①暴马丁香成熟胚外植体培养30 d可见直接体胚发生、60 d可见子叶型体胚;②BA在愈伤组织诱导过程中起到了主导作用,在0.5 mg·mL^-1BA和5~6 mg·mL^-1NAA组合下,愈伤组织诱导率可达100%;在0.5 mg·mL^-1BA和5 mg·mL^-1NAA组合体胚诱导率可达8%;③多酚含量在愈伤组织形成初期急剧上升且在培养过程中保持较高水平,子叶型胚期PAL和POD活性升高、MDA和SOD活性略下降。     

蝴蝶兰胚性愈伤组织诱导的氧化还原动态

离体培养蝴蝶兰（Phalaenopsis spp.）类原球茎切块诱导胚性愈伤组织,在培养第11～18 d时出现胚性细胞,第25～32 d观察到幼嫩的球形胚。在培养第0～11 d,外植体.OH相对含量波动不大,O2.生成速率持续提高并达到高峰,H2O2水平明显提高,抗氧化能力在低位波动,推测外植体可能经受较强的氧化胁迫,以利细胞脱分化和获得胚性潜能。此外,此期间SOD活性持续提高,POD和CAT活性达到一个高峰。在愈伤组织诱导全过程中,总抗氧化能力、DPPH自由基清除率、H2O2水平基本上都持续提高,与SOD活性均极显著正相关,SOD是影响培养物清除自由基能力和提高抗氧化水平的主要因素。综合分析实验结果表明,在蝴蝶兰胚性愈伤组织诱导过程中,SOD、O2.和H2O2起到关键的作用。     

黄花蒿组织培养研究

目的：筛选黄花蒿组织培养的配方,以寻求黄花蒿生长的最佳自然条件。方法：在MS基本培养基中添加不同激素,制备不同培养配方,筛选黄花蒿愈伤组织诱导与分化的最适培养基配方。结果与结论：诱导黄花蒿愈伤组织形成的最佳培养基配方为MS＋6-苄氨基嘌呤（6-BA）（2．0mg／L）＋激动素（KT）（2．0mg／L）;诱导黄花蒿愈伤组织分化的最佳培养基配方为MS＋6-BA（1．0mg／L）＋吲哚乙酸（IAA）（1．0mg／L）。叶片诱导愈伤组织出愈时间最慢,出愈率较低,但其愈伤组织为胚性愈伤组织,后期愈伤组织的分化率、出苗率都很高,且不易产生褐变;带腋芽的茎段诱导愈伤组织形成最快,但分化率不及叶片愈伤组织,易褐变,适于快速转入生根培养基中直接用于快繁;花序基本不形成肉眼可见愈伤组织,直接形成幼苗。     

连香树胚性与非胚性愈伤组织生理生化差异及同工酶分析

本研究以连香树未成熟种子子叶胚诱导产生的胚性愈伤组织与非胚性愈伤组织为原材料,测定了两者的可溶性蛋白质、可溶性糖及游离脯氨酸的含量,并利用非变性聚丙烯酰胺凝胶电泳技术(polyacrylamide gelelectrophoresis, PAGE)对两者的过氧化物酶(peroxidase, POD)、超氧化物歧化酶(superoxide dismutase,SOD)、淀粉酶(amylase, AMY)同工酶进行了分析。比较发现胚性愈伤组织在可溶性蛋白质和游离脯氨酸的含量上均极显著高于非胚性愈伤组织。在可溶性糖含量上,胚性愈伤组织也显著地高于非胚性愈伤组织。胚性愈伤组织具有较高的代谢活性,为进一步胚性分化提供物质和能量基础。POD、SOD和AMY同工酶在连香树胚性愈伤组织与非胚性愈伤组织中酶带差异明显,表明这3种酶的活性与胚性愈伤组织的诱导密切相关,酶谱的差异可作为体细胞胚胎发生的参考标记。     

HZ-841吸附树脂精制银杏叶总黄酮

本文研究了用HZ-841吸附树脂精制银杏叶总黄酮的工艺。用10 BV 70％的乙醇分三次提取脱脂银杏叶粉中的银杏叶总黄酮,其收得率为4．8％,纯度为21．7％;用30BV纯净水、微波解冻提取30min,银杏叶总黄酮的收得率及纯度分别是2．63％和13．4％。HZ-841树脂对银杏叶总黄酮的动态吸附容量在pH=7．0时为0．265g／mL,树脂,动态吸附平衡时间为10min。酸度对HZ-841树脂吸附银杏叶总黄酮有显著影响,当pH=5．0时,其静态吸附量可达到0．322g／mL。吸附了银杏叶总黄酮的HZ-841树脂可用乙醇洗脱,当洗脱液pH=9．0、乙醇浓度为90％、洗脱流速3BV／h时,5BV洗脱液的收得率为1．8％。用无水乙醇洗脱的银杏叶总黄酮经过真空浓缩、干燥,获得的浅黄色粉末中银杏叶总黄酮含量为37．3％,产品收得率为2．41％。     

珍稀濒危植物珙桐超氧化物歧化酶活性

采用比色法测定了不同年龄和部位珙桐叶的超氧化物歧化酶(SOD)活性,以探讨不同年龄级珙桐的生理生态适应性。结果表明:不同年龄同一生长期珙桐叶的SOD活性不同,同年龄级同层珙桐叶的SOD活性随叶位增加大致呈先升后降的变化;同叶位不同层及同层不同叶位的珙桐叶的SOD活性不同。珙桐叶SOD活性对生长期敏感,其活性随叶片的生长、衰老呈先升后降变化。即珙桐叶SOD活性受生物和非生物因子共同影响,且不同年龄级、叶位、层次均存在差异。     

Biological activities of Ginkgo extracts

The biological activity of methanolic the extracts of leaves, roots, leaf-derived callus, root-derived callus, ginkolide A, ginkgolide B, bilobalide and a commercial Ginkgo product (Tanakan) was assessed. Bioassays consisted of the Agrobacterium tumefaciens-induced potato tumor assay and a Kirby-Bauer microbial sensitivity assay with pure strains of Escherichia coli, Klebsiella pneumoniae, Pseudomonas aeruginosa, Staphylococcus aureus, Staphylococcus epidermidis and Streptococcus pyogenes. Methanolic extracts of leaves, leaf-derived callus, root-derived callus, bilobalide and Tanakan inhibited tumor formation significantly, but more weakly than the positive control, camptothecin. No activity against E. coli was detected, but extracts from both callus types inhibited the growth of K. pneumonia, P. aeruginosa, S. aureus, S. epidermidis and S. pyogenes. All extracts and reference compounds inhibited the growth of S. pyogenes. Leaf and root tissues contained the highest levels of ginkgolide A, as compared to the callus tissues; leaf tissue contained more of all three marker compounds than the callus tissues.     

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     

小麦原生质体分离过程中生理状态的变化

酶解处理使小麦对肉原生质体膜流动性降低,膜脂过氧化产物丙二醛（ＭＤＡ）积累,说明脱璧过程对细胞有伤害作用,损伤位点可能发生在膜上。胚性愈伤组织的具有分裂能力的原生质体,不表现上述变化。酶解脱壁还使超氧化物歧化酶（ＳＯＤ）和过氧化氢酶（ＣＡＴ）活性上升;过氧化物酶（ＰＯＸ）在叶肉原生质体中活性下降,在胚性愈伤组织来源的原生质体中活性上升。以上结果表明：在原生质体分离过程中,细胞的生理特性发生了变化;膜损伤的发生可能与原生质体能否进入正常分裂状态有关。     

杂交稻及其三系叶片衰老过程中SOD、CAT活性和MDA含量的变化

对杂交水稻及其三系主茎第11叶叶片自然衰老过程中超氧物歧化酶(SOD)、过氧化氢酶(CAT)和丙二醛(MDA)含量的变化进行了研究,结果表明:叶片衰老过程中,SOD和CAT活性下降,MDA的含量增加,可作为衰老特征的叶绿素和可溶性蛋白质含量明显下降;SOD的活性和MDA的含量变化相对应;CAT活性大幅度下降与SOD之间的不平衡,致使O_2~-代谢中间产物累积而引起膜的损伤。不育系的衰老进程比杂交水稻、恢复系和保持系慢,其SOD和CAT活性明显高于其它三者,可能是不育系不易早衰的原因之一。     

The Effect of Light and Hormones on Leaf Senescence

With wheat leaves as material, the changes of superoxide dismutase (SOD), lipid peroxi-dation and membrane permeability during leaf senescence in light or dark, and treated withphytohormones (KT or ABA) have been studied. The changes of chlorophyll content, lipidperoxidation and fine structure of spinach chloroplasts senescing in light or dark have alsobeen studied. When leaves senesce in light, the activity of SOD increased at first then decreased. The increase of SOD activity was able to result from the synthesis of new protein. Lightwas found to delay the leaf senescence obviously but also accelerate leaf senescence by causinglipid peroxidation when prolonged the illumination time. The delay or acceleration of leafsenescence by exogenous hormones were observed, it may be due to the control of lipid peroxi-dation by adjusting the activity of SOD. O2-participated the chlorophyll decomposition andlipid peroxidation during chloroplasts senesce in light. A favourable role of light in mainta-lng the fine structure of isolated chloroplasts was clear.     

Evidence for the expression of morphological and biochemical characteristics of C3-photosynthesis in chlorohyllous callus cultures of Zea mays

A Zea mays callus culture containing chlorophyll was established and grown photomixotrophically. Cell chloroplast structure, and pigment and soluble protein contents were examined. Expression of some key enzymes of C₄ carbon metabolism was compared with that of etiolated (heterotrophic) and green photoautotrophic leaves. Chlorophyll content of the callus was 15–20% that of green leaves. Soluble protein content of callus was half that of leaf cells. Electron microscopic observations showed that green callus cells contained only typical granal chloroplasts. Ribulose-1,5-bisphosphate carboxylase/oxygenase (Rubisco, EC 4.1.1.38) activities in green callus were ca 30% those of green leaves but 2–3 times higher than in etiolated leaves. Quantitative enzyme protein determination, using antibodies specific to maize leaf Rubisco showed that the chloroplastic carboxylase represented about 7% of total soluble protein in green callus, in parallel to its low chlorophyll content. The specific activity of Rubisco in callus and leaves was unchanged. Phosphoenolpyruvate carboxylase (PEPC, EC 4.1.1.31) activity in green callus was about 20% that of green leaves and similar to that measured in etiolated leaves. Apparent K_m (PEP) values (0.08 mM) for PEPC isolated from green callus and etiolated leaves were very different from values (0.5 mM) obtained with PEPC from green leaves. These kinetic characteristics together with the absence of inhibition by malate and activation by glucose-6-phosphate suggest that the properties of PEPC isolated from green callus and etiolated maize leaves are very similar to those of PEPPC from C₃ plants. Using PEPC antibodies specific to green maize leaf enzyme, immunotitration of PEPC preparations containing identical enzyme units allowed complete precipitation of the green leaf enzyme with increasing antibody volumes. In contrast, 60–70% of the activity of PEPC from etiolated and green callus was inhibited, suggesting low affinity for the maize green leaf PEPC antiserum (typical C₄ form). Ouchterlony double diffusion tests revealed only partial recognition of PEPC in green callus and etiolated leaves. NAD-malate dehydrogenase (NAD-MDH, EC 1.1.1.37) activity in callus was 2 and 3 times higher, respectively, than in etiolated and green leaves. NADP-malic enzyme (NADP-ME, EC 1.1.1.40) activity in callus cultures was much lower than in green leaves. All our data support the hypothesis that cultures of fully dedifferentiated chlorophyllous tissues of Zea mays possess a C₃-like metabolism.     

西花蓟马取食与机械损伤对菜豆叶片抗氧化系统的影响

本文研究了西花蓟马Frankliniella occidentalis(Pergande)取食和机械损伤诱导对菜豆抗氧化系统的影响,比较了不同损伤形式诱导的抗氧化酶活力和抗氧化物质含量的变化差异。结果表明,西花蓟马取食和机械损伤均可诱导菜豆叶片内过氧化物酶(POD)、超氧化物歧化酶(SOD)和过氧化氢酶(CAT)3种抗氧化酶活性有不同程度的升高,但两种处理诱导的抗氧化酶活性变化规律不完全相同,各种酶活性达到最高点时间不同,西花蓟马取食对抗氧化酶活力的诱导作用大于机械损伤的。两种处理诱导下的类胡萝卜素的含量变化不大,但不论西花蓟马取食还是机械损伤均导致菜豆叶片内类黄酮和总酚含量整体呈下降趋势,西花蓟马的取食诱导的下降幅度大于机械损伤的。因此,西花蓟马取食诱导明显高于机械损伤对菜豆抗氧化系统的影响。     

Subcellular distribution of superoxide dismutase in leaves of ureide-producing leguminous plants

The subcellular localization of superoxide dismutase (SOD; EC. 1.15.1.1) was studied in leaves of two ureide-producing leguminous plants ( Phaseolus vulgaris L. cv. Contender and Vigna unguiculata [L.] Walp). In leaves of Vigna and Phaseolus , three superoxide dismutases were found, an Mn-SOD and two Cu, Zn-containing SODs (I and II). Chloroplasts, mitochondria, and peroxisomes were purified by differential and density-gradient centrifugation using either Percoll or sucrose gradients. The yields obtained in intact chloroplasts and peroxisomes from Vigna were considerably higher than those achieved for Phaseolus . Purified chloroplasts only contained the Cu, Zn-SOD II isozyme, but in mitochondria both Mn-SOD and Cu, Zn-SOD I isozymes were present. In purified peroxisomes no SOD activity was detected. The absence of SOD activity in leaf peroxisomes from Vigna contrasts with results reported for the amide-metabolizing legume Pisum sativum L. where the occurrence of Mn-SOD was demonstrated in leaf peroxisomes (del Río et al. 1983. Planta 158: 216–224; Sandalio et al. 1987. Plant Sci. 51: 1–8). This suggests that in leaf peroxisomes from Vigna plants the generation of O₂^- radicals under normal conditions probably does not take place.