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药用植物半夏细胞悬浮培养和植株再生体系建立研究 总被引:3,自引:0,他引:3
为了建立甘肃产药用植物半夏的细胞悬浮培养和植株再生体系,通过对其不同外质体筛选、愈伤组织诱导和悬浮培养的愈伤组织类型、激素水平、起始密度、震荡速率等影响因素以及细胞生长状态、生长曲线和pH值变化等指标的研究.结果表明:以叶柄为外质体,选择"松散型"愈伤组织和2×105个/mL的起始培养密度,在附加2,4-D 2.0 mg/L BA 0.5 mg/L NAA0.5 mg/L的1/2 MS培养基上培养,温度(23±2)℃,转速90 rpm,pH值6.0,黑暗培养为最适悬浮培养体系;经检测细胞生长曲线呈"S"型,培养液pH随着细胞的生长先上升后下降,18 d时细胞基数达到最大值;悬浮继代培养后,通过降低或停止震荡转速,在添加BA 0.5 mg/L NAA0.5 mg/L的MS液体和固体培养基上可诱导分化出芽和根,进一步形成完整的植株. 相似文献
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研究了植物生长调节剂和接种方式对怀地黄叶片愈伤组织诱导的影响和细胞悬浮培养体系的建立。结果表明:将叶片正接在MS+2,4-D 1mg/L+6-BA 0.4mg/L培养基上,愈伤组织生长良好,诱导率达100%;在该培养基上多次继代,可形成3种类型的愈伤组织,其中Ⅰ型愈伤组织淡黄色、颗粒透亮、分散性好且疏松易脆,适合进行细胞悬浮培养;在MS+2,4-D 3mg/L+6-BA 0.5mg/L培养基中,悬浮培养细胞生长曲线呈"S"型,培养15d时细胞干重达最大值,为4.94g/L。 相似文献
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以北沙参叶片和茎段为外植体,采用MS和B5培养基对其进行愈伤组织诱导,继代培养后进行悬浮培养,研究了不同浓度碳源、6-BA和NAA对其细胞生长的影响,以期为进一步建立北沙参细胞培养体系以及提高其次生代谢物产量奠定试验基础。结果表明:1/2MS+0.4mg/L 6-BA+1.5mg/L NAA琼脂培养基为北沙参愈伤组织诱导的较适宜培养基;1/2MS+0.2mg/L 6-BA+1.2mg/L NAA为适宜的继代培养基;在细胞悬浮培养过程中,分别添加0.2mg/L 6-BA、1.6mg/L NAA和20g/L蔗糖有利于细胞的生长。 相似文献
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建立鸡屎藤愈伤组织的诱导、增殖继代培养及其细胞悬浮培养的技术体系。研究不同植物激素组合对叶片、茎段和带腋芽茎段愈伤组织诱导的影响;采用正交试验研究不同处理对叶片愈伤组织增殖的影响;采用液体振荡培养法测定叶片愈伤组织细胞悬浮生长曲线。结果表明:叶片最易诱导产生愈伤组织,最佳激素配比为MS+6-BA1.0mg/L+NAA0.5mg/L,诱导率可达100%。叶片愈伤组织最佳增殖培养基为MS+2,4-D0.5mg/L+NAA0.3mg/L+6-BA0.5mg/L,pH5.8。由叶片愈伤组织建立的细胞系生长呈"S"型曲线,培养24d达到最大生长量,最适继代周期为18~20d。以叶片为外植体经愈伤诱导、增殖培养可建立良好的细胞悬浮系。 相似文献
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以水南一号龙眼为试材,在龙眼树四周打根洞,用蚯蚓粪作根洞填充物,研究了在根洞填充物中加入不同植物生长调节剂对根系生长和果实品质的影响。结果发现,与对照(清水处理)相比,IBA 5 mg/L+NAA 6 mg/L诱导的根数、根粗、根鲜干重比分别提高了317.01%、31.53%、4.98%,果实可溶性固形物含量、可食率、百粒重、产量分别提高了9.00%、4.39%、18.39%、41.10%;IBA 5 mg/L+NAA 5 mg/L诱导的根数、根粗分别提高了549.11%、62.57%,而诱导的根鲜干重比差异不显著,果实可溶性固形物含量、可食率、百粒重、产量分别提高了6.63%、4.14%、29.40%、54.79%;IBA 4 mg/L+NAA 6 mg/L诱导的根数、根粗、根鲜干重比分别提高了54.80%、50.55%、12.11%,果实可溶性固形物含量、可食率、产量分别提高了12.95%、4.75%、19.19%,而百粒重差异不显著。 相似文献
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以灵芝真菌为试材,研究了10L发酵罐发酵培养好氧型灵芝真菌条件下,溶氧控制条件对生产灵芝酸产量的影响。结果表明:溶氧对灵芝真菌发酵生产灵芝酸的影响较大,高通气量有利于缩短发酵时间,适宜的通气量为0.4L·L-1·min-1,此条件下,菌体和灵芝酸的最大产量分别为8.27g/L和171.26mg/L;相对初始发酵条件,菌体和灵芝酸最大产量分别提高13.13%和13.28%;相对于低通气量,菌体和灵芝酸达到最大产量的时间缩短12h;表明0.4L·L-1·min-1的通气量较适宜于10L发酵罐发酵生产灵芝酸。 相似文献
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以苹果试管苗叶片的再生不定芽嫩叶为试材,对苹果体细胞悬浮系的建立及植株再生的影响因素进行了研究.结果表明:在MS+NAA 0.5 mg/L+BA 2.0 mg/L培养基上可诱导获得高活力的愈伤组织.将该愈伤组织转入MS+2,4-D 2.0 mg/L+BA 1.0 mg/L液体培养基中培养,采用无菌筛网分离获得含单个细胞和少于8~10个细胞的小细胞团进行继代培养,建立体悬浮细胞培养系.将悬浮细胞转到MS+2,4-D 2.0 mg/L+BA 1.0 mg/L的固体增殖培养基上暗培养25 d后,可形成微型愈伤组织,将该愈伤组织转到MS+BA 5.0 mg/L+IAA 0.5 mg/L的植株再生培养基上暗培养30 d后转入光下,光照培养30 d后53%的愈伤组织可再生植株.该研究建立的苹果体细胞悬浮培养技术,不仅可用于细胞融合及遗传转化过程中杂种细胞和转化细胞的分离及植株再生研究,而且对于利用组织培养技术筛选变异株系也具有重要意义. 相似文献
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Taking into account the recalcitrance of date palm (Phoenix dactylifera L.) to tissue culture, establishment and proliferation of embryogenic suspension cultures in two date palm cultivars, namely Boufeggouss (BFG) and Bouskri (BSK) was implemented using liquid medium with different concentrations of the 6-benzylaminopurine (BAP) (0.3, 0.4, 0.5 mg/l) and various subculturing times (7 days, 15 days and 20 days). Total phenols and peroxidase activities have been determined and oxidative browning process checked in different cultures. The liquid medium with 0.3 mg/l BAP allowed cell suspension cultures to produce the greater globular embryos and the highest number of somatic embryos in comparison with the other treatment for BFG and BSK date palm cultivars. The low rate of culture transfers (every 15 or 20 days) tended to increase the phenolic contents and peroxidase activities in plant tissue leading to an enhancement of tissues/cells browning and then a decrease in the proliferation of embryogenic cells. The transfer of cultures on fresh medium every 7 days allowed a substantial reduction of tissue/cell oxidative browning, which is related to reduction of phenolic compounds and decrease in peroxidase activities with a resultant increase in the proliferation of embryogenic cells. This result suggests that oxidative browning is mainly peroxidase-based in date palm suspension cultures. Relationships between low concentrations of BAP as cytokinin, high growth of embryogenic cells, low contents of phenolics, decrease of peroxidase activities and oxidative browning process, are discussed, in order to optimize date palm multiplication through somatic embryogenesis and solve recalcitrance of many date palm cultivars to tissue culture. 相似文献
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在1/2 MS基本培养基中,分别加入不同种类、剂量的由野生春兰根部分离到的内生真菌所制成的干菌丝诱导子,对由春兰种子萌发成的原球茎、根状茎等组培物进行诱导培养.结果表明,4 种真菌诱导子均可不同程度的促进春兰组培物生长量的增加及不定芽的分化,其诱导作用依次为:CF11>CF3>CF1>CF8>CK.加入CF11、CF3号诱导子的处理与对照相比,鲜重分别增加了74.38%、73.93%,诱导效果最好.此外,春兰组培物的鲜重随着CF1号诱导子加入量的增大而增加,其诱导作用依次为20 g/L>10 g/L>5 g/L>0 g/L,加入量为20 g/L时,其鲜重与对照相比达到了显著水平. 相似文献
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Maria Skłodowska Ewa Gajewska Elżbieta Kuźniak Artur Mikiciński Piotr Sobiczewski 《Scientia Horticulturae》2010
BTH (S-methylbenzo-1,2,3-thiadiazole-7-carbothiate), an active compound of the commercial preparation Bion, has been studied as an elicitor of resistance to fire blight (Erwinia amylovora) in apple. However, the biochemical mechanisms of its action are not fully elucidated. Our study indicated that BTH at the best time of its protection activity (2–14 days after application) induced changes in prooxidant–antioxidant balance in the leaves of apple trees, but in different ways in the enzymatic and nonenzymatic antioxidants. Glutathione as low molecular antioxidant as well as superoxide anion radical and lipid peroxides as oxidants exhibited changes at the early phase of BTH action. Glutathione-dependent enzymes were strongly affected by the elicitor used. On the 2nd day glutathione transferase (GST) and glutathione peroxidase (GSH-Px) activities increased by about 70% and 30% above the control, respectively. GST activity normalized about the 14th day but GSH-Px at the same time showed 27% of the control value. Among enzymes utilising hydrogen peroxide only catalase showed increase (37%) at the early phase of experiment. Compared with the control, BTH-treated plants did not show changes in ascorbate peroxidase and phenylalanine ammonia-lyase activities. Tocopherol (TOC) level diminished starting from the 7th day after BTH treatment and on the 14th day it was only 28% of the control. It is proposed that extinguishing of BTH-mediated signal resulted from TOC and glutathione action. The diminished ascorbate level at all examined times may play a crucial role in BTH-mediated cell growth regulation. The direct influence of BTH on lipid metabolism should be also taken into consideration. 相似文献
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白鹤芋胚性细胞悬浮培养和高效植株再生体系的建立 总被引:1,自引:0,他引:1
以白鹤芋(Spathiphyllum cannifolium)试管苗叶柄为外植体诱导获得胚性愈伤组织,建立了胚性细胞悬浮培养系并高频率再生出植株。结果表明,最优的悬浮培养条件为:装有20 mL液体培养基的100 mL三角瓶中接种0.3 g胚性细胞团,悬浮培养基为MS + 0.5 mg ? L-1 TDZ + 1.0 mg ? L-1 2,4-D + 30 g ? L-1蔗糖或麦芽糖,pH 5.8;继代间隔14 d;每个继代周期,胚性细胞团可增殖至接种量的5倍以上;植株再生最优的分化培养基为1/2MS + 0.3 mg ? L-1 6-BA + 30 g ? L-1蔗糖 + 8.0 g ? L-1琼脂粉,pH 5.8,平均每个胚性细胞团可分化再生出25.1株小植株;胚性细胞的快速增殖和高频率植株再生的状态可保持24周。 相似文献
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Fusarium mairei, a paclitaxel-producing fungal endophyte, its effects on taxoid synthesis in Taxus cells were investigated via adding the fungal endophyte culture supernatants (FECS) to the suspension cultures of Taxus cuspidata. The concentration of FECS was determined on its total carbohydrate. When 100 mL of Taxus cell suspension cultures were treated with several dosages of FECS (4, 6 and 8 g) at day10, the cultures treated with 6 g FECS produced the highest yield of paclitaxel (5.84 mg L−1), which was 1.8-fold the yield from controls (3.14 mg L−1). The major elicitor element in FECS was an oligosaccharide of 2 kDa. In addition, the cultures treated with 6 g FECS resulted in 25.86 mg L−1 of the precursor 10-deacetylbaccatin III (10-DAB) accumulation, which was 11 times that of control cultures (2.32 mg L−1), and 4.7 times higher than that of cultures treated with 200 μM methyl jasmonate (MJ) (5.43 mg L−1). These results indicate that FECS favors to stimulate 10-DAB accumulation more effectively than paclitaxel accumulation. 相似文献