新疆雪莲愈伤组织诱导及总黄酮的测定

以新疆雪莲植株叶片为外植体,研究了不同植物激素和培养时间对新疆雪莲愈伤组织生长、总黄酮含量和产量的影响.结果表明:新疆雪莲愈伤组织最佳诱导培养基为MS+2.00 mg/L NAA+1.00 mg/L 6-BA,其愈伤组织诱导率最高(97.0%);继代培养基以MS+2.00 mg/L NAA+1.00 mg/L 6-BA中愈伤组织生长量和总黄酮产量最高,分别达4.58 g和12.24 mg;以MS+4.00 mg/L NAA+2.00 mg/L 6-BA中的愈伤组织总黄酮含量最高,达2.17 mg/g;愈伤组织的生长量随继代培养时间的延长而增加,而其总黄酮含量不稳定,于继代培养第30天达到最高(2.7 mg/g),此时总黄酮产量最高可达26.66 mg.     

培养基成分对杜仲愈伤组织生长及次生代谢产物含量的影响

以Bs＋0．5mg／L NAA＋0．5mg／L BA为基本培养基,研究了B5培养基中8种主要无机盐浓度对杜仲愈伤组织生长及绿原酸和总黄酮两种次生代谢产物含量的影响。结果表明：在1000～5000mg／L范围内增加培养基中KNO3的含量有利于愈伤组织生长,B5培养基中当KNO3的浓度达到2／3时,绿原酸和总黄酮含量及产量最高;(NH4)2SO4以4／3原浓度时对愈伤组织生长量、总黄酮含量及产量最高,对绿原酸的含量则是其为原浓度的1／3时最高;MgSO4以2／3浓度对生长量及1／3浓度对绿原酸、总黄酮积累最高;NaH2PO4、CaCl2和MnSO4以原浓度的愈伤组织生长和次生代谢产物合成最好;ZnSO4和FeSO4的原浓度愈伤组织的生长量最大,而1／3浓度的绿原酸和总黄酮含量最高。     

培养基及培养条件对杜仲愈伤组织生长及次生代谢产物含量的影响

以MS、LS、B5、N6、H、Nitsch、White、1／2MS为基本培养基,分别添加0．5mg／L NAA和0．5mg／L BA,分析不同类型培养基对杜仲愈伤组织生长及次生代谢产物含量的影响,并以B5培养基进行光照条件、碳源、蔗糖浓度试验。结果表明：B5培养基不仅有利于愈伤组织生长,也有利于总黄酮的形成,而1／2MS培养基有利于绿原酸的积累;12h／d光照对愈伤组织的生长及绿原酸和总黄酮的合成有明显的促进作用,黑暗不影响愈伤组织的生长,但却抑制绿原酸和总黄酮的形成;3种碳源中,愈伤组织的增长量、绿原酸和总黄酮的含量均以蔗糖为碳源时最高,葡萄糖最低;蔗糖浓度在10～50g／L范围内绿原酸的含量随着糖浓度的升高而升高,40g／L时愈伤组织的增长量和总黄酮的含量最高。     

新疆紫草的组织培养及其染色体分析

新疆紫草(Arnebia euchroma)的胚轴、子叶、根都产生愈伤组织。不同来源的外植体产生苗的能力不同,胚轴来源的愈伤组织分化苗最多,而且苗生长旺盛。根的愈伤组织只产生不定根。新疆紫草的核型是 K(2n)=14=2m 8sm 2sm(SAT) 2st。随着继代培养代数增多,染色体变异的种类和频率以及染色体数目显著增加。如果继代培养时间较长,染色体数目变异的范围更广,而且出现染色体结构变异。     

新疆紫草的组织培养及其染色体分析

新疆紫草(Arnebia euchroma)的胚轴、子叶、根都产生愈伤组织。不同来源的外植体产生苗的能力不同,胚轴来源的愈伤组织分化苗最多,而且苗生长旺盛。根的愈伤组织只产生不定根。新疆紫草的核型是K(2n)=14=2m+8sm+2sm(SAT)+2st。随着继代培养代数增 多,染色体变异的种类和频率以及染色体数目显著增加。如果继代培养时间较长,染色体数目变异的范围更广,而且出现染色体结构变异。     

印楝愈伤组织形成及其印楝素含量测定

取印楝（AzadirachtaindicaA．Juss）不同器官作外植体,研究培养基和继代次数对愈伤组织生长及其印楝素（Azadirachtin）生物合成的影响．印楝的不同器官（根、叶、茎及皮）均能诱导出愈伤组织,这些愈伤组织均有合成印楝素的能力．其中以叶诱导的愈伤组织生长速率及印楝素含量为最高．含有较低按盐的B5培养基有利于细胞生长,含有较高铵盐的MS培养基有利于印楝素积累,不含铰盐的White培养基对两者均不利．愈伤组织继代2—3代,有利于愈伤组织生长和印楝素合成。     

茉莉酸甲酯和水杨酸对西洋参愈伤组织生长和皂苷形成的影响CSCD

本文旨在探究不同诱导子对西洋参愈伤组织生长、相关酶活性及其人参皂苷含量的影响,在西洋参愈伤组织中,应用HPLC检测了添加诱导子茉莉酸甲酯(methyl jasmonate,MeJA)和水杨酸(salicylic acid,SA)后西洋参愈伤组织皂苷生物合成的变化。结果显示,MeJA抑制生长,但SA对其生长影响较小;2种诱导子均可以显著激活西洋参愈伤组织中超氧化物歧化酶(superoxide dismutase,SOD)、过氧化氢酶(catalase,CAT)、过氧化物酶(peroxidase,POD)活性和丙二醛(malondialdehyde,MDA)含量;培养基中MeJA浓度为100μmol/L时,愈伤组织中总皂苷含量和产量均达到最大值,人参皂苷Rg_(1)、Re、Rb_(1)、Rc的含量最高,在SA诱导子试验组中,当SA浓度为300μmol/L时,西洋参愈伤组织中总皂苷含量和产量均达到最大值,人参皂苷Rb_(1)的含量最高。结果表明,在西洋参愈伤组织中添加适当浓度MeJA和SA诱导子会提高人参皂苷化合物含量,其中以MeJA诱导人参总皂苷的效果最好,同时也会影响西洋参愈伤组织的生长量。说明在培养过程中添加外源诱导子,有利于提高西洋参愈伤组织中皂苷含量。     

烟草愈伤组织分化和芽原基形成期间呼吸代谢途径的改变

接种在继代培养基上的柳叶烟草愈伤组织,未观察到组织分化和芽原基形成。在分化培养基上生长的愈伤组织,接种后第6天可见拟分生组织和管胞分化,9—12天有芽原基形成,15—18天可观察到苗端结构。根据碘乙酸、Na_3PO_4和丙二酸抑制试验,以及3-磷酸甘油醛脱氢酶与琥珀酸脱氢酶活性测定结果,初步表明烟草愈伤组织呼吸中存在有EMP、HMP和TCAC代谢途径.在发生输导组织和芽原基分化的愈伤组织中(接种后第6—12天),HMP途径的运行程度较高;而芽原基的继续生长(培养12天以后),则与EMP途径的增加有关;分化培养基上生长的愈伤组织,始终较继代培养愈伤组织具有较高的FCAC活性水平。     

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.     

烟草愈伤组织继代培养和分化期间蛋白质代谢的比较研究

柳叶烟草(Nicotiana tabacum L.)愈伤组织在分化和芽原基形成期间,与继代增殖培养物比较,蛋白质含量与蛋白水解酶活性均缓慢上升;组分Ⅱ(水溶性蛋白与酶蛋白)蛋白质和核糖体组蛋白的合成速率都明显高于继代培养物;分化组织中总核糖体水平,特别是执行蛋白质合成功能的多聚核糖体的构建也都高于继代培养物。显然,分化培养愈伤组织较继代培养物有更高的蛋白质合成作用;而且组织分化和芽原基形成所需合成的新蛋白组分是与继代增殖生长的不同。继代培养后期,蛋白水解酶活性迅速升高,多聚核糖体相对量与蛋白质合成速率均明显下降,蛋白质含量急剧降低,表现出继代培养物开始衰老的代谢特征。此时期的分化培养组织,随着芽原基的继续生长,虽然蛋白质合成速率、多聚核糖体水平与蛋白质含量较前期有所降低,但都明显高于同时期的继代培养愈伤组织。     

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%。     

苦丁茶愈伤组织的诱导与褐变抑制

以叶片为材料,对苦丁茶愈伤组织的诱导,继代培养及褐变调控的研究表明：（１）苦丁茶愈伤组织的诱导及继代培养均以ＭＳ附加ＢＡ ２．０ｍｇＬ＾－１和ＮＡＡ ４．０ｍｇＬ＾－１的培养基效果最好;（２）０．１％的植酸可明显促进愈伤组织生长、抑制褐变,而硫代硫酸钠效果最差;（３）连续培养４０－５０ｄ愈伤组织增长倍数达到最大值;（４）继代２７次以后愈伤组织生长速度开始下降。这些条件为下一步细胞培养生产苦丁茶甙等     

Morphogenesis in leaf,hypocotyl and root explants of Digitalis thapsi L. cultured in vitro

The effects of the auxins 2,4-D, NAA and IAA either alone or in combination with kinetin or BA were investigated to assess the morphogenetic potential of leaf, root and hypocotyl explants of Digitalis thapsi. Calluses were obtained from the three explants in basal medium without the addition of growth regulators and in leaves, the calluses formed roots. Application of 2,4-D, NAA or BA increased callus formation. The presence of NAA induced root formation and that of BA induced shoot formation via callus interphase. Indole-3-acetic acid alone only induced the generation of roots in the hypocotyl callus. Kinetin was ineffective in all the explants tested. Combinations of NAA with kinetin or BA were more effective in inducing organogenesis in leaf explants. Optimum responses were obtained in hypocotyl and root explants by using IAA in combination with BA, the highest rate of shoot regeneration being observed in hypocotyl explants.Rooting of the differentiated shoots was readily achieved in media without growth regulators. Regenerated plantlets were transferred to soil and grew with a survival rate of 70%.Abbreviations BA benzyladenine - 2,4-D 2,4-dichlorophenoxyacetic acid - IAA indoleacetic acid, Kin-kinetin - NAA naphthaleneacetic acid     

The Influence of Different Factors on Cell Growth and α-tocopherol Content of Culture from Carthamus tinctorius

Calli were induced from root, hypocotyl, cotyledon and flower bud of Carthamtus tinctorius. All calli had the capability to synthesize α-tocopherol. Among these calli, the hvpocotylcallus was better than others in cell growth rate and α-toeopherol content. Culture conditions could intensively influence the growth rate and α-tocopherol production of callus from Carthamus tinctorius. Sucrose (30g/L) was good for callus growth and glucose (30g/L) was good for α-tocopherol accumulation. High concentration (0.55%) of inositol could obviously stimulate both growth rate and α-tocopherol synthesis of callus. The inoculum quantity for best callus growth was 0.035 to 0.067g dry wt/flask (50ml volume). In addition, α-tocopherol content was effectively increased by culture callus in high CO2 concentration. Studies on optimum cuhure conditions of callus culture showed that the callus growth rate, α-tocopherol content and yield were 1.88, 2.03 and 3.30 times respectively as high as of the control by administration of 0.45%–0.55% inositol, 10% coconut milk, 0.1–0.5% casamino acid, 30g/L sucrose and 10g/L glucose.     

Production of adventitious root biomass and secondary metabolites of Hypericum perforatum L. in a balloon type airlift reactor

The effects of inoculum density, aeration volume and culture period on accumulation of biomass and secondary metabolites in adventitious roots of Hypericum perforatum in balloon type airlift bioreactors (3 l capacity) were investigated. The greatest increment of biomass as well as metabolite content occurred at an inoculum density of 3 g l⁻¹ and an aeration volume of 0.1 vvm. After 6 weeks of culture, an approximately 50-fold increase in biomass was recorded containing 60.11 mg g⁻¹ dry weight (DW) of phenolics, 42.7 mg g⁻¹ DW of flavonoids and 0.80 mg g⁻¹ DW of chlorogenic acid. Liquid chromatography–mass spectroscopy/mass spectroscopy demonstrated that the presence of quercetin and hyperoside in adventitious roots at a level of 1.33 and 14.01 μg g⁻¹ DW, respectively after 6 weeks of culture. The results suggest scale-up of adventitious root culture of H. perforatum for the production of chlorogenic acid, quercetin and hyperoside is feasible.     

怀牛膝细胞悬浮培养条件的优化

为进一步优化怀牛膝(Achyranthes bidentata)细胞悬浮培养条件,对接种量、继代周期、pH、光照及Cu2+等多种影响因子的作用效果进行了研究,以提高怀牛膝细胞生长量及牛膝多糖含量。结果显示,接种量50 g·L^–1、继代周期14天,pH5–6和光照培养可以使细胞保持良好的生长状态及多糖合成能力;添加50μmol·L^–1 Cu^2+,细胞的干重最大,可达44.63 g·L^–1,多糖含量也最高,为4.02 mg·g^–1。     

多效唑和缩节胺对现蕾前忍冬枝叶生长及花蕾性状和有效成分含量的影响

以忍冬品种‘九丰一号’(Lonicera japonica‘Jiufeng 1’)为实验材料,采用叶面喷施方法研究了不同质量浓度多效唑和缩节胺对现蕾前(抽枝生长初期)枝叶生长和叶片叶绿素含量以及花蕾性状和花蕾中绿原酸和总黄酮含量的影响。结果显示:分别喷施100、400、700和1 000 mg·L-1多效唑和50、100、150和200 mg·L-1缩节胺后,多数处理组的开花枝条数、着花节数和叶绿素含量较对照CK1(水)有不同程度提高,但叶面积差异不明显。随处理时间延长,各处理组枝条节间长度总体上呈逐渐增加的趋势,其中多数处理组枝条节间长度增长缓慢。各处理的花蕾长度、百蕾鲜质量和干质量总体上小于CK1,而花蕾折干率、总黄酮含量和绿原酸含量显著或不显著高于CK1。此外,在喷施多效唑和缩节胺的同时喷施质量体积分数1.0%尿素和质量体积分数0.1%硼砂,忍冬的叶面积、着花节数、花蕾长度、折干率、百蕾鲜质量和干质量总体上有所提高,而绿原酸含量降低,但各指标的差异总体较小。研究结果表明:喷施适量多效唑和缩节胺可调控忍冬枝条生长,并能提高花蕾中总黄酮和绿原酸含量。     

Shoot organogenesis and plantlet regeneration from hypocotyl-derived cell suspensions of a tree legume, Dalbergia sissoo Roxb

Summary A complete and efficient protocol is presented for plant regeneration from cell-suspension cultures of Dalbergia sissoo Roxb., an economically important leguminous tree. Factors influencing callus initiation, establishment of cell-suspension culture, callus formation from embredded microcolonies, and shoot organogenesis from suspension-derived callus were identified. Of the two different auxins tested, callus induction was better on a medium containing naphthalene acetic acid (NAA). The percentage of callus induction increased considerably when NAA at 2.0 mg l⁻¹ (10.8 μM) was added in conjunction with 0.5 mg l⁻¹ (2.2 μM) N⁶-benzyladenine (BA). Of the three different explants evaluated for callus induction, hypocotyl segments were most responsive. Friable hypocotyl-derived callus from the second subculture passage was used to initiate the cell-suspension culture. Optimum growth of the cell suspension was observed in MS medium supplemented with the same growth regulators as described above for callus induction, with an initial inoculum cell density of 1%. The plating efficiency of the microcolonies was greatly influenced by harvesting time and the gelling agent used for plating. Efficiency was highest (93%) with cells harvested at their exponential growth phase and plated in 1.2 g l⁻¹ Phytagel. Shoot organogenesis from callus cultures was higher on a medium supplemented with a combination of BA and NAA than on BA alone. Seventy-one per cent of cultures exhibited shoot-bud differentiation on a medium containing 3.0 mg l⁻¹ (13.3 μM) BA and 0.5 mg l⁻¹ (2.7 μM) NAA. Regenerated shoots were rooted on half-strength MS medium containing 1 mg l⁻¹ each of indole-3-acetic acid (5.7 μM), indole-3-butyric acid (4.9 μM) and indole-3-propionic acid (5.3 μM). Plantlets were acclimated and established in soil.