杂景天愈伤组织诱导及红景天苷测定

以杂景天的子叶、胚轴为外植体,接种在附加不同激素组合的MS培养基上诱导愈伤组织产生,愈伤组织经继代培养后,用高效液相色谱检测红景天苷含量.结果显示,杂景天的子叶是诱导愈伤组织的理想外植体,子叶在MS 1 mg/L BA 0.5 mg/L 2,4-D和MS 1 mg/L BA 0.5 mg/L NAA培养基上的愈伤组织诱导率较高(81.8%、80%).愈伤组织有红、绿2种类型.HPLC检测显示红色愈伤组织不含红景天苷,绿色愈伤组织红景天苷含量为0.288 6%.表明利用组织培养生产红景天苷是可行的.     

西藏红景天组织培养研究

以西藏红景天嫩叶和幼茎为材料,研究西藏红景天不同外植体的离体培养技术。结果表明：西藏红景天嫩叶是诱导愈伤组织和芽的理想外植体,诱导松散型愈伤组织有两种培养基,分别是黑暗培养下MS 6-BA2．0 mg／L NAA0．2mg／L和MS 6-BA3．0mg／L NAA0．3mg／L;生芽培养基为MS 6-BA2．0mg／L IAA0．25mg／L;生根培养基为MS IAA0/5～1．0mg／L。     

植物生长物质对大花红景天愈伤组织诱导及苗再生的影响

以大花红景天植物的茎、叶、芽、种子为材料,采用DPS软件正交设计法,通过添加不同植物生长物质,考察其对大花红景天愈伤组织形成及苗再生的影响,为低海拔条件下建立大花红景天植株再生体系提供数据基础。结果表明：种子和芽是诱导愈伤组织最适宜的外植体;植物生长物质对愈伤组织形成的影响大小为6-BA〉KT〉2,4-D〉NAA〉IAA,愈伤组织形成的最佳培养基为MS＋6-BA 4.0 mg.L-1＋KT 0.1 mg.L-1＋NAA 0.1 mg.L-1＋IAA 0.2 mg.L-1;不定芽分化的最佳培养基为MS＋TDZ 0.5 mg.L-1＋NAA 0.5 mg.L-1,诱导率达63.14%,不定芽数平均为11.4。本试验获得的最佳培养基配方,适宜在低海拔条件下进行快速诱导大花红景天的愈伤组织和植株再生。     

大海林地区不同生境高山红景天根部红景天甙含量的差异

在黑龙江省大海林地区海拔１４００－１６２０ｍ的高山上,依据高山红景天的生境于岳桦林内、高山草甸、苔原上选择了６个样地,测定了高山红景天根部的红景天甙含量。生境不同的高山红景天,根部红景天甙的含量有明显差异,红景天甙含量范围内０．１０１％－０．６８４％。从测定结果看,光照条件好、土壤肥沃的生境下高山红景天根部的红景天甙含量较高。     

大海林地区不同生境高山红景天根部红景天甙含量的差异

在黑龙江省大海林地区海拔1400~1620m的高山上,依据高山红景天的生境于岳桦林内、高山草甸、苔原上选择了6个样地,测定了高山红景天根部的红景天甙含量。生境不同的高山红景天,根部红景天甙的含量有明显差异,红景天甙含量范围为0.101%~0.684%。从测定结果看,光照条件好、土壤肥沃的生境下高山红景天根部的红景天甙含量较高。     

不同来源高山红景天材料中有效成分的HPLC分析

本文用HPLC测定、比较了5种不同来源的高山红景天材料中有效成分—红景天甙及其甙无酪醇的含量,并建立了高山红景天有效成分分析的样品制备方法,结果表明:固体培养、液体培养及反应器培养的高山红景天组织中均含有上述两种有效成分.其中固体培养的材料中有效成分的含量最高.三种提取方法比较表明,用索氏提取器连续回流提取明显优于冷浸提取和批式回流提取.     

红景天甙的生物合成及其关键代谢酶研究

红景天甙是红景天属植物的主要药效成分,是一种很有前途的环境适应药物,具有抗疲劳、抗衰老、抗微波辐射、抗病毒及抗肿瘤等特异功效,尤其在军事、航天、运动及保健医学上具重要应用价值,近年来备受关注。本文在介绍了红景天属植物的开发利用价值及资源现状的基础上,重点探索了红景天甙生物合成的可能途径。认为其甙元酪醇是经由莽草酸途径合成,再由UDP葡萄糖基转移酶（Glucosyltransferase）催化葡萄糖和酪醇合成红景天甙,而红景天甙又可能在β-D-葡萄糖苷酶作用下降解为甙元酪醇和葡萄糖。文章阐述了参与红景天甙代谢的上述两个关键酶的研究现状及前景。     

固定化β-葡萄糖苷酶催化合成红景天甙的研究

目的:利用海藻酸钠和壳聚糖固定β-葡萄糖苷酶,催化合成红景天甙,并对固定化条件的选择、固定化酶催化合成红景天甙的条件优化进行研究.方法:采用正交实验方法寻求最佳固定化条件,以转化率为指标对合成条件进行优化.结果:固定β-葡萄糖苷酶的最佳条件为:壳聚糖浓度1.5%,吸附时间9h,交联时间12h,戊二醛浓度1.0%,吸附温度O℃,酶活力回收率达74.38%.催化合成红景天甙的最佳条件为:反应介质pH 5.8醋酸缓冲液/叔丁醇(1:9),底物浓度酩醇5g/L,D-葡萄糖与酪醇摩尔比为1:1,反应时间50h,反应温度50℃,红景天甙的转化率最高可达到71.9%.结论:固定化酶催化合成红景天甙的转化率得到较大的提高,为工业化生产提供了可靠的理论依据.     

高山红景天愈伤颗粒组织悬浮培养动力学及工艺的研究

研究了高山红景天愈伤组织颗粒悬浮培养过程的生长及红景天甙合成的规律,发现红景天甙合成与细胞生长偶联,两者之间的关系表示为q=5．38×10^-3μ。其产率系数Yx/s及维持系数m分别为0．66g·g^-1和0．033g·(g·d)^-1。在3.5L气升式反应器中研究了培养过程的操作特性,发现“发泡”现象几乎不存在,氧传递系数K1a的变化不同常规植物细胞的悬浮培养．愈伤组织颗粒在培养过程中经历一个由小变大,后叉变小的过程。     

4种红景天植物的组织培养研究

以大花红景天、云南红景天、长鞭红景天和库页红景天的茎和叶为外植体进行组织培养,结果表明：大花红景天以茎为外植体诱导芽效果最好,其它3种红景天以叶为外植体诱导芽效果最好。云南红景天和长鞭红景天适合的芽诱导激素组合是0．1mg／L NAA和2．5mg／L 6-BA的组合,在该激素水平下两种红景天的出芽频率分别达到71％和84％;大花红景天和库叶红景天适合的芽诱导激素组合是0．5mg／L NAA和2．5mg／L 6-BA的组合,在该激素水平下两种红景天的出芽频率均达到80％。长鞭红景天和库叶红景天在添加IBA的培养基上容易生根形成完整植株,生根率分别达到87％和73％;经过炼苗后,长鞭红景天再生苗能够成功移栽,成活率达66％。     

梨枣叶片和茎段再生体系的建立

用不同浓度配比的生长素和细胞分裂素诱导梨枣叶片和茎段愈伤组织的产生,并研究了不定芽诱导的最佳配方,建立了梨枣叶片和茎段的再生体系.结果表明,梨枣叶片愈伤组织诱导的最佳培养基为MS+2,4-D 1.5 mg·L-1+6-BA 0.5 mg·L-1;茎段为MS+2,4-D 1.0 mg·L-1+6-BA 0.5 mg·L-1.叶片不定芽诱导的最佳培养基为MS+IBA 0.1 mg·L-1+6-BA 1.5 mg·L-1. AgNO3能阻止叶片外植体褐化并有效地促进叶片愈伤组织分化.茎段能在同一培养基上产生愈伤组织并直接分化出不定芽.     

干旱胁迫对紫花苜蓿黄酮类化合物含量及其合成途径关键酶活性的影响

以3个紫花苜蓿(Medicago sativa)品种为试验材料,利用不同浓度(0、5%、10%、15％、20%、25%)的PEG 6000溶液模拟干旱胁迫,探究干旱胁迫对不同品种紫花苜蓿黄酮类化合物合成上游3个关键酶活性以及黄酮类化合物含量的影响。结果表明：(1)随着PEG 6000胁迫浓度的增加,3种紫花苜蓿叶片中PAL(苯丙氨酸解氨酶)、C4H(肉桂酸 4 羟基化酶)和4CL(4 香豆酸辅酶A连接酶)活性均呈先升高后降低的趋势,但不同酶类之间响应存在差异,即PAL活性在10% PEG 6000胁迫浓度下最高,C4H和4CL活性则在15% PEG 6000胁迫浓度下最高,且均显著高于相应对照。(2)3种紫花苜蓿植株地上部总黄酮含量和8种黄酮类化合物含量均随着PEG 6000胁迫浓度的增加呈先上升后下降的趋势,且均在10%~15% PEG 6000胁迫浓度时达到最高值,并显著高于相应对照。(3)各品种紫花苜蓿叶片黄酮类化合物含量与其关键酶活性呈不同程度的相关性,即紫花苜蓿黄酮类化合物合成途径上游3个关键酶活性与其地上部黄酮类化合物含量存在密切的关系。研究认为,不同程度的干旱胁迫可以通过促进黄酮类化合物合成途径上游关键酶活性的变化来调节紫花苜蓿植株中黄酮类化合物的合成,且适度干旱胁迫能显著促进相关酶活性增强和黄酮类化合物含量增加。     

A Novel Burdock Fructooligosaccharide Induces Changes in the Production of Salicylates, Activates Defence Enzymes and Induces Systemic Acquired Resistance to Colletotrichum orbiculare in Cucumber Seedlings

The ability of burdock fructooligosaccharide (BFO), a type of linear fructooligosaccharide extracted and isolated from the roots of Arctium lappa , to induce systemic acquired resistance (SAR) was studied in cucumber seedlings. BFO strongly induced changes in salicylic acid (SA) and SA-glucoside (SAG) in BFO-treated leaves, and similar changes of SA and SAG were also found in untreated leaves of the same seedling. The level of SA in the first leaves sprayed with BFO (5.0 g/l) increased by 3.6 times after 24 h and then gradually declined from 48 to 96 h and finally decreased to a nadir at 120 h. The SAG level increased by 2.1 times at 24 h and then continued to increase to about 10.0 times as much as that in control from 96 to 120 h. The levels of SA in the untreated leaves of the same seedling only increased by 1.6–1.9 times during the period of 24–72 h followed by a decrease at 120 h, while SAG increased by 1.1 times at 24 h but steadily continued to increase to its maximum from 24 to 120 h. In summary, the patterns of expression of SA and SAG in the untreated leaf were similar to that of the treated leaf of the same seedling, while the pattern of expression of SAG was quite different from that of SA both in the treated and untreated leaves. Pretreatment with BFO reduced the lesions caused by Colletotrichum orbiculare by 56.8%. Additionally, the amount of lignin and the activities of some defensive enzymes including peroxidase, superoxide dismutase, polyphenoloxidase and β-1,3-glucanase significantly increased in the first leaves pretreated with BFO and followed with C. orbiculare inoculation. These results demonstrate that BFO can enhance the contents of endogenous SA, the resistance against C. orbiculare , and the activities of defensive enzymes of cucumber seedlings.     

植物生长调节剂对南方红豆杉愈伤组织培养和紫杉醇合成的影响

通过不同种类和水平植物生长调节剂对南方红豆杉(Taxus chinensisvar.mairei)愈伤组织诱导、生长和紫杉醇合成能力影响的研究发现:诱导培养初期,以无植物生长调节剂的MS为基本培养基,在附加不同植物生长调节剂组合作用下愈伤组织产生的时间和生长、在相同植物生长调节剂组合作用下不同外植体愈伤组织的产生时间和生长均表现出较显著差异,2,4-D/NAA高于0.4时,不利于南方红豆杉愈伤组织的诱导。转换到附加不同植物生长调节剂组合的B5培养基上后,随培养继代次数的增加,生长差异逐渐缩小,直至不显著,表明参考不同文献报道最优配方所设计的各植物生长调节剂组合对南方红豆杉愈伤组织的生长均较适宜,有利南方红豆杉愈伤组织生长的植物生长调节剂优化组合没有唯一性。但不同调节剂组合作用下的同源愈伤组织中、相同调节剂作用下不同源愈伤组织中紫杉醇含量均存在着极显著差异,适当水平(2 mg/L)的2,4-D单用,或与适当水平的KT、6-BA、KT GA配合使用,对南方红豆杉愈伤组织紫杉醇的合成较有利,NAA则不太有利,幼茎和叶愈伤组织产紫杉醇的水平较其它愈伤组织为高。     

Chitinase and β-1,3-glucanase activities in chickpea (Cicer arietinum). Induction of different isoenzymes in response to wounding and ethephon

Chitinase and β-1,3-glucanase activities were assayed in roots, stems and leaves of 12-day-old chickpea ( Cicer arietinum L.) plants. While glucanase activity was higher in roots than in the aerial parts of the plant, leaves had higher Chitinase activity. Both glucanase and chitinase activities were induced in roots and stems in response to wounding (excision into 1-cm pieces), with activity increasing 6 h after treatment, reaching a maximum between 24 and 48 h, and thereafter remaining nearly constant up to 72 h. Ethephon treatment also induced β-1,3-glucanase and chitinase activities in stems but not in roots. Both enzymes occurred in root and stem tissues as a complex mixture of isoenzymes. At least four different peaks with glucanase and chitinase activities could be resolved by gel filtration chromatography on Sephacryl S-200 and chromatofocusing on PBE 94 (pH 4–7). Following ammonium sulfate precipitation and ion exchange on CM- and DEAE-Trisacryl, three β-1,3-glucanase and chitinase fractions, referred to as basic, neutral and acidic, were separated on the basis of their chromatographic behaviour. Most of the total protein (75%) of stem extracts was found in the acidic fraction, whereas the major glucanase (53%) and chitinase (62%) activities were in the basic and neutral fractions, respectively. While wounding resulted in an increase in the neutral glucanase and chitinase activities, the activities of the acidic fractions were promoted by ethephon.     

SOMATIC EMBRYOGENESIS AND PLANT REGENERATION IN HYOSCYAMUS NIGER

Culture of segments of leaves, petioles, and stems and of germinated seeds and whole ovaries enclosing unpollinated ovules of Hyoscyamus niger in a medium containg 2, 4-dichlorophen-oxyacetic acid caused massive growth of calluses. In cultured germinated seeds and ovaries, calluses were formed from the hypocotyl and placenta, respectively. Leaf and stem explants generally gave rise to hard, white, and compact calluses. Upon transfer of these calluses to an auxin-free medium they regenerated roots or buds but showed no further signs of morphogenesis. Calluses formed on petioles, germinated seeds, and ovaries were a mixture of the white, compact and brown, friable types. Subsequent morphogenesis of the friable calluses in a medium lacking auxin was characterized by the formation of somatic embryos whose pattern of development was similar to that of zygotic embryos.     

A comparative study of somatic embryogenesis in Secale vavilovii

Summary The ability of immature embryos, inflorescences and leaves of Secale vavilovii to form embryogenic callus was tested on Murashige and Skoog (1962) medium supplemented with different concentrations of 2,4-D. All cultured immature embryos formed calluses. The highest percentage of embryogenic callus production was from 1–2 mm embryos. Young leaves also formed calluses, mainly from the 10–15 mm basal segment, the percentages of embryogenic calluses being higher when cultures were maintained in darkness. Embryogenic calluses were obtained also from all the cultured immature inflorescences, in the three cases, rooted green plants were obtained and grown in soil. Comparison of the responses of the three explants used indicates that immature inflorescence is the most useful explant for obtaining regenerated plants in Secale vavilovii.     

杂交水稻金优63幼苗期SOD和POD特性研究

对杂交水稻金优63幼苗不同时期的根、茎、叶进行SOD同工酶电泳分析,并测定SOD、POD活性。结果表明,自播种后第7天到第13天,幼苗的SOD同工酶在根、茎、叶中有明显的器官特异性,且SOD活性叶 >茎 >根。相同器官不同时期的SOD同工酶电泳谱带条数及SOD活性都有变化,且SOD活性强弱与SOD同工酶电泳谱带中有无Mn-SOD同工酶带有一定的关系。幼苗的POD活性在根、茎、叶中也有明显的器官特异性,茎中POD活性明显高于根和叶,且POD活性变化与SOD活性变化有一定的关系。     

板栗扦插生根过程的解剖结构和生理变化研究

以5年生和30年生板栗插穗以及30年生板栗母树不同月份(5、6、7月)插穗为材料,进行不同浓度(0、200和500mg/L)吲哚丁酸(IBA)处理,观测各类处理材料的生根情况,并对扦插前插穗(初始茎)和扦插后插穗愈伤组织的碳氮代谢物质、褐化相关物质和解剖学结构进行分析观察,探讨不同年龄母树及不同月份插穗扦插生根能力,为板栗扦插生根技术体系奠定理论基础。结果表明:(1)5年生板栗的初始茎和愈伤组织中碳氮代谢相关物质淀粉和可溶性糖含量明显高于30年生板栗,而其可溶性蛋白含量30年生高于5年生板栗,但其γ-氨基丁酸(GABA)含量在不同年龄母树间无明显变化;30年生板栗插穗初始茎中可溶性蛋白、GABA和可溶性糖含量均随着月份的增加而逐渐降低,但是在愈伤组织中GABA、可溶性糖和淀粉含量随着月份增加而逐渐升高。(2)板栗插穗初始茎和愈伤组织的褐化相关物质单宁、酚类和类黄酮含量均表现为30年生不同程度低于5年生;除类黄酮含量在7月份的愈伤组织中有显著升高外,不同月份板栗的单宁、酚类、类黄酮含量均没有明显变化。(3)生根状况及解剖学观察结果显示,5年生板栗插穗的愈伤率和生根率高于30年生的插穗,且30年生插穗6月份扦插的愈伤率最高;板栗插穗属于诱导生根类型,皮层中存在环状厚壁组织;5年生板栗插穗的木质素含量低于30年生板栗,但形成层细胞层数高于30年生。研究认为,板栗扦插生根过程中插穗褐化现象严重,生根率较低;不同年龄母树和不同月份的板栗插穗在扦插过程中的碳氮代谢相关化合物含量都有明显的差异,而褐化相关物质含量仅在不同年龄母树间有较大的差异,不同月份间大部分无明显差异。     

干旱胁迫对3种灌木不同器官化学计量特征的影响

采用盆栽控水试验,研究了3个水分处理,即田间持水量(FC)的(75±5)%、(55±5)%和(35±5)%,对丁香、黄刺梅、连翘的叶、茎、极细根(0～1 mm)、细根(1～2 mm)和粗根(>2 mm)化学计量特征的影响。结果表明: 3种灌木相同器官间氮(N)和磷(P)含量以及C∶N、C∶P、N∶P均存在显著差异。随着干旱胁迫的加剧,3种灌木各器官C含量总体上无显著变化;叶片N含量呈增加趋势,茎N含量逐渐下降,极细根和细根N含量均呈先上升后下降的趋势;叶片、茎P 含量呈降低趋势,极细根和细根P含量呈先增加后下降的趋势。3种灌木在干旱胁迫下叶C∶N呈降低趋势,叶和茎C∶P、N∶P均呈增加趋势。干旱胁迫对极细根C∶N和叶C∶P、N∶P影响最大,而对粗根C∶N、N∶P以及极细根C∶P的影响最小。土壤C、N含量与灌木各器官中C、N、P含量无显著相关性,但土壤P含量与叶和根系C、N、P含量均显著相关。土壤中相对缺乏的P是影响灌木器官化学计量特征的最重要因素。干旱对不同灌木不同器官化学计量的影响不同,叶和极细根的化学计量较其他器官对干旱胁迫更加敏感。干旱可能主要通过影响灌木对土壤中养分P的吸收和在不同器官中的分配,进而影响灌木不同器官的化学计量特征,尤其是与P相关的化学计量特征。